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2006 Ministry of Health - Brazil. All rights reserved.

This work may be partially or totally reproduced, provided that it mentions the source and such reproduction is not for sales or any other commercial purpose. The technical area is responsible for copyrights of texts and images herein. The Ministry of Health - Brazil institutional collection can be accessed at the Biblioteca Virtual em Sade of the Ministry of Health - Brazil: http://www.saude.gov.br/bvs Series D. Meetings and Conferences Circulation: 1st edition 2006 500 issues Edition, distribution and inqualification: MINISTRY OF HEALTH - BRAZIL Secretariat for Health Surveillance Department of Epidemiological Surveillance Esplanada dos Ministrios, bloco G, Edifcio-sede, 1 andar, sala 134 ZIP: 70.058-900, Brasilia DF E-mail: svs@saude.gov.br Internet adress: www.saude.gov.br/svs Printed in Brazil

Catalog Record
___________________________________________________________________________________________________ International Conference Rabies in the Amricas (17. : 2006 Oct. : Brasilia, Brazil) RITA XVII : Rabies in the Amricas : Annals / Ministry of Health - Brazil = RITA XVII : Reunin Internacional de Rabia en las Amricas : Anales / Ministerio de la Salud de Brasil = RITA XVII : Reunio Internacional de Raiva nas Amricas : anais / Ministerio da Sade. Brasilia : Ministry of Health - Brazil, 2006. 220 p. (Series D. Meetings and Conferences) Brasilia, Brazil, 15th-20th October / 2006 ISBN 85-334-1292-4 1. Rabies. 2. Epidemiologic Surveillance. I. Title. II. Series. NLM WC 550 _______________________________________________________________________________________________________ Catalog record in the source General-Coordination of Documentation and Information Editora MS OS 2006/1251

Index titles: Portuguese: Reunio Internacional de Raiva nas Amricas: Anais Spanish: Reunin Internacional de Rabia en las Amricas: Anales

MINISTRY OF HEALTH - BRAZIL


SECRETARIAT FOR HEALTH SURVEILLANCE DEPARTMENT OF EPIDEMIOLOGICAL SURVEILLANCE

Brasilia, Brazil, 15th-20th October / 2006

Series D. Meetings and Conferences Brasilia DF 2006

INTERNA TIONAL STEERING COMMITTEE/ COMIT CIENTFICO INTERNA CIONAL/COMIT INTERNATIONAL INTERNACIONAL/COMIT CIENTFICO INTERNACIONAL
Hilary Koprowski - Thomas Jefferson University, Philadelphia, Pennsylvania, USA Charles Rupprecht - Center of Diseases Control & Prevention, Atlanta, Georgia, USA Susan Nadin-Davis - Canadian Food Inspection Agency, Nepean, Ontario, Canada Dennis Donovan - Ontario Ministry of Natural Resources, Trent University, Canada Laura Robinson - Texas Department of Health, Harlinger, Texas, USA Alan C. Jackson - Queens University, Kingston, Ontario, Canada Fernando Vargas Pino - Secretara de Prevencin y Proteccin a la Salud, Mxico, D.F ., Mxico George Baer - Mxico, D.F., Mxico Dr. Didoro Batalla Campero - National Center for Diagnostic Services in Animal Health, Mexico Albino Jos Belotto - Pan American Health Organization, Washington, D.C USA Rosely Cerqueira de Oliveira (chairperson XVII RITA) - Ministrio da Sade, Braslia, Brasil

NA TIONAL SCIENTIFIC COMMITTEE/COMIT CIENTFICO NA CIONAL/COMIT CIENTFICO NATIONAL NACIONAL/COMIT NACIONAL


Rosely Cerqueira de Oliveira - Ministrio da Sade, Braslia, Brasil Expedito Luna - Ministrio da Sade, Braslia, Brasil Eduardo Hage Carmo - Ministrio da Sade, Braslia, Brasil Lucia Regina Montebello Pereira - Ministrio da Sade, Braslia, Brasil Marcelo Yoshito Wada - Ministrio da Sade, Braslia, Brasil Carlos Roberto Zanetti - Universidade Federal de Santa Catarina, Florianpolis, Santa Catarina, Brasil Fumio Homna Ito - Universidade de So Paulo, So Paulo, Brasil Ivanete Kotait - Instituto Pasteur, So Paulo, Brasil Neide Yumie Takaoka - Instituto Pasteur, So Paulo, Brasil Ruben Figueroa - OPAS/Brasil, Braslia, Brasil

Neusa Maria Frazatti Gallinas - Instituto Butantan, So Paulo, Brasil Marcelo Simo Ferreira - Sociedade Brasileira de Medicina Tropical, Brasil

EXECUTIVE OFFICE/COMISSO ORGANIZADORA/COMISIN ORGANIZADORA


Eunice de Lima - Ministrio da Sade, Brasil Lucia Regina Montebello Pereira - Ministrio da Sade, Brasil Rosely Cerqueira de Oliveira - Ministrio da Sade, Brasil Ana Maria Lacerda Kuchembuck - Instituto Pasteur, So Paulo, Brasil Ivanete Kotait - Instituto Pasteur, So Paulo, Brasil Sabrina Mendes - Ministrio da Sade, Brasil Marcelo Yoshito Wada - Ministrio da Sade, Brasil Fbio Camilo - Ministrio da Sade, Brasil Carlos Estnio - Ministrio da Sade, Brasil Tatiana Portela - Ministrio da Sade, Brasil Otvio Guimares - Ministrio da Sade, Brasil Mrcia Maria de Oliveira - Instituto Pasteur, So Paulo, Brasil Carla da Silva Goulart - Ministrio da Agricultura, Brasil

SUMRIO COMPENDIUM SUMARIO

SUMRIO / COMPENDIUM / SUMARIO


APRESENTAO / INTRODUCTION / PRESENTACIN ............................................................ 18 FIRST SESSION: HUMAN RABIES EPIDEMIOL OG Y, PROPHYLAXIS AND TREA TMENT EPIDEMIOLOG OGY TREATMENT Clinical and Epidemiological Features of Human Rabies Cases at San Lazaro Hospital, Philippines 1987 to 2006 ..................................................................................................................................... 23 Analyze the Possible Factors of Human Rabies Epidemic in China in 2005 ......................................... 24 Epidemiology of Human Rabies in the USA, 1980-2006 ................................................................... 25 Cambios del Perfil Epidemiolgico de la Rabia en Brasil: Estudios Antignicos y Genticos ................. 26 Control de la Rabia: 30 Aos de Trabajo Intersectorial de en las Amricas ......................................... 27 Description of Human Rabies Transmitted by Wild Animals - Brazil - 1996 to 2005 ......................... 28 Human Rabies : A Survival From the Past ............................................................................................ 29 Clinical Experience With Purified Vero Cell Rabies Vaccine (PVRV) for Pre-Exposure Immunization in Children ........................................................................................................................................... 30 Cross-Neutralization of Purified Chick Embryo Cell Culture Vaccine (PCECV) Against Non-Classical Rabies Virus Strains .............................................................................................................................. 31 Evaluation of the Information System of the Program of the Rabies Epidemiological Surveillance, Brasil - 2001 to 2005 ......................................................................................................................... 32 Post-Exposure Prophylaxis (PEP) Against Thai Street Rabies Virus And Eurasian Bat Lyssaviruses With Purified Chick Embryo Vaccine, Pepsin-Digested Erig, Human Rabies Immune Globulin and a Human Monoclonal Antibody (MAB 17C7) ....................................................................................... 33 Comparative Study on Immunogenicity, Safety and Tolerence of Purified Duck Embryo Vaccines (PDEV) Manufactured in India (VAXIRAB) and Switzerland (LYSSAVAC) ............................................ 34 Evaluation of Prophylaxis Treatment for Rabies During a Human Rabies Outbreak Due to Vampire Bat Exposure Turia Municipalty, Maranho State, Brazil, 2005 .................................................... 35 Limitations of F(AB)2 Fragments in Rabies Post-Exposure Prophylaxis ............................................... 36 Evaluation of The Indexes Of Rabies Prophylaxis And Control In The State Of Amazonas - 1996 To 2005 .............................................................................................................................................. 37 Biopterin Deficiency In Human Rabies ................................................................................................ 38

Therapy With Minocycline Aggravates Experimental Rabies In Mice .................................................... 39 siRNA-Directed Inhibition of Rabies Virus and Protection from Rabies Infection in Mouse Models..... 40 Lack of Efficacy of Ketamine Therapy in Experimental Rabies in Mice .................................................. 41 Antiviral Strategies Against the Lyssavirus Replication Complex: Peptides Mimicking the NH2 Terminus of the Rabies P Protein Have Antiviral Activity ...................................................................... 42 Evaluation of Human Monoclonal Antibodies CR57 and CR4098 Against a Global Panel of Rabies Viruses ................................................................................................................................................ 43 Web-Based Rabies Treatment Registry With Downloadable Protocol [www.chw.org/rabies] ........... 44 Prevalence Study on Vampire-Bat (Desmodus Rotundus) Bites in a Rural Population Following an Outbreak of Rabies-Related Deaths - Maranho State, Brazil, 2005 ................................................... 45 Description of Human Rabies Case in Veterinary, Minas Gerais State, 2006 ..................................... 46 Hbitos Alimenticios de la Comunidad hah y Riesgo de Exposicin a la Rabia Silvestre ............... 47 Manejo de Pacientes con Lesiones Graves Provocadas por Perro en reas con Ausencia de Casos de Rabia Canina, en el Estado de Chiapas Mxico ............................................................................... 48 Outbreak of Vampire-Bats Bites in Humans in Vila Velha-ES, 2006 .................................................... 49 Attacks of the Common Vampire Bats at Indigenous Villages From Northern Brazil ........................... 50 Anthropological Study of Rabies Outbreak in the City of Portel (PA/Amaznia/Brazil): Conceptions and Representations of the Illness for the Family of the Victims .......................................................... 51 Rabies Prevention and Global Awareness ........................................................................................... 52 SECOND SESSION: RABIES P ATHOGENESIS PA Identification of Pathogenicity Determinants of Rabies Virus (RV) and Assessment of the Role of the Rate of Virus Replication/Transcription in Pathogenesis ........................................................................ 55 Dominance of the Rabies Virus G Protein Carrying a Non-Pathogenic Determinant over the G Protein Carrying a Pathogenic Determinant ......................................................................................... 56 Consequences of the Innate Immune Response to Rabies Virus Infection ......................................... 57 Early Spread of Rabies Virus and the Development of Immunity ......................................................... 58 Rabies Pathogenesis and the Blood-Brain Barrier ................................................................................. 59 Clearance of Rabies Virus from the Central Nervous System ............................................................. 60 Host Response to European Bat Lyssavirus (EBLV) Infection, Part I: Susceptibility of Eptesicus Fuscus to EBLV-1 Following Different Routes of Infection .................................................................. 61 Differences between EBLV-1 and EBLV-2 Pathogenesis: Sheep as a Model of Disease Recovery ...... 62

A New Model to Study an Alternative Pathway for Rabies Transmission ............................................. 63 Non-Bite Transmission of Rabies Virus .............................................................................................. 64 Aislamiento de Virus Rabico en Glandulas Salivales de Murcielagos Insectivoros ................................ 65 Isolation of Rabies Virus in Mice and N2A Cells from Naturally Infected Bats of the State of So Paulo, Southern Brazil ......................................................................................................................... 66 Investigacin del Virus de la Rabia en Glndulas Salivares de Cnidos Silvestres (Cerdocyon Thous), Infectados Naturalmente, en el Estado de Piau, Brasil ........................................................................ 67 THIRD SESSION: DIA GNOSTIC, CHARA CTERIZA TION AND MOLECULAR EPIDEMIOL OG Y DIAGNOSTIC, CHARACTERIZA CTERIZATION EPIDEMIOLOG OGY Research Trends on Rabies in the FGI Federal Centre For Animal Health ....................................... 71 Laboratorial Diagnostic of Rabies: Comparation between Rabies Tissue Culture Inoculation, Mouse Inoculation and Fluorescent Antibody Tests, Using Bovine and Equine Specimens ............................. 72 Human Rabies Transmitted by Vampire Bats: Antigenic and Genetic Characterization of Rabies Virus Isolates from Ecuador and Brazil ......................................................................................................... 73 Molecular Epidemiological Study of Rabies Virus Isolates from Trinidad and Comparison with Isolates from the South American Continent ...................................................................................... 74 Molecular Epidemiology of Rabies Viruses from Domestic and Wild Mammalian of Brazil ................ 75 Rabies in Cerdocyon Thous (Crab Eating Fox), Northeastern Brazil: Genetic Identification and Fox Rabies Association with Dogs and Cats .............................................................................................. 76 Antigenic and Molecular Characterization of Positive Canine Samples to Rabies in Mexico from 1997 To 2006 ................................................................................................................................... 77 Antigenic Characterization of Rabid Animal Isolates, Northern Brazil, 2005 / 2006 ........................... 78 Molecular Analysis of the Aerial Cycle of the Rabies Virus in Mexico .................................................. 79 A Comparison Study of International Rabies Immunoglobulin Reference Standards ........................... 80 A Quantitative Indirect Elisa Test to Monitor the Effectiveness of Rabies Vaccination in Domestic and Wild Carnivores .................................................................................................................................. 81 Competitive Enzyme-Linked Immunosorbent Assays (CELISA) for the Detection of Anti-Rabies Antibodies in Wildlife Sera - Development, Validation and Practical Applications ................................. 82 Rabies Proficiency Tests: A Global Analysis of the 13 Ring Tests Performed from November 1999 to October 2004 ................................................................................................................................ 83 Assuring the Quality of Test Results .................................................................................................... 84 Recent Epidemiology of Southern African Lyssaviruses ....................................................................... 85

FOUR TH SESSION: RABIES IN TERRESTRIAL WILDLIFE: EPIDEMIOL OG Y AND CONTROL FOURTH EPIDEMIOLOG OGY CONTROL Epidemiological Surveillance of Rabies in Wild Animals: Road Kills from the Highways and/or Found In Urban Areas of the Municipalities of Jacare, Santa Branca and Salespolis, Vale Do Paraba -Sp, Southeastern Brazil .............................................................................................................................. 89 Rabies in Wild Species in Cear - Brazil, Epidemic and Enviroment Factors ......................................... 90 Spatial - Temporal Variation in Rabies Specimen Submission and Positivity during a Raccoon-Rabies Epizootic: Preliminary Results .............................................................................................................. 91 Effectiveness of Oral Rabies Vaccine to Manage Rabies in Striped Skunks (Mephitis Mephitis).............. 92 Trap-Vaccinate-Release and Rabies Management Programs to Manage Rabies Outbreaks in Striped Skunks (Mephitis Mephitis) In Flagstaff, Arizona ................................................................................... 93 Rabies DNA Vaccination Through Intradermal Into Ear Pinnae, Immunochemistry Study ................... 94 Tracking the Attenuation and Origin of Sad Derived Attenuated Rabies Virus Vaccines ......................... 95 ERA Vaccine-Derived Cases of Rabies in Wildlife and Domestic Animals in Ontario, Canada during the Period of 1989 To 2004 .............................................................................................................. 96 Development of a Recombinant Canine Adenovirus 2 - Rabies Virus G Protein Vaccine (CAV2-RV G)..... 97 Safety Studies of Rabies Strain ERA G 333 as a Candidate for Oral Rabies Vaccine ............................. 98 A Benefit-Cost Analysis of the Ontario Raccoon Oral Rabies Vaccination Program ............................. 99 The Raccoon - The Ultimate Wild Carnivore Challenge Facing Oral Rabies Vaccination ..................... 100 Oral Vaccination of Skunks with a Canine Adenovirus Type 2-Rabies Recombinant Virus Vaccine ....... 101 Evaluation of Two Candidate Vaccines for Oral Vaccination of Skunks against Rabies ........................... 102 Evaluation of Oral Wildlife Rabies Vaccination: Interpretation of Field Serology and Results of Challenge in Captivity ........................................................................................................................... 103 Risk-Based Cost Modeling of Oral Rabies Vaccine Interventions for Raccoon Rabies ......................... 104 FIFTH SESSION: RABIES IN BA TS BATS Vampire Myths, Vampire Bats and the T yranny of Theories ................................................................ 107 Primer Relato de Murcilago Hematofago (Desmodus Rotundus) Infectado por el Virus de la Rabia, Encontrado en un Centro Urbano, SP , Brasil ....................................................................................... 108 Primera Epizootia de Rabia en Desmodus Rotundus en la Costa del Per ......................................... 109 Rabia en Murcilago Hematfago Diphylla Ecaudata y Desmodus Rotundus en el Estado de Piau, Nordeste del Brasil .............................................................................................................................. 110 Epidemiologa de la Rabia en Chile, 1989 - 2005 ............................................................................... 111

Genetic Characterization of Brazilian Bat Rabies Viruses. Evaluation of the Pathogenicity and Cross Protection in Mice ................................................................................................................................. 112 Vigilancia Epidemiolgica de La Rabia en Murcilagos Realizada en Braslia, DF , Brasil ............................ 113 Rabia en Murcilagos en el rea Urbana de Belo Horizonte, Minas Gerais, Brasil: Identificacin de Especies y Estudios Antignicos ............................................................................................................. 114 Bats Rabies in the Period between 2000 - 2006, in Ribeiro Preto County, So Paulo State, Brazil .... 115 Bat Rabies in So Jos do Rio Preto, Southeast of Brazil ....................................................................... 116 Virus Rabia en Murcilagos Hematofagos y No Hematofagos de Areas Urbanas y Rurales en el Departamento del Valle, Colombia ....................................................................................................... 117 SIXTH SESSION: SURVEILLANCE AND CONTROL OF ANIMAL RABIES Identification of Shortcommings in Rabies Control Methods of Kwazulu-Natal ..................................... 121 Rabies Surveillance in The United States, 2005-06 ............................................................................... 122 Management of Illegal Introduction of Pet Carnivores: The Cases Recorded in Italy in 2005 and 2006 .................................................................................................................................................... 123 Quantification of Rabies Neutralizing Antibodies in Dogs from a Rabies Endemic Area in Northern Colombia 2004 .................................................................................................................................... 124 Canine Rabies Elimination from the United States: An Epidemiological and Virological Perspective ....... 125 Relato de un Caso de Rabia Canina, en el Municpio de Corumb, Mato Grosso do Sul, Brasil, en 2006 ..................................................................................................................................................... 126 Reintroduccin de la Rabia Urbana en Lima Metropolitana y el Callao - Per ........................................ 127 Estrategia para el Control de la Rabia de los Herbvoros en el Estado de So Paulo - Brasil, 2000 a 2005 ..................................................................................................................................................... 128 Capture and Control of Bats and Herbivorous Vaccination during an Outbreak of Human Rabies Trasmitted by Vampire-Bats, Maranho State, Brazil, 2005 ................................................................... 129 Estimated Dog Population Analysis to Perform Epidemiological Surveillance and Procedures of Dogs Antirabies Vaccination to Enhance Tasks in State Of So Paulo, Brazil, During 1996 to 2005 ............... 130 Video para la Prevencion y el Control de la Rabia Paralitica Bovina ........................................................ 131 SEVENTH SESSION: STRA TEGIES FOR THE CONTROL OF ANIMAL POPULA TION STRATEGIES POPULATION Dinmica de la Poblacin Canina con Dueo en reas Urbanas y Rurales del Estado de Mxico, 1979-1999 .......................................................................................................................................... 135

Modelo de Prospeccin de Poblacin Canina para la Planificacin de Acciones Sanitarias ...................136 Resultados del Proyecto Masivo de Esterilizacin Qumica con Gluconato de Zinc Neutralizado con Arginina en Perros Machos Residentes de los Estados de: Hidalgo, Mxico y Puebla ......................... 137 The Impact of the Program of Ambient Health in the Control of the Urban Rabies in the City of Recife ...................................................................................................................................................139 POSTER I Estudio Epidemiolgico de la Encefalitis Rbica Humana en el Estado de Mxico, 1979- 2003 ......... 143 Relato de Caso: Avaliacin de los Tratamientos Antirbicos en Personas que Tubieron Contacto con Bovino Positivo para Rabia ................................................................................................................... 144 Epidemiologics Aspects about Human Rabies in Pernambuco .............................................................146 Epidemiologic Control Program of Rabies in Pernambuco: Epidemiologic and Operational Aspects ...147 Difficulties Faced for Professionals to Persuade Persons to Realize Prophylaxis for Rabies ................ 148 Immunopathologic in Rabies Infection in Lines of Mice Selected for High or Low Acute Inflammatory Reaction ...............................................................................................................................................149 Aspects of Rabies Prophylaxis, Teresina Municipality, Piau State, 2005 .............................................. 150 Importance of Preexposition Prophylaxis of Rabies in High-Risk Groups ........................................... 151 Dogs Bite in a Veterinary Staff on Veterinary Hospital of University of Brasilia (UnB) - Case Report .......152 Ecology and Management of Vampire Bat Attacks upon Human Beings in the Poor Villages from the Amazonas State, Northern Brazil .........................................................................................................153 Epidemiologic Control Program of Rabies in Pernambuco: Epidemiologic and Operational Aspects .......154 Anlisis Comparativa de los Programas de Control y Prevencin de la Rabia en Amrica Latina .........155 Development of an Imunoperoxidase Inhibition Assay (IIA) For Rabies Antibody Detection ............. 156 Rabies Virus Stability after Twenty Passages in Mice ............................................................................. 157 A Collaborative Study on the Testing of Equine Antirabies Serum by a Virus Neutralization Test in BHK- 2 1 Cells .................................................................................................................................... 158 Antigenic and Genetic Characterization of First Rabies Virus Isolated From Eumops Perotis Bat of Brazil .................................................................................................................................................... 159 Rabies Virus Neutralizing Antibodies: Comparison of Two Evaluation Test in Cell Culture ................. 160 Comparison of Rffit Performed in Lab-Tek and in 96-Well Microtitre Plates ........................................161 The So Paulo State/Brasil Rabies Control Program, Under the Vision of Its Coordination (1996 2005) .................................................................................................................................................162

Polymerase Chain Reaction (RT-PCR) and Heminested-RT PCR (HN RT-PCR) Evaluation in Fresh and Decomposed Central Nervous System Samples for Rabies Diagnosis ........................................ 163 Evaluation of the Human Antirabic Service, Minas Gerais, Brazil, 1999 To 2004 ................................. 164 Health Education Program for Rabies, Maranho State, Brazil, 2006 ................................................... 165 Influence of Different Serum-Free Media in the Rabies Virus Production ............................................. 166 Analysis of the Last 16 Years (1990 - 2005) - Post Exposure Prophylaxis So Paulo (Brasil) State ..... 167 Laboratorial Profile of the Rabies Diagnosis in Evandro Chagas Institute, 2003 - 2005 ....................... 168 The Prophylaxis Profile of the Human Rabies in the State of Cear ..................................................... 169 Epidemiology of Human Rabies Post-Exposure Prophylaxis and Relationship between Rabies Control Strategies in Centro Sul Area of Rio De Janeiro, Brazil ......................................................................... 170 Implementacin de una RT-PCR Anidada como Alternativa para la Deteccin del Virus de la Rabia en Muestras Infectadas .............................................................................................................................. 171 The Profile of the Prophylaxis of Human Rabies in the Regional Cell of Health of Caucaia - CE .......... 172 Evaluation of the Anti-Rabies Reexposure Treatments in the Clinic of the Pasteur Institute of So Paulo, Brazil .......................................................................................................................................... 173 Analysis of Information Related to Prophylactic Treatment against Rabies Given to Residents of Jaboticabal, So Paulo, Brazil ................................................................................................................. 174 Measurement of the Immune Response to Rabies Vaccination Using a Novel and Traditional Assays ...... 175 Principal Risk Factors for Human Rabies Transmitted by Vampire Bats, Maranho State, Brazil, 2005 ...... 176 Descript of Two Human Rabies Cases through Canine Transmition in the State Of Maranho in 2006 and the Importance of Revaccinate the Dog Which Were Already Vaccinated .............................177 Budgetary and Financial Resources Versus Human and Canine Rabies Control in the Municipal District of Rio Branco - Acre - Brazil, From 1997 To 2002 .................................................................178 Human and Canine Rabies Control Program Evaluation in the Acre State, Brazil, From 1996 To 2002 .. 179 Human Rabies and their Animal Exposures: Descriptive Analysis from Transmission Cycles in Twenty Years ......................................................................................................................................... 180

POSTER II Dog Rabies Prevention Proposal by the State Health Secretary for the Counties of Rio de Janeiro ...... 183 Acciones de Vigilancia Epidemiologica que Permiten al Estado de Baja California (Mexico) Mantenerse por 24 Aos Sin Casos de Rabia Urbana 2001-2006 ........................................................................ 184

Contribution of Zoonosis Diagnosis Service, Unesp-Botucatu/Sp-Brazil, to Rabies Diagnosis in the Period of 1991 to 2005 ................................................................................................................... 185 Rabid Bats and Pets ............................................................................................................................ 186 Fire, Bats and the Risk of Rabies Transmisson .................................................................................... 187 Macroscopic and Microscopic Finding in a Bovine Rabies Case at Distrito Federal (Brazil) ................. 188 Body Contact on Bat Species Harboring in the Same Day Roost: A Possible Way to Interspecific Rabies Transmission among Bats ....................................................................................................... 189 Presence of Rabies Virus in Fruitful Bat of the Species Artibeus Lituratus in Vitria-ES ........................ 190 Immune Response to BHK-21 Cell Rabies Vaccine Produced at Instituto Butantan in Bovines .......... 191 Situation of Animal Rabies in Brazil, 2005 ........................................................................................... 192 Characterization of Dog and Cat Bites in Porto Alegre/Brazil in the Year of 2005 and Its Role in The Surveillance of Rabies .......................................................................................................................... 193 Clinical Aspects of Rabies in Bovine in the Centro-Oeste of Brazil ..................................................... 194 Cerebrospinal Fluid Evaluation in Bovine with Rabies .......................................................................... 195 Differential Diagnostic of Bovine Rabies in the West-Center Region of Brazil ..................................... 196 First Detection of Ehv-4 in an Equine Case of Suspected Rabies in Rio de Janeiro State, Brazil .......... 197 Descriptive Analyze of Rabies Distribution, in the Year of 2005, in Rio De Janeiro State, Brazil ......... 198 Evaluation of Rabies Virus Neutralizing Antibodies Titres of Pets That Moves to the Europe Union from Brazil .......................................................................................................................................... 199 Epidemiological Profile of the Confirmed Cases of Animal Rabies between 2000 and 2005 ........... 200 Evaluation of the Rabies Activities Realized In Centers of Control of Zoonosis - Brazil ...................... 201 Dog Rabies Transmitted By a Bat in Urban Area of Teresina Municipality-Piau State-Brazil-2006. Reported Case .................................................................................................................................... 202 Chiroptera Monitoring In a Cave Northwest of Minas Gerais, 2004-2005 ...................................... 203 The Experience of Activities of Blockade of Rabies in Bats in Botucatu - Sp, Brazil .............................. 204 Survellaince of Rabies in Bats in the City of Botucatu - SP ................................................................... 205 Rabia en Murcilagos No-Hematfagos en la Area Urbana de Caapava, So Paulo, Brasil ............... 206 Course of Wild Rabies - An Alternative to Implement Actions ............................................................ 207 Caucaia-CE: The Scenery of the Rage in The World .......................................................................... 208 Animal Rabies in Pernambuco from 2000 To 2005 .......................................................................... 209 Zoonosis Control Versus Social Pleas ................................................................................................ 210

Rabies in the Northwestern Sao Paulo State, Brazil: Antigenic and Genetic Characterization of Virus Isolates ............................................................................................................................................... 211 Implementacin de un Sistema de Vigilancia de Rabia en Vampiros en la Costa del Departamento de Lima - Peru ......................................................................................................................................... 212 Eficacia a Campo del Vampiricid Gel Para el Control Selectivo Indirecto de los Murcilagos Hematfagos (Desmodus Rotundus - E. Geoffroy, 1810) en Brasil .....................................................213 A Canine Kept in Observation after Contact with a Rabies Positive Tadarida Brasiliensis ......................214 Gray Fox Research to Support Oral Rabies Vaccination Programs in Texas: An Overview .................... 215 Poblaciones con Alta Consanguinidad como Factor Desencadenante de Brotes de Rabia en las Poblaciones de Vampiros ....................................................................................................................... 216 Vaccinating Cattle to Protect Aganist Rabies Transmission from Rabid Bat Populations en Mexicos Endemic Zones: An Analysis Using Economic Models of the Benefits and Cost ................................... 218 Population Genetic Study of Desmodus Rotundus in an Area of High Bat Rabies Incidence in Cattle, San Luis Potos State, Mexico .................................................................................................................219 Platelia Rabies Ii: A Helpful Tool for Monitoring Raccoons Rabies Vaccination Programs by Antibodies Titration ................................................................................................................................................. 220

APRESENTAO INTRODUCTION PRESENTACIN

APRESENTAO

A Reunio Internacional de Raiva nas Amricas - RITA vem sendo, desde 1990, um dos mais importantes fruns de atualizao cientfica sobre a raiva. Neste ano o Ministrio da Sade do Brasil tem a honra de sediar a XVII RITA. Participam conosco importantes pesquisadores dos mais diversos pases, alm de profissionais da rea da sade, agricultura, meio ambiente, universidades, trazendo tambm suas experincias de campo, na vigilncia e controle da raiva. A Comisso Cientfica Nacional procurou distribuir os 180 trabalhos inscritos de forma racional e objetiva, abordando os grandes temas que compem o estado da arte da raiva. A Comisso Organizadora trabalhou ativamente, para proporcionar toda a infra-estrutura do evento e as atividades culturais e sociais, tornando ainda mais agradvel este nosso encontro. Agradecemos a todos que contriburam para o xito da XVII RITA, instituies pblicas e privadas, ressaltando que a raiva uma doena cujo controle s ser possvel a partir de uma viso multidisciplinar e uma ao multiinstitucional. Ao Comit Cientfico Internacional, o nosso agradecimento pelo apoio constante. Esperamos que todos os participantes tenham um debate proveitoso, nesses dias que se seguem.

Rosely Cerqueira de Oliveira Presidente da Comisso Cientfica

18

INTRODUCTION

The International Meeting on Rabies in the Americas - RITA has been one of the most important forums for scientific updating on rabies since 1990. This year, the Brazilian Ministry of Health has the honor to receive the XVII RITA. Important researchers, from the most diverse countries, participate with us, besides professionals from the areas of public health, agriculture, the environment and universities, also bringing us their experience on their field activities The National Scientific Committee aimed to distribute the one hundred and eighty (180) papers submitted in a rational and objective manner, approaching the great questions, which compose the state of the art in rabies. The Organizational Committee worked actively to prepare the infrastructure for this event and the cultural and social activities, making your stay even more enjoyable. Our thanks to the International Scientific Committee for the constant support. We hope all participants will take the best advantage of their stay with us

Rosely Cerqueira de Oliveira President of the Scientific Committee

19

PRESENTACIN

La Reunin Internacional de la Rabia en las Amricas - RITA, desde 1990, constituye oportunidad singular para la actualizacin cientfica de la comunidad internacional sobre esta zoonosis. En este ao el Ministerio de Salud de Brasil tiene el honor de ser sede de la XVII RITA. Participan en este evento internacional importantes investigadores de diversos pases y de las diferentes reas de especializacin: salud, agricultura, medio ambiente, universidades, entre otras, para compartir sus experiencias de campo, relacionadas con la investigacin, la vigilancia epidemiolgica y el control de la rabia. La Comisin Cientfica Nacional procur distribuir los 180 trabajos inscriptos, de manera racional y objetiva, alrededor de los grandes ejes temticos que configuran el estado de la arte de la rabia. La Comisin Organizadora trabaj activamente para proporcionar toda la infraestructura y comodidades logsticas para el desarrollo del evento cientfico, as como de la programacin cultural y social, en un ambiente amigable, de unin y solidaridad continental. Gracias a todos los que contribuyeron para el xito de la RITA XVII, tanto a las instituciones pblicas como privadas participantes, en cuyo marco institucional ha sido posible destacar que la rabia es una enfermedad cuyo control solo ser posible a partir de una visin multidisciplinaria y de una accin multinstitucional. A la Comisin Cientfica Internacional, nuestras gracias por su incansable apoyo a la organizacin y desarrollo de la reunin. A todos los participantes anticipamos nuestro reconocimiento por su activa participacin, su actitud positiva y sus valiosas contribuciones para un eficiente abordaje de la rabia, como problema que configura los perfiles de salud de las poblaciones ms vulnerables de los pases de las Amricas.

Rosely Cerqueira de Oliveira Presidente de la Comisin Cientifica Nacional

20

FIRST SESSION
HUMAN RABIES EPIDEMIOLOGY, PROPHYLAXIS AND TREATMENT

21

HUMAN RABIES EPIDEMIOLOGY, PROPHYLAXIS AND TREATMENT

22

FIRST SESSION

CLINICAL AND EPIDEMIOLOGICAL FEATURES OF HUMAN RABIES CASES AT SAN LAZARO HOSPITAL, PHILIPPINES 1987 TO 2006
Efren M. Dimaano 1, Stephen J. Scholand 2, Maria Theresa P . Alera 1
1) San Lazaro Hospital, Santa Cruz Manila, Philippines, 2) A Rabies Free World, Philadelphia, PA, USA

Dr. Maria Theresa P . Alera, MD - tetalera@yahoo.com

Phillipines

Rabies continues to be a threat in the Philippines where the disease is highly endemic. San Lazaro Hospital is the national referral center for infectious diseases where we care for human rabies patients at our Central Nervous System Pavilion and treat rabies exposed patients in our Animal Bite Treatment Center. From January 1987 through June 2006, 1,839 human rabies patients were admitted. We present results from a retrospective chart review of these cases. Diagnoses were made clinically by experienced physicians. Clinical features on presentation included inspiratory spasm due to phobic stimuli (air and water) (100%), agitation (16%), and fever (9%). The youngest patient was aged 2 while the oldest was 89. Adults outnumbered children (age <20) by 2 to 1 (66% to 34%) and male gender predominated (69% to 31%). Our data show a clinical correlation between location of bite site and duration of incubation period. The shortest incubation was 6 days with multiple head and neck bites while the longest incubation was 5 years with bites to the lower extremities. All patients were bitten by either unvaccinated house pets or stray dogs/cats except for a few cases which revealed no history of injury. 97.05% of rabies cases were dog related, 2.90% were cat related, and 0.05% was unknown. Of the 1,839 rabies patients, only 136 (7.4%) had sought medical attention prior to disease onset. Patients largely derived from lower socioeconomic groups with 96% considered totally indigent and 51% unemployed. Knowledge of these clinical and epidemiological characteristics will help future efforts to reduce rabies suffering in the Philippines.

23

HUMAN RABIES EPIDEMIOLOGY, PROPHYLAXIS AND TREATMENT

ANALYZE THE POSSIBLE FACTORS OF HUMAN RABIES EPIDEMIC IN CHINA IN 2005


T. Qing 1, Song Miao 1, Li Hao 1, Liang Guodong 1
1) Chinese Center for Disease Control and Prevention, Beijing, China

Dr. Tang Qing - q_gtang22@yahoo.com

China

To collect epidemic data from national rabies surveillance systemretrospect investigation on human rabies cases and patients who receive post exposure treatment, laboratory investigate the collected samples of the cases and animal hosts of rabies, analyze all the above data occurred in 2005 in order to understand the possible factors that cause rabies epidemic in south part of China in recent years. Human rabies case number is 2548 in 2005. 885 of human cases were investigated, 60.56%, 49.04%, 96.16% have not received wound treatment, vaccination and immunoglobulin injection respectively. Among 278 post exposure treatment cases, 86% and 45% of them should receive vaccination and immunoglobulin, but actually only 16.92% and 10.77% of them received vaccination and immunoglobulin. Among the exposed patients, 89.95% received wound treatment, 94.93% were immunized, and 8.56% received immunoglobulin; and 94.86% of them accomplished the full rabies post-exposure prophylaxis course, but 8.56% of them didnt accomplish it. 88.50% human cases and 92% exposure persons were bitten by dogs. Dog density is about 3.20 to 13.37 per 100 persons. Dog vaccination coverage rate is totally different in different areas, lower from 5.31% to high 75.11%. The investigation of animal host infection of human rabies was done by both of DFA for rabies virus antigen detection and RT-PCR for rabies virus RNA detectionchecked by Real-time PCR in three highest epidemic provinces and showed the rabies infectivity of the animal host in these areas were 2.93% to 6.40%. The percentage of post-exposure prophylaxis of human rabies cases is lower. Part of the cases and the exposed patients dont accomplish full rabies post-exposure prophylaxis course. Most of the cases and exposure patients were bitten by dogs. Dog density and rabies virus infectivity of the animal host in high epidemic areas are higher. Dog vaccination coverage is lower in most of the epidemic areas. All of these factors are related with the human rabies epidemic in China in 2005.

24

FIRST SESSION

EPIDEMIOLOGY OF HUMAN RABIES IN THE USA, 1980-2006


Cathleen A. Hanlon 1, Alice S.Chapman, Lilian Orciari 1, Michael Niezgoda 1, Pamela Yager 1, Eric J. Mandel, John W. Krebs 1, Charles E. Rupprecht 1
1) Centers for Disease Control and Prevention, Atlanta, GA, USA

Dr. Cathleen Hanlon - chanlon@cdc.gov

USA

Rabies is an acute, rapidly progressive, fatal viral encephalitis. In the USA, the virus is maintained in a variety of wildlife reservoirs, including raccoons, skunks, foxes, and numerous bat species. Whereas rabies is enzootic among wildlife, human rabies is rarely diagnosed in the USA. In addition to reviewing the last published summary of cases from 1980 to 1996, this study examines the epidemiology of human rabies from 1997 through 2006. More than half of the cases are diagnosed postmortem, suggesting that clinical suspicion remains low even in the presence of compatible clinical signs. Specific patterns of suspect clinical signs observed in patients (e.g. nonspecific prodrome lasting 3-4 days, followed by rapid onset of encephalitis) should prompt clinicians to consider rabies in the differential diagnosis even in the absence of a definite animal exposure history. A majority of human rabies cases acquired indigenously in the USA are caused by rabies virus variants associated with insectivorous bats. Epidemiologic trends revealed among reported cases from 1980 through 2006 indicate a transmission patterns consistent with unrecognized encounters with bats or a dismissal of the risks from a seemingly minor encounter. To date, no patients had sought post-exposure prophylaxis prior to becoming ill, apparently as a result of not recognizing a potential exposure. Given the hope of future experimental treatment, current epidemiological trends underscore the need for higher clinical suspicion and rapid antemortem diagnosis.

25

HUMAN RABIES EPIDEMIOLOGY, PROPHYLAXIS AND TREATMENT

CAMBIOS DEL PERFIL EPIDEMIOLGICO DE LA RABIA EN BRASIL: ESTUDIOS ANTIGNICOS Y GENTICOS


Rosely Cerqueira de Oliveira 1, Marcelo Y. Wada 1, Lucia Regina Montebello Pereira1, Rosngela Rosely Cerqueira de Oliveira 1, Marcelo Y. Wada 1, Lucia R. Montebello 1, Rosngela Machado 1, Pedro Carnieli Jr 2, Juliana G. Castilho 2, Maria Luiza Carrieri 2, Ivanete Kotait 2
1) Secretaria de Vigilncia em Sade, Ministrio da Sade, DF, Brasil 2) Instituto Pasteur, So Paulo, SP , Brasil

Dr. Rosely Cerqueira de Oliveira - oliveira.rosely@gmail.com

Brasil

El objetivo fundamental de los programas gubernamentales de control de la rabia canina est asociado al decrecimiento del nmero de casos de rabia en humanos, puesto que la especie canina es evidenciada como el principal reservorio del virus de la rabia en reas urbanas. En Brasil, de 175 casos humanos (151 transmitidos por perros) registrados en 1980, disminuyeron a 17 casos (14 transmitidos por perros) en el 2003. Sin embargo, a partir de 2004 y 2005 este nmero volvi a incrementarse a 30 y 44 casos, respectivamente (22 y 42, transmitidos por murcilagos hematfagos). Estos fallecimientos ocurrieron en la regin norte y nordeste del pas, en 7 municipios, en los cuales la poblacin estaba expuesta al riesgo de agresin por murcilagos hematfagos. La disminucin de los casos de rabia canina permiti una vigilancia de la rabia de los animales silvestres, y el cambio del perfil fue identificado por la aplicacin rutinaria de mtodos de biologa molecular, tales como la tipificacin antignica, utilizando un panel de anticuerpos monoclonales cedido por el CDC/Atlanta/EUA y la realizacin de la RT-PCR y del secuenciamiento genmico. En el ciclo areo qued evidente la participacin del Desmodus rotundus como el ms importante reservorio de la rabia silvestre del pas. La identificacin de la variante (AgV3) fue realizada en diversas especies domsticas y silvestres, murcilagos del gnero Artibeus sp y Molossus sp. Estos aislamientos demostraron la ocurrencia frecuente de spillover en nuestro medio. Las especies de murcilagos insectvoros, como Tadarida brasiliensis e Lasiurus ega generalmente presentan una variante especie-especfica, identificadas como variantes 4 e 6, respectivamente. Estos ciclos e otros, en los cuales hay la participacin de los murcilagos insectvoros, han sido encontrados en las regiones sudeste y centro-oeste del pas. En la regin nordeste, no obstante, hay otros ciclos silvestres mantenido por cnidos del gnero Cerdocyon sp., cuyo virus tiene origen probable de cnidos domsticos, y del mono Callithrix jacchus, que se mantn entre estos animales y ocasionalmente ha infectado el hombre. Tanto el ciclo areo, que ocurre frecuentemente en centros urbanos, como los silvestres terrestres, representan riesgos para la salud pblica, dadas las caractersticas sinantrpicas de los animales comprometidos, principalmente murcilagos del gnero Artibeus sp. y cnidos silvestres, del gnero Cerdocyon sp.

26

FIRST SESSION

CONTROL DE LA RABIA: 30 AOS DE TRABAJO INTERSECTORIAL DE EN LAS AMRICAS


M.C. Schneider 1, A. Belotto,1, D. Fernandes 1, F.L. Leanes, 2, M.A. Genovese 2
1) Pan American Health Organization, Washington DC, USA, 2) PANAFTOSA, Rio de Janeiro, Brazil

Dr. Rosely Cerqueira de Oliveira - oliveira.rosely@gmail.com

USA

Los pases de las Amricas tienen una tradicin de ms de 30 aos de trabajo intersectorial entre salud y agricultura en el control de la rabia. Desde 1968 la OPS convoca a los Ministerios de Agricultura para discutir temas comunes, entre estos estuvo la decisin poltica de crear el Programa Regional de Eliminacin de la Rabia Transmitida por Perros en Amrica Latina y el Sistema de Informacin Regional de Vigilancia Epidemiolgica de la Rabia en las Amricas (SIRVERA). Desde 1969 los pases de la Regin integran el SIRVERA, quien colecta informacin referente a la rabia humana y animal. La informacin recopilada se publica en boletines epidemiolgicos semanales que son distribuidos a los Ministerios de Salud y Agricultura, y al alcance pblico en la pgina electrnica de PANAFTOSA. En base a esta informacin, hubo una reduccin del 93% en casos de rabia humana transmitidos por perros en Amrica Latina, entre 1990 y 2005 solo 11 casos. Este logro, fue uno de los temas tratados en la ltima Reunin Interamericana a Nivel Ministerial en Salud y Agricultura (RIMSA 14), realizada en la ciudad de Mxico en 2005. El nmero de casos humanos transmitidos por murcilagos hematfagos fue ms del doble de los transmitidos por perros, entre 2004 y 2005 se detectaron 51 casos. La actuacin intersectorial trabajando por el control de la rabia es de suma importancia, para lograr la erradicacin de la rabia en las Americas. Con este propsito se est convocando a los representantes de agricultura a participar de la prxima Reunin de Directores de Programas Nacionales de Control de la Rabia (REDIPRA) y se est organizando una reunin intersectorial de expertos sobre rabia human transmitida por murcilagos, ambas sern realizadas en octubre de 2006. Varios pases de la Regin vienen realizando reuniones, talleres y acciones de control de brotes de rabia humana y animal de forma conjunta entre salud y agricultura, tanto a nivel nacional como local. Se sugiere que este tipo de accin se incremente desde el nivel internacional hasta el local.

27

HUMAN RABIES EPIDEMIOLOGY, PROPHYLAXIS AND TREATMENT

DESCRIPTION OF HUMAN RABIES TRANSMITTED BY WILD ANIMALS - BRAZIL 1996 TO 2005


Marcelo Yoshito Wada 1, Lucia Regina Montebello 1, Sabrina Lzaro Mendes 1, Rosely Cerqueira de Oliveira 1
1) Secretaria de Vigilncia em Sade, Ministrio da Sade, DF, Brasil

Dr. Marcelo Yoshito Wada - marcelo.wada@saude.gov.br

Brasil

Introduction. Rabies is a preventable viral zoonosis that infects the central nervous system of mammals, causing encephalopathy with a mortality rate of 100%.Human rabies transmitted by dogs are decreasing in Brazil and wild animals are being identifying as source of human rabies. Methods. This study describes human rabies cases reported to federal level in Brazil from 1996 to 2005. There were evaluated reporting forms of the human rabies cases(HRC) and a standard questionnaire was applied during the outbreaks to collect data about signals and symptoms. Medical records were analyzed. Results. A total of 254 HRC were reported, 34% were transmitted by wild animals, 90% by bats, 9% marmosets, 1% others species. Among HRC transmitted by wild animals, 53% were female, median age was 11 ranging from 1 to 62 years. North region contributed with 48% of the cases, southern 7% and middle west 1%. Exposure in urban area: 4%. Median incubation period from exposure date to onset of symptoms was 38 ranging from 16 to 244 days. Median evolution to death was 7 ranging from 2 to 25 days. Among 30 HRC who were applied the standard questionnaire, the principal symptoms were: 90% fever, 83 paralysis, 73% myalgia, 75% dysphagia, 67% dyspnea, 63% hypersalivation, 63% agitation, 63% headache, 63% paresthesia, 57% pain throat, 53% abdominal distension, 53% urinary retention, 50% disorientation. Conclusions. HRC transmitted by wild animals are being a reemergence challenge, where vampire-bats are identified as source of outbreaks in areas with hard access and poor knowledge. Wild animals are responsible to keep rabies viruses in these areas and needs to create prevention and control measures, including health education, prophylaxis for rabies and control of animals. It is important to alert physician to suspect rabies when symptoms include paralytic disease without furious symptoms.

28

FIRST SESSION

HUMAN RABIES : A SURVIVAL FROM THE PAST


Yolande Rotivel 1, Maryvonne Goudal 1
1) Institute Pasteur, Paris, France

Dr. Yolande Rotivel - yrotivel@pasteur.fr

France

Even in areas where canine rabies has been eliminated, human deaths still occur. As shown in Western Europe, to be efficient, eradication of canine rabies should be at least regional, or even better global. Rabies epidemiology in animals as well as in humans is changing. Rabies deaths after organ transplantations were only diagnosed because of the donor state on the patient. What if they had not been considered as potential donors? It should be considered that a number of human rabies cases are still unknown, even in outstanding health care systems. In this situation, rabies immunization, whether pre or postexposure, is the ultimate weapon. Current rabies vaccines are safe and potent. They are getting more and more available all over the world. New concepts of preexposure immunization associated with simple booster injections schedules validated by extensive clinical trials on the field as well as accurate immunological studies are bitterly needed. Moreover, preexposure immunization avoids the use of RIG, which is both rare and costly. Preexposure immunization should also decrease the number of treatment failures possibly caused by the absence of RIG injection within a suitable period of time after the exposure. Potential adverse effects of RIG are also to be considered. As far as cell-culture rabies vaccines are concerned, availability is no more the major difficulty. Actually, distribution networks, knowledge of prescriptors and education of public are the true limitations. Rabies might not be eradicated as reservoirs seem to be multiple, especially in chiropters. Eradicating rabies in wildlife might be wrong from an ecological point of view. Lyssaviruses might only be controlled. In the end, what are ours goals? First, eradicate human disease; second, eradicate rabies in domestic animals, whether pets or not. The second goal has been achieved by immunization in many places. Why not succeed in the first one?

29

HUMAN RABIES EPIDEMIOLOGY, PROPHYLAXIS AND TREATMENT

CLINICAL EXPERIENCE WITH PURIFIED VERO CELL RABIES VACCINE (PVRV) FOR PRE-EXPOSURE IMMUNIZATION IN CHILDREN
A. Rassouli 1, C. Weinberger 1, G. Houillon 2
1) Medical Affairs International, 2) Medical Affairs, Sanofi Pasteur, Lyon, France

Dr. Anvar Rassouli - anvar.rassouli@sanofipasteur.com

France

Introduction: Children have a high risk of exposure to rabies in countries where the disease is endemic. Preexposure anti-rabies vaccination offers increased protection against any type of exposure and simplifies postexposure treatment if required, with a reduced number of doses and lowering the need for immunoglobulin treatment. Methods: Data is available from three randomized studies. Purified vero-cell rabies vaccine (PVRV, Verorab, Sanofi Pasteur) was given intramuscularly (IM) at 2, 4 and 12 months of age, simultaneously with DTPIPV vaccine (N=84). A similar study evaluated three intradermal (ID) injections at 2, 3 and 4 months or two IM doses at 2 and 4 months, both with boosters at 1 and 5 years (N=235). A third study evaluated IM and ID administration at days 0, 7, 28 and a booster at 1 year in children 5-12 years of age (N=190). Results: All infants receiving intramuscular PVRV at 2 and 4 months of age (two studies) had anti-rabies titres 0.5 IU/mL 1 month after primary vaccination and 75-97% had titres 0.5 IU/mL at 1 year. Seroprotection rates decreased from 100% after booster vaccination to 63.3-80% at 5 years. All children given intradermal PVRV at 2, 3, 4 months (N=117), or intramuscular PVRV at 2 and 4 months of age (n=118) were seroprotected 1 month after the last dose; GMTs were 30.6 IU/mL in the IM group and 12.0 IU/mL in the ID group. Five years after booster, 80% of children in the IM and 54% in the ID group had titers 0.5 IU/mL, but all were seroprotected, following a second booster. In children 5-12 years of age, 99% given ID, and 100% given IM vaccine had GMTs e0.5 IU/mL at 56 days. A booster dose at day 365 provoked a strong anamnestic response in both groups, with all children seroprotected throughout 3 years of follow-up. GMTs at 180 and at 365 days were higher in the children given PVRV intramuscularly. Conclusions: PVRV administered concomitantly with DTP-IPV or DTP and OPV, was highly immunogenic, without clinically significant vaccine interactions and protected a majority of children for 5 years after booster vaccination. In young children, pre-exposure administration or four doses following a 0, 7, 28, 365 day schedule resulted in durable protection over at least 3 years.

30

FIRST SESSION

CROSS-NEUTRALIZATION OF PURIFIED CHICK EMBRYO CELL CULTURE VACCINE (PCECV) AGAINST NON-CLASSICAL RABIES VIRUS STRAINS
Claudius Malerczyk 1, Nol Tordo 2, Thomas Mller 3
1) Novartis Vaccines, Marburg, Germany, 2) Institut Pasteur, Paris, France, 3) Friedrich-Loeffler-Institut, Germany

Dr. Claudius Malerczyk - claudius_malerczyk@chiron.com

Germany

Background: Human rabies remains an invariably deadly disease, accounting for an estimated 55,000 deaths annually. While the majority of human cases occur in Africa and Asia after exposure to classical rabies virus (RABV), in Europe and Australia deaths have occurred after exposure to bat transmitted non-classical rabies strains like European bat lyssavirus (EBL1 and EBL2) and Australian bat lyssavirus (ABL). Undoubtedly, modern tissue culture vaccines like PCECV convey protection against RABV via pre-exposure prophylaxis, or in combination with passive immunization via post-exposure prophylaxis (PEP). A serum level of 0.5 IU/mL antibody concentration, as determined by a virus neutralizing test (e.g. rapid fluorescent focus inhibition test, [RFFIT]) is regarded as adequate for offering protection against RABV. Objective: The question remains to what extent protection against ABL and EBL virus variants exists and whether there is a correlation between the RVNA concentration against classical rabies strain and these non-classical rabies virus strains. Methods: To answer these questions, a crossneutralization study was carried out, using sera from a clinical trial with PCECV administered in a PEP regimen. A set of 100 sera was selected to cover the range between 0.5 IU/mL and 500 IU/mL. A RFFIT was performed to determine the neutralizing antibody concentrations against classical RABV CVS-11 challenge strain as well as ABL, EBL1 and EBL2. Results: The RVNA concentrations against CVS-11 correlate well with RVNA concentrations against ABL, EBL1, and EBL2. For all strains, a positive close correlation was seen: The higher the RVNA concentration against CVS-11, the higher the RVNA concentration against the respective bat strain. While, for EBL1 the results were a little lower compared to CVS-11 (less than a log-scale), results were almost identical for CVS-11, EBL2 and ABLV. Conclusion: This study demonstrates that after vaccination with PCECV adequate virus neutralizing antibody concentrations against the non-classical rabies virus strains ABL, EBL1, and EBL2 are achieved in the majority of cases. Against these virus strains a virus neutralizing antibody concentration of equal to or above 0.5 IU/mL as obtained by standard RFFIT using the classical RABV CVS-11 is indicative for adequate protection.

31

HUMAN RABIES EPIDEMIOLOGY, PROPHYLAXIS AND TREATMENT

EVALUATION OF THE INFORMATION SYSTEM OF THE PROGRAM OF THE RABIES EPIDEMIOLOGICAL SURVEILLANCE, BRASIL - 2001 TO 2005
L.R. Montebello 1, Rosely C. Oliveira 1, M.L.N.S. Arsky 1, V.L.C. Silva 1, W.G. Almeida1
1) Secretaria de Vigilncia em Sade, Ministrio da Sade, Brasil

Dr. Lcia Montebello - lucia.montebello@saude.gov.br

Brasil

Information system is decentralized in 5.560 counties of the country. It has purpose of subsidizing the process of decision. Municipality is the instance responsible for feeding the data-base and execution of the actions and services. This system captures early alterations that may happen in morbi-mortality of the disease in certain rea. It allows to analyze the covering of activities developed by the epidemiological surveillance. Objective: To describe the system, evaluate its qualitative, quantitative attributes, usefulness, propose recommendations. Methodology: descriptive study, historical series 2001 - 2005. They were used for comparative analysis of systems: rabies program data-base; SINAN (System of Agraves Information); SIM (System of Mortality Information), Tabwin; Epi-Info;Guidelines for Evaluation Systems of surveillance .Results: It is useful,, esteems magnitude of the problem (people, time, place), determines tendencies groups and risk factors. Addresses surveillance actions, control and prevention opportunely. Qualitative attributes: the system is complex, involves many professionals; notification record possesses many fields; little flexible for changes; the acceptability by professionals is high. Quantitative attributes: it is sensitive; captures cases early; positive representativeness; identifies cases; it is opportune to compulsory and immediate notification up to 72 hours,as well as, the control actions and prevention; detects outbreaks and evaluates impact of the control measures. Conclusion: Is important to establish appropriate strategies for quality improvement of data ( personnel development) .Otimizes the opportunity and the representativeness turning it better, the diagnosis suspicion (detection of the viral circulation) ; to revise fields notification record; to establish gold pattern for validation of data; to clean SINAN data-base ; to stimulate the operational technical capacity to follow up the several SUS (Unique Health System) information systems ; to propose interinstitucional plan of joint action for the control surveillance ( wild cycle), to stimulate scientific research.

32

FIRST SESSION

POST-EXPOSURE PROPHYLAXIS (PEP) AGAINST THAI STREET RABIES VIRUS AND EURASIAN BAT LYSSAVIRUSES WITH PURIFIED CHICK EMBRYO VACCINE, PEPSIN-DIGESTED ERIG, HUMAN RABIES IMMUNE GLOBULIN AND A HUMAN MONOCLONAL ANTIBODY (MAB 17C7)
Boonlert Lumlertdacha 2, Joshua Self 2, Cathleen Hanlon 2, Susan Sloan 1, William Thomas 1, Donna Ambrosino 1, Charles E. Rupprecht 2
1) University of Massachusetts Medical School, Jamaica Plain, MA, USA, 2) Centers for Disease Control and Prevention, Atlanta, GA, USA

Dr. Boonlert Lumlertdacha - ero8@cdc.gov / Qsmibld@yahoo.com

USA

Questions remain about the utility of locally produced immune globulins in PEP against newly described lyssaviruses, and classical rabies viruses in severe exposure settings. The objective of this experiment was to compare the efficacy of different biologicals in an experimental animal PEP model. Syrian hamsters were infected with rabies virus isolated from a Thai dog (TRV), Irkut virus (IKV), or Aravan virus (ARV). Six hours later, we used the following vaccine and biologicals: Purified Chick Embryo Cell Culture Vaccine (PCECV, RabAvert, Chiron Vaccines); Purified and pepsin-digested rabies immune globulin of equine origin (ERIG) and of human origin (HRIG), both manufactured by the Thai Red Cross, as well as an experimental human monoclonal antibody (Mab 17C7). Animals were divided into the following groups with 9 animals in each and received: 1) intramuscular doses of PCECV alone, on days 0, 3, 7, 14, 28 (Essen protocol); 2) ERIG; 40 iu/kg, on day 0 alone; 3) HRIG; 20 iu/kg, on day 0 alone; 4) ERIG and PCECV/Essen; 5) HRIG and PCECV/Essen; 6) Mab 17C7 20 iu/kg, on day 0 alone; 7) Mab 17C7 and PCECV/Essen; 8) Negative control group, with no PEP . Animals infected with IKV experienced 100% mortality, regardless of group. The majority (89-100%) of ARV and TRV-infected animals succumbed, with the exception of groups 6 and 7. However, ARV and TRV groups given Mab 17C7 alone, or Mab 17C7 plus vaccine, demonstrated protection of 89-100%. We conclude that Mab 17C7 may be of greater efficacy than the biologicals used in the first experiments. We repeated the experiments after TRV infection using the same Essen PCECV regimens, but repeated the ERIG, by adding another dose on day 3, or on day 7. These animals demonstrated an improved survival rate of 83-100%, in comparison to the first experiment which showed no survival from TRV infection. Different manufacturing methods of RIG result in variable biological properties. We suggest that a second application of equine RIG fractions 3-7 days after the initial application on day 0 may result in increased protection. Such suggestions may be of particular importance when pepsin-digested fragments of immune globulin are used, especially in severe exposures.

33

HUMAN RABIES EPIDEMIOLOGY, PROPHYLAXIS AND TREATMENT

COMPARATIVE STUDY ON IMMUNOGENICITY, SAFETY AND TOLERENCE OF PURIFIED DUCK EMBRYO VACCINES (PDEV) MANUFACTURED IN INDIA (VAXIRAB) AND SWITZERLAND (LYSSAVAC)
B.J. Mahendra 1, S.N. Madhusudana 2, D.H. Aswath Narayana 1, G. Sampath 3, S.S. Dutta 4, S. Manjula 5, K. Muhamuda 2, G.M. Venkatesh, M.K. Sudarshan 1, Gangaboriah 1
1) Institute of Medical Sciences, Bangalore, India, 2) National Institute of Mental Health and Neurosciences (NIMHANS), Bangalore, India 3) Institute of Preventive Medicine, Hyderabad, India 4) Pasteur Institute, Kolkata, India 5) Cadila Healthcare ltd, Ahmedabad, India

Dr. Madhusudana Shampur - snmadhu@nimhan.kar.nc.in

ndia

Rabies and rabies prophylaxis is a major health concern in Asian Countries. In India alone 20,000 people die of rabies and nearly 8 million people undergo post exposure prophylaxis due to animal bites. Recently, the production and use of Semple vaccine has been discontinued. Consequently, there is a huge demand for modern rabies vaccines. In this context the introduction of a purified duck embryo vaccine (PDEV) manufactured in India with technology transfer from Berna Biotech, Switzerland is a welcome step. This study was done to assess the safety, immunogenicity and tolerance of PDEV manufactured in India (VaxiRab) in comparison to that manufactured at Switzerland (Lyssavac). Healthy volunteers from three centers (Bangalore, Hyderabad and Kolkata) were recruited for the study. One hundred and twelve persons were administered VaxiRab and 109 were administered Lyssavac by intramuscular route using Essen regimen. The subjects were followed for 6 months and adverse reactions were recorded. Antibody titers were estimated by rapid fluorescent focus inhibition test (RFFIT) on days 0 (before vaccination) 14, 30, 90 and 180 post vaccination. None of the subjects had neutralizing antibody titers on day 0. All subjects in both the groups developed adequate titers by day 14. Both the groups had more than adequate titers on day 180.The Geometric mean titers on different days for VaxiRab was 10.3, 14.3, 7.3 and 3.7 IU/ml and for Lyssavac was 10.9, 15.1, 10.2 and 5.3 IU/ml respectively. There was no significant difference between the titers on days 14 and 30 but the titers with Lyssavac were significantly higher on days 90 and 180 (P <0.0001). The total adverse reactions to VaxiRab were 13% that was significantly lower when compared to Lyssavac that was 18.5% (P<0.02). To conclude, this study has shown that PDEV (VaxiRab) manufactured in India is highly immunogenic, safe, well tolerated and comparable to PDEV (Lyssavac) manufactured at Switzerland. This vaccine along with other modern vaccines will go a long way in meeting the ever-increasing demand for safe and effective rabies vaccines in India and other Asian countries.

34

FIRST SESSION

EVALUATION OF PROPHYLAXIS TREATMENT FOR RABIES DURING A HUMAN RABIES OUTBREAK DUE TO VAMPIRE BAT EXPOSURE TURIA MUNICIPALTY, MARANHO STATE, BRAZIL, 2005
Marcelo Yoshito Wada 1, Henrique Jorge dos Santos 2, Mildes Mendes Pereira 2, Jakeline Trinta Rios 2, Jos Orlando Martins 2, Manuel Cludio Hiplito 3, Francisco Nolio Fernandes 3, Gracilia do Livramento Domingues 3, Rosely Cerqueira de Oliveira 1
1) Secretaria de Vigilncia em Sade, Ministrio da Sade, DF, Brasil 2) Departamento de Epidemiologia da Secretaria Estadual, MA, Brasil 3) Secretaria Municipal de Sade, Turia, MA, Brasil

Dr. Marcelo Yoshito Wada - marcelo.wada@saude.gov.br

Brasil

Introduction. Rabies is a preventable viral zoonosis that infects the central nervous system of mammals, causing encephalopathy with a mortality rate of 100%. During 2004 and 2005, a reemergence of human rabies outbreaks transmitted by vampire bat were identified in Amazonic Region, including Par and Maranho State from Brazil. Following seventeen deaths due to human rabies in Turia municipality, Maranho State, Brazil, an investigation was conducted to evaluate control and prevention measures. Methods. This study describes prophylaxis for rabies adopted in this municipality from September to December, 2005. Active-case-finding of aggressions by vampirebats was conducted in health facilities and the community. Person who was bitten by vampire-bat until one year and lived in areas with difficult access received prophylaxis. Using excel, it was created a standard sheet to accomplish complete prophylaxis. Results. A total of 17 human rabies case-patients were identified in two rural communities; none(0%) had received pre-exposure prophylaxis(PRE) or pos-exposure prophylaxis(POS) for rabies or immunoglobulin at an opportune time. A total of 696 persons realized prophylaxis, 80%PRE and 20%POS. Among POS, persons who were bitten by vampire-bats 73%, dogs 13%; marmoset 5%; cats 3%; ignore 3%, 1% other species. Persons who have done complete POS were 54% and PRE 68%. Over than 2,530 doses of vaccine were applied to attend at least 38 localities of Turia municipality. Conclusions. This large outbreak was due to vampire bat bites. The population was informed about risks associated with bat exposure, offered PEP , and encouraged to promptly report any contact with bats. This study demonstrate the importance of PRE for prevention, and the need for prompt reporting and standardized treatment of bat exposure/bites with human diploid cell vaccine and the alert for surveillance to monitor the prophylaxis, because the tax of release of treatment was high.

35

HUMAN RABIES EPIDEMIOLOGY, PROPHYLAXIS AND TREATMENT

LIMITATIONS OF F(ab)2 FRAGMENTS IN RABIES POST-EXPOSURE PROPHYLAXIS


R. Franka 1, C. A. Hanlon 1, J.S. Self 1, C. E. Rupprecht 1
1) Centers for Disease Control and Prevention, Atlanta, GA, USA

Dr. Richard Franka - rpf5@cdc.gov

USA

Modern post-exposure prophylaxis (PEP) using effective biologicals relies upon neutralization of rabies virions by passive antibody administration and further clearance of rabies virus (RABV) through induced active immunity. To minimize adverse effects from heterologous immune serum products, some equine immune globulins have been refined by digestion to F(ab)2 fragments. These fragments effectively neutralize RABV in cell culture, however, their use in relevant animal models has demonstrated significant limitations in efficacy. The objective of our study was to uncover mechanisms underlying the limited efficiency of F(ab)2 fragments, using a Syrian hamster model. We studied the dynamics of a dog/coyote RABV variant spread and the effect of PEP with immune serum (HRIG - human immune globulin, or F(ab)2 fragments of equine immune globulin) alone or in combination with HDCV vaccine, using a standard scheme for PEP in this animal model. In control animals (no PEP), RABV reached the lumbar portion of the spinal cord by day 4 postinfection (p.i.) and the brain by day 6 p.i., with animals succumbing on days 10 - 14 p.i. after virus inoculation into the gastrocnemius muscle. Use of vaccine alone was unable to limit the spread of RABV. Despite neutralizing antibodies detected as early as day 6 p.i. (5th day of PEP), all vaccineonly animals succumbed to infection at the same time as controls. However, HRIG given as a single dose on day 1 p.i. had a long biological half life (15 days), retarded RABV spread, and resulted in protection in 14 of 15 animals. Conversely, F(ab)2 fragments had a significantly shorter biological half life (8h) and only partially hampered RABV spread. Virus reached the spinal cord and brain 12 - 22 days p.i. Vaccine improved survivorship when used in combination with immune globulin. Clearly, early passive immunization with effective immune globulins, having an adequate biological half life in circulation, is a critical part of PEP .

36

FIRST SESSION

EVALUATION OF THE INDEXES OF RABIES PROPHYLAXIS AND CONTROL IN THE STATE OF AMAZONAS - 1996 TO 2005
A.C.R Campos 1, N.R. Reinehr 1
1) Fundao de Vigilncia em Sade, Manaus, AM, Brasil

Dr. Ana Cristina Campos - anacristina@vivax.com.br

Brasil

Rabies is a zoonosis transmitted to humans by inoculation of the virus from infected animals, and preventive measures to assist people exposed to the virus are very costly. Urban rabies is still a grave problem in public health, and dogs are the main source of disease infection. This work describes the improvement obtained by the State of Amazonas in the last ten years in human-rabies control and prophylaxis with respect to the indexes used by the Brazilian Ministry of Health. The epidemiological study was divided into two groups: Vigilance of human rabies and vigilance of canine rabies. Seven indexes were evaluated in total. We considered an improvement whenever the distance between the observed index and the pre-established target was reduced. For the vigilance of human rabies, there was a decrease in the rate of abandonment of treatment from 26.5% to levels close to zero, and an increase from 54.0% to 100.0% in the number of the municipalities with trained professionals and human antirabies vaccine available to the public. For the vigilance of canine rabies, there was an increase from 4.8% to 87.1% in the number of municipalities that kept minimum vaccination coverage in two consecutive years; from 14.5% to 91.9% in the number of municipalities with annual anti-rabies vaccination equal to or greater than the established target; from 1.6% to 62.9% in the number of municipalities that sent dog samples for laboratory analysis, and zero cases of canine rabies in 2004 and 2005. During the period of the study, there was a constant improvement in the indexes due to a more intense production of educative material for public awareness, to an increase in the number of professional trainings, and to the annual supervision of 80% of the municipalities in the State of Amazonas.

37

HUMAN RABIES EPIDEMIOLOGY, PROPHYLAXIS AND TREATMENT

BIOPTERIN DEFICIENCY IN HUMAN RABIES


R.E. Willoughby, T. Maier, C.E. Rupprecht, W.J. Rhead, K. Hyland, G.F. Hoffmann, J. Hoyer - Medical College of Wisconsin, Wisconsin, USA, Philipps University, Marburg, Germany, Centers for Disease Control and Prevention, Atlanta, GA, USA, Horizon Molecular Medicine, Atlanta, Georgia, USA, Heidelberg University, Heidelberg, Germany) Dr. Rodney Willoughby, MD - rewillou@mcw.edu USA

Rabies is a highly neurotropic and universally fatal infection in man. Pathogenesis is distinctive with severe symptoms with little neuronal loss or inflammation, followed by loss of encephalogram (EEG) amplitude and sometimes shock. An American survived rabies in 2004 using an improvised protocol. Deficiencies in coenzyme Q10 and cerebrospinal fluid (CSF) tetrahydrobiopterin (BH4) were noted during her convalescence. A German patient developed rabies from transplanted organs. The improvised treatment protocol was modified for his immune deficiency. He died after 56 days with low brain electrical activity and low intracranial blood flow, without simultaneous edema by neuroimaging. He received coenzyme Q10 but developed severe BH4 deficiency, retrospectively present from the second week of rabies. BH4 is an essential cofactor for synthesis of monoamine neurotransmitters (NT) and nitric oxide (NO). Both patients had severe declines in NT metabolites but not NO metabolites reflecting BH4 deficiency in CSF. Central but not peripheral nervous system symptoms of rabies overlap with congenital BH4 deficiencies. NT loss may explain EEG loss and adrenal medullary failure, while NO loss reduces brain flow to 10-50% of normal. BH4 can be supplemented orally, and may directly and specifically remedy the biochemical pathogenesis of rabies. NO may extend brain injury acutely, so BH4 supplementation must follow clear proof of deficiency, preferably in animal studies and formal clinical trials.

38

FIRST SESSION

THERAPY WITH MINOCYCLINE AGGRAVATES EXPERIMENTAL RABIES IN MICE


A.C Jackson 1, J. Owen, C.A. Scott 1, S.C. Weli 1, J.P . Rossiter 1
1) Queens University, Kingston, ON, Canada

Dr. Alan C. Jackson - jacksona@post.queensu.ca

Canada

Minocycline is a tetracycline-derivative with anti-apoptotic and anti-inflammatory properties, and the drug has been shown to have beneficial effects in neurodegenerative models and in experimental reovirus encephalitis. In this study minocycline improved the viability of CVS-infected embryonic mouse cortical and hippocampal neurons in vitro using trypan blue exclusion. To investigate the potential in vivo neuroprotective effects of minocycline, two-day-old ICR mice were inoculated in the right hindlimb thigh muscle with the attenuated SAD-D29 strain of rabies virus and they received daily subcutaneous injections of either 50 mg/kg minocycline or vehicle (PBS). Infected minocycline-treated mice experienced an earlier onset of neurologic signs and greater mortality (83% vs. 50%) than those receiving vehicle (p=0.002 and p=0.003, log rank test). Immunohistochemical analysis of rabies virus antigen distribution was performed at early time points and in moribund mice. There was no significant difference between the numbers of infected neurons in regional brain areas in the two treatment groups. There was more apoptosis (p=0.01) and caspase 3 immunostaining (p=0.0008) in the midbrains of mice treated with vehicle than with minocycline, consistent with a neuroprotective role of neuronal apoptosis. Reduced infiltration of CD3+ T-cells was observed in the pons/medulla of moribund mice that received minocycline therapy (p=0.008), suggesting that the anti-inflammatory actions of minocycline may intensify the neurologic disease. These findings indicate that minocycline has important detrimental effects in the therapy of experimental rabies, and empirical therapy with minocycline should be approached with caution in human rabies and possibly other viral encephalitides until more experimental data become available.

39

HUMAN RABIES EPIDEMIOLOGY, PROPHYLAXIS AND TREATMENT

siRNA-DIRECTED INHIBITION OF RABIES VIRUS AND PROTECTION FROM RABIES INFECTION IN MOUSE MODELS
Rongliang Hu 1, Shoufeng Zhang 1, Anthony R. Fooks 2, Zhong Li 1, Qingzhu Li 1, Ye Liu 1, Fei Zhang 1, Jinzhong Li 3, Quan Liu 1, Zhengfang Fu 4
1) Academy of Military Medical Science, China, 2) Veterinary Laboratories AgencyCWeybridge, Surrey, UK, 3) Department of Neurological Surgery, University of Virginia Health System, Charlottesville, USA, 4) College of Veterinary Medicine, University of Georgia, Athens, USA

Dr. Professor Rongliang Hu - hurongliang@hotmail.com

China

Rabies virus is lethal to humans once the virus reaches the brain via axonal retrograde transport where active virus replication occurs. At this stage death is an inevitable outcome of infection. One possible therapeutic intervention is to stop virus replication at the site of exposure, and prevent the virus from penetrating the peripheral nervous system. Inhibition of the virus replication at the biting site may delay the incubation period or stop the occurrence of rabies. RNAi would be an efficient strategy to prevent the virus replication at the exposure site. In this study, a total of nine siRNA molecules directed to the glycoprotein and the nucleoprotein RNA were designed, synthesized in oligo form and cloned into plasmid pSilencer2.1-U6-Hygro. The recombinant plasmids were transfected into BHK-21 cells, the cells were screened under hygromycin B and individual cell clones were established. Inhibition effect of the transformed BHK-21 cells on rabies virus replication was observed in different cell clones with variations of inhibition efficacy from 10-99%. Two siRNA transformed cell clones were demonstrated to have the ability to inhibit about 95% and 99% of 1000 TCID50 rabies virus CVS-11 replication. The transformed cell lysate, the transient transfection with a high amount of the recombinant plasmids harboring the siRNA, and the corresponding in vitro synthesized siRNA molecules exhibited the same inhibitory characteristics to rabies virus in BHK-21 cells. Injection of the siRNA expression plasmids against the glycoprotein, rather than against the nucleoprotein, delayed the appearance of clinical signs, with the increasing of amount of plasmids, the infection was stopped, 60% mice were protected when the amount of plasmids reached 300g/mice. This study indicated that in a murine model, siRNAs can interrupt rabies virus spread and prevent the virus from penetrating the peripheral/ central nervous system and subsequently prevent the virus from reaching the brain.

40

FIRST SESSION

LACK OF EFFICACY OF KETAMINE THERAPY IN EXPERIMENTAL RABIES IN MICE


C.A .Scott 1, S.C. Weli 1, A.C. Jackson
1) Queens University at Kingston, Canada

Dr. Alan C. Jackson - jacksona@post.queensu.ca

Canad

Ketamine, a non-competitive N-methyl-D-aspartate (NMDA) receptor antagonist, was recommended as a therapeutic option for the management of rabies on the basis of studies performed by Tsiang and co-workers at Institut Pasteur in Paris. Recently, a patient survived rabies in Milwaukee, and ketamine was included in the therapeutic regimen. We found no benefit of ketamine on the viability of CVS-infected mouse embryonic neurons from the cerebral cortex and hippocampus as assessed by trypan blue exclusion. In this study, mice were inoculated intracerebrally or in the right hindlimb footpad with CVS and treated with ketamine (60 mg/kg) or vehicle (PBS) intraperitoneally every 12 hours. Mice inoculated intracerebrally or in the hindlimb footpad that received ketamine or vehicle showed no difference in the time of onset or progression of disease (log rank test, p=0.30 and 0.54, respectively) or in the mortality rate (p=0.50 and 0.53, respectively). Mice inoculated in the hindlimb footpad showed similar numbers of infected neurons between treatment groups on days 3-6 post-inoculation (p.i.), as demonstrated by immunohistochemistry (p>0.05). On day 5 p.i. viral infectivity assays showed a similar amount of infectious virus in the brain and spinal cord of mice that received either treatment. In moribund mice inoculated in the footpad, there was no difference in the number of infected neurons in the cerebral cortex or hippocampus, while there were more infected neurons in the midbrain of mice treated with ketamine as compared to vehicle (p=0.0009). Mice inoculated intracerebrally also showed no difference in the numbers of infected neurons between treatment groups (p>0.05). Furthermore, there were similar amounts of infectious virus in the brain at day 5 p.i. Histopathological analysis of intracerebrally inoculated mice showed neuronal apoptosis and a loss of pyramidal neurons in the hippocampus with both treatments and no significant difference between the groups (p>0.05). These findings do not support a role for excitotoxicity in rabies virus infection. Further experimental studies are necessary to determine the efficacy of ketamine before it becomes a standard therapy for human rabies

41

HUMAN RABIES EPIDEMIOLOGY, PROPHYLAXIS AND TREATMENT

ANTIVIRAL STRATEGIES AGAINST THE LYSSAVIRUS REPLICATION COMPLEX: PEPTIDES MIMICKING THE NH2 TERMINUS OF THE RABIES P PROTEIN HAVE ANTIVIRAL ACTIVITY
G. Castel 1, S. Mehouas 1, E. Ral 1, C. Jallet 1, M. Mavrakis 2, RWH Ruigrok 2, N. Tordo 1
1) Institut Pasteur, Paris, France, 2) Institut de Virologie Molculaire et Structurale & EMBL Outstation Grenoble, France

Dr. Noel Tordo - ntordo@pasteur.fr

Despite the remarkable publication of Willoughtby et al. (N Engl J Med. 2005, 352:2508-14) the therapeutic approach against rabies has been largely neglected up to now in comparison to more conventional tools like postexposure vaccination and immunotherapy. However, the rather long incubation period of rabies (2 months in average) could be advantageously used to develop antiviral strategy. Our approach consists in targeting the different components of the lyssavirus replication complex (L-P polymerase and N-RNA template). Because of the strong conservation of these targets, we prospect at designing large spectrum antiviral drugs interfering with the replicase of lyssaviruses as well as other Negative Strand RNA Viruses (NSRV). Using suitable assays we are screening inhibitors by two complementary experimental approaches: (1) A random approach taking advantage of high throughput technologies (2-hybrid ; phage display, minireplicons) to discover molecules interacting and inhibiting the polymerase activity (presented to the RITA XVI - Ottawa) (2) A cognitive approach by investigating the structure/function relationships and interactions within the replication complex.The latter approach has been applied to the P phosphoprotein. P is important in positioning L onto the N-RNA template as well as in keeping N soluble (N) and ready for proper viral RNA encapsidation. The N terminus of P is particularly interactive, involved in both L binding (residues 1-19) and N binding (residues 4-40). It was thus designed as a promising target for antiviral strategy aiming at destabilizing the replication complex. Plasmids encoding the first 42 (P42) and 60 (P60) residues of P were co-transfected in vT7-BHK-21 cells together with a rabies minigenome encoding luciferase. Their inhibitory effect on luciferase expression was evaluated and both peptides exhibited an interesting inhibitory potential. This inhibitory effect of P60 was however stronger and longer lasting that of P42. Specific inhibition on viral encapsidation, transcription, replication and production have been analysed. The results will be discussed in light of the P structure/function relationships. This strategy is applicable more largely to other NSRV either on the same or on other targets.

42

FIRST SESSION

EVALUATION OF HUMAN MONOCLONAL ANTIBODIES CR57 AND CR4098 AGAINST A GLOBAL PANEL OF RABIES VIRUSES
I. V. Kuzmin 1, W.C. Weldon 1, A.B.H. Bakker 2, W. E. Marissen 2, C.E. Rupprecht 1
1) Centers for Disease Control and Prevention, Atlanta, GA, USA 2) Crucell Holland BV, Netherlands

Dr. Ivan Kuzmin - ibk3@cdc.gov

USA

A cocktail of human anti-rabies virus (RV) monoclonal antibodies (MAbs) CR57 and CR4098, manufactured by Crucell Holland BV (The Netherlands), was previously shown to neutralize a panel of RV representatives, predominantly of New World origin. A linear epitope at the aa position 226-231 of the RV glycoprotein (G) was identified as a binding site for CR57, whereas aa position 336 has been implicated in the binding site for CR4098. In the present study we extended the virus panel with representatives of major RV lineages circulating worldwide; generated partial G sequences to cover the CR57 binding site; and performed modified rabies rapid fluorescent focus inhibition tests for individual MAbs and the cocktail. The majority of the RVs were neutralized by individual MAbs. The CR57 binding site was conserved as well, with 0 to 1 amino acid substitutions. A few exceptions to these observations were obtained. A south-central skunk RV isolate (1 critical amino acid substitution at position 226) was incompletely neutralized by this Mab. Furthermore, two RV, a North American Eptesicus fuscus-Myotis-associated RV and a Sri Lanka dog/cow RV, were not neutralized by the CR57 MAb. Both these RV held no substitutions within the 226-231 site, indicating that other parts of the G glycoprotein can indirectly influence the binding of this Mab. Non-conserved substitutions at aa 336 or 338 led to incomplete neutralization by MAb CR4098, confirming the critical role of aa 336 in the conformational binding site of CR4098. When the extended RV panel was tested by the MAb cocktail, all RVs tested to date were neutralized.

43

HUMAN RABIES EPIDEMIOLOGY, PROPHYLAXIS AND TREATMENT

WEB-BASED RABIES TREATMENT REGISTRY WITH DOWNLOADABLE PROTOCOL [WWW.CHW.ORG/RABIES]


R.E. Willoughby 1, J. Hayes 1, S. Sather 1, B. Medina 1, P . Overholt 1
1) Medical College of Wisconsin and Childrens Hospital of Wisconsin, Milwaukee, Wisconsin, USA

Dr. Rodney Willoughby, MD - rewillou@mcw.edu

USA

BACKGROUND: Rabies is rapidly and universally fatal for 50,000 people each year. Survival from rabies is so extraordinary that proof of therapeutic efficacy requires only 2-3 survivors. We reported the first survivor of rabies without receipt of prophylaxis in 2004. We then received calls and emails by medical staff and families seeking access to the Milwaukee protocol (MP). Most communications involved multiple time zones that required 2-3 days to fully complete, and were too late. Publication of MP [NEJM (2005) 352:2508] markedly reduced urgent contacts, but there is published skepticism of MP and frequent requests for clarification. We have further experience with MP and new laboratory findings to share. AIM:(1) Expedite availability and updating of MP (2) Provide online, real-time access to a Rabies Treatment Registry (RTR). METHODS: MP is updated as online Adobe Acrobat document. RTR supports web entry and real-time survival statistic. Data are restricted to age, country, laboratory confirmation of rabies, treatments including individual MP drugs, and simple outcome of survival. We can censor (hackers) or correct data with auditable record. RTR is capable of future online queries of outcome sorted by drugs of interest. RESULTS: We recommend no MP before diagnosis is confirmed. Uses of MP included advance planning by intensivists pending confirmatory diagnosis, actual rabies care, and trial design. Ketamine worries many. Of 4 known MP attempts, only 2 are registered. Both included full MP plus immunologic boosting. Unregistered MP attempts occurred despite frequent communication during care of rabies patients. CONCLUSIONS: Slow accrual severely limits current usefulness of online MP and RTR, despite online open-science format.

44

FIRST SESSION

PREVALENCE STUDY ON VAMPIRE-BAT (DESMODUS ROTUNDUS) BITES IN A RURAL POPULATION FOLLOWING AN OUTBREAK OF RABIES-RELATED DEATHS MARANHO STATE, BRAZIL, 2005
L.V. de Knegt 1, E.I.M. Renoiner 1, W.N. Arajo 1, M.Y. Wada 1, M.A.B. Almeida 1, H.J. Santos 2, M.G. Lirio 3, J.M.T. Rios 4, D.L. Hatch 5
1) Secretaria de Vigilncia em Sade, Ministrio da Sade, DF Brasil 2) Superintendncia de Epidemiologia, MA, Brasil 3) Departamento de Vigilncia Epidemiolgica, MA, Brasil 4) Departamento de Zoonoses, MA, Brasil, 5) Centers for Disease Control and Prevention, Atlanta, GA, USA

Dr. Leonardo Vctor de Knegt - leonardo.knegt@saude.gov.br

Brasil

Background: Rabies is a vaccine-preventable viral encephalitis with a 100% mortality rate. Prior to 2004, in Brazil the disease was mainly caused by rabid domesticated carnivores; Since 2004, vampire-bat bite became the leading cause. After six rabies deaths due to suspected vampire-bat exposure occurred between July and September 2005 in the Alto Turi Region, Maranho State, an investigation was conducted to determine risk factors for vampire-bat exposures. Methods: Using a cross-sectional study design, interviews were conducted with headsof-households in three areas of Alto Turi; information on selected behaviors, vampire-bat bite(s), housing, and vaccination was collected. Results: Interviews were completed in 174 residences (participation rate=100%). Vampire-bat bite(s) were reported by 87 (42%) of 205 family members during their lifetime; the foot was the most common location (86%). No person with vampire-bat exposure reported being vaccinated. Factors associated with vampire bites included a residence with an open, unscreened area (jirau) for food preparation (Prevalence Ratio [PR]=3.3;95% Confidence Interval [95%CI]=1.4-7.9;p=0.001), residing <5 kilometers from pastureland (PR=4.5;95%CI=1.1-17.4;p=0.008), sleeping in hammocks (PR=1.6;95%CI=1.1-2.3;p=0.007) or without bednets (PR=1.9;95%CI=1.1-3.1;p=0.02). Bats captured at residences using mist nets were identified as vampire-bats (Desmodus rotundus). Conclusions: Lack of vaccination was due to several factors, such as vampire-bat bites common in the population, limited access and inadequate use of vaccine by health staff. Vampire-bats normally feed on sylvatic mammals, but encroaching human settlement in these remote areas may have changed the epidemiology of disease. Because rabies may occur up to one year after exposure, rabies antiserum and five doses of Vero-cell rabies vaccine was administered to residents with vampire-bat bite(s) in the prior year. To prevent rabies in persons without exposure in last year; three doses of vaccine was administered. Health education was conducted about risk of contact with unvaccinated mammals, and bednets were distributed.

45

HUMAN RABIES EPIDEMIOLOGY, PROPHYLAXIS AND TREATMENT

DESCRIPTION OF HUMAN RABIES CASE IN VETERINARY, MINAS GERAIS STATE, 2006


T.L.Chamone 1, M.B. Gontijo 1, F. L. Lemos 1, P .P . Botelho 1, F .J. Silva 2, M.Y. Wada 3, A B. Miranda 4
1) Secretaria de Estado da Sade MG,Brasil, 2) Gerncia Regional de Sade So Joo Del Rei, MG,Brasil, 3) Secretaria de Vigilncia em Sade, Ministrio da Sade, DF, Brasil 4) Hospital Eduardo de Menezes, MG, Brasil

Dr. Talita Leal Chamone - talita.chamone@gmail.com

Brasil

Introduction: Minas Gerais (MG) state has been presenting cases of rabies in dispersed herbivores for whole the State. In 2006, a human rabies case occurred in a veterinary transmitted by a herbivores. Objective: To describe clinical and epidemiological aspects of the human rabies case in this veterinary. Methods: A descriptive study was done, including an epidemiological investigation of the case. Members of the family were interview and medical records were analyzed. Results: On 15 May, 2006, the SES-MG was reported of a suspected human rabies case interned at the reference hospital of the state. The suspected case was a male, 27 years, coming from the area of the Campo das Vertentes municipality, MG. He was a veterinary who rendered their services in farms of the area. In February, 2006, there were registered rabid herbivorous confirmed by laboratorial tests and this veterinary was responsible to attend these animals. The patient hadnt pre-exposure prophylaxis or pos-exposure prophylaxis. On 07 May, 2006, the patient presented headache, parasthesis where had exposure with herbivores (finger). He developed mental confusion and depression of the level of conscious. He was sedated in Ramsay 6 with ketamine and midazolam, it was administered anti-viral and support treatment was offered. Clinical evolution include intense poliuria, hiponatremy and episodes of arrhythmias heart that were controlled with medication. The death happened on 26 May, 2006, due to attack heart. The case was confirmed by PCR in fragments of hair and saliva. Genetic sequence was performed and demonstrated variant compatible with Desmodus rotundus. There was also confirmed by fluorescent antibody test in fragments of the central nervous system. Conclusions: This investigation showed the high importance of pre-exposure prophylaxis in professionals who has high possibility to infect by rabid animals. Otherwise if persons had exposure to rabid suspect animals, they have to realize pos-exposure prophylaxis.

46

FIRST SESSION

HBITOS ALIMENTICIOS DE LA COMUNIDAD HAH Y RIESGO DE EXPOSICIN A LA RABIA SILVESTRE


J.C.Lozano-Olivo 1, E. Meraz-Romero, 1, 2, J. Bautista-Urbano 1, J.I. Olave-Leyva1, 3, S. Soto-Simental 1, L. A GmezMendieta.4, A Gonzlez-Origel.4, Z. Arellano-Balderrama, 1, A Aguilar-Setin 5, M. Jimnez-Prez 1, M. Hernndez-San Agustn 1, A L. Pereira-Surez. 1, J. Ocampo-Lpez. 1, J. A Montao-Hirose 1
1) Instituto de Ciencias Agropecuarias de la Universidad Autnoma del Estado de Hidalgo, Mxico 2) Ganadera, Colegio de Postgraduados en Ciencias Agrcolas, Mexico 3) Facultad de Medicina Veterinaria y Zootecnia, Universidad Nacional Autnoma de Mxico, Mexico 4) Secretara del Medio Ambiente y Recursos Naturales, Mexico 5) Instituto Mexicano del Seguro Social, Mexico

Dr. Juan A. Montao - juan_montano@virologiahoy.org

Mxico

El pueblo hah conserva costumbres de sus antepasados, como alimentarse de animales silvestres. Se cazan con trampas, y ocasionalmente, cuando de regreso a casa se encuentran con un animal lo cazan a mano con la ayuda de recursos que tengan o encuentren disponibles en el momento, como una bolsa de mandado, una pala, una piedra o un palo. La caza la realizan en la noche. Aunque no se han notificado casos de rabia en humanos en la regin, s se ha identificado en zorrillos. Por ello existe el riesgo de exposicin a la rabia, debido a sus hbitos alimenticios relacionados con la captura, sacrificio y preparacin de los animales. La comunidad hah habita desde el Valle de Toluca, en Mxico, hasta la Sierra Gorda, pasando por el Valle del Mezquital. El Valle del Mezquital abarca 27 municipios. En algunos de ellos se inici la colecta de datos sobre los hbitos alimenticios de los hahs, en base a un muestreo simple aleatorio. Hasta el comento se han completado la cuota de encuestas calculada para el municipio de Santiago de Anaya. Adems, se han colectado datos de comunidades no hahs. Para obtener el nmero de encuestas a aplicar por comunidad, se utiliz la siguiente frmula (Taro Yamane, 1967): ni = n (Ni/N) donde: ni = nmero de encuestas a ser aplicadas n = nmero de encuestas propuestas por el investigador. Ni = nmero de unidades experimentales en la muestra (casas/municipios). N = total de de unidades experimentales en el rea de estudio. El consumo de zorrillo en las casas en Santiago de Anaya, fue de 62 (54%) y las casas donde no se consume fueron 53 (46%). En la regin la gente lo consume espordicamente. Los resultados muestran que los hbitos alimenticios de la comunidad hah presentan un riesgo de exposicin a la rabia, cuya evaluacin debe ser completada con monitoreo de la fauna silvestre.

47

HUMAN RABIES EPIDEMIOLOGY, PROPHYLAXIS AND TREATMENT

MANEJO DE PACIENTES CON LESIONES GRAVES PROVOCADAS POR PERRO EN REAS CON AUSENCIA DE CASOS DE RABIA CANINA, EN EL ESTADO DE CHIAPAS MXICO
Raul Betanzos Lopez 1, Hermilo Dominguez Zarate 1, James Gomez Montes 1, Luz Maria Yanez Gamboa 1
1) Secretaria de Salud, Chiapas, Mexico

Dr. Raul Betanzos Lopez - zoonosischiapas@salud.gob.mx

Mxico

Manejo de pacientes con lesiones graves provocadas por perro en reas con ausencia de rabia canina, en el Estado de Chiapas Mxico. Introduccin. Hace dos dcadas la incidencia de rabia canina en Chiapas registraba niveles de positividad superiores al 75%; situacin que mantena alto riesgo de transmisin a los humanos y por ende elevada demanda de tratamientos. El trabajo sanitarista sostenido durante las recientes dcadas, ha favorecido una evolucin notablemente satisfactoria del programa rabia en las principales ciudades de la entidad; destacan dentro de las actividades: las elevadas coberturas de vacunacin antirrbica canina, capacitacin del personal operativo y la participacin de la sociedad organizada. ha medida que disminuyo la positividad en perros, ha disminuido de manera importante el inicio de tratamientos a razn de 7% en estas regiones; situacin que ha permitido un criterio conservador en el manejo de lesiones graves. Objetivos. Describir el impacto favorable de la correcta aplicacin de los lineamientos normativos para la prevencin de la rabia y manejo de pacientes con lesiones graves. Mtodo. El presente trabajo comprende el estudio de 28 pacientes con mordedura de perro en diversas partes del cuerpo; 11 femeninos y 17 masculinos, promedio de edad de 22 aos, se les practic lavado exhaustivo, debridacin de tejido necrtico y sutura. Un paciente fue amputado del dedo pulgar, 19 requirieron de ciruga plstica, se manejaron con antibiticos y en ningn caso se manejo tratamiento antirrbico. Los casos son residentes de localidades con ausencia de rabia y en todos se realiz estricta vigilancia del animal agresor. Resultados. Las condiciones de ausencia de rabia en las localidades de los 28 pacientes estudiados, el plan teraputico empleado fue de medidas generales de las lesiones y atencin quirrgica especializada en 19 casos; simultneamente manejo mdico de los casos, observacin estricta del animal agresor durante 10 das a partir de la fecha de agresin. En ningn caso se utiliz terapia antirrbica. Conclusin. Las heridas causadas por animal identificado, vacunado y habitante de reas con ausencia de rabia, deben ser suturadas de primera intencin previo lavado mecnico y debridacin del rea, posteriormente manejadas con antibiticos va sistmica, este plan teraputico favorece un mejor resultado cosmtico y menor porcentaje de complicaciones.

48

FIRST SESSION

OUTBREAK OF VAMPIRE-BATS BITES IN HUMANS IN VILA VELHA-ES, 2006


Virgnia Teixeira do Carmo Emerich 1, Carla Cavalcanti Teixeira Oliver 1, Augusto Marchon Zago 2, Luiz Carlos Barboza Tavares 3, Marcelo Yoshito Wada 4
1) Centro de Controle de Zoonoses, Vila Velha, ES, Brasil, 2) Secretaria da Sade, ES, Brasil, 3) Instituto de Defesa Agropecuria Florestal do Esprito Santo, SES, ES, Brasil 4)Secretaria de Vigilncia em Sade, Ministrio da Sade, DF, Brasil

Dr. Carla Cavalcanti - carlacvet@yahoo.com.br

Brasil

Introduction: Bats are related with rabies transmission since last century. In Brazil, an effective control of rabies in domestical animals have contributed for a decline in human rabies cases, but in 20042005 occurred outbreaks of human rabies transmitted by vampire-bats in Amazonic region, demonstrating the importance of bats on rabies transmission. In an urban area of Vila Velha municipality, Espirito Santo State, there were identified bites in humans between JanuaryMarch, 2006. Methods: A descriptive study was done using a questionnaire to describe the outbreak and active search was performed. Capture and control of vampire-bats was realized using a fog net and vampiricide anticoagulant. Prophylaxis was offered to persons who were bitten by bats. Health education was done. Results: There were identified ten bites in humans, being in five different residences in a buffer of twenty to fifty meters located in the edge of the river Aribiri. All patients received prophylaxis for rabies and related bites during the night, while they were sleeping. All of them were bitten on the foot and related that they left windows opened or slept outside of the house. Residences are done with mansory with holes in the floor, wall and roof. Around three kilometers from the houses was identified a cave where there were captured 28 vampire-bats (Desmodus rotundus) and five were treated with vampiricide and 23 were sent for rabies diagnosis and all were negative. A vaccination of domestic animals were performed. Conclusions: The cave is being monitored. An inquiry to identify bites was implemented as routine. Education health is important, mainly to look for assistance when bite occurs. After capture and control of vampire-bats, there wasnt any other bites in humans. All actions to prevent human rabies cases were conducted at opportune time and no human rabies cases occurs.

49

HUMAN RABIES EPIDEMIOLOGY, PROPHYLAXIS AND TREATMENT

ATTACKS OF THE COMMON VAMPIRE BATS AT INDIGENOUS VILLAGES FROM NORTHERN BRAZIL
Wilson Uieda 1, Rmulo Henrique da Cruz 2, Marina Albuquerque 2, Pedro Carneiro Junior 2
1) Instituto de Biocincias da Universidade Estadual Paulista, Botucatu, SP , Brasil; 2) Departamento de Sade Indgena, Fundao Nacional de Sade

Dr. Wilson Uieda - wilson.uieda@funasa.gov.br

Brasil

Recent outbreaks of human rabies (2004 and 2005) transmitted by the vampire bat Desmodus rotundus took place in the Amazonian region and involved poor people from Par and Maranho states. Similar outbreaks involving indigenous people are not known in Brazil, but there is an unpublished and unconfirmed report of rabies in two young native Apinajs, apparently transmitted by D. rotundus, in Tocantins state in 1998. On the other hand, attacks by vampire bats upon indians were already reported several times. In March 2005, we started a survey on the occurrence of attacks at Brazilian indigenous villages and we verified that they are concentrated in the Amazonian region. Until March 2005, 356 natives from the states of Tocantins (60.4%), Rondnia (31.1%), Par (4.8%), Roraima (3.4%) and Amap (0.3%) were bled at least once in their life. No data from other Brazilian states was obtained. Between July and November 2005, an aggression outbreak occurred at three Kaiap villages in the Municipalities of Ourilndia do Norte and So Felix do Xingu (Par), where more than 200 natives were bled by vampires. Between December 2005 and May 2006, vampire bats were netted at seven indigenous villages of Altamira and Anap, but their attacks were observed only at the Arara, Xipaia and Kaiap villages. At a Kaiap village, where many aggressions were frequently observed, the population growth of D. rotundus must be related to cattle raising by the natives themselves. Many cows rested every night at the village and this could have allowed bats to get food from both sources: cows and humans. Later, when the cattle was taken off the village, natives turned the main blood source to vampires. This situation is well known on outbreaks of aggressions and of rabies on humans by D. rotundus at small and poor villages in Latin America.

50

FIRST SESSION

ANTHROPOLOGICAL STUDY OF RABIES OUTBREAK IN THE CITY OF PORTEL (PA/ AMAZNIA/BRAZIL): CONCEPTIONS AND REPRESENTATIONS OF THE ILLNESS FOR THE FAMILY OF THE VICTIMS
P .M. Bezerra 1
1) Universidade Federal do Par / Universidade Federal da Amaznia / FIOCRUZ, AM, Brasil

Dr. Patrcia Bezerra - patriciamelob@yahoo.com.br

Brasil

All the epidemic phenomenon lived deeply has a construction of the motive now empirical, now science, now the interaction of these factors. The cognitive knowledge of the family resident in the margins of the river Acutipereira in Portel city of Par they attacked by the most greatest outbreak of human rabies (15 cases confirmed) it recorded and accompanied in the word, is the objective of this work. The approach utilized was of qualitative character, interviews do not structured, with collection of narratives the which were carried out join to two family (a couple) of six of the victims and other two more family of different communities after entreaty by the community in what play to the dialogue with the researcher about the subject of the research. The conceptions of the motives of these family they are of the order of the divine: when consider the outbreak as a punishment by a fisherman of the community it to killed a boto in the locality; an action man-made harmful: where a new timber it evacuate poison in the waters of the Acutipereira river; of the nature attributing to a very strong wind that contaminated the bats; mystic being the vector bat the one that strip the blood and afterwards kills. The empirical knowledge of an phenomenon epidemic it is prominent in the measure that influences the mobilization of efforts in the prevention. The identification of the cultural cause is part of the process therapeutic. That initial work this inserted in a research of bigger carriage than is being carried out by the researcher join to the masters in Health, Society and Endemic Disease in the Amaznia (UFPA/UFAM/FIOCRUZ).

51

HUMAN RABIES EPIDEMIOLOGY, PROPHYLAXIS AND TREATMENT

RABIES PREVENTION AND GLOBAL AWARENESS


D.J. Briggs 1
1) Kansas State University, Manhattan, Kansas, USA

Dr. Deborah J Briggs - briggs@vet.ksu.edu

USA

Rabies is a very neglected totally preventable disease with the highest case fatality rate of any infectious disease known to man and 45 to 60 % of all human deaths occur in children. Unfortunately, rabies continues to receive very little attention on a global scale. Even the progress made to reduce, and in some cases eliminate, canine rabies from countries in Latin America has not received the international attention that it should have received. The reasons for the lack of international attention have always focused on the fact that the responsibility for rabies control is split between human health professionals and agricultural entities and the failure of governments in developing countries to recognize and support rabies prevention programs, both of which have lead to a lack of support and funds from international organizations. Although these are valid points, it is time for all of us in the rabies community to join together globally and devise a different strategy for increasing global awareness of rabies focusing on the fact that rabies can be totally eliminated from the animal population. Rabies has many unique features that make its elimination a challenge but by using these challenges as a focal point for awareness we can dramatically increase global attention and consequently increase funding support for its elimination. Through the help of WHO, we are proposing a World Rabies Day wherein students of veterinary medicine will join forces with rabies experts in canine rabies endemic countries to increase the awareness of rabies throughout the world. The strategy for moving this program forward is already underway in several countries. Additionally, the rabies community needs to utilize the fact that rabies is a disease of children and that by eliminating rabies in the canine population rabies in the human population, especially in children will also decrease.

52

FIRST SESSION

SECOND SESSION
RABIES PATHOGENESIS

53

HUMAN RABIES EPIDEMIOLOGY, PROPHYLAXIS AND TREATMENT

54

SECOND SESSION

IDENTIFICATION OF PATHOGENICITY DETERMINANTS OF RABIES VIRUS (RV) AND ASSESSMENT OF THE ROLE OF THE RATE OF VIRUS REPLICATION/ TRANSCRIPTION IN PATHOGENESIS
Milosz Faber 1, Rojjanaporn Pulmanausahakul, Amy Papaneri, Marie-Luise Faber 2, Jianwei Li, Matthias J. Schnell, and Bernhard Dietzschold 2
1) Molecular Targenting Technologies, Philladelphia, PA, USA, 2) Thomaz Jefferson University, Philladelphia, PA, USA

Dr. Milosz Faber - Milosz.Faber@jefferson.edu

USA

We derived a recombinant clone, SB0, of the highly pathogenic silver-haired bat-associated RV strain (SHBRV) that completely mimics the pathogenic phenotype of SHBRV and is the first infectious clone of a street RV. In vitro and in vivo experiments showed that SB0 has an increased ability to spread from cell to cell and to penetrate cells faster than the highly attenuated SN0 strain. This suggests that the more pathogenic RVs escape the host responses by a faster virus uptake and virus spread. Phenotypic analysis of chimeric RVs assembled from SB0 and the RV vaccine strain SN0 demonstrated that, although RV pathogenicity is a multigenic trait involving different RV encoded proteins and transcriptional elements, the RV G and M protein both play a predominant role in the ability of SB0 to invade the CNS from a peripheral site. Analysis of chimeric RVs also revealed that the RV G protein determines the neuroinvasiveness of a RV by facilitating fast virus entry and efficient virus spread via interaction with the RV M, and by regulating viral transcription/replication together with the RV M. Our data show that the pathogenicity of an RV correlates inversely with the rate of virus growth and viral transcription / replication in vitro, supporting the concept that the replication of an RV must be tightly regulated so that it can accomplish a complete life cycle.

55

RABIES PATHOGENESIS

DOMINANCE OF THE RABIES VIRUS G PROTEIN CARRYING A NON-PATHOGENIC DETERMINANT OVER THE G PROTEIN CARRYING A PATHOGENIC DETERMINANT
Marie-Luise Faber 1, Milosz Faber 2, Jianwei Li, Amy Rice, Jeffrey Mattis, Koon Yan Pak, Matthias Schnell, Bernhard Dietzschold 1
1) Thomaz Jefferson University, Philladelphia, PA, USA, 2) Molecular Targenting Technologies, Philladelphia, PA, USA

Dr. Marie-Luise Faber - mfaber@mtarget.com

USA

RV G protein is not only the major RV antigen responsible for induction of protective immunity but also the major contributor to RV pathogenesis. Several rabies virus (RV) vaccine strains containing aspartic acid (Asp) or glutamic acid (Glu) instead of arginine (Arg) at position 333 of the RV glycoprotein (GAN) are apathogenic in immunocompetent mice. This makes these variants suitable as live-attenuated vaccines. A recombinant RV carrying two identical Arg333!Glu333 G genes (SPBNGAN-GAN) over-expressed the RV G and conferred protective immunity in a variety of animals, superior to that of a RV variant carrying only a single GAN protein (SPBNGAN). A major problem with the use of live attenuated RV vaccines is reversion to the pathogenic phenotype. It has been shown that a Asn194! Lys194 mutation in G (GAK) was solely responsible for the emergence of the pathogenic phenotype. To investigate whether the presence of two G genes can actually increase the possibility for reversion to pathogenicity, we constructed RVs that contain both the GAN and GAK gene (SPBNGAN-GAK or SPBNGAKGAN). Additionally, we constructed a RV containing two GAK genes (SPBNGAK-GAK). While SPBNGAK-GAK was pathogenic, both SPBNGAN-GAK and SPBNGAK-GAN were non-pathogenic after i.c. infection of adult mice. This indicates that the non-pathogenic phenotype determined by GAN is dominant over the pathogenic phenotype associated with GAK. Most interestingly, virus growth and RV RNA transcription/replication rates were significantly higher in SPBNGAN- as compared to SPBNGAK-infected neuroblastoma cells. In cells infected with SPBNGAN-GAK or SPBNGAK-GAN, virus growth kinetics and RV RNA transcription rates were similar to those seen SPBNGAN-infected cells, indicating that the pathogenicity of RV correlates inversely with its transcription/ replication rate. From these data we conclude that an rRV carrying two identical attenuated G genes is not only more immunogenic but also exhibits a lower risk for reversion to the pathogenic phenotype.

56

SECOND SESSION

CONSEQUENCES OF THE INNATE IMMUNE RESPONSE TO RABIES VIRUS INFECTION


Marzena Fabis 1, Timothy W. Phares 1, Hilary Koprowski 1, D. Craig Hooper 1
1) Thomas Jefferson University, Philladelphia, PA, USA

Dr. Marzena Fabis - Marzena.J.Fabis@jefferson.edu

USA

Infection with pathogenic rabies virus (RV) is generally associated with the development of neurological disease. However, there are a number of avirulent RV strains, such as CVS-F3, that do not cause disease in immunocompetent animals. Attenuated RV spread to the CNS tissues of normal mice but are efficiently cleared when the following occur: 1/ an innate immune response by CNS resident cells; 2/ RV-specific adaptive immunity which originates in the periphery; 3/ increased blood-brain barrier (BBB) permeability; and 4/ accumulation of the appropriate effectors of adaptive immunity in the CNS tissues. The innate immune response, which is essential to attract immune effectors into the CNS tissues, can be triggered by virus infection or by the consequences of the RV-specific immune response including enhanced BBB permeability and immune cell invasion. Elements of the innate, ICAM-1, IP-10, MCP-1,aresponse that are triggered by the infection include TNF, and RANTES. While the elevated expression of theseb, MIP-1aMIP-1 pro-inflammatory mediators does not cause disease in CVS-F3 infected mice, it is expected that they could contribute to the induction of immune-mediated disease if they attract inappropriate effectors into the CNS tissues. To determine if this is the case mice immunized with antigens associated with myelin were infected with CVS-F3. Immunization of mice with these antigens generally causes the development of an autoimmune disease resembling multiple sclerosis, experimental allergic encephalitis (EAE). CVS-F3 infection increases the incidence and severity of EAE in conventional mice. Moreover, when mice are immunized with myelin antigens they rarely develop severeadeficient in TNF- -/-aEAE. Infection with CVS-F3 overcomes this deficit and myelin-immunized TNF- mice then develop EAE with the incidence and severity of normal mice. We conclude that infection with rabies virus drives a CNS pro-inflammatory response that is essential to attract RV-specific immune effectors into the tissues and clear the virus. However, this comes with an increased risk of drawing immune effectors with other specificities into the CNS tissues and causing, for example, CNS autoimmunity. Supported by National Institutes of Health Grant AI09706.

57

RABIES PATHOGENESIS

EARLY SPREAD OF RABIES VIRUS AND THE DEVELOPMENT OF IMMUNITY


Rhonda B. Kean 1, Timothy W. Phares 1, Anirban Roy 1, Mikhail Prosniak 1, Hilary Koprowski 1, D. Craig Hooper 1
1) Thomas Jefferson University, Philladelphia, PA, USA

Dr. Rhonda Kean - Rhonda.kean@jefferson.edu

USA

While considered highly neurotrophic, most rabies viruses (RV) replicate in non-neuronal tissues in vivo, including at the site of infection. Certain virus strains, such as those associated with silver-haired bats (SHBRV) have an enhanced capacity to replicate in non-neuronal cells in vitro that may be relevant to their infectivity in vivo. For example, virus spread to non-neuronal tissues such as the spleen and lymph nodes may either positively or negatively impact the development of anti-viral immunity. To provide insight into the relationships between pathogenesis, the spread of RV to different sites and the development of immunity, we compared these parameters in mice infected with attenuated, vaccine RV strains versus highly pathogenic isolates. Mice were infected either intra-muscularly (gastrocnemius) or intra-nasally and various tissues collected for RT-PCR analysis of rabies nucleoprotein and glycoprotein mRNAs at time intervals from 24 to 72 hours afterwards. Parameters of immunity including RV-specific T cell reactivity and antibody titers were also assessed. Nucleoprotein mRNA of all RV tested, whether pathogenic in conventional mice or not, becomes detectable in the CNS tissues of immunocompromised mice as early as 24 hours post infection for pathogenic strains and 48 hours for apathogenic variants. However, nucleoprotein mRNA also appears in spleen and lymph node over a similar time period. Studies of cell subsets isolated from the spleens of RV-infected mice identify the cells positive for nucleoprotein mRNA as non-T cells, likely dendritic or B cells. We consider that this is likely to be relevant to the development of antiviral immunity as, regardless of the pathogenicity of the virus, nucleoprotein specific T cells can be detected in lymph nodes 48 hours after infection. The failure of these cells to respond to viral glycoprotein suggests that they are specific for newly synthesized nucleoprotein rather than the intact virus. We speculate that the peripheral replication of viral proteins in lymphoid organs is important in the early development of anti-viral immunity and this does not differ significantly between different virus isolates. Supported by National Institutes of Health Grant AI09706.

58

SECOND SESSION

RABIES PATHOGENESIS AND THE BLOOD-BRAIN BARRIER


D.C .Hooper 1, T.W. Phares 1, H. Koprowski 1, A. Roy 1
1) Thomas Jefferson University, Philladelphia, PA, USA

Dr. Craig Hooper - douglas.hooper@jefferson.edu

USA

Rabies post-exposure prophylaxis of infected individuals generally fails once clinical evidence of the infection appears. In animal models the development of clinical signs of rabies infection is associated with the spread of the virus to the CNS. While all rabies viruses can invade the CNS tissues of mice, only attenuated, vaccine strains of the virus are efficiently cleared by rabies virus-specific immune mechanisms. In addition, the CNS inflammatory response associated with the clearance of attenuated rabies viruses from CNS tissues is not seen in mice infected with pathogenic isolates. These observations suggest that there may be a selective deficit in the development of immunity to pathogenic rabies viruses. To establish whether this is the case we compared the development of immunity to a highly pathogenic rabies virus isolate associated with the silver-haired bat, SHBRV-17, with an attenuated, laboratory strain CVS-F3. Measures of innate as well as peripheral anti-viral immunity are comparable or higher in SHBRV-17 infected mice by comparison with CVS-F3 infected animals at the time virus clearance begins in the latter. However, immune effectors including CD4 T cells and B cells fail to invade the CNS tissues of SHBRV-17 infected mice and clear the virus. Moreover, the enhanced blood-brain barrier (BBB) permeability seen in the cerebellum when CVS-F3 is cleared from CNS tissues does not develop in SHBRV-17 infected animals. Adoptive transfer experiments reveal that immune cells from SHBRV-17 infected mice are fully functional as they can clear CVS-F3 from the CNS tissues of immunocompromised recipients. We conclude that as a consequence of SHBRV-17 infection the BBB becomes refractory to the permeability changes associated with immune effector delivery to CNS tissues and virus clearance. We speculate that once the capacity to open the BBB has been lost, conventional post-exposure prophylaxis will fail. Further understanding of the mechanism utilized by the immune response to enhance BBB permeability and how this can be inhibited is necessary to develop treatment approaches to clear rabies virus after it has reached CNS tissues. Supported by National Institutes of Health Grant AI09706.

59

RABIES PATHOGENESIS

CLEARANCE OF RABIES VIRUS FROM THE CENTRAL NERVOUS SYSTEM


Timothy Phares 1, Anirban Roy 1, Hilary Koprowski 1, D. Craig Hooper 1
1) Thomas Jefferson University, Philadelphia PA, USA

Dr. Timothy Phares - djfrict@yahoo.com

USA l

Infection of humans with rabies virus (RV) is generally lethal in the absence of post-exposure prophylaxis consisting of both the administration of rabies-specific virus neutralizing antibodies (VNA) and active rabies immunization. Passively transferred antibodies contribute to recovery from infection by neutralizing RV while active immunization, in addition to providing the stimulus for long-term production of more RV-specific antibody likely has some additional therapeutic effect. We believe that an active RV-specific immune response is required to deliver immune effectors to infected tissues and clear the virus, which is expected to be particularly difficult if the virus has spread to nervous tissue where it is protected from immune effectors by the blood-brain barrier (BBB). To test this hypothesis we have studied the clearance of the apathogenic RV, CVS-F3 from the CNS. Like other avirulent RV, CVS-F3 spreads to the CNS of normal adult mice before it is cleared without sequelae by the naturally developing immune response. Clearance follows the CD4 T cell-dependent increase in the permeability of the BBB. Interestingly, the increase in BBB permeability allows only molecules smaller that antibodies to infiltrate CNS tissues and occurs several days prior to peak serum levels of rabies-specific antibody. Moreover, the reduction in virus replication in the CNS tissues begins after restoration of BBB integrity. The onset of BBB permeability is associated with the appearance of T and B cells in the CNS -lightktissues and the clearance of virus with the expression of high levels of chain mRNA in the CNS. Furthermore, the passive administration of VNA does not clear RV from the CNS of mice lacking B cells. These data argue that the in situ production of VNA is more critical for the clearance of RV from the CNS rather than the leakage of serum antibody from the circulation into CNS tissues. We speculate that this may also be the case for the clearance of RV from other tissues. Supported by National Institutes of Health Grant AI09706.

60

SECOND SESSION

HOST RESPONSE TO EUROPEAN BAT LYSSAVIRUS (EBLV) INFECTION, PART I: SUSCEPTIBILITY OF EPTESICUS FUSCUS TO EBLV-1 FOLLOWING DIFFERENT ROUTES OF INFECTION
A. Vos 1, N. Johnson 2, T. Mueller 3, L. Neubert 1, I. Kaipf 4, K. Mansfield 2, C. Freuling 3, R. Franka 5, A. Denzinger 4, C.E. Rupprecht 5, A.R. Fooks 2
1) Impfstoffwerk Dessau - Tornau GmbH, Rosslau, Germany, 2) Veterinary Laboratories Agency - Weybridge, Surrey, United Kingdom, 3) Friedrich-Loeffler-Institute, Wusterhausen, Germany, 4) Zoological Institute, Eberhard Karls University, Tuebingen, Germany, 5) Centers for Disease Control and Prevention, Atlanta, GA, U.S.A.

Dr. Adriaan Vos - ad.vos@idt-direct.de

Germany

The rabies situation in Europe has changed dramatically in the past 25 years. As a result of oral vaccination, terrestrial rabies due to genotype 1 Lyssavirus (RABV) has been eradicated from large areas of Europe. Meanwhile, two other lyssaviruses have been identified in many European countries: European Bat Lyssavirus type 1 and 2 (EBLV-1 and EBLV-2). Unfortunately, only limited knowledge on the susceptibility of the natural EBLV-hosts, insectivorous bats, to EBLV is available. As part of an ongoing project to investigate the host response to EBLVinfection, a preliminary study to evaluate the susceptibility and pathology associated with an EBLV-1 infection in Eptesicus fuscus was studied. The following groups were investigated; Group 1: negative control (mouse brain suspension) - i.m., n=7; Group 2: i.n., 10 3.2 MLD50, n=7; Group 3: i.c., 10 3.2 MLD50, n=6; Group 4: i.m. 10 3.2 MLD50, n=7; Group 5: p.o. 10 3.2 MLD50, n=7; Group 6: i.m. 10 2.2 MLD50, n=7. The animals were observed for 90 days post-infection. Blood - and saliva samples were collected from all animals on a monthly and weekly basis. At the end of the study, tissues were collected from all animals and examined for the presence of viral antigen. The results indicated that the pathogenesis of EBLV-1 in their natural hosts differs markedly from RABV in terrestrial wildlife. For example, no mortality in the p.o. and i.n. inoculated animals was observed, but also 2 of 6 bats inoculated i.c. did not succumb to infection. Furthermore, only two surviving animals, one in each i.m.-inoculated group, had detectable levels of virus neutralizing antibodies. Limited virus shedding was demonstrated in the saliva by virus isolation and PCR in some animals. We conclude from this preliminary study that EBLV-1 is transmitted predominantly by the bite of an infected bat, possible other routes of (horizontal) transmission does not seem to be of any relevance.

61

RABIES PATHOGENESIS

DIFFERENCES BETWEEN EBLV-1 AND EBLV-2 PATHOGENESIS: SHEEP AS A MODEL OF DISEASE RECOVERY
Sharon M. Brookes 1, Robert Klopfleisch 2, Thomas Muller 2, Derek M. Healey 1, Jens Teifke 2, Elke Lange 2, Janette Kliemt 2, Nicholas Johnson1, Christopher Finnegan 1, Linda Johnson 1, Volker Kaden 2, Aian Vos 3, Anthony R. Fooks 1
1) Veterinary Laboratories Agency, Surrey, UK, 2) Friedrich Loeffler Institut, Germany, 3) Impstoffwerk Dessau Tornau, Germany

Dr. Sharon Brooks - s.brookes@vla.defra.defra.gsi.gov.uk

U.K.

European bat lyssaviruses (EBLVs) have been known to cross the species barrier from their native bat host to other terrestrial mammals. EBLV type-1 has been shown to infect humans, sheep and a stone marten in the field. Other susceptible animals include cats, dogs, foxes and rodents following natural peripheral inoculation under experimental conditions. EBLV type-2 appears to be less virulent both in bats and in its ability to spill-over in to other species; two human cases and experimentally infected rodents only. Our studies have confirmed EBLV-1 and -2 susceptibility in sheep (Ovis ammon) following direct introduction of virus to the central nervous system. Notably, mild clinical disease was also observed in those peripherally exposed to virus via the intramuscular (IM) route. However, 75% of animals infected with EBLV-1 and 100% of those that had received EBLV-2 developed rabies clinical signs and then recovered, this re-occurred between 4-70 dpi during the 94-day observation period. The disease pathogenesis scenario also varied substantially between the two viruses. EBLV-1 resulted in per-acute clinical signs that were suggestive of motor neurone involvement, the induction of an antibody response and less inflammatory infiltrate and antigen in the brain in comparison to that in EBLV-2 infected sheep. The latter involved a more protracted development of disease that was behaviour orientated, however, the virus neutralising antibody response was not evident and there was more viral antigen and inflammation in the brain than that of EBLV-1. It is clear that a high infectious dose is required to establish EBLV infection under experimental conditions (>5.0 logs/ml), the infectious dose in field cases remains unknown. EBLVs represent a low but real risk to human and animal health across Eurasia where the hazard is high if exposure goes untreated.

62

SECOND SESSION

A NEW MODEL TO STUDY AN ALTERNATIVE PATHWAY FOR RABIES TRANSMISSION


Silvana R. Favoretto 1, 6, Marilene F . Almeida2, 6, Luzia F. A . Martorelli 2, 6, Caroline C. Aires 3, 6, Marcelo B. Labruna 4, Dbora R. V. Sacramento 5, 6, Edison L. Durigon 6 Hilary Koprowski 7
1) Instituto Pasteur, So Paulo, Brasil 2) Centro de Controle de Zoonoses, Municpio de So Paulo, SP , Brasil 3) Museu de Zoologia, Universidade de So Paulo, SP , Brasil 4) Faculdade de Medicina Veterinria da Universidade de So Paulo, SP , Brasil 5) Genomic Engenharia Molecular Ltda 6) Instituto de Cincias Biomdicas da Universidade de So Paulo, SP , Brasil 7) Thomas Jefferson University, PA, USA

Dr. Silvana Regina Favoretto Lazarini - srfavoretto@usp.br

Brasil

During the last years, countries as the USA, Finland, Australia, Mexico, Chile and Canada have been described several humans cases that were not possible to detect the rabies transmission way or the possible transmitter animal. The question concerns the link between bats and rabies victim in the absence of skin lesions attributable to a bat bite or without documented history of it. In Brazil, besides one rabies virus maintained by dog population and an arboreal cycle transmitting the rabies directly to humans by marmosets, there are different epidemiological cycles with several chiropteran species acting as reservoirs and/or transmitters. Considering these questions, we developed a protocol for the study of alternatives pathways of rabies transmission with a multidisciplinary professional team and the utilization of the appropriated facilities. This project considers the possible transmission of rabies to several animal specie including humans by a different way involving the presence of the ectoparasites as a vector in this process. Literature datas showed that ticks from Ornitodorus Order are found as ectoparasites of different bats species in the Americas. This protocol included the utilization of the ticks, dipterous and bats captured in their habitat and maintained in laboratory Nowadays, the standardization of molecular techniques have been developed having in mind the presence of controls to detect ectoparasites genes and also mammals genes. Different methods for RNA extraction are also tested.

63

RABIES PATHOGENESIS

NON-BITE TRANSMISSION OF RABIES VIRUS


A.R. Fooks 1, N. Johnson 1, R. Phillpotts 2, A. Vos 3, T. Mller 4, C. E. Rupprecht 5
1) Veterinary Laboratories Agency, UK, 2) Defense, Science and Technology Laboratory, UK, 3) Impfstoffwerk Dessau Tornau GmbH, Streetzerweg, Germany, 4) Friedrich Loeffler Institute, Germany, 5) Centers for Disease Control and Prevention, Atlanta, GA, USA

Dr. Tony Fooks - t.fooks@vla.defra.gsi.gov.uk

U.K.

A Scottish bat conservationist developed rabies and subsequently died from infection caused by a bat variant of rabies virus: European bat lyssavirus type-2 (EBLV-2). The source of this infection and the means of transmission have not been confirmed, although contact with infected saliva from a bat by the small bite hypothesis was the most probable route of exposure. We have further tested the hypothesis that lyssaviruses, particularly rabies virus and EBLV-2, might be transmitted via the airborne route. Mice were challenged via direct introduction of lyssavirus into the nasal passages. Two hours after intranasal challenge with a mouse-adapted strain of rabies virus (Challenge Virus Standard), viral RNA was detectable in the tongue, lung and stomach. We have also observed that genomic RNA was detectable within non-CNS tissue including the tongue, bladder, and stomach of naturally infected Daubentons bats. All of the mice challenged with CVS by direct intranasal inoculation developed disease signs by 7 days post-infection. Two of five mice challenged by direct intranasal inoculation of EBLV-2 also developed disease between 16 and 19 days post-infection. Mice challenged through aerosol exposure were highly susceptible to infection with CVS but not EBLV-2 by inhalation. In a second study, 1 of seven bats (Eptesicus fuscus) that was used as an experimental control in a pathogenesis study became sick and died whilst being housed in a unit alongside bats infected with EBLV-1. The dead bat was not in direct contact with the infected bats thereby eliminating the bite route as a source of transmission in this case, and suggesting cryptic transmission. These data support the hypothesis that under rare circumstances, lyssaviruses can be transmitted by non-bite transmission.

64

SECOND SESSION

AISLAMIENTO DE VIRUS RABICO EN GLANDULAS SALIVALES DE MURCIELAGOS INSECTIVOROS


Federico Gury Domen 1, Fernando Beltran 1
1) Instituto de Zoonosis Louis Pasteur, Buenos Aires, Argentina

Dr. Federico Gury Dohmen - guryfe@hotmail.com

Argentina

La rabia urbana fue controlada a partir del ao 1981, mediante los programas de prevencin y control de la rabia llevados a cabo en la Ciudad de Buenos Aires y alrededores, pero a partir de 1990 adquiri una real importancia epidemiolgica el ciclo areo debido a la presencia de casos positivos en murcilagos insectvoros y por el aumento del nmero de contactos con o sin mordeduras con el hombre y los animales domsticos. La especie predominante fue Tadarida brasiliensis y en menor grado, Lasiurus cinereus y Lasiurus borealis. La prevalencia observada en la ciudad de Buenos Aires para estas especies es cercana al 3 %. Nuestro objetivo fue determinar la presencia de virus rabico, su titulo en glndulas salivales, caracterizacin antignica y el riesgo probable de transmisin por contacto con animales domsticos y el hombre. En el estudio realizado se seleccionaron 26 muestras positivas en cerebro correspondientes al perodo 1999-2005 en el que se produjeron 20 contactos con y sin mordeduras para realizar aislamientos por inoculacin intracerebral en ratones lactantes de 1 a 3 das a partir de sus glndulas salivales. Tambin se escogieron 20 muestras negativas en cerebro como lote testigo. Las tipificaciones se realizaron utilizando un panel de 19 anticuerpos monoclonales dirigidos contra la nucleoprotena viral cedidos por el CDC de Atlanta, Georgia (USA); se usaron como testigos cepas de las variantes 2-4-6-ERA y DR19, como as tambin improntas de cerebro normal. Los resultados obtenidos mostraron un alto porcentaje de aislamientos (76,9%) en glndulas salivales, con un ttulo promedio de 4.02 (expresado como Log DL50). En un lote de muestras positivas en cerebro y glndulas salivales, se compararon sus ttulos promedio, siendo sus valores 4.75 y 3.81 respectivamente. En el lote testigo negativo no pudo aislarse virus en glndulas salivales. Los virus aislados correspondieron principalmente a las variantes 4 para Tadarida brasiliensis, 6 para Lasiurus cinereus y borealis y sus respectivas subvariantes. Para concluir, de la comparacin de ambos lotes de murcilagos al igual que en otros animales, la diseminacin siempre se produce desde el cerebro hacia las glndulas salivales. Los ttulos hallados en cerebros no fueron muy altos, o bien por las condiciones en que fueron enviadas las muestras o probablemente por una infeccin con ttulo bajo, lo que permitira en la mayora de los casos, la diseminacin hacia las glndulas salivales determinando su aislamiento en las mismas. El riesgo probable de transmisin expresado como el N de contactos/N de murcilagos positivos en sus glndulas, fue del 50% para el perodo en estudio, indicando que de un lote de 20 de aislamientos positivos en glndulas, solo 10 ejemplares pudo estar en condiciones de transmitir la enfermedad.

65

RABIES PATHOGENESIS

ISOLATION OF RABIES VIRUS IN MICE AND N2A CELLS FROM NATURALLY INFECTED BATS OF THE STATE OF SO PAULO, SOUTHERN BRAZIL
K.C. Scheffer 1, M.L. Carrieri 1, H.C.P . Santos 1, I. Kotait 1, F .H. Ito 2
1) Instituto Pasteur, So Paulo, SP Brasil 2) Faculdade de Medicina Veterinria e Zootecnia da Universidade de So Paulo, SP , Brasil

Dr. Karin Corra Scheffer Ferreira - ksferreira@pasteur.saude.sp.gov.br

Brasil

The Diagnostic Rabies Section of the Pasteur Institute, So Paulo-SP , received 4,393 bats from the State of So Paulo, in the period of 2002-2003. Classified into vampires, frugivorous, and insectivorous, and 10 distinct genera, 82 bats were positive for rabies by the Fluorescent Antibody Test. Viral isolation from different tissues was performed in mice and in N2A cells, by inoculating 20% (weight/volume) suspensions prepared from brain, salivary gland, tongue, urinary bladder, kidneys, stomach, pectoral muscle, brown fat, lung, heart, and genital tract. The rabies-positive mice brains were submitted to a panel of monoclonal antibodies (Mabs). Antigenic typing revealed that viruses were divided into two groups: viruses common to Desmodus rotundus vampire bats and fruit-eating Artibeus sp, sharing the antigenic variant 3 (AgV3); and viruses from insectivorous bats presented profiles of AgV 4 and 6. Another group of viruses from insectivorous bats was not compatible with the isolates defined by the Mabs panel. The results of viral isolation were analyzed by the Kruskal-Wallis nonparametric ANOVA test, applied for the positive (scored = 1) and negative (scored = 0) results. Significant results were found when brain materials of the vampire and frugivorous bats were compared to heart, urinary bladder, kidney, pectoral muscle, genital tract, and stomach. The salivary gland was significant when compared to kidney, and stomach, using = 0.05 and P<0.001. Similarly, specimens from the insectivorous bats were significant with P<0.001, when results of brain were matched with those of the lung, tongue, heart, urinary bladder, kidney, brown fat, pectoral muscle, genital tract, and stomach. For the vampire and frugivorous bats, the submission of brain, salivary gland, lung, tongue, and heart were the most adequate for viral isolation in N2A cells, and for the insectivorous bats, the brain, salivary gland, brown fat, lung, and urinary bladder were the most suitable.

66

SECOND SESSION

INVESTIGACIN DEL VIRUS DE LA RABIA EN GLNDULAS SALIVARES DE CNIDOS SILVESTRES (CERDOCYON THOUS), INFECTADOS NATURALMENTE, EN EL ESTADO DE PIAU, BRASIL
Romualdo Oliveira Spindola 1, Joo Pereira da Silva 1, Rosangela Cavalcante de Carvalho 1, Vnia Maria Alves Carvalho 1, Samira M. Achkar 2, Juliana Galera Castilho 2, Rodrigo Stefani Ceretta 2, Ivanete Kotait 2
1) Centro de Controle de Zoonoses de Teresina, Piau, Brasil, 2) Instituto Pasteur, So Paulo, SP , Brasil

Dr. Romualdo Spndola - romualdospindola@yahoo.com.br

Brasil

La reduccin de la rabia en el perro, en Brasil, ha derivado en una mayor atencin de las acciones de vigilancia epidemiolgica de esta enfermedad en mamferos silvestres, particularmente en murcilagos y en cnidos. En el nordeste del pas, el ciclo de la rabia en cnidos viene mantenindose desde hace algunos aos, habiendo sido los silvestres responsables de varias muertes humanas. En el 2005, en el Estado de Piau, fueron identificados varios animales, de la especie Cerdocyon thous, infectados con el virus de la rabia. La amplia distribucin de esta especie animal en Brasil, los hbitos sinatrpicos de estos animales y la evidente asociacin entre la rabia de los cnidos domsticos y silvestres, ya relatada por pesquisadores del Instituto Pasteur, con base en estudios antignicos y genticos, evidencian ser un nuevo desafo para los profesionales de la salud pblica. A travs de la immunofluorescencia directa y del aislamiento viral en ratones y clulas N2A, se investig la presencia del virus de la rabia en glndulas salivares (sublingual y submandibular) en especimenes de Cerdocyon thous, naturalmente infectados con el virus de la rabia. La positividad fue del 27,3% en las glndulas sublinguales y del 80% en las submandibulares, destacndose la importancia epidemiolgica de estos animales en el mantenimiento de la rabia en la regin y la necesidad de estudios que avizoren el establecimiento de uno programa de vacunacin oral para los cnidos silvestres de la regin.

67

THIRD SESSION
DIAGNOSTIC, CHARACTERIZATION AND MOLECULAR EPIDEMIOLOGY

DIAGNOSTIC, CHARACTERIZATION AND MOLECULAR EPIDEMIOLOGY

70

THIRD SESSION

RESEARCH TRENDS ON RABIES IN THE FGI FEDERAL CENTRE FOR ANIMAL HEALTH
S. Rybakov 1, K. Gruzdev 1, A. Metlin 1
1) Federal Centre for Animal Health, Russia

Dr. Sergey Rybakov - s.s.rybakov@mail.ru

Rssia

Rabies is constantly endemic in many regions of the Russian Federation and rabies cases in different species of wild, farm and domestic animals appear every year. In the year of 2005 more than five thousand animal rabies cases were recorded. The FGI Federal Centre for Animal Health (FGI ARRIAH) is an all-Russia research centre in rabies and it conducts animal rabies monitoring, diagnosis, epidemiological studies, improvement of rabies diagnostic techniques, evaluation of rabies vaccine safety and potency and investigation of genetic characteristics of the field isolates and vaccine strains of rabies virus. Because the live attenuated oral rabies vaccine aimed at the prevention of rabies in wild animals as well as inactivated rabies vaccines for domestic and farm animals are produced in the FGI ARRIAH, centre conducts studies on efficacy and safety of anti-rabies vaccines. The following techniques are used for the laboratory diagnosis of rabies: fluorescent antibody test (FAT), mice inoculation test, virus isolation in cell cultures, ELISA, latex agglutination, histological and immunohistochemical technologies. The polymerase chain reaction followed by the PCR-product sequencing is used for the genetic studies. A rabies diagnostic kit in FAT was developed consisting of all the components needed for carrying out the test including positive and negative controls. Direct liquid-phase FAT method for rabies diagnosis in brain samples conserved in formalin have been developed and patented recently. In the framework of the Finnish-Russian collaboration programme to control rabies in wild animals, we have studied the molecular and biological characteristics of 40 field isolates from different parts of Russia, Finland, Estonia. Thus, the work focused on diagnostic methods improvement, epidemiology, prevention and control of rabies is carried out in the FGI ARRIAH.

71

DIAGNOSTIC, CHARACTERIZATION AND MOLECULAR EPIDEMIOLOGY

LABORATORIAL DIAGNOSTIC OF RABIES: COMPARATION BETWEEN RABIES TISSUE CULTURE INOCULATION, MOUSE INOCULATION AND FLUORESCENT ANTIBODY TESTS, USING BOVINE AND EQUINE SPECIMENS
Adolorata Ap. Bianco Carvalho 1, Eduardo Rigo Deberaldini 1, Marli Francisca C. Santos 1, Roseli Mendes Resende 1, I Fumio Honma Ito 2
1) Universidade Estadual Paulista, Jaboticabal, SP , Brasil, 2) Universidade de So Paulo, SP , Brasil

Dr. Adolorata Aparecida Bianco Carvalho - adbianco@fcav.unesp.br

Brasil l

The fluorescent antibody test (FAT) has proven to be a fast and most accurate microscopic test available for the routine diagnosis of rabies. For being not infallible (false-negative and false-positive results may occur), the WHOs current recommendation is to use it in parallel with the mouse inoculation test (MIT), but nowadays researchers are seeking other modern systems to replace the MIT. For this purpose, the rabies tissue culture inoculation test (RTCIT), using mouse neuroblastoma cells is recommended by the French National Reference Center for Rabies. In countries like Brazil, due to many technical and economic limitations, the MIT is still the most widely used test, but claims to replace the MIT are increasing. The proposal of the present work is to compare the sensitivity of RTCIT and MIT, using bovine and equine specimens, and to assess the test characteristic and the safety of the RTCIT. A total of 358 brain specimens rabies positive were used (12 samples originally FATweakly positive). The brain suspensions were inoculated, in parallel, intracerebrally into mice and in neuroblastoma cells. RTCIT was performed using g of DEAE-D for 1mL of medium containing 10% fetal bovine serum,m15 3.5x105cells/mL, viral suspensions diluted at 10-2 and an antinucleocapside conjugate solution 1:25; incubation in a 34oC chamber incubator with 5% CO2, for 96h. The sensitivity of the tests was determined and the agreement values were established by using the KAPPA indicator: MIT X RTCIT 98,32%; RTCIT X FAT 96,37%; MIT X FAT 96,37%. The results showed a great agreement between the MIT X RTCIT, suggesting that the RTCIT test can be comparable to MIT in routine conditions, especially for evaluation of FAT-weakly positive samples. The advantage of the RTCIT resides in the fact that results are obtained within two or four days, as compared to 30 days in the MIT.

72

THIRD SESSION

HUMAN RABIES TRANSMITTED BY VAMPIRE BATS: ANTIGENIC AND GENETIC CHARACTERIZATION OF RABIES VIRUS ISOLATES FROM ECUADOR AND BRAZIL
Juliana G. Castilho 1, Elizabeth Salb Travassos da Rosa 2, Anbal Mantilla 3, Ekaterina Durymanova 1, Maria Luiza Carrieri 1, Ivanete Kotait 1
1) Instituto Pasteur, So Paulo, SP Brasil 2) Instituto Evandro Chagas, Belm, Par, Brasil 3) Instituto Nacional de Higiene Leopoldo Izquieta Perez, Quito, Ecuador

Dr. Juliana Galera Castilho - juliana.castilho@uol.com.br

Brasil

Since 2004, the main transmitter of human rabies in Latin America has been the vampire bat (Desmodus rotundus). According to data from PANAFTOSA/OPAS, 145 deaths by rabies were registered in the region between 2004 and 2005. Of these, 98 (67.6%) were caused by transmission by vampire bats (46 in 2004 and 52 in 2005). The highest number of cases during this period (64) occurred in Brazil. Ecuador registered two cases in 2005, in Pastaza Province. Deaths by rabies in other countries with continued deforestation in the Amazon region were also reported. These deaths were confirmed by laboratory diagnosis or by an epidemiological link with aggression by vampire bats in populations living in vulnerable dwellings in areas where the animal population was small. These factors constitute the characteristics of risk areas. The main objective of this study is to compare and classify the antigenic and genetic characterization of the 29 human rabies-virus isolates from different areas of the Amazon region. Two isolates were from Ecuador and 27 from Brazil (11 from the municipalities of Carutapera, Godofredo Viana and Turiau in Maranho State and 16 from the municipalities of Portel, Viseu and Augusto Corra in Par State). All the samples were submitted to antigenic typing using a panel of eight monoclonal antibodies provided by CDC Atlanta, USA, and were characterized as antigenic variant 3 (AgV3), which is compatible with the variant frequently isolated from Desmodus rotundus. The partial N gene (nt 105-1477) of the 29 isolates was sequenced. The sequences were aligned with homologous sequences derived from GenBank using the CLUSTAL/W method and used to build a neighbor-joining distance tree with the K-2-P model. The 29 human isolates were genetically grouped into the Desmodus rotundus cluster, which was divided into three subclusters. Two subclusters were composed of isolates from Brazil; one of these subclusters was composed of isolates from different municipalities (Carutapera, Godofredo Viana, Turiau, Viseu and Augusto Corra), and the other isolates from the municipality of Portel. The third subcluster consisted of isolates from Ecuador. The results of this study show that the isolates belong to the same rabies cycle as that in the vampire bat D. rotundus. However, the division of subclusters for variant AgV3 suggests that there are geographic differences between isolates obtained over a short period of time.

73

DIAGNOSTIC, CHARACTERIZATION AND MOLECULAR EPIDEMIOLOGY

MOLECULAR EPIDEMIOLOGICAL STUDY OF RABIES VIRUS ISOLATES FROM TRINIDAD AND COMPARISON WITH ISOLATES FROM THE SOUTH AMERICAN CONTINENT
D.A. Marston 1, C. Ezeokoli 2, A. Velasco-Villa 3, L.A. Orciari 3, L.M. McElhinney 1, A. Newaj-Fyzul 2, J.D. Watkins 2, S. Titus 2, J. Ryan 2, C.E. Rupprecht 3, A.R. Fooks 1
1) Veterinary Laboratories Agency, UK, 2) The University of the West Indies, Trinidad and Tobago - 3) Centers for Disease Control and Prevention, Atlanta, GA,USA

Dr. Denise Marston - d.marston@vla.defra.gsi.gov.uk

U.K.

The island of Trinidad provides an interesting platform to study sporadic outbreaks of rabies in domestic livestock in the apparent absence of canine rabies. Here, we describe a molecular epidemiological study of a discrete panel of rabies isolates from an outbreak in livestock in Trinidad between 1997 and 2000. During the outbreak, one of 51 vampire bats tested for rabies by FAT was confirmed positive, implicating this vector in disease transmission. However, since the early 1970s, intensive trapping and testing of vampire bats failed to detect rabies virus by FAT, raising the possibility that the initial bat vector in this outbreak was from a neighboring country on mainland South America. Venezuela and Guyana are Trinidads closest neighbors with Venezuela being only 9 miles south of Trinidad at the closest point. Using partial N gene sequence, we have successfully compared the Trinidad outbreak isolates with isolates from neighboring countries. Our phylogenetic data supports the hypothesis that the outbreak was due to a bat rabies variant, likely to be from vampire bats. In addition, the phylogenetic data suggests there were two independent foci, one being closely associated with isolates from Guyana.

74

THIRD SESSION

MOLECULAR EPIDEMIOLOGY OF RABIES VIRUSES FROM DOMESTIC AND WILD MAMMALIAN OF BRAZIL
Leda Maria Silva Kimura 1, Joeler Vargas Dantas Jr 2, Wlamir Correa Moura 3, Ivanete Kotait 4, Victor Augustus Marin 2, Paulo Eduardo Brando 4
1) Laboratrio de Biologia Animal da Empresa de Pesquisa Agropecuria do Estado do Rio de Janeiro, Brasil 2) Instituto Nacional de Controle de Qualidade em Sade- Fundao Instituto Oswaldo Cruz, RJ, Brasil 3) Instituto Pasteur de So Paulo, So Paulo, Brasil 4) Faculdade de Medicina Veterinria e Zootecnia - Universidade de So Paulo, So Paulo, Brasil

Dr. Leda Maria Silva Kimura - ledavet@globo.com

Brasil

Rabies is one of the most feared zoonosis, once it always results in the death of the affected patient, what makes rabies morbidity equal to its mortality. Furthermore, economic losses due to rabies epidemics in cattle, the economic impact in agrobusiness derived from decreased milk and meat production and implications in public health must also to be taken into account. Using classic and traditional techniques, the present research aimed to study the molecular diversity of rabies virus strains from domestic and wild animals that circulate in different Brazilian regions (North, Northeastern, Southeastern, South and Center-Western) comparing the strains amongst them and with strains from a municipality from Rio de Janeiro State (Porcincula) based on the gene coding for the nucleoprotein. The RT-PCR was applied to 32 samples of central nervous system tissue of rabies-suspected animals, showing an agreement of 100% with the classic tests, allowing a positive diagnosis even in decomposed samples. Thirteen out of these 32 samples were submitted to partial sequencing resulting in the expected groups of rabies virus variants, i. e., antigenic variants 2, 3, fixed strains and marmoset strains. A regional pattern was found regarding the variant 3 of rabies virus. Data obtained from DNA sequencing allow a better understanding of the molecular diversity of the rabies virus strains circulating in the regions under study, a fundamental step for the generation of information to be used in the molecular epidemiology of rabies, as the determination of sources of infection, origins of outbreaks and phylogeographic relationships among rabies virus strains from different species. KEY WORDS - Rabies, RT-PCR, diagnosis, molecular epidemiology.

75

DIAGNOSTIC, CHARACTERIZATION AND MOLECULAR EPIDEMIOLOGY

RABIES IN CERDOCYON THOUS (CRAB EATING FOX), NORTHEASTERN BRAZIL: GENETIC IDENTIFICATION AND FOX RABIES ASSOCIATION WITH DOGS AND CATS
Pedro Carnieli Junior 1, Willian de Oliveira Fahl 1, Juliana Galera Castilho 1, Rafael de Novaes Oliveira 1, Eduardo Eizirik 2, Rodrigo S. P . Jorge 3, Ronaldo G. Morato 3, Romualdo O. Spndola 4, Lindenberg M. Machado 5, Jos E. ngar de S 6, Enock L. V. da Silva 7, Maria Luiza Carrieri 1, Ivanete Kotait 1.
1) Instituto Pasteur, So Paulo, Brasil 2) Faculdade de Biocincias - Pontificia Universidade Catlica, RS, Brasil 3) CENAP/IBAMA,So Paulo, SP , Brasil 4) Fundao Municipal de Sade, Teresina, PI, Brasil 5) Laboratrio de Apoio Animal, PE, Brasil 6) LACEN - BA, Brasil 7) LACEN - SE, Brasil

Dr. Pedro Carnieli - pecarniel@yahoo.com.br

Brasil

According to data of the Ministry of Health/Brazil, rabies in wild animals has presented a gradual increase, particularly in bats and wild-canids, and these last animals have been responsible for 7,9% of the human deaths caused by wild animals from 1986 to 2006. Regarding wild-canids, it is noteworthy, in Brazil; the identification of species has been carried out in a subjective and empirical way, according to which two species stand out: Cerdocyon thous and Pseudalopex vetulus. The main objectives of this study have been to genetically identify the wild-canids involved in the cycle of rabies in the northeast of Brazil, as well as establish a connection between the rabies of these animals and that of cats and dogs, by the genetic characterization of the isolated viruses in the region. The genetic sequencing of 570 nucleotides of the mitochondria DNA, extracted from the brain of wildcanids suspected of being infected with the rabies virus, has been analyzed and the specie of the animal could be identified. The generated genetic sequences, when compared with others of the Pseudalopex and Canis genera, have allowed the identification of the species of the analyzed animals as being Cerdocyon thous. The sequencing of 1390 nucleotides of gene N of the rabies virus, obtained from 30 samples of the central nervous system of Cerdocyon thous, 15 of dogs and 5 of cats, all of which from the northeast of Brazil, have originated two clusters, one of dogs and cats and another one of Cerdocyon thous. Also, different subclusters have been formed within the two clusters, which confirm the existence of regional groupings of the rabies virus among the wild-canids of the northeast of Brazil, as well as among dogs and cats. It has also been observed, in the tree of generated identity, that a dog was segregated in the Cerdocyon thous cluster. Seeing that the two animals belong to the same geographical area, it is possible to suggest that the dog had been infected by a wild animal, which ultimately reveals the connection between the rabies of dogs and that of wild-canids, as well as the need to adopt specific measures of control of rabies in wild species.

76

THIRD SESSION

ANTIGENIC AND MOLECULAR CHARACTERIZATION OF POSITIVE CANINE SAMPLES TO RABIES IN MEXICO FROM 1997 TO 2006
A. Melndez Flix 1, 2, I.L. Padilla Medina 1, R.A. Trejo Rodrguez 1, M. Gmez Sierra 1, G. Hernndez-Rodrguez 1, D. Martnez Sols 1, A. Carrera Terrazas 1, A. Garca Guerrero1, A. Sandoval Borja 1, B. Escamilla Ros 1, I. Animas Vargas 1, O. Hernndez Monter 1, I.F. Villaseor Ruz 1 J.C. Gudio Rosales 1, F. Vargas Pino 3 G. Ziga Bermdez 2 & R.M. RibasAparicio 2.
1) Instituto de Diagnstico y Referencia Epidemiolgicos, Secretara de Salud, Mxico, 2) Escuela Nacional de Ciencias Biolgicas, Instituto Politcnico Nacional, Mexico, 3) Centro Nacional de Vigilancia Epidemiolgica y Control de Enfermedades, Secretara de Salud, Mexico.

Dr. Alejandra Melndez Flix - alejandramelendezfelix@yahoo.com.mx

Mxico

Since 1990 The Secretary of Health in Mexico carry out massive dog rabies vaccination campaigns to eliminate this zoonosis in the domestic canine population and to reduce with it the risk of transmission to human beings. Millions dose of the antirabic vaccine are applied every year, increasing annually (7.1 in 1990 up to 16.1 in 2005). Thus, the reduction of canine rabies cases is significant, from 3045 in 1990 to 125 in the 2005. In 1996 the laboratory works began with the antigenic characterization by using a panel of eight monoclonal antibodies kindly provided by the Rabies Section of rabies of the Centers Disease Control and Prevention of the United States (CDC). At the moment 287 dog samples are identified as positive to rabies with the following variants: V-1, V-3, V-7, V-9 and V-11. The persistence of cases of canine rabies in some places of the country, in spite of being observed in smaller scale, determines the necessity of an accurate identification by means of genetic approaches such as sequencing analysis of the 3semivariable region of the N gene. Our results agree with the presence of the lineages D5, D6 and D8, previously reported by de CDC. A politomy was observed in a group of 61 viral strains that have circulated in the Valley of Mexico from 1999 to the date, mainly at the east area, and involving two cases of human rabies. Four rabies strains isolated from dogs and a pig segregated independently, all of them coming from the municipalities of Ecatepec and Teotihuacan. Another homogeneous group involved dog samples coming from the center-south of Puebla and Tlaxcala suggesting that these viral strain circulated in this area from 1999 to 2004.

77

DIAGNOSTIC, CHARACTERIZATION AND MOLECULAR EPIDEMIOLOGY

ANTIGENIC CHARACTERIZATION OF RABID ANIMAL ISOLATES, NORTHERN BRAZIL, 2005 / 2006


L.M.N. Casseb 1, T.F. Barbosa 1, A.S. Pereira 1, H.C. Peixoto 2, A.L. Begot 3, R.J.S. Lima 3, M.M. Souza 4, P .F .C. Vasconcelos 1, E.S. Travassos da Rosa 1
1) Instituto Evandro Chagas, Ministrio da Sade, PA, Brasil 2) Universidade Federal da Agricultura da Amaznia, AM, Brasil 3) Ncleo de Doenas Endmicas, PA, Brasil 4) Laboratrio Central de Sade Pblica, RO, Brasil

Dr. Elizabeth Salb T. da Rosa - elizabethsalbe@superig.com.br

Brasil

IIntroduction: The identification of the antigenic variants of rabies virus furnish important information on the origin of the circulating rabies virus in a determined region and, in consequence, the existing cycles and the involved species on them. Aiming to extend the knowledge of the epidemiology of the rabies in northern Brazil, we incorporated the technique of antigenic characterization for the laboratorial diagnosis of the rabies. Objective: To determine the epidemiologic profile of the animal rabies in northern Brazil, during the period of January 2005 through July 2006. Method: A total of 31 rabies virus isolates from wild and domestic animals were used for antigenic characterization as follows: 22 dogs (13 from Par and 9 from Rondnia); two cats, a swine, and 6 bats all of them from Par State. The Indirect Imunofluorescence Assay (IIFA) was performed using a panel of monoclonal antibodies against the rabies virus nucleoprotein (produced by CDC/Atlanta, USA). Results: All 22 dog isolates and one cat sample were identified as Variant 2, strain commonly found among dogs; the other feline, the swine strain and the 6 bat rabid samples (3 vampires and 3 non-hematophagous) were identified as Variant 3, whose reservoir is the hematophagous bat Desmodus rotundus. Conclusions: These results suggest that in the northern region of Brazil the antigenic variant 2 continues to be the prevalent in dogs; the other strains were antigenically characterized as belong to the variant 3 (from vampire bats). It is interesting that the swine and cat strains coming from the municipality of Augusto Corra, northeastern region of Par State, where an important bat-transmitted rabies outbreak was reported in 2005. The genetic characterizations of all studied strains are on going and should complement this study. Financial support: IEC/SVS/MS

78

THIRD SESSION

MOLECULAR ANALYSIS OF THE AERIAL CYCLE OF THE RABIES VIRUS IN MEXICO


A Melndez-Flix, 1, 2, I.L. Padilla-Medina 1, R.A Trejo-Roguez 1, M Gmez-Sierra 1, G. Hernndez-Rodriguez, 1, D. Martnez Sols 1, A Carrera-Terrazas 1, A Garca-Guerrero1, A Sandoval-Borja, 1 B. Escamilla-Ros 1 I. Animas-Vargas 1, O Hernndez-Monter 1, I.F. Villaseor-Ruz, 1, J.C. Gudino-Rosales 1, F . Vargas-Pino 3, G. Ziga Bermdez, 2 & R.M. Ribas-Aparicio 2
1) Instituto de Diagnostico Y Referencia Epidemiolgicos, Mxico, DF, Mxico, 2) Escuela Nacional de Cincias Biolgicas, Instituto Politcnico Nacional, Mxico 3) Centro Nacional de Vigilancia Epidemiolgica y Control de Enfermedades, Secretara de Salud, Mexico

Dr. Alejandra Melndez Flix - alejandramelendezfelix@yahoo.com.mx

Mxico

At present in Mexico the bats cause more than 50% of human rabies being a serious problem of public health, as well as the transmission to terrestrial animals mainly to the bovine livestock causing considerable economic losses at national level. In this work, samples coming from diverse states and species of our country were analyzed and found infected by different reservoirs from the aerial cycle, through their antigenic identification with a panel of eight monoclonal antibodies, and the nucleotide sequencing of the 3' semivariable region (267pb) of the N gene. The phylogenetic analysis showed 7 different groups belonging to the antigenic variants V-11, V-9, V-6, V-5 and V-8. A homogeneous phylogenetic group of hematophagous bats was identified as V-11 (Desmodus rotundus) coming from cattle and chiropters at the near states to the coast of the Gulf of Mexico between 2002 and 2005, contrasting with a small subgroup of the same variant in this area from the period 2003 to 2004. Another phylogenetic group also isolated from cattle, segregated in independent form as V-8 coming from the coast of the Pacific in 2005, this group had a narrow genetic relationship with atypical viral strains identified by genetic characterization, as well as with the group of rabies strains V-3 from bats hematophagous coming from the country southeast, in spite of being identified antigenically as skunk strains. And finally a group of rabies strains from insectivorous bats characterized as V-6 and V-9 of diverse domestic points. The information generated in this study has been carried out at the Rabies Laboratory of the InDRE in collaboration with other laboratories and institutions in order to establish patterns of the rabies virus dissemination in areas of high risk in Mexico to help national authorities the implementation of appropriate strategies that allow them to maintain a system of active surveillance.

79

DIAGNOSTIC, CHARACTERIZATION AND MOLECULAR EPIDEMIOLOGY

A COMPARISON STUDY OF INTERNATIONAL RABIES IMMUNOGLOBULIN REFERENCE STANDARDS


Susan Moore 1, Teri Ricke 1
1) Kansas State University, Manhattan, KS, USA

Dr. Susan Moore - smoore@vet.ksu.edu

USA

International rabies immunoglobulin (RIG) standards are used throughout the world to provide a means of ensuring uniformity in the potency determination RIG products used for prophylaxis, individual vaccine response or disease diagnosis by expressing the level of rabies antibody present in a serum sample international units (IU/mL). The first International Standard for anti-rabies serum (IARS) of equine origin was replaced in 1984 by the WHO International Standard for Rabies Immunoglobulin of human origin (WHO RIG 1). This preparation was evaluated at six international laboratories using the RFFIT and assigned at potency of 59 IU. In 1993, due to dwindling supply of WHO RIG 1, a replacement preparation was evaluated at nine international laboratories using both the MNT and RFFIT and was approved by WHO as the second International Standard for Rabies Antibodies (WHO RIG 2). The mean potency was determined from the RFFIT data to be 30 IU/mL. In the United States, the most recent comparison of the two WHO RIG standards was in 1997 at three laboratories (FDA, CDC, and KSU) and at that time WHO RIG 1 was estimated to have lost approximately 2.5% of its potency versus WHO RIG 2. Currently new RIG products for prophylaxis are being evaluated by rabies serologic methods using these international standards. The importance of consistent and accurate assignment of IU/mL values cannot be overstated. To determine whether any further change in potency had occurred since 1997, a repeat potency comparison of the two WHO RIG standards was performed at KSU. Results of indicate the relative potency of WHO RIG 1 is now decreased by an average of 14%. A decreased potency was verified by another neutralization method (FAVN). Assignment of IU/mL values to RIG preparations based on the lower potency WHO 1 would affect the calculated dose per kg. Though potency differences between the two WHO standards should be evaluated by another interlaboratory study, based on these findings there is need of a standardized approach for the evaluation of RIG products, not only for choice of international standard, but also in the complete testing protocol used.

80

THIRD SESSION

A QUANTITATIVE INDIRECT ELISA TEST TO MONITOR THE EFFECTIVENESS OF RABIES VACCINATION IN DOMESTIC AND WILD CARNIVORES
A. Servat.1, M. Feyssaguet 2, I. Blanchard 2, F. Boue 1, F . Cliquet 1
1) Laboratoire dEtudes et de Recherches sur la Rage et la Pathologie des Animaux Sauvages, Nancy, France 2) Bio-Rad, Marnes-LaCoquette, France

Dr. Alexandre Servat - a.servat@afssa.fr

France

This review reports a new ELISA test to measure the level of rabies anti-glycoprotein G antibodies after vaccination. The PLATELIA RABIES II kit was evaluated on defined populations of samples of dog, cat and fox origin. For each target species, serums from nave, unvaccinated and vaccinated animals were tested. PLATELIA RABIES II results were compared to those obtained using the reference FAVN test (for dogs and cats) and to a published in house ELISA test (for foxes). PLATELIA RABIES II test demonstrated to be highly specific whatever the species considered (more than 98%) using a cut-off value of 0.5 EU/mL. The indexes of sensitivity ranged between 92.4 and 94.5 for fox samples, and reached 83% for domestic carnivores population. Data collected by testing field samples populations revealed that the rate of false negative results ranged between 8.9 and 11.1 % and that the rate of false positive results ranged between 1 and 2 % for dog and cat population. Inter-laboratory trial study revealed that the performance of the kit is highly reproducible. The PLATELIA RABIES II test would be a good candidate for the routine detection of rabies antibodies not only in domestic carnivores (in the frame of international trades and follow up of the efficiency of dogs mass parenteral rabies vaccination) but also in foxes for the follow up of rabies oral vaccination campaigns.

81

DIAGNOSTIC, CHARACTERIZATION AND MOLECULAR EPIDEMIOLOGY

COMPETITIVE ENZYME-LINKED IMMUNOSORBENT ASSAYS (CELISA) FOR THE DETECTION OF ANTI-RABIES ANTIBODIES IN WILDLIFE SERA - DEVELOPMENT, VALIDATION AND PRACTICAL APPLICATIONS
Christine Fehlner-Gardiner 1, M. Kim Knowles 1, Andrea Clark 1, Lise Murphy 1, Natalia Barkar 1, Lindsay Elmgren 1, Alexander Wandeler 1
1) Canadian Food Inspection Agency, Ottawa, ON, Canada

Dr. Christine Fehlner-Gardiner - fehlnergardiner@inspection.gc.ca

Canad

We previously reported the development of a competitive ELISA for the detection of anti-glycoprotein (G) antibodies in wildlife sera (Laboratory Techniques in Rabies, 4th Ed., WHO, 1996). The basis of this assay is the inhibition of binding of a peroxidase-labelled, G-specific, neutralizing monoclonal antibody (mAb) to immobilized ERA virus by antibodies present in animal serum. Initial validation experiments showed very good agreement between cELISA and rabies serum neutralization assays (RSNA) on fox and skunk serum samples. The sensitivity and specificity of the rabies G cELISA were found to exceed 98% when compared against results from RSNA. At the time it was assumed that this level of specificity and sensitivity would apply to any sera tested in the rabies G cELISA, regardless of source. This assumption was not upheld in the case of raccoon sera, where both sensitivity and specificity were significantly decreased. The discordance in results between cELISA and RSNA was reproducible and could not be overcome by changing the assay format, the affinity or specificity of the competing mAb or the nature of the coating antigen (whole virus vs. recombinant protein). Based on these observations, we have made numerous changes to the rabies G cELISA as originally reported, including improved antigen preparation methods, the introduction of species-specific control sera, implementation of test acceptance criteria and establishment of species-specific cut-off values for positive samples. In addition to the rabies G cELISA test improvements, we have developed cELISAs for the detection of nucleoprotein (N)- and phosphoprotein (P)-specific antibodies in serum. These assays for G-, N- and P-specific antibodies are being utilized in studies of immune response to rabies vaccines in both wildlife and laboratory animals and in viral protein epitope mapping studies. Aspects of the development and validation of the three cELISAs, and their practical applications will be discussed.

82

THIRD SESSION

RABIES PROFICIENCY TESTS: A GLOBAL ANALYSIS OF THE 13 RING TESTS PERFORMED FROM NOVEMBER 1999 TO OCTOBER 2004
M. Wasniewski 1, A. Servat 1, F. Cliquet 1
1) Centre for Research and Management in Zoonoses Control, Nancy, France

Dr. Marine Wasniewski - m.wasniewski@afssa.fr

France

For many years, quarantine confinement has proven to be an efficient method against rabies introduction in rabies free territories, however it was unsatisfactory for animal health and for the owner. Between 1993 and 2000, many rabies free countries have alleviated their quarantine measures and adopted a scheme requiring a rabies vaccination followed by a serological control. This alternative measure allows to guarantee the safety of free movements of pets and preserves the rabies free status of the countries. For laboratories willing to carry out the rabies serological controls, the European Commission decided to establish a system of community approval of such laboratories in order to guarantee an effective control system. The European Commission has designated a Community Reference Institute to coordinate the approval of the laboratories. The main task of this institute is to organize rabies proficiency tests for laboratories already agreed or willing to be agreed to perform rabies serological controls. These proficiency tests allow to determine the performance of individual laboratories for specific tests and to evaluate the laboratories continuing performance. We have conducted a global analysis of the rabies proficiency tests performed from November 1999 to October 2004 (representing 13 tests and 315 appraisals). A specific analysis for each criterion (specificity, intralaboratory repetability and consistency, interlaboratory consistency) has also been performed for this period. The data obtained from these studies are highly satisfactory. Indeed after 13 rabies proficiency tests, 95,9 % of laboratories obtained satisfactory results and 98,74% of participating laboratories succeeded for their first participation in rabies proficiency tests whatever the OIE prescribed method used (RFFIT or FAVN test). It should be noted that failures obtained during the 13 tests have never been attributed to the specificity but to a lack of intra-laboratory consistency. The data obtained from these studies will be presented and discussed.

83

DIAGNOSTIC, CHARACTERIZATION AND MOLECULAR EPIDEMIOLOGY

ASSURING THE QUALITY OF TEST RESULTS


C. Gordon 1
1) Kansas State University, Manhattan, Kansas, USA

Dr. Chandra Gordon - gordon@vet.ksu.edu

USA

Most laboratories have long held the opinion or misconception that monitoring the accuracy and precision of their test results by the inclusion of a positive and negative control standard was the best means of verifying the quality of the laboratorys work. This approach has well become an antiquated system due to the varying complexities and ease of modifying current test methodologies in use. Not only should control standards be certified reference materials that are traceable to a known international standard but just as equally as important is the ability of the laboratory to record this date in order to detect trends or subtle shifts over time. Most laboratories will use statistical techniques such as control charts relying on the two standard deviation bench mark to determine the success or failure of their work. Accessing the quality of the test results must be a more encompassing approach not just limited to monitoring the standards results, but should also include participation in inter-laboratory comparison or proficiency testing program, assessment for the competency of personnel, review of patient results against known medical history, and replicate tests using the same or different test methodologies for conformation of results.

84

THIRD SESSION

RECENT EPIDEMIOLOGY OF SOUTHERN AFRICAN LYSSAVIRUSES


Louis H. Nel 1, J. Randles, J. T. Paweska, W. Markotter, P . Coetzee, C. T. Sabeta
1) University of Pretoria, South Africa

Dr. Louis Nel - louis.nel@up.ac.za

frica do Sul

All four lyssavirus genotypes known to be present in Africa have been isolated from South Africa in the year 2006. These are: (1) Duvenhage virus (genotype 4), isolated from a human fatality for the first time in 36 years; (2) Mokola virus (genotype 3), isolated from a dog and cat for the first time since 1998; (3) Lagos bat virus (genotype 2) continually isolated from Epomophorus fruit bats and from incidental terrestrial animals during the past 3 years; (4) Rabies virus (genotype 1) - with two virus biotypes being endemic in mongooses and in canines (domestic and wildlife), respectively. In the case of dog rabies, the virus continues to radiate into new geographical areas or hosts and the disease have reached serious epidemic proportions in two large geographical areas, viz. the adjacent provinces of Kwazulu-Natal and Eastern Cape and to the north, in the Limpopo province (during 2006). Correspondingly, there has been a drastic increase in the cases of human fatalities. Analyses of the abovementioned viruses will be presented and the implications of findings will be discussed.

85

FOURTH SESSION
RABIES IN TERRESTRIAL WILDLIFE: EPIDEMIOLOGY AND CONTROL

RABIES IN TERRESTRIAL WILDLIFE: EPIDEMIOLOGY AND CONTROL

88

FOURTH SESSION

EPIDEMIOLOGICAL SURVEILLANCE OF RABIES IN WILD ANIMALS: ROAD KILS FROM THE HIGHWAYS AND/OR FOUND IN URBAN AREAS OF THE MUNICIPALITIES OF JACARE, SANTA BRANCA AND SALESPOLIS, VALE DO PARABA -SP , SOUTHEASTERN BRAZIL
M.C.A.M. Souza 1, R.N. Oliveira 2, F.H. Ito 3, J.J. Ferrari 4
1) Secretaria de Agricultura e Abastecimento, SP , Brasil. 2) Instituto Pasteur, So Paulo, SP , Brasil 3) Faculdade de Medicina Veterinria e Zootecnia, Universidade de So Paulo, SP , Brasil 4) Secretaria Municipal de Agricultura, Jacare, SP , Brasil

Dr. Maria Conceio A. M. Souza - cessasouza@uol.com.br

Brasil

Rabies in wild animals has been a constant danger to man and domestic animals. In 2005, vampire-bat-transmitted human rabies numbered 44 cases in Northern and Northeastern states of Brazil, although canine rabies has decreased drastically in late years. In some Northeastern states, wild canids are important reservoirs for rabies, but few cases are diagnosed at rabies diagnostic laboratories. In this work, 52 road kills from the vicinity of the city of Jacare, Santa Branca, and Salespolis, located at the Paraba do Sul River Valley, in the state of So Paulo were collected and submitted for rabies diagnosis, during the period of January 2004 to April 2006. The wildlife collected were represented by Chiroptera (56.0%), Marsupials (28.7%) and wild canids (15.3%). Brain fragments and the non-nervous tissues were submitted to the direct Fluorescent Antibody (dFA) test, mouse inoculation test (MIT), and Reverse Trancriptase - Polymerase Chain Reaction (RT-PCR) technique. None of the materials examined by these techniques have been found positive for rabies although rabies in herbivores is still prevalent in the Vale do Paraba. It has been suggested the need to apply a continuous rabies surveillance program to wild animals, since the region has had great topographic modifications in the last years puts at risk the health of population. Key words: Epidemiology of rabies, Wildlife. Vampire bats. Marsupials. Road kills, Rabies diagnosis.

89

RABIES IN TERRESTRIAL WILDLIFE: EPIDEMIOLOGY AND CONTROL

RABIES IN WILD SPECIES IN CEAR - BRAZIL, EPIDEMIC AND ENVIROMENT FACTORS


David Caldas 1
1) Secretaria de Sade, Cear, Brasil

Dr. David Caldas - davi@saude.ce.gov.br

Brasil

INTRODUCTION: The wild Rabies is an important challenge in the area of the Health in Cear for the significant occurrence of cases, variety of the affected species, space distribution in several areas, climate and ecological units . OBJECTIVE: It was intended to analyze in the period from 1990 to 2005 the occurrence of cases related to the animals and humans. MATERIAL AND METHODS: Collected data of the records VE-7, Reports of the Program of Control of the Rabies in Cear and of the Institute of Research and Economical Strategy of Cear. RESULTS: In the analyzed period happened 179 cases in foxes, 29 in marmmosets and from 2001 to 2005, seven cases in raccoons. From 1990 2005 14 cases of human Rabies originating from wild cycle were registered, 10 for marmmosets, 3 for bats and one for raccoon. Cear is divided in 33 geographical microregions, 11 units ecological units and 05 climatic types. The Rabies in foxes has been happening in all the areas of the state, however they are evidenced with larger tendency in areas of high altitude and humid forests, mountainous and climate tropical humid and warm. Detaching the metropolitan area of the capital Fortaleza and the south of the state. The cases of Rabies in marmmosets, as well as the humans, originated by these species has a predominance in coastal areas of the state, that corresponds an extension of 543km, in the Solid of Baturit, where it prevails humid forests, mountainous characterized by climate tropical humid and warm. In humans the largest concentration of cases was in the period from 1991 to 1998 (13 cases), in most of them as much the victim as the families they ignored the risk of the transmission of Rabies for these species, fact that generated the absence of the search of the medical attendance. CONCLUSION: Before this epidemic and environmental picture Cear comes as a vulnerable area and high risk for occurrence of cases of animal Rabies among carnivorous wild and urban, primates marmmosets, bats and herbivores and accidentally the man for the absence of an environmental education

90

FOURTH SESSION

SPATIAL - TEMPORAL VARIATION IN RABIES SPECIMEN SUBMISSION AND POSITIVITY DURING A RACCOON-RABIES EPIZOOTIC: PRELIMINARY RESULTS
T.P . Algeo 1; B.M. Bjorklund 1; X. Wang 2; D. Hennigan 2; K. Von Hone 3; M.D. Chandler 1; D.M. Grove 4; A. Robbins 5; D.J. Wilda 1; C.E. Rupprecht 6; D. Slate 1
1) USDA, APHIS, Wildlife Services, 2) Massachusetts Department of Public Health Laboratory Institute, 3) Yarmouth Division of Natural Resources/Barnstable County Rabies Task Force, 4) Cape Wildlife Center, 5) Tufts University Cummings School of Veterinary Medicine, 6) Centers for Disease Control and Prevention, Atlanta, GA, USA

Dr. Timothy P . Algeo - timothy.p.algeo@aphis.usda.gov

USA

Raccoon-variant rabies was detected on the ocean side of the Cape Cod Canal for the first time in March 2004, after ten years of rabies-free status, while raccoon rabies spread throughout the remainder of Massachusetts. Despite attempts to control the rabies epizootic through an extensive trap-vaccinate-release program, followed by multiple oral rabies vaccination campaigns, raccoon variant rabies spread to the extreme end of Cape Cod, and was detected there in 2006. Cape Cod municipalities, The Massachusetts Department of Public Health, and USDA, APHIS, Wildlife Services engaged in enhanced rabies surveillance to track the spread of rabies down the Cape. During March 2004-December 2005, raccoon rabies suspect submissions ranged from 3-74/month, and skunk submissions ranged from 0-25/month. Rabies among Cape Cod raccoons submitted for testing ranged from 18-67%/month. We characterize the Cape Cod rabies epizootic by presenting preliminary data on spatialtemporal variation in rabies surveillance results, quantify rabies surveillance efforts, and plan to improve upon existing rabies surveillance strategies through use of a direct rapid immunohistochemical test in collaboration with CDC and MADPH to optimize the use of available resources.

91

RABIES IN TERRESTRIAL WILDLIFE: EPIDEMIOLOGY AND CONTROL

EFFECTIVENESS OF ORAL RABIES VACCINE TO MANAGE RABIES IN STRIPED SKUNKS (MEPHITIS MEPHITIS)
Krista Wenning 1, David Bergman 1, Dennis Slate 1, Charles Rupprecht 2, Chad Heuser 1, Barbara Worgess 3, Elisabeth Lawaczeck 4, Mark Weissinger 1
1) USDA/APHIS/WS, 2) Centers for Disease Control and Prevention, Atlanta, GA, USA 3) Coconino County Health Department, USA 4) Arizona Department of Health Services, USA

Dr. Krista Wenning - krista.m.wenning@aphis.usda.gov

USA

Oral rabies vaccination (ORV) has been under field investigation in the United States for use in raccoons (Procyon lotor), gray fox (Urocyon cinereoargenteus) and coyote (Canis latrans) since 1990. However, there is no licensed oral vaccine that is immunogenic in striped skunks (Mephitis mephitis) for use in the United States. A multiagency coalition consisting primarily of USDA-APHIS-WS, the Centers for Disease Control and Prevention, the Coconino County Health Department, and the Arizona Department of Health Services initiated a field study to test the effectiveness of the Raboral V-RG oral vaccine in striped skunks. This was the first field trial of its kind targeting striped skunks. In April of 2005, 1,875 coated sachet baits containing Raboral V-RG vaccine were distributed by hand at a density of 75 baits per km2 throughout the vaccine study area in Flagstaff, Arizona. Cage trapping began 5 weeks post-baiting and 12 striped skunks were captured from the vaccine study site: 11 had rabies titers of 0.04 International Units (IU) and 1 had a titer of 0.05 IU. All animals were confirmed negative for rabies using the dFA test. Results from this study will aid in the development of an effective oral vaccine for use with striped skunks.

92

FOURTH SESSION

TRAP-VACCINATE-RELEASE AND RABIES MANAGEMENT PROGRAMS TO MANAGE RABIES OUTBREAKS IN STRIPED SKUNKS (MEPHITIS MEPHITIS) IN FLAGSTAFF, ARIZONA.
David Bergman 1, Mark Weissinger 1, Elisabeth Lawaczeck 2, Barbara Worgess 3, Krista Wenning 1
1) USDA/APHIS/WS, 2) Arizona Department of Health Services, 3) Coconino County Health Department

Dr. David Bergman - david.l.bergman@aphis.usda.gov

USA

A bat variant of the rabies virus, common in big brown bats (Eptesicus fuscus), occurs throughout the state of Arizona. In 2001, a striped skunk (Mephitis mephitis) was infected with a bat variant causing an unprecedented rabies outbreak in Flagstaff, Arizona. By the end of 2001, this rare spillover had infected 19 skunks and was the first documented event of a bat variant of the rabies virus being transmitted and maintained in skunks. A multiagency coalition consisting of USDA-APHIS-WS, the Arizona Department of Health Services, Coconino County Health Department, the Centers for Disease Control and Prevention, and others responded to this outbreak by initiating a trap-vaccinate-release (TVR) campaign for striped skunks in the area. Approximately 300 striped skunks have been vaccinated during the ongoing campaign. There were no reported rabies cases in terrestrial wildlife consistent with the big brown bat variant for 3 years following the initial TVR campaign. However, the spillover reoccurred in late 2004 when 5 striped skunks and 1 gray fox (Urocyon cinereoargenteus) were reported positive for the bat variant of rabies. Additionally, 1 striped skunk, 1 domestic cat (Felis cattus) and 1 gray fox were reported positive in 2005. At present, several rabies management programs, including the TVR campaign, are in place to manage rabies outbreaks in striped skunks.

93

RABIES IN TERRESTRIAL WILDLIFE: EPIDEMIOLOGY AND CONTROL

RABIES DNA VACCINATION THROUGH INTRADERMAL INTO EAR PINNAE, IMMUNOCHEMISTRY STUDY
E. Tesoro Cruz 1, R. Caldern Rodrguez, R. Hernndez Gonzlez, A. Prez, F . Blanco Favela, A. Aguilar Setin
1) Instituto Mexicano del Seguro Social, Mexico, DF, Mexico

Dr. Emiliano Tesoro Cruz - emiliano_tesoro@hotmail.com

Mxico

Domestic animal rabies (especially dogs) has been controlled in industrialized countries using traditional parenteral vaccination, but wild animal rabies has been more difficult to control due to host inaccessibility. It has, thus, remained an important problem for the public health. Cats are considered company animals and they may transmit rabies to humans. In fact, in Quintana Roo, Mexico, in 2004, a case of a human death was reported when a wild cat bit a child. The diagnostic was confirmed postmortem by the Reference Laboratory of Ministry of Health (INDRE, SSA), and the autopsy confirmed an encephalitis caused by the rabies virus belonging to the antigenic variant V-3, which corresponds to a bat specific variant. The DNA vaccine against rabies consists of a mammalian expression vector pCl-neo, into which a glycoprotein-encoding rabies gene from a Mexican isolation was inserted (pGQH), previously reported by Tordo. The Gfp is a reporter plasmid that express a fluorescent green protein and it was used in this experiment. Different inoculation routes for DNA vaccines have been explored, but efficacy varies with the used animal model and inoculation route. In this work, only one dose of pGQH was administrated ID-EP to cats, in order to know the neutralizing antibodies titers and to study if dendritic cells were involved in the immune response in a cat. Five female one year old cats were used in this experiment, all were inoculated through the ID-PO route. Three of 5 received 100 mg of pGQH, one received 100 mg of GFp. and one more received 100 ml of PBS. Sera samples from cats (1, 2, 3, and 5) were obtained on days 0, 15, 30, 60, 90, 120, 150, and 180. Virus neutralizing antibody titers (VNAb) were determined in all sera, using the rapid fluorescent focus inhibition test (RFFIT) and the results were expressed in International Units per ml. The fourth cat was sacrificed at 72 h, after GFp was delivered and different organ samples were taken (ear, spleen, lung, liver, and cervical and popitleo ganglions) to study the green protein fluorescence expression, and also dendritic cells detection, using CD1A antibodies. VNAb titers from cats 1, 2 and 3 were >1.5 IU on day 15, >6 IU 4 months later, and finally >3 IU six months later. The fifth cat, negative control, lacked anti-rabies antibodies. Fluorescent green protein was detected in ear, spleen, and cervical ganglion, and DCs were evidenced in ear and cervical ganglion. The ID-EP was revealed as an excellent route to deliver DNA vaccines. The success of the ID-EP route could be due to represent a vast vascular draining site, also the DCs participation are implicated, setting up the possibility that detected DCs could arrive to nearer ganglions and trigger the immune response.

94

FOURTH SESSION

TRACKING THE ATTENUATION AND ORIGIN OF SAD DERIVED ATTENUATED RABIES VIRUS VACCINES
L. Geue 1, S. Schares 1, J. Kliemt 1, M. Beckert 1, B. Hoffmann 1, C. Freuling 1, R. Zanoni 2, D. Marston 3, L. McElhinney 3, A. Fooks 3, T. Mller 1
1) Friedrich-Loeffler-Institut, Germany, 2) Institute of Veterinary Virology, University of Bern, Bern, Switzerland, 3) Veterinary Laboratories Agency, Surrey, UK

Dr. Thomas Muller - thomas.mueller@fli.bund.de

Germany

All attenuated rabies virus vaccines except the vaccinia rabies glycoprotein recombinant vaccine (VRG) used for oral vaccination of wildlife descend from a common ancestor; the Street Alabama Dufferin (SAD) field strain of rabies virus isolated in North America in 1935. However, after more than 30 years only scant information is available on how the numerous attenuated SAD derivatives can be genetically distinguished from each other and what the genetic basis for their attenuation is. In a recent project, we aimed at the complete nucleotide sequencing of 4 commercially available SAD vaccine viruses currently licensed in Germany. By further including other SAD derivatives such as the original SAD Bern, the ERA and a SAD 1-3670 Wistar strain originating from 1975, 1978 and 1965, respectively, and a SAD Bern strain currently used in the Czech Republic, it allowed us for the first time to track down the attenuation of SAD derivatives on a genetic basis. Whereas the SAD1-3670 Wistar and ERA strain differ by single mutations in the N-, P-, G- and L-gene, the ERA strain and the original SAD Bern strain share still a higher sequence homology. Interestingly, the SAD1-3670 Wistar and the SAD Bern strain both were a problem for sequencing of the M-G and G-L intergenic region. Cloning of the respective parts of the genome into E.coli and subsequent sequencing revealed that both strains were not genetically pure virus strains but consist of at least two different virus variants. The largest attenuation step happened to occur between SAD Bern and the currently widely used SAD vaccine viruses with considerable nucleotide mutations and amino acid exchanges in the N-, P- and partially in the G- gene and leader sequence (LDR). Phylogenitic analysis gave surprising insights into the derivation of SAD B19, SAD P5/88, SAD VA1, and SAG2. Two commercially available vaccine viruses were identified as not being the designated SAD strains.

95

RABIES IN TERRESTRIAL WILDLIFE: EPIDEMIOLOGY AND CONTROL

ERA VACCINE-DERIVED CASES OF RABIES IN WILDLIFE AND DOMESTIC ANIMALS IN ONTARIO, CANADA DURING THE PERIOD OF 1989 TO 2004
Christine Fehlner-Gardiner 1, Susan Nadin-Davis 1, Frances Muldoon 1, Janet Armstrong 1, Peter Bachmann 2, Alexander Wandeler 1
1) Canadian Food Inspection Agency, Ottawa, ON Canada, 2) Ontario Ministry of Natural Resources, Trent University, Peterborough, ON Canada

Dr. Christine Fehlner-Gardiner - fehlnergardiner@inspection.gc.ca

Canad

A vaccination program for the control of terrestrial rabies in the province of Ontario, Canada began in 1989. During the period between 1989 and 2004, over 13 million baits containing the live, attenuated rabies virus ERABHK21 were distributed across the province, with the aim of immunizing foxes by the oral route. Animals recovered from bait distribution areas were assayed by fluorescent antibody test for rabies virus infection. Immunoreactivity with a panel of monoclonal antibodies that discriminate between ERA and rabies virus variants known to circulate in Ontario, and molecular genetic analyses were used to identify animals infected with ERA. Nine cases of ERA variant rabies were identified over the 16 year period of study; these did not appear to stratify by species, year of discovery, or location of capture. ERA positive animals were found across the province in eight counties, all of which had been baited in the year of case discovery. Four red foxes, two raccoons, two striped skunks and one bovine calf were among the positive cases discovered. Molecular phylogenetic analyses of the partial N gene sequences generated from these isolates indicated that these nine cases were due to infection with the ERA variant. Nucleotide sequencing of the glycoprotein genes and alignment of their predicted amino acid sequences with those of two laboratory stock ERA viruses revealed the mutation of residue 120 from arginine to histidine in all but one isolate. At present, the significance of this mutation is unclear. Our data demonstrate that the ERA vaccine distributed in Ontario carried residual pathogenicity, however, there does not appear to be any evidence of establishment of ERA in wildlife populations over the 16 years studied. These results are consistent with previous reports of the rare detection of ERA vaccine-induced rabies, as well as with laboratory studies of ERA pathogenicity.

96

FOURTH SESSION

DEVELOPMENT OF A RECOMBINANT CANINE ADENOVIRUS 2 - RABIES VIRUS G PROTEIN VACCINE (CAV2-RV G)


Bernhard Dietzschold 1, Jianwei Li 1, Milosz Faber 1, Amy Papaneri 1, Marie-Luise Faber 2, James P . McGettigan, Matthias J. Schnell 1
1) Thomaz Jefferson University, Philladelphia, PA, USA, 2) Molecular Targenting Technologies, Philladelphia, PA, USA

Dr. Bernhard Dietzschold - bernhard.dietzschold@jefferson.edu

USA

Rabies vaccines based on live attenuated rabies viruses or recombinant pox viruses expressing the rabies virus (RV) glycoprotein (G) hold the greatest promise of safety and efficacy, particularly for oral immunization of wildlife. However, while these vaccines induce protective immunity in foxes, they are less effective in other animals, and safety concerns have been raised for some of these vaccines. Because canine adenovirus 2 (CAV2) is licensed for use as a live vaccine for dogs and has an excellent efficacy and safety record, we used this virus as an expression vector for the RV G. The recombinant CAV2-RV G produces virus titers similar to those produced by wild-type CAV2, indicating that the RV G gene does not affect virus replication. Comparison of RV G expressed by CAV2RV G with that of vaccinia-RV G recombinant virus (V-RG) revealed similar amounts of RV G on the cell surface. A single intramuscular or intranasal immunization of mice with CAV2-RV G induced protective immunity in a dose-dependent manner, with no clinical signs or discomfort from the virus infection regardless of the route of administration or the amount of virus.

97

RABIES IN TERRESTRIAL WILDLIFE: EPIDEMIOLOGY AND CONTROL

SAFETY STUDIES OF RABIES STRAIN ERA G 333 AS A CANDIDATE FOR ORAL RABIES VACCINE
D. Bankovsky 1, G. Safonov 1
1) Pokrov Biologics Plant, Vladimir, Russia

Dr. Denis Bankovskiy - den_bankovskiy@rambler.ru

Rssia

Russia is the historically unsuccessful country on rabies. The most important role in rabies distribution have wild carnivorous (foxes, raccoon dogs) and stray dogs and cats. ORV campaigns spent now have not led to reduction of number of rabies cases. The reasons of it are not only insufficient quantity of spread baits in the limited territories, but also that applied rabies strains are not safe and have a residual virulence. The given fact can be the reason of occurrence of vaccinal rabies in areas of carrying out of oral vaccination. The experiences on safety studying of strain ERA G 333 for some species of animals have been carried out. 10 foxes, 10 polar foxes, 8 dogs, 5 kittens, 20 guinea-pigs, 30 adult mice, 30 young mice, 72 newborn mice were exposed to rabies strain ERA G 333 which past 10 passages into BSR cells. Foxes, arctic foxes and dogs received virus intracerebraly and intramuscularly. Kittens were infected intramuscularly. Three routes of virus introduction have been used in guinea-pigs: intracerebral, intramuscular and subcutaneous. Adult mice were infected intracerebraly, young mice and newborn mice were infected intracerebraly, intramuscularly and subcutaneously. All animals except newborn mice survived during 60 days after virus inoculation. Newborn mice died after all used routes of introduction. Thus, ERA G 333 innocuous for foxes, arctic foxes, dogs, cats, guinea-pigs, adult and young mice in our study after different routes of introduction. The given strain is safe and can be used in further immunogenicity studying.

98

FOURTH SESSION

A BENEFIT-COST ANALYSIS OF THE ONTARIO RACCOON ORAL RABIES VACCINATION PROGRAM


Christopher Nunan 1, Stefanie Shwiff, Rick Rosatte
1) Peterborough, ON, Canada

Dr. Christopher Nunan - cnunan@cogeco.ca

Canad

Continuing with the work begun with last years presentation of the costs and benefits of the red fox Oral Rabies Control program, we now turn our attention to the other rabies control program in Ontario that deals with the influx of raccoon rabies from the United States. The two programs are profoundly different. In comparison to the endemic fox rabies outbreak, raccoon rabies is invading the province. This resulted in researchers using different strategies to deal with the invading disease. Following detection of each case of raccoon rabies the animal was dispatched, the brain was sent to Ottawa for examination and confirmation and the site was surrounded by concentric rings of depopulation, trap- vaccinate-release and oral rabies baiting. Since it first entered the province, rabies has not spread as quickly nor has it spread as far, nor has as many cases been detected as had been predicted from experiences in the northern United States. This has been not attributed to quick and decisive action by the Ontario Ministry of Natural Resources. This paper will discuss the benefits and costs of this approach to rabies control.

99

RABIES IN TERRESTRIAL WILDLIFE: EPIDEMIOLOGY AND CONTROL

THE RACCOON - THE ULTIMATE WILD CARNIVORE CHALLENGE FACING ORAL RABIES VACCINATION
D. Slate1, C. Rupprecht 2, B. Dietzschold 3, D. Donovan 4
1) USDA, APHIS, Wildlife Services, NH, USA, 2) Centers for Disease Control and Prevention, Atlanta, GA. USA, 3) Thomaz Jefferson University, Philladelphia, PA, USA, 4) Ontario Ministry of Natural resources, Trend University, Canada

Dr. Denis Slate - denis.slate@aphis.usda.gov

USA

Oral rabies vaccination successes have been documented for red foxes in Europe and Canada and gray foxes and coyotes in the United States. While specific ORV successes can be identified for raccoons, myriad factors point to daunting challenges for virus containment and elimination. Extrinsic factors such as translocation and intrinsic factors such as population structure, density and dynamics are among several variables that must be considered in formulating control strategies. A more comprehensive understanding of inter-specific interactions among mesocarnivores such as skunks, foxes and raccoons may be important to developing more effective rabies control strategies. In addition, a better understanding of the independent or interaction effects of the timing, pattern and density for bait distribution; the spatial-temporal habitat preferences of raccoons and nontarget competitors for baits; and the contribution of natural and human-made features to the barrier effect in preventing rabies spread are also important in strategy development and refinement. While developing and implementing effective raccoon rabies control represents a complex multivariate challenge, the immediate emphasis should remain on the most obvious limiting factors to long-term success. Clearly, field performance data for currently available oral vaccine and baits point to the need for improved or new technology for long-term success. Delay in requisite laboratory and field testing toward full implementation of new or improved technology could impede progress in real time such that public support could weaken for raccoon rabies control programs, or more broadly for ORV. We identify and discuss the complex of potentially important variables and the critical needs to better ensure longterm success for raccoon rabies control.

100

FOURTH SESSION

ORAL VACCINATION OF SKUNKS WITH A CANINE ADENOVIRUS TYPE 2-RABIES RECOMBINANT VIRUS VACCINE
C.E. Rupprecht 1, C.A. Hanlon 1, J. Self 1, J. Blanton 1, L. Greenberg 1, M.L. Faber 2, J.A. Mattis 2, K.Y. Pak 2, J. Li 3, M. Faber 3, B. Dietzschold 3
1) Centers for Disease Control and Prevention, Atlanta, GA, USA, 2) Molecular Targenting Technologies, Philladelphia, PA, USA, 3) Thomaz Jefferson University, Philladelphia, PA, USA

Dr. Charles Rupprecht - cyr5@cdc.gov

USA

From a continental perspective, skunks (Order Carnivora, Family Mephitidae) are important rabies reservoirs throughout North America. Unlike other significant mesocarnivores, such as raccoons, foxes, and coyotes, biologicals for oral rabies vaccination (ORV) have not been licensed for skunks. The objective of this experiment was to test the innocuity and effectiveness in skunks of a canine adenovirus type 2 recombinant virus vaccine (CAV2R), expressing the rabies virus glycoprotein. Animals consisted of 36 captive-bred, unvaccinated adult striped skunks (Mephitis mephitis). All animals were negative for evidence of rabies virus neutralizing antibodies (VNA) prior to study inclusion, with 6 groups of six animals per group. In all cases, the experimental vaccine or placebo volume was 1 ml. Negative control skunks received parental CAV2 vaccine per os. Two groups received CAV2R per os (108 or 109 TCID50). Two groups received an experimental recombinant rabies virus vaccine (SPBNGASGAS) developed by reverse genetics per os (108 or 109 TCID50). One group received CAV2R by the intramuscular (IM) route (108 TCID50). All animals remained well, regardless of group. Approximately 5 weeks post-vaccination, skunks were infected IM with a street rabies virus of skunk origin. Within 1 week post-challenge of rabies virus, all animals had developed detectable rabies VNA, with the exception of controls, and 1-2 animals each in the groups receiving the lower doses of SPBNGASGAS and CAV2R, respectively. The challenge phase is ongoing, and survivorship data will be updated ad hoc. Beyond this preliminary proof of concept of ORV using CAV2R, future research should include comparative safety testing, the potential ability for intra-specific transmission, the evaluation of minimal effective doses per os, and efficacy via baits, as well as extension to other significant target species.

101

RABIES IN TERRESTRIAL WILDLIFE: EPIDEMIOLOGY AND CONTROL

EVALUATION OF TWO CANDIDATE VACCINES FOR ORAL VACCINATION OF SKUNKS AGAINST RABIES
J. Self 1, J. Blanton 1, C. Hanlon 1, J. Maki, D. Grosenbaugh, CE. Rupprecht. 1
1) Centers for Disease Control and Prevention, Atlanta, GA, USA

Dr. Joshua S. Self - dnz0@cdc.gov

USA

The striped skunk (Mephitis mephitis) is the second leading animal diagnosed with rabies in the USA. A potential control measure for rabies in wildlife is vaccination through the distribution of baits containing vaccine. At present, there is a need for a safe and effective vaccine for skunks. A vaccinia-rabies glycoprotein (V-RG) recombinant virus vaccine is licensed for programs targeting wildlife, such as raccoons, gray foxes, and coyotes. However, inconsistent oral efficacy has been demonstrated with this vaccine in skunks. In this study, the oral efficacy of a highly attenuated rabies virus recombinant vaccine, constructed via reverse genetics, and designated SPBN-GASGAS, was compared with that of the V-RG vaccine in skunks. Three groups of six animals received 1.5 ml of either VRG (108.0 TCID50), SPBN-GASGAS (107.3 TCID50), or a placebo (phosphate buffered saline) in the oral cavity while under sedation. Six weeks later, the skunks were inoculated with street rabies virus (salivary gland homogenate from a naturally infected skunk from California) administered intramuscularly in the right and left masseter muscle (0.5 ml each site). The majority of the V-RG vaccine recipients developed rabies virus neutralizing antibodies by two weeks post-vaccination. In contrast, none in the SPBN-GASGAS group demonstrated antibody until seven days post challenge when three of the six animals demonstrated an apparent anamnestic response along with all the V-RG vaccinated animals. At the first definitive signs of rabies, animals were sedated and euthanized. Three of six skunks in the SPBN-GASGAS group and one of six in the V-RG group developed clinical signs and were euthanized. Rabies was confirmed by the direct fluorescent antibody test. In view of earlier studies that produced conflicting outcomes, these data require further corroboration, but add to a growing body of evidence supporting both the needs and opportunities involved towards the development of a safe and effective oral rabies vaccine for skunks.

102

FOURTH SESSION

EVALUATION OF ORAL WILDLIFE RABIES VACCINATION: INTERPRETATION OF FIELD SEROLOGY AND RESULTS OF CHALLENGE IN CAPTIVITY
Cathleen A. Hanlon 1, Craig B. Swope1, Jason Suckow 1, Michael Niezgoda 1, William Weldon, Jesse Blanton, Joshua Self 1, Dennis Slate 2, Charles E. Rupprecht 1
1) Centers for Disease Control and Prevention, Atlanta, GA. USA, 2) USDA, APHIS, Wildlife Services, NH, USA

Dr. Cathleen Hanlon - chanlon@cdc.gov

USA

In the USA, the area considered enzootic for raccoon rabies overlaps with a large and dense human population along the eastern seaboard, along the Appalachian Mountains and inland to Ohio. Beyond prevention efforts, including public education and human rabies post-exposure prophylaxis, oral rabies vaccination is a major tactic for wildlife rabies control. As part of the continuing evaluation of the oral rabies vaccination baiting program intended for free-ranging raccoons, animals from baited areas of Pennsylvania and unbaited areas in Michigan were livetrapped and transferred to captivity for rabies virus challenge studies. All 20 sero-negative (<0.05 IU/ml), rabiesnave raccoons succumbed to rabies. A group of seven raccoons were considered low-positive (0.05-0.10 IU/ml) on the basis of rabies virus neutralizing antibody serology at the time of capture. Upon challenge 4-6 months later, 3/7 (43%) survived lethal rabies virus inoculation. Among a medium-positive (>0.10-0.45) rabies antibody group, 6/11 (55%) survived challenged. The highest level of survivorship (8/11, 83%) was achieved among a group of raccoons with rabies antibody titers > 0.45 IU/ml at the time of capture, 4-5 months prior to challenge. All animals demonstrated a drop in detectable antibody levels during the period when they first arrived and the start of challenge. An overall survivorship of 63% (19/30) was observed among raccoons from all sero-positive groups (low-, med-, and high-). The determination of rabies virus neutralizing antibody levels is a valuable tool for the assessment of oral wildlife rabies vaccination efficacy, as it is an estimate of herd immunity. Many factors affect antibody levels in a free-ranging population. A critical consideration is the timeliness of sample collection as antibody kinetics from oral vaccination may lead to a narrow window of detection. The presence of antibody indicates a reasonable probability of immunity to a lethal rabies virus challenge and, conversely, a lack of rabies virus neutralizing antibody suggests a very high (but not absolute) susceptibility.

103

RABIES IN TERRESTRIAL WILDLIFE: EPIDEMIOLOGY AND CONTROL

RISK-BASED COST MODELING OF ORAL RABIES VACCINE INTERVENTIONS FOR RACCOON RABIES
Sergio Recuento 1, Millicent Eidson, Bryan Cherry, Glen Jonson
1) New York State Department of Health, Albany, NY, USA

Dr. Sergio Recuenco MD, DrPH - sergio.recuenco@gmail.com

USA

Oral rabies vaccine (ORV) is a potentially effective but costly strategy currently used to control raccoon rabies. Due to high costs, ORV for raccoon rabies in the U.S. has been limited to epizootic areas, with few and brief attempts to use ORV in enzootic areas, leaving extensive raccoon rabies regions without any ORV intervention. The number of rabid terrestrial animals in New York State (NYS) was modeled with a Poisson regression including land elevation, land use type, human population density, presence of large rivers/lakes, major roads, year, ecoregion, and proximity to ORV areas as covariates for the model. Several cost scenarios for ORV application in raccoon rabies enzootic and epizootic regions were modeled in NYS to obtain the total cost of ORV baits per scenario and potential savings compared with uniform ORV baiting strategies. These cost scenarios modeled application of ORV baits at different densities according to levels of risk defined by the observed number of raccoon rabies cases per km2, and the expected number of cases per km2 using estimates calculated with a Poisson model, at the census tract level. Use of these differential bait densities is projected to result in lower bait purchase costs than uniform baiting, for both the NYS enzootic region and the Long Island epizootic zone. The proportion of savings for the NYS enzootic region could be as much as 29.57% (cost scenario based on expected number of cases per km2). The proportion of savings for the Long Island epizootic zone could be as much as 38.9% (cost scenario based on expected number of cases per km2). Cost savings are contingent on the assumption that risk-based application of bait is at least as effective in controlling rabies as uniform baiting.

104

FOURTH SESSION

FIFTH SESSION
RABIES IN BATS

105

RABIES IN BATS

106

FIFTH SESSION

VAMPIRE MYTHS, VAMPIRE BATS AND THE TYRANNY OF THEORIES


Christopher Nunan 1
1) Peterborough, ON, Canada

Dr. Christopher Nunan - cnunan@cogeco.ca

Canad

Previously, I have indicated that the vampire legends came out of the East to infect a very susceptible Europe. But now, we are faced with a dilemma-where did the East get its vampires. Authors have previously looked towards a variety of gods, demons and devils (for example: Kali) to explain vampire legend origins. I suggest a simpler explanation- the myths began with Eastern explorers returning from bat infested areas of the globe Vampire bats had a profound effect upon early European explorers of Mid-America, from Columbus onward. But vampire bats are native only to Mid-America. How then could these bats begin the Eastern vampire mythos? Work by previous researchers in various fields has suggested that, unlike the standard North American Land Bridge Theory of human settlement/evolution in North America, people were visiting North, Central and South America using ships long before the land Bridge theory suggests they were on the landmass. It is these explorers who I suggest were encountering vampire bats and returning with the beginnings of the vampire legends. The theory of pre-Columbian oceanic transport is gaining acceptance. Hopefully, the vampire story will add to the growing body of evidence being amassed by the diffusionists.

107

RABIES IN BATS

PRIMER RELATO DE MURCILAGO HEMATOFAGO (DESMODUS ROTUNDUS) INFECTADO POR EL VIRUS DE LA RABIA, ENCONTRADO EN UN CENTRO URBANO, SP , BRASIL
Claudia Ferraz 1, Samira M. Achkar 2 e Ivanete Kotait 2
1) Centro de Controle de Zoonoses - Ubatuba/SP , 2) Instituto Pasteur, So Paulo, SP , Brasil

Dr. Samira Achkar - samira_achkar@yahoo.com.br

Brasil

Los murcilagos hematfagos son, en la actualidad, los principales reservorios del virus de la rabia en nuestro medio, particularmente el Desmodus rotundus (vampiro comn). Es reconocido, internacionalmente, que estE animal adems de causar grandes prdidas econmicas directas, como transmisor de la rabia a los herbvoros, ocasiona prdidas originadas del ataque per. se: anemia, disminucin de peso, depreciacin del cuero e infecciones secundarias. Desde el ao 2004, segn relatos de los Coordinadores Nacionales de los Programas de la Rabia en los pases de la Amrica Latina, ellos son, tambin, los principales transmisores de la rabia a los humanos, en especial en la regin de la Amazona. Otro aspecto importante esta relacionado a los animales de estimacin (perros y gatos) que, en reas urbanas consideradas controladas, presentan rabia que, a los estudios antignicos y genticos, revelan infeccin por la variante Desmodus rotundus (AgV3). Como espcimen del genero Artibeus sp han sido, con frecuencia, encontrados con la AgV3, perros y gatos, por contacto con los murcilagos frugvoros, de hbitos sinantropicos. En mayo del 2006, uno morador de la regin central de Ubatuba, encontr en el rea externa de su residencia, un murcilago caminando en el piso, a las diez de la maana. El Centro del Control de Zoonosis, al atender la denuncia, verific que el murcilago estaba inquieto, procuraba esconderse y, durante su captura, dio seales de agresividad. El animal fue encaminado al Laboratorio de Diagnostico del Instituto Pasteur, que lo identific como Desmodus rotundus. Cuando fueron realizadas las pruebas clasicas de diagnstico de rabia por imnufluorescencia directa y aislamiento viral, las dos resultaron positivas. Este es el primer relato de Desmodus rotundus, infectado por el virus de la rabia, encontrado en un centro urbano, del que los autores tienen conocimiento. La identificacin de espcimen de Desmodus rotundus, infectados por el virus de la rabia, s frecuente en reas rurales, en abrigos naturales y artificiales. En reas urbanas, todava su identificacin es rara, pero ya fue encontrado en centros de algunas capitales del BrasilSalvador, So Paulo, Belo Horizonte, Rio de Janeiro y Recife), con resultados negativos a las pruebas diagnsticas.

108

FIFTH SESSION

PRIMERA EPIZOOTIA DE RABIA EN DESMODUS ROTUNDUS EN LA COSTA DEL PER


Jorge Gmez-Benavides 1, Andrs Velasco-Villa 2, Ana Escudero-Quintana 3, Adelia Cahuina 4, MagaliChun 4, Mery Salvador 4, Ivn Vargas 4, Mnica Villanueva 5, Luis Suarez 1, Charles Rupprecht 2
1) Direction of Epidemiology of Ministry of Health, Lima, Peru, 2) Centers for Disease Control and Prevention, Atlanta, GA, USA, 3) Direction of Health II Lima Sur, Peru, 4) Direction of Health III Lima, Peru, 5) Centre Anti Rabies of Lima - Direction of Health V, Lima, Peru

Dr. Jorge Luis Gomez Benavides - jgomez@oge.sld.pe

Peru

Introduccin: A pesar de la amplia distribucin del Desmodus rotundus, en el Per, generalmente los brotes de rabia se presentaron en la selva. La Cordillera de los Andes, ha sido una barrera natural para el desplazamiento de la rabia silvestre a la costa. En los valles costeros se ha identificado al D. rotudus, por ms de 30 aos, pero la primera epizootia de rabia en vampiros fue notificada en julio del 2004, en el valle del Ro Mala, a 80 Km al sur de Lima, cuando se detecta un mono Cebus apella y un vampiro con VAg 3. Objetivo: describir la epizootia de rabia y alertar sobre el desplazamiento accidental de vampiros a la ciudad de Lima. Mtodos Se revisaron los informes de las epizootias de rabia silvestre desde julio 2004 hasta julio 2006. Resultados Se determin que el mono fue desplazado de la selva y haba permanecido en el valle por ms de 3 aos, haba mordido a una nia el 18/04/04, no evidenciaba signos de rabia, la IFD fue negativa, la prueba biolgica fue positiva, inicindose vacuna VERO 54 das despus de la exposicin. Actualmente, la nia est saludable. No se identificaron ms personas expuestas al mono. En el valle, haba mordeduras por murcilagos en el ganado, pero no se inform de muerte de ganado. Se realiz captura de murcilagos, encontrndose que los tneles de canales de regado era un lugar ideal para refugios de vampiros. Desde mayo del 2004 hasta julio del 2005, se han presentado 25 casos de rabia (18 vampiros, 1 murcilago frugvoro, 4 monos, 1 gato y 1 can) distribuidos en forma dispersa en los valles costeros de Mala y Chancay y en Lima Metropolitana, 8 causaron mordeduras de personas y 17 fueron capturados por vigilancia. En 1 can, 2 monos y 2 vampiros se encontr VAg relacionado con vampiros. Conclusin Se reporta por primera vez una epizootia de rabia en D. rotundus en 2 valles de la costa peruana, con desplazamiento accidental del reservorio infectado a la ciudad de Lima y transmisin a canes y monos.

109

RABIES IN BATS

RABIA EN MURCILAGO HEMATFAGO DIPHYLLA ECAUDATA Y DESMODUS ROTUNDUS EN EL ESTADO DE PIAU, NORDESTE DEL BRASIL
Juliana G. Castillo 1, Rosangela Cavalcante 2, Antonio Augusto Santana 3, Wellington L.G.Alencar Rosa 3, Maria Luiza Carrieri 1, Ivanete Kotait 1
1) Instituto Pasteur, So Paulo, SP , Brasil 2) Laboratorio de Patologa Animal, Teresina, PI, Brasil, 3) Secretaria de Desenvolvimento Rural, Teresina, PI, Brasil

Dr. Juliana Galera Castilho - juliana.castilho@uol.com.br

Brasil

La fauna de los murcilagos brasileos est constituida por cerca de 150 espcies y representa, aproximadamente, 30% de las especies de mamferos existentes en Brasil. De estas especies, solamente tres (3) poseen hbitos hematofgicos: el Desmodus rotundus - murcilago hematfago comn, que se alimenta preferentemente de los mamferos; el Diaemus youngii - que prefiere las aves - y el Diphylla ecaudata - que se alimenta de aves y ocasionalmente de mamferos. Sin embargo el Desmodus rotundus es considerado el principal reservorio silvestre del virus de la rabia en nuestro medio, rarsimos son los aislamientos de este virus en las otras dos especies. En el ao 2004, el Laboratorio de Patologa Animal (ADAPI/Teresina) despus del diagnstico de la rabia, en dos espcimenes de murcilagos hematfagos (uno Desmodus rotundus y otro Diphylla ecaudata), por las tcnicas de inmunofluorescencia directa y aislamiento viral, encamin las muestras al Instituto Pasteur de So Paulo, donde fueron realizados estudios antignicos y genticos. La tipificacin antignica, a travs de la inmunofluorescencia indirecta con el panel de anticuerpos monoclonales del CDC/Atlanta/USA result, en los dos aislados, Variante Antignica 3 (AgV3), compatible con la variante frecuentemente aislada de Desmodus rotundus. El estudio gentico fue realizado por secuenciamento, a partir de una regin de 249pb, correspondiente al gen de la nucleoprotena. Las secuencias finales de 165 nucletidos de la 3 terminal del gen N, localizadas entre los nucletidos 1318 y 1482 fueron aliadas con secuencias homlogas presentes en el GenBank. El anlisis filogentico demostr que los aislados presentaron 99,3% de identidad entre ellos y segregaron juntamente con otros aislados de murcilagos hematfagos (Desmodus rotundus), de diferentes regiones del Brasil. Este es el primer aislamiento del virus de la rabia en Divilla ecaudata, en Brasil, desde 1934, cuando fue aislado en el sur del pas (Estado de Santa Catarina). Los anlisis antignico y filogentico sugieren que las especies hematfagas co-habitan los mismos abrigos.

110

FIFTH SESSION

EPIDEMIOLOGA DE LA RABIA EN CHILE, 1989 - 2005


Myriam Favi Corts 1, Luis Rodrguez Aguayo 1, Carla Espinosa Martnez 1
1) Instituto de Salud Pblica, Ministerio de la Salud, Santiago, Chile

Dr. Myriam Favi Corts - mfavi@ispch.cl

Chile

Introduccin - La importancia de los animales silvestres en la trasmisin de la rabia en Chile fue reconocida en el ao 1985,cuando se detect por primera vez rabia en murcilagos insectvoros de la especie Tadarida brasiliensis. A partir de entonces el patrn epidemiolgico de la rabia en Chile se ha caracterizado por una endemia en quirpteros algunos casos en animales domsticos y un caso humano cuya fuente de infeccin fueron murcilagos insectvoros. Objetivo Describir la evolucin que ha tenido la rabia en Chile entre los aos 1989 al 2005, mediante un estudio epidemiolgico de tipo retrospectivo Mtodo Se utilizaron los datos existentes (39751 registros), incorporndolos al programa Excel, utilizando tablas dinmicas de ste para resumir o cruzar la informacin, y Statgraphics versin 5.0 y Stata versin 6.0 para las pruebas de hiptesis. Resultados La distribucin porcentual del total de murcilagos positivos por regiones en el periodo estudiado fue especialmente alta para las Regiones: Metropolitana,VIII y V (zona centro sur del pas) .La especie Tadarida brasiliensis es la que obtuvo el mayor porcentaje de positividad en comparacin con especies como Laciurus spp,Myotis chiloensis e Histiotus macrotus. Se detect una inesperada tendencia creciente de la proporcin de murcilagos positivos en el perodo estudiado usando test de Mann-Kendall (p= 0.024). Adems, usando la prueba exacta de Fisher para los 17 aos estudiados, se verific que el muestreo de murcilagos por captura (Vigilancia) rindi una proporcin bajsima de animales positivos en comparacin con lo observado en las muestras recibidas por sospecha de Rabia. Tal diferencia result altamente significativa con la nica excepcin del ao 1990 (p=0.736). Los meses con mayor proporcin de murcilagos positivos correspondieron al perodo: Octubre-Marzo.

111

RABIES IN BATS

GENETIC CHARACTERIZATION OF BRAZILIAN BAT RABIES VIRUSES. EVALUATION OF THE PATHOGENICITY AND CROSS PROTECTION IN MICE
E. M. S. Cunha 1, A.F .C. Nassar 1, C.S.H. Lara 1, E.C.M. Villalobos 1, T. Itou 3, T. Sakai 3, Y. Shoji 3, F .H. Ito 2
1) Instituto Biolgico, So Paulo, SP , Brasil, 2) Faculdade de Medicina Veterinria e Zootecnia, Universidade de So Paulo, SP , Brasil, 3) College Nihon University, Fujisawa, Kanagawa, Japan

Dr. Elenice Maria Sequetin Cunha - sequetin_cunha@hotmail.com

Brasil

The N-protein of twenty-three bat-rabies viruses was genetically sequenced. Four lineages were identified: those related to the vampire bats and insectivorous bats 1, 2, and 3. The insectivorous bats lineage 1 was from the Eptesicus furinalis; lineage 2 included isolates from Molosssus spp, and the lineage 3, from the Nictinomops laticaudatus. The identity among viruses of the lineages 1, 2, and 3 were greater than 99%, 97% and 99%, respectively. Vampire bat lineages were represented by three isolates from Desmodus rotundus; five from the Artibeus lituratus and Artibeus planirostris; two from insectivorous bats, and two from unidentified species. The pathogenicity of five bat isolates and one dog isolate were assessed in mice through intracerebral (IC) and intramuscular (IM) inoculation. All the isolates inoculated via IC were pathogenic, provoking death in 4 - 14 days. Mice inoculated with 500 ICLD50 with same isolates via IM showed varying death rates: 60.0% (D. rotundus); 50.0% (dog and N. laticaudatus); 40.0% (A. lituratus); 9.5% (M. molossus), and 5.2% (E. furinalis). These isolates were submitted for the cross protection test using a commercial vaccine of veterinary use, by vaccinating the mice subcutaneously and administrating one or two shots, and challenged through IC and IM routes. Mice receiving two shots and challenged intracerebrally were protected against all the isolates, and mice receiving one shot were partially protected against the PV strain and the dog isolate. Mice challenged intramuscularly showed 100.0% of protection, with the exception of those vaccinated with one dose and challenged with PV strain, with 66.0% of survivors. These results suggest the circulation of rabies virus variants in different species of bats, and that the vampires, frugivorous, and insectivorous bats share the same lineage of rabies viruses. The mice resisted against the challenge, suggesting that the PV vaccine was effective against those bat-rabies isolates.

112

FIFTH SESSION

VIGILANCIA EPIDEMIOLGICA DE LA RABIA EN MURCILAGOS REALIZADA EN BRASLIA, DF, BRASIL


Pricles Massunaga 1, Maria Luiza Carrieri 2, Angelika Bredt 1, Maria Isabel Rao Boffil 1, Rodrigo Mena Barreto Rodrigues 1, Ivanete Kotait 2
1) Diretoria de Vigilncia Ambiental, DF, Brasil, 2) Instituto Pasteur de So Paulo, SP , Brasil

Dr. Pricles Massunaga - pericles@saude.df.gov.br

Brasil

La vigilancia epidemiolgica de la rabia en murcilagos, en reas urbanas y rurales de Braslia, DF, fue implantada en el ao 1988, a partir del diagnstico de los especimenes enviados al laboratorio, ya sea por demanda espontnea o por capturas realizadas por el equipo de la Direccin de Vigilancia Ambiental (DIVAL). A pesar de que este trabajo se vena ejecutado desde hace varios aos atrs, solamente en el ao 2001 fueron obtenidos resultados positivos para la rabia, mediante la aplicacin de las tcnicas de inmunofluorescencia directa y aislamiento viral en ratn. Desde entonces fueron encontrados 16 murcilagos positivos: seis insectvoros, en reas urbanas centrales distintas (cinco Nyctinomops laticaudatus y uno Lasiurus blossevilli): nueve hematfagos en reas rurales (Desmodus rotundus) y un frugvoro, tambin en rea urbana (Artibeus lituratus). La identificacin de las especies fue realizada segn la Clave para Determinacin de Quirpteros Brasileros (Visotto & Taddei). Todos los aislados de murcilagos insectvoros, del murcilago frugvoro y de cinco murcilagos hematfagos fueron sometidos a la tipificacin antignica, por inmunofluorescencia indirecta, utilizando el panel de anticuerpos monoclonales, cedido por CDC/ Atlanta, para lo estudio de variantes antignicas aisladas en las Amricas. Todos los aislados de Desmodus rotundus y de Artibeus lituratus tuvieron padrn de reaccin compatible con la variante antignica 3 (AgV 3), como ya fue demostrado en otros estudios antignicos y genticos realizados en Brasil. El padrn de reaccin del aislado de Lasiurus blossevilli fue compatible con la variante antignica 6 (AgV6), propia del gnero Lasiurus sp. Los cinco aislados de Nyctinomops laticaudatus no mostraron compatibilidad con los perfiles definidos por el panel de anticuerpos monoclonales utilizado, exhibiendo dos perfiles antignicos distintos, contrariamente a lo que ha sido relatado en estudios recientes, que sugieren la existencia de un solo padrn antignico especie-especfico, entre los murcilagos insectvoros. Estudios genticos estn siendo realizados para la confirmacin de stos resultados.

113

RABIES IN BATS

RABIA EN MURCILAGOS EN EL REA URBANA DE BELO HORIZONTE, MINAS GERAIS, BRASIL: IDENTIFICACIN DE ESPECIES Y ESTUDIOS ANTIGNICOS
Adamastor Santos Busolotti 1, Ana Paula Machado Tancredi de Araujo 1, Maria do Carmo de Araujo Ramos 1, Varlei Antnio de Almeida Gomes 1, Francisco Elias Nogueira da Gama 1, Marlia de Oliveira Cavalieri 2, Talita Leal Chamone 3, Maria Luiza Carrieri 4, Ivanete Kotait 4
1) Centro de Controle de Zoonoses, Belo Horizonte, MG, Brasil, 2) Instituto Mineiro de Agropecuria, SEA, MG, Brasil, 3) Secretaria de Estado da Sade de Minas Gerais, MG, Brasil, 4) Instituto Pasteur, So Paulo, SP , Brasil

Dr. Maria do Carmo de Arajo Ramos - cczsmsa@pbh.gov.br

Brasil

En el 2002, la Secretara Municipal de Salud, inici en la Regin Metropolitana de Belo Horizonte, un Programa de Vigilancia de la Rabia en Murcilagos, mediante el envo de los especimenes encontrados en situaciones nohabituales para el Laboratorio de Diagnstico local. La ciudad de Belo Horizonte, desde el 1989, no relataba circulacin del virus de la rabia. El laboratorio de diagnstico realiz las pruebas clasicas (immunofluorescncia directa e aislamiento viral) en 704 especimenes, enviados en el periodo 2002 - julio 2006, habiendo 29 resultado positivos para el virus de la rabia. De estos animales, 21 fueron frugvoros (Artibeus lituratus) y 8 insectvoros (3 Nyctinomops laticaudatus, 2 Myotis sp, 1 Histiotus velatus, 1 Dasypterus sp e 1 Cynomops sp). Los aislados fueron, entonces, encaminados al Instituto Pasteur, para la tipificacin antignica, a travs de uso de anticuerpos monoclonales, siendo identificada la variante 3 (AgV3), propia del Desmodus rotundus, en 16 espcimen de Artibeus lituratus, como haba sido observado en otros centros urbanos del pas, particularmente de la regin sudeste. En 5 murcilagos insectvoros fueron identificadas variantes no compatibles con los padrones antignicos definidos por lo panel del CDC de Atlanta. Estos estudios preliminares refuerzan la necesidad del mantenimiento del Programa de Vigilancia de la Rabia en Murcilagos, para la proteccin de la salud pblica.

114

FIFTH SESSION

BATS RABIES IN THE PERIOD BETWEEN 2000 - 2006, IN RIBEIRO PRETO COUNTY, SO PAULO STATE, BRAZIL
Maria Luiza Carrieri 1, Rafael de Novaes Oliveira 1, Flvia M. Carnelo 2, Juliana Galera Castilho 1, Pedro Carnieli Jr.1, Samira Achkar 1, Karin Scheffer 1, Carla Izabel Macedo1, Paulo E.Brando 3, Ivanete Kotait 1
1) Instituto Pasteur, So Paulo, SP , Brasil 2) Centro de Controle de Zoonoses, Ribeiro Preto, SP , Brasil

Dr. Maria Luiza Carrieri - mlcarrieri@pasteur.saude.sp.gov.br

Brasil

With the control of dog-related rabies in the southern and some areas of southeastern Brazil achieved in the last two decades, rabies from wild animals, including bat rabies, has received special interest from researchers and public health authorities. Among Brazilian cities in which rabies in non-haematophagous bats is a more serious problem, the Ribeiro Preto county is the one where the highest frequency of bats rabies occurs. In the period between 2000 and July 2006, 4,057 brain samples from bats found in urban area were processed by FAT and MIT or N2A cells culture isolation. Of these, 64 were positives (1, in 2000; 3, in 2002; 15, in 2003; 1, in 2004; 28, in 2005 and 16 in 2006 (July). The positive species identified had frugivorous and insectivorous habits: Artibeus sp, Myotis sp, Molossus rufus, Eptesicus furinalis, Lasiurus ega, Nyctinomops laticaudatus, Myotis nigricans, Eumops perotis and, Cynomops abrasus. The antigenic studies made with the panel of 8 monoclonal antibodies, provided by CDC/ Atlanta/USA, identified antigenic variants 3 and 6 (AgV3 and AgV6), and two other with an indefinable pattern. The partial N gene (nt 124 to 1472) of 20 rabies virus isolates from these bats was sequenced, being 10 from frugivorous (Artibeus lituratus) and 10 from insectivorous species from the genera Nyctinomops, Eptesicus, Molossus, Myotis, Cynomops and Eumops. The sequences were aligned with homologous sequences retrieved from the Genbank using the CLUSTAL/W method and used to build a neighbor-joining distance tree with the K2-P model. The sequences from A. lituratus segregated with the rabies virus variant associated in Brazil with the vampire bat Desmodus rotundus, while those isolates from the insectivorous bats segregated in the following clusters: one isolate from the species Eptesicus furinalis segregated in the Eptesicus furinalis cluster; six isolates from N. laticaudatus segregated in the cluster classically associated with this bat species; one isolate from a Cynomops abrasus bat segregated in the cluster related to the genus Molossus-associated rabies virus and finally two isolates from a Myotis nigricans and Myotis sp segregated in the Myotis spp specific cluster. These results show that non-vampire bats, mainly the insectivorous ones, are hosts for a wide diverse range of rabies virus strains.

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RABIES IN BATS

BAT RABIES IN SO JOS DO RIO PRETO, SOUTHEAST OF BRAZIL


L.F.A. Martorelli 1, 3, M.F. Almeida 1, S.R. Favoretto 2, 3, J. Trezza-Netto 1, M.M. So 1, L.F .V. Amaral 4, A.P .A.G. Kataoka 1, E.L. Durigon 3
1) Centro de Controle de Zoonoses, So Paulo, SP , Brasil 2) Instituto Pasteur, So Paulo, SP , Brasil 3) Instituto de Cincias Biomdicas, Universidade de So Paulo, SP , Brasil 4) Centro de Controle de Zoonoses, So Jos do Rio Preto, SP , Brasil

Dr. Luzia Martorelli - lumartorelli@yahoo.com

Brasil

In So Paulo State - Brazil, rabies programs actions have progressively reduced the number of rabies cases in domestic animals. Bats have emerged as the most important rabies reservoir. So Jos do Rio Preto, situated in northwestern of So Paulo State, the actions of rabies control are well established, with special care to educational and community mobilization to the importance the presencee of rabid bats in urban area and rabies risk to humans and domestic animals by bats contact. During the years 2004 and 2005, 1069 bats were sent to rabies diagnosis and species identification. These bats were sent to rabies laboratory after contact with human or domestic animal or when they are find in local and time unusual to the normal behavior of the animal. The insectivorous bats constituted the majority of the sample (70%), 23% were frugivorous bats, and 7% of other alimentary habits. The diagnostic was made by Direct Immunofluorescent Test (FA) and Mouse Inoculation Test (MIT). Fourteen bats were positive to rabies (1,3%). The antigenic characterization was made by a monoclonal antibodies panel against the viral nucleoprotein, from CDC, Atlanta, USA. These analyses identified rabies antigenic variants associated with Desmodus rotundus, Tadarida brasiliensis, and showed a different profile not pre established at the panel. The genetic characterization of rabies virus isolates was made by direct sequencing and molecular analysis of a 320pb fragment of the N gene. The phylogenetic analysis revealed the presence of different genetic variants from insectivorous and one variant from hematophagous found in frugivorous bats. The identification of the same genetic variant in isolates obtained from different bats species underscores the complexity of rabies epidemiology in the Order Chiroptera and to reinforce the possibility of inter-species transmission or the maintenance of the same rabies variant by two or more different species.

116

FIFTH SESSION

VIRUS RABIA EN MURCILAGOS HEMATOFAGOS Y NO HEMATOFAGOS DE AREAS URBANAS Y RURALES EN EL DEPARTAMENTO DEL VALLE, COLOMBIA
C.L. Nez Meja1
1) Universidad del Valle, Calli, Colombia

Dr. Constanza L. Nuez Mejia - colenume@yahoo.com

Colmbia

La intervencin del hombre al medio ambiente ha provocado migracin de los murcilagos a las ciudades, convirtindose en un factor de riesgo para la transmisin del virus rabia silvestre al hombre y animales domsticos. El vampiro (Desmodus rotundus), el frutero Carollia perspicillata y los murcilagos insectvoros (Molossus molossus y Eptesicus brasiliensis) han salido positivos a virus rabia y han ocasionado mordeduras en humanos y mortalidad en animales domsticos en Colombia. El objetivo principal de la vigilancia de la rabia transmitida por murcilagos de hbitos caseros y silvestres en el departamento del Valle, es examinar algunos factores de riesgo que estn asociados con la transmisin de rabia por murcilagos en reas urbanas y rurales. La frecuencia de infestacin de murcilagos al interior y exterior de las edificaciones, las especies de murcilagos y la infestacin con el virus rabia fueron estudiadas desde el ao 2000 hasta el 2005 en los municipios del Valle. Se realizaron las capturas tomando la informacin epidemiolgica de quejas de mordeduras y de invasin en edificaciones. Los murcilagos fueron capturados con redes de niebla para hacer bsqueda del virus rabia en el cerebro mediante inmunofluorescencia directa e inoculacin en cerebros de ratones lactantes. Cuando se encontraba un murcilago positivo se realiz tipificacin viral por inmunofluorescencia indirecta usando anticuerpos monoclonales especficos. La positividad para rabia en murcilagos fue de 0.1%(1/890) en el 2000, 0%(0/319) 2001, 0.7%(1/145) en 2002, 0%(0/347) en 2003, 0%(0/178) en 2004 y 0%(0/3) en 2005. El estudio ofreci servicios de asesora a la comunidad sobre el desalojo de murcilagos de los techos y en el control para evitar que estos vuelvan a invadir las viviendas. Los resultados muestran que los murcilagos son reservorios importantes del virus rabia. Se recomienda continuar la vigilancia para prevenir la rabia transmitida por murcilagos en el rea urbana de Colombia.

117

SIXTH SESSION
SURVEILLANCE AND CONTROL OF ANIMAL RABIES

SURVEILLANCE AND CONTROL OF ANIMAL RABIES

120

SIXTH SESSION

IDENTIFICATION OF SHORTCOMMINGS IN RABIES CONTROL METHODS OF KWAZULU-NATAL


Kevin Le Roux 1
1) South Africa Veterinary Services, Kwazulu-Natal, South Africa

Dr. Kevin Le Roux - kleroux@allerton.kzntl.gov.za

frica do Sul

Abstract: Kwazulu-Natal (KZN), our most densely populated province, is located on the eastern seaboard of South Africa. This province also has a very large dog population. KZN Health Services spend R17 million a year ($3m) on post exposure prophylaxis alone, while the Veterinary Services of KZN attempt to carry out rabies control with a limited budget and infrastructure. Massed annual parenteral immunization campaigns have failed to consistently reduce the numbers of recorded cases in dogs over the past 10 years. In an attempt to understand this, a small pilot research project was conducted. Preliminary findings indicate that only 38% of dogs are being vaccinated. Some of the reasons identified include: (a) An increasing stray (feral) dog population (currently 10%) fueled by socioeconomic factors such as poverty, poor education, and changes in family structures due to HIV& AIDS. (b) Upward of 20% of owners are not at home during these massed campaigns. (c) Up to 25% of dogs can simply not be controlled. (d) With no veterinary input, breeding is uncontrolled and has many spin-offs in terms of movement of animals, and abandoned dogs. Our research will be expanded to cover all community types in KZN and control methods are reviewed to overcome all the identified shortcomings. Among our primary objectives are improved co-operation with Health services as well as the application of Bait vaccines as an adjunct to parenteral vaccination.

121

SURVEILLANCE AND CONTROL OF ANIMAL RABIES

RABIES SURVEILLANCE IN THE UNITED STATES, 2005-06


Jesse D. Blanton 1, John W. Krebs 1, Cathleen A. Hanlon 1, Charles E. Rupprecht 1
1) Centers for Disease Control and Prevention, Atlanta, GA, USA

Dr. Jesse D. Blanton - asi5@cdc.gov

USA

During 2005, 6,417 cases of rabies in animals and 1 case in a human were reported to CDC, representing a 6.2% decrease from the 6,844 cases reported in 2004. Approximately 92.3% of the cases were in wildlife and 7.7% were in domestic animals (compared with 92% and 8%, respectively in 2004). Relative contributions by the major animal groups were 2,534 raccoons (39.5%), 1,478 skunks (23.0%), 1,408 bats (21.9%), 376 foxes (5.9%), 269 cats (4.2%), 93 cattle (1.4%), and 76 dogs (1.2%). Compared with the numbers of reported cases in 2004, cases in 2005 decreased among all groups except bats. One human rabies case occurred in Mississippi during 2005. Samples from the patient were submitted by the State Department of Health to CDCs unexplained deaths project. Rabies was diagnosed through antibody detection in paired serum and CSF . Viral characterization was not possible from the available samples. A subsequent epidemiologic investigation indicated a bat as the probable source of infection. The number of rabies cases reported by 20 states during the first 6 months of 2006 has continued to show a decreasing trend: the overall number of reported cases reported by these states (1,403) was 1.2% lower than that reported by the same states during the same period for 2005 (1,420). As of July, one human rabies case had been reported from Texas. The rabies virus variant associated with Tadarida brasiliensis (Mexican free-tailed bat) has been implicated in this case.

122

SIXTH SESSION

MANAGEMENT OF ILLEGAL INTRODUCTION OF PET CARNIVORES: THE CASES RECORDED IN ITALY IN 2005 AND 2006
F. Mutinelli 1, P . Grandi 1, P . Pavan 1, L. Piccoli 1
1) National Reference Laboratory for Rabies, IZS delle Venezie, Italy

Dr. Franco Mutinelli - fmutinelli@izsvenezie.it

Italy

Harmonisation of animal health requirements applicable to the non-commercial movement of pet animals between Member States and from third countries is necessary and only measures adopted at Community level can enable that objective to be achieved. Regulation (EC) no 998/2003 on the animal health requirements applicable to the non-commercial movement of pet animals and amending Council Directive 92/65/EEC has then been issued. Some of its provisions, in particular concerning rabies, have as their direct objective the protection of public health. Over the past 10 years the rabies situation has improved spectacularly throughout the Community following the implementation of programmes for the oral vaccination of foxes in regions affected by the sylvatic rabies epidemic. This improvement has led some Member Countries to abandon the system of quarantine which they applied for decades, in favour of an alternative, less restrictive system providing an equivalent level of safety. Cases of rabies observed in pet carnivores in the Community now mainly affect animals originating in third countries where urban rabies is endemic or community animals that became infected in a third country. The animal health requirements generally applicable hitherto by the Member States to pet carnivores introduced from such third countries should accordingly be made more stringent. Two cases of illegal introduction of pet animals were recorded in Italy after the new EU regulation came into force. A 5-month-old and a 10-month-old dogs were illegally imported from Morocco and Ukraine into Piemonte Region in August and September 2005 respectively and a 3-month-old cat from Lebanon into Veneto Region in April 2006. An epidemiological investigation was carried out by local Veterinary Services in order to gather information about the history of the pets and apply the more appropriate preventative measures to guarantee the protection of public health. Both the dogs and the cat were quarantined at home with special restrictions, the dogs had in their history a rabies vaccination and the antibody titre tested after introduction was greater than 0.5 IU/ml. After nine months the dog is well. The cat is still observing a 6-month-quarantine, no vaccination was reported and is presently clinically healthy. The rules governing movement of pet animals from third countries and the measures adopted in case of illegal introduction are discussed.

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SURVEILLANCE AND CONTROL OF ANIMAL RABIES

QUANTIFICATION OF RABIES NEUTRALIZING ANTIBODIES IN DOGS FROM A RABIES ENDEMIC AREA IN NORTHERN COLOMBIA 2004
Andrs Pez Martnez 1, Javier Gomez Meza, Jos Fernndez Manrique, Mara Fernanda Mndez, Oscar Francisco Franco. 1) La Salle University, Bogot, Colombia Dr. Andrs Pez-Martnez - apaez@lasalle.edu.co Colmbia

Dog rabies has been a threat for human health in northern Colombia for more than 20 years. Magdalena department has been the most threatened region with over 90% of the total of rabies cases in Colombia in years 20032006 despite the massive rabies vaccination campaigns. Pivijay and Cienaga are two towns in Magdalena that had most of the rabies cases in years 2003-2004. These two towns were therefore chosen for measuring rabies neutralizing antibodies in the dog population, and to evaluate the impact of the rabies vaccination campaigns in this area. Blood samples where taken from 408 dogs. 218 in Cienaga and 190 in Pivijay. Serum was separated by centrifugation and rabies antibodies were measured using glicoprotein coated ELISA plates. 88% of dogs in Cinaga and 84.9% in Pivijay had rabies neutralizing antibodies. 70% of dogs in Cinaga and 64% in Pivijay had protective concentrations of rabies neutralizing antibodies (over 0.5 UI/ml). The rabies antibody concentration was independent of gender and increased gradually with age which is expected in regions with periodical vaccination campaigns. Dogs over 8 years old had decreased antibody concentrations which suggests that aging may lower the immune response in these animals. Dogs between 6 months and 1 year old were classified as of high risk of getting rabies due to the low rate of seroprotected individuals. It is therefore advisable the rabies vaccination campaigns are carried out at least every six months rather than annually. Overall the rabies vaccination campaigns in the study area have been efficient in the sense that most vaccinated dogs become protected against the disease. In fact no dog rabies was reported in Cienaga and Pivijay in 2005 and 2006 indicating that the protection rates of 64% to 70% reported in this study are sufficient to stop rabies transmission.

124

SIXTH SESSION

CANINE RABIES ELIMINATION FROM THE UNITED STATES: AN EPIDEMIOLOGICAL AND VIROLOGICAL PERSPECTIVE
Andrs Velasco-Villa 1, Lillian A. Orciari 1, Charles E Rupprecht. 1
1) Centers for Disease Control and Prevention, Atlanta, GA, USA

Dr. Lllian Orciari - LOrciari@uga.edu

USA

During the first half of the 20th century, canine rabies was enzootic throughout the United States, with more than 9,000 dog rabies cases reported during 1944 alone. By the end of World War II, a comprehensive plan of dog rabies vaccination and stray animal removal began. By the mid-1950s, reported cases were reduced by more than 50%, but rabies associated with dogs remained of greatest concern in the southeastern and southcentral states through the 1960s. Within the next decade, the major dog rabies virus variant had been eliminated. However, during the late 1980s, an epizootic of canine rabies, maintained by coyotes, but also involving domestic dogs, began in southern Texas along the border with Mexico. Early on, considerations circulated that this new epizootic was related to a long term dog rabies enzootic located in northwestern Mexico (the so-called Sonora dog variant). Historical and recent samples associated with several dog rabies foci throughout Mexico (1961-2004), as well as samples obtained from the dog-coyote rabies focus circulating at the Mexico border were analyzed by molecular methods. The dog-coyote focus does not have strong phylogenetic relationships with the prior Sonora dog rabies virus variant. Conversely, both major enzootic foci located at the border between Mexico and the United States (Sonora dog focus, dog-coyote focus) appear related to long term dog rabies enzootics circumscribed in central Mexico. Thus, multiple rabies viruses, some now extinct, have contributed to canine rabies maintenance in the United States over the past century. These preliminary results highlight the ongoing needs for multi-national collaborations in the program to eliminate canine rabies in this Hemisphere, as well as the key role of canine rabies viruses as an operative factor for host shifts into other important carnivore populations, such as gray foxes.

125

SURVEILLANCE AND CONTROL OF ANIMAL RABIES

RELATO DE UN CASO DE RABIA CANINA, EN EL MUNICPIO DE CORUMB, MATO GROSSO DO SUL, BRASIL, EN 2006
Ademar Etiro Mori 1, Danielle Ahad das Neves 1, Gislaine Coelho Brando 2, Veronique M. C. L. Cortada 1, Maria Luiza Carrieri 3
1) Agncia de Defesa Sanitria Animal e Vegetal, MS, Brasil 2) Secretaria de Sade, MS, Brasil 3) Instituto Pasteur, So Paulo, SP ,Brasil

Dr. Vronique Louvet Cortada - cortadalouvetvmc@yahoo.com.br

Brasil

Una de las acciones del Programa Nacional de Control de la Rabia es el envo sistemtico de muestras de perros y gatos para el diagnstico laboratorial, con el objetivo de verificar la circulacin viral en la poblacin animal. En Enero de 2006, fue enviado por el Centro de Control de Zoonosis de Corumb, al Laboratrio de Diagnstico de Enfermedades Animales - LADDAN perteneciente a la Agencia Estatal de Defensa Sanitaria Animal y Vegetal de MS - IAGRO, una muestra del sistema nervioso central de un perro errante, de aproximadamente cinco aos, que no presentaba sintomatologa nerviosa y que fue sometido a eutanasia. La muestra fue procesada, presentando positividad en las pruebas de immunofluorescencia directa y prueba de aislamiento viral. Esta muestra fue encaminada al Instituto Pasteur para la realizacin de la tipificacin antignica, utilizando el panel de anticuerpos monoclonales producidos por lo CDC/Atlanta. El resultado obtenido fue variante 1 (AgV-1), propia del perro, nunca antes identificada en Brasil. El barrio donde el animal fue encontrado, pertenece a una regin prxima a la frontera con Bolivia. Despus de la investigacin realizada, tanto por la Secretaria Municipal de Salud de Corumb, como por la Gerencia de la Red de Salud - Provincia German Busch - Bolivia, se concluy que el perro fue originario de Bolivia, donde agredi a una persona, la cual oportunamente recibi tratamiento profilctico. Las acciones para el control de la Rabia en Corumb fueron realizadas coordinadamente con los servicios locales de salud de Bolivia. Acciones semejantes a estas fueron adoptadas en todo el Estado de Mato Grosso do Sul. El ltimo caso de rabia humano registrado en el Estado de Mato Grosso do Sul fue en 1994 y de rabia canina en julio de 2001 en el municipio de Ponta Por, frontera con Paraguay, en un perro errante del Paraguay.

126

SIXTH SESSION

REINTRODUCCIN DE LA RABIA URBANA EN LIMA METROPOLITANA Y EL CALLAO - PER


Jorge Gmez-Benavides 1, Andrs Velasco-Villa 2, Ivn Vargas 3, Mnica Villanueva 4, Albina 5, Charles Rupprecht 2
1) Direccin General de Epidemiologa del Ministerio de Salud, 2) Rabies Section. Centers for Disease Control and Prevention, Atlanta, GA, USA, 3) Direccin de Salud III Lima Norte, 4) Centro Antirrbico de Lima - Direccin de Salud V Lima Ciudad

Dr. Jorge Luis Gomez Benavides - jgomez@oge.sld.pe

Peru

Introduccin - El control de la rabia urbana en el Per, ha reducido drsticamente el nmero de casos. En Lima y Callao, principales metrpolis del pas, los ltimos casos de rabia canina fueron en 1999. Despus del ao 2000, se reducen las coberturas de vacunacin antirrbica canina, debido a la reduccin en el financiamiento. El movimiento de mascotas es libre. Lima y Callao, comprenden 49 distritos y su extensin de 2,799.75 km2, urbanizado en su mayor parte y su poblacin estimada es 8325,615 habitantes. En julio del 2004, se inicia la epizootia con casos de rabia canina, confirmada por IFD, en el distrito de San Juan de Lurigancho. Objetivo: Describir la epizootia de rabia urbana en Lima y Callao, desde julio 2004 hasta julio 2006. Mtodos: Se revisaron todos los informes de control de focos de rabia remitidos a epidemiologa. Resultados En Lima y Callao, se presentaron 16 casos de rabia canina y 4 de rabia en gato, distribuidos en 9 distritos, 15 estuvieron relacionados con accidentes de mordedura y 5 fueron recolectados en la va pblica por vigilancia activa de virus rbico. Se emitieron alertas epidemiolgicas al personal de salud, se investigaron todos los casos, se buscaron personas expuestas y contactos, que iniciaron tratamiento antirrbico; se eliminaron canes y gatos expuestos a los animales rabiosos; vacunacin antirrbica canina en el rea focal y se brind educacin sanitaria a la comunidad. Se abasteci con biolgicos antirrbicos a los servicios de salud, con limitaciones en la disponibilidad de suero heterlogo. En 3 canes y 2 gatos se encontr la VAg 1. No hay casos de rabia humana. Conclusiones Se reporta por primera vez la reintroduccin de rabia urbana en Lima, despus de 5 aos de silencio epidemiolgico, relacionado con coberturas de vacunacin antirrbica menores del 70%.

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SURVEILLANCE AND CONTROL OF ANIMAL RABIES

ESTRATEGIA PARA EL CONTROL DE LA RABIA DE LOS HERBVOROS EN EL ESTADO DE SO PAULO - BRASIL, 2000 A 2005
V.S. Nogueira 1, C.A. Gonalves 1, M.N. Gomes 1
1) Coordenadoria de Defesa Agropecuria, Campinas, SP , Brasil

Dr. Vladimir Nogueira - vnogueira@cda.sp.gov.br

Brasil

Introduccin: En el ao de 2000 el estado de So Paulo tuvo 773 propiedades con diagnstico positivo de Rabia, provocando prdidas econmicas y riesgo a la salud pblica. La rabia en herbvoros se transmite por murcilagos hematfagos. La vacunacin apenas evita que el animal contraiga la enfermedad y, aun as, esta puede suceder cuando el desafo viral superar la proteccin inmunolgica. Observando esa informacin la Coordinacin de Defensa Agropecuaria prepar un plan de trabajo con enfoque en el control del transmisor (Desmodus rotundus). Objetivos: La reduccin de la ocurrencia de Rabia de los herbvoros con enfoque en el control del transmisor en todo el estado, visando una convivencia de tal modo que el impacto econmico y el riesgo a la salud pblica sea lo menor posible. Mtodos: Se seleccionaron auxiliares tcnicos para el control poblacional de Desmodus rotundus y veterinarios para supervisar, controlar focos y suplir las necesidades de insumos. Esos funcionarios reciben equipamientos de seguridad individual (EPIs) para la captura de los murcilagos. Participan desde la captura hasta la planificacin de las actividades. Reciben tratamiento profilctico preexposicin, capacitaciones de actualizacin y realizan valoracin de anticuerpos anualmente. Para el control de Desmodus rotundus la metodologa utilizada fue mutiro. Los equipos actan en conjunto en los municipios de una regional en la bsqueda de animales agredidos por el Desmodus rotundus. Abrigos con vestigios de murcilagos hematfagos son georeferenciados y a travs de mtodos selectivos la poblacin de Desmodus rotundus es controlada. Cuando no se encuentran abrigos, la captura se realiza en la fuente de alimentacin. El inicio de ese trabajo se dio a partir del regional de Piracicaba, en el sentido contrario al desplazamiento de la Rabia. En el foco de rabia se hace control minucioso del transmisor en un rayo de hasta 12 Km., teniendo como fin eliminar colonias contaminadas, pues ocurrencias de Rabia confirmadas en laboratorio ponen de manifiesto la presencia del virus en el medio ambiente y la existencia de actividad viral en el transmisor. En las reas de riesgo la vacunacin es obligatoria. La participacin de la comunidad ha sido fundamental en ese proceso. Resultados: Se observ una reduccin significativa de 773 propiedades con animales con diagnstico positivo para la Rabia en 2000 para 536, 203, 119, 64 y 62 en 2001, 2002, 2003, 2004 y 2005, respectivamente. Conclusin: La estructuracin del Programa con enfoque en el control de la poblacin del transmisor, fue efectiva en la reducin de la Rabia de los herbvoros en el Estado de So Paulo.

128

SIXTH SESSION

CAPTURE AND CONTROL OF BATS ANF HERBIVOROUS VACCINATION DURING AN OUTBREAK OF HUMAN RABIES TRASMITTED BY VAMPIRE-BATS, MARANHO STATES, BRAZIL, 2005
Sonivalde Santana Monte 1, Valrio do Carmo Melo 1, Paulo Andr Sampaio Ferreira 1, Jos de Nazar Ramos de Sousa 1, Carlos Celso de Mendona Ramos 1, Roberto Carlos Negreiros de Arruda 2, Vladimir Nogueira Filho 3, Marcelo Yoshito Wada 4
1) Agncia Estadual de Defesa Agropecuria, MA, Brasil 2) Superintendncia Federal de Agricultura, MA, Brasil 3) Secretaria de Agricultura, SP , Brasil 4)Secretaria de Vigilncia em Sade, Ministrio da Sade, Brasil

Dr. Sonivalde Santana Monte - sonivalde@yahoo.com.br

Brasil

Introduction: Rabies is a zoonotic disease with a mortality rate of 100%. During 2005, an outbreaks of human rabies transmitted by vampire-bats occurred in four municipalities of Maranho state. Agriculture Department of Maranho State(AGED-MA) is the institution that is responsible to take actions as capture and control of vampirebats and vaccination of herbivorous which are essential for prevent human rabies cases. Methods: This is a descriptive study about actions of the AGED-MA to control rabies in Maranho during 2005. Bats were captured using a fog net around hen houses and cattle. They were classified in vampire-bat and non-hematophagous bat. Among vampire-bat, the specie Desmodus rotundus was treated with an anticoagulant substance to control population and others species were released. It was offered vaccination of herbivorous in the communities. Results: A total of 24 municipalities realized actions to control vampire-bats during the study period. Around 644 captures of bats were done, and 1,589 vampire-bats were captured and treated with the anticoagulant. Human rabies were identified in four municipalities (Godofredo Viana(GV), Carutapera(CA), Cndido Mendes(CM) and Turia(TU)). TU, GV and CA had the higher number of vampire-bats captured, 16.5%, 15.4% and 8.9% respectively. TU, Cedral and GV were the municipalities where had more captures realized, 11.3%; 10.9% and 9.1% respectively. Among vaccination of herbivores, 24 focus of rabies were identified, being 20 bovine, 1 equine, 1 pig, 1 ovine and 1 asinine. Over than 1,380,800 animals was vaccinated, mainly bovines. Conclusions: This study shows the importance of actions of capture and control of vampire-bats and vaccination of herbivores as one of the activities to prevent human rabies cases. All described actions need to be continued as a routine in municipal level. Education in health is also performed and others actions to complement prevention and control measures were taken by the Secretariat of Health.

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ESTIMATED DOGS POPULATION ANALYSIS TO PERFORM EPIDEMIOLOGICAL SURVEILLANCE AND PROCEDURES OF DOGS ANTIRABIES VACCINATION TO ENHANCE TASKS IN STATE OF SO PAULO, BRAZIL, DURING 1996 TO 2005
Maria de Lourdes A. B. Reichmann 1, Maria Helena Dib Amado 1, Vania de Ftima Plaza Nunes 2, Neide Yumie Takaoka 1
1) Instituto Pasteur, So Paulo, SP , Brasil 2) Prefeitura do Municpio de Jundia, SP , Brasil

Dr. Maria de Lourdes Aguiar Bonadia Reichmann - mreichmann@saude.sp.gov.br

Brasil

The mass vaccination of dogs is recommended by the World Health Organization, Pan-American Health Organization, Health Ministry and Health Office of the State of So Paulo. The results have been demonstrating efficacy since the beginning of the works in 1976 in the State of So Paulo. During 1996 to 2000 all the municipalities developed the systematic mass antirabies vaccination and it has been considered the rate estimative of 1 dog to 10 until 1 dog to 7 inhabitants, according the official recommendations. The evaluation of the periodic results has indicated the under estimation of the dogs population because the final values were over 100% of the estimated population. This situation has been clearer when the Pasteur Institute of the State of So Paulo had begun the official coordination of the program. Since 2000 it has been offer a new system to estimate the dogs population of each of the municipalities. The second step was the development of the measurement of the canine population in the interior cities of So Paulo State witch results in a 1 to 4 inhabitant proportion (2002). In the comparison of the two methods it was evidenced for some municipalities with less than 10 thousand inhabitants the number of 1 dog to 3 or to 1 person, when it was calculated the 80% ratio of dog population vaccination. Nevertheless some municipalities are presenting less than 80% ratio of dog population vaccination if it is considered the number that was defined in the research. These better populations estimative of each municipality have been used to improve a better Epidemiological Surveillance system of a lot of injuries, to evaluate the number of canine and feline samples direct to laboratory diagnostic and to analyze the ratio of antirabies vaccination.

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VIDEO PARA LA PREVENCION Y EL CONTROL DE LA RABIA PARALITICA BOVINA


M.D. Manzano-Martnez 1, J.A. Jimnez-Ramrez 2, J.A. Montao-Hirose 3, R. Ramrez-Hernndez 1, A. AlmanzaVzquez 4, M. Orozco-Islas 5, E. Meraz-Romero 6, A. De Vega-Garca 1,7
1) Comit de Fomento y Salud Animal del Estado de Puebla, 2) Campaa Nacional contra la Rabia Paraltica Bovina- SENASICA, 3) Instituto de Ciencias Agropecuarias - UAEH, 4) Eklipse Publicidad y Comunicacin, 5) Unin Ganadera Regional de la Sierra Oriente de Puebla, 6) Colegio de Posgraduados en Ciencias Agrcolas, 7) Unin Ganadera Regional del Norte de Puebla, Mexico

Dr. Mara Dolores Manzano Martinez - lolismanzano@yahoo.es

Mxico

INTRODUCCION Los Estados de Puebla, Hidalgo y Veracruz conforman una de las regiones de mayor incidencia de casos de Rabia Paraltica Bovina a nivel nacional, causando grandes prdidas econmicas y representando un riesgo de salud pblica y salud animal. La Rabia en los humanos, es una enfermedad que tradicionalmente se ha asociado a las clases econmicamente desfavorecidas, siendo en realidad el factor ms importante en su prevencin, la educacin de los individuos y no su posicin econmica o social. En este contexto, la informacin y capacitacin que reciban autoridades gubernamentales, productores, organizados e independientes, as como la poblacin en general, ser fundamental en la prevencin de esta enfermedad, en dos de sus presentaciones: en los humanos y en el ganado. Una excelente herramienta para efectuar una capacitacin y difusin de medidas preventivas y de control es el video, que hasta la fecha, existen pocos sobre el tema y en ocasiones son de dificil adquisicin. OBJETIVO Fomentar en la sociedad rural, en los ganaderos, autoridades y poblacin en general, una cultura de prevencin, a travs de la difusin y capacitacin, sobre los riesgos de rabia en humanos y casos de rabia en ganado en zonas de riesgo. As como poner a disposicin de la comunidad internacional un video sobre las medidas preventivas y de control de la Rabia Paraltica Bovina, dirigido a productores de Mxico y aplicable a Amrica Latina. METODO El video se realiz en los municipios endmicos de Rabia de la Sierra Norte del Estado de Puebla, donde las personas persisten con hbitos de contacto directo sin proteccin, con animales sospechosos y positivos a esta enfermedad. Se cont con el apoyo financiero de la Fundacin Produce Puebla A.C., agrupaciones ganaderas, profesionales en comunicacin agropecuaria y responsables de la Campaa de Rabia Paraltica, quienes a partir de un diagnstico oficial por parte de las autoridades sanitarias se dieron a la tarea de producir un video en formato DVD con la finalidad de promocionar y difundir las acciones a la poblacin. El video se estructur en dos partes. La primera, narra con un lenguaje propio de los ganaderos de la regin, el caso de un brote de rabia, y comprende cuatro secuencias audiovisuales:reporte del caso, atencin de foco rbico, seguimiento y medidas

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preventivas y la segunda, incluye cpsulas informativas sobre red de fro, vacunacin, toma y envo de muestras al laboratorio, clasificacin de murcilagos. CONCLUSIONES Realizacin de un video en formato DVD, con calidad digital profesional y con duracin de 50 minutos sobre las medidas preventivas y de control de Rabia Paraltica Bovina en Mxico, para la capacitacin de productores, tcnicos, autoridades y poblacin en general, aplicable a los pases de America Latina que padecen esta enfermedad.

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SEVENTH SESSION
STRATEGIES FOR THE CONTROL OF ANIMAL POPULATION

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134

SEVENTH SESSION

DINMICA DE LA POBLACIN CANINA CON DUEO EN REAS URBANAS Y RURALES DEL ESTADO DE MXICO, 1979-1999
S.L. Prez. 1, R.V. De la Rosa 2, A. Neri 2
1) Instituto de Salud del Estado de Mxico, Mexico 2) Facultad de Medicina Veterinaria y Zootecnia de la Universidad Autnoma del Estado de Mxico, Mexico

Dr.Luis S. Prez Sotelo - luis_07@yahoo.com.mx

Mxico

Introduccin: no se conoce la dinmica de poblacin canina en el Estado de Mxico, indispensable para el control de la Rabia. Objetivo: conocer la dinmica de la poblacin canina para la planeacin de la Campaa de vacunacin. Material y Mtodo: fue un proyecto de intervencin en salud, epidemiolgico descriptivo y prospectivo de 1997 a 1999, piloteando el censo-encuesta, aplicado en las 19 jurisdicciones y 122 municipios. Resultados: en 1997, participaron 13 jurisdicciones, se aplic el censo a 9,339 viviendas en 318 localidades, se registraron 59, 072 habitantes, 14, 336 caninos, 2,106 gatos, existiendo un canino por 4.12 habitantes y un cachorro por 3.25 viviendas. En 1998, participaron 9 jurisdicciones, se visitaron 34,235 viviendas y 940 localidades, se registraron 212,400 habitantes, 48,837 caninos, 8,004 gatos, con relacin de un canino por 4.35 habitantes y un cachorro por 4.97 viviendas. En 1999, participaron 13 jurisdicciones, se visitaron 50,558 viviendas en 1,310 localidades con 276,984 habitantes, 78,387 caninos, 12,840 gatos, la relacin es un perro por cada 3.53 habitantes y un cachorro por 4.90 viviendas. Conclusiones: se registraron 141,560 caninos, 22,050 gatos, 548,456 habitantes con 94,138 viviendas en 2,568 localidades la relacin fue un canino por cada 3.87 habitantes, se inmuniz contra la rabia a el 79.86% de caninos y 83.44% de gatos, las jurisdicciones con mayor numero de caninos son Texcoco, Nezahualcoyotl, Tlanepantla y Naucalpan. Sugerencias, continuar con la encuesta adoptndola en el programa respectivo, revacunar cachorros menores de 3 meses y controlar el crecimiento de la poblacin canina por medios qumicos. Palabras clave: Poblacin, Canina, Dinmica.

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MODELO DE PROSPECCIN DE POBLACIN CANINA PARA LA PLANIFICACIN DE ACCIONES SANITARIAS


A. Alfieri 1, J. Arango, A. Seghesso, A. Lapalma, L. Bel, B. Perez
1) Universidad Nacional de Rosrio, Santa F, Argentina

Dr. Arsenio Alfieri - jumagu@arnet.com.ar

Argentina

Los servicios de control de zoonosis urbanas que incluyen programas de rabia, enfrentan la toma de decisiones en la asignacin de recursos generalmente escasos, confrontando la investigacin del problema en su totalidad o la cobertura de las demandas sociales sentidas respecto a su seguridad frente a esta enfermedad. La opcin en los niveles de decisin poltica tiene slo una respuesta, satisfacer la demanda social. La clave pasa entonces por la estimacin del tamao y estructura de la poblacin bajo programa y el desafo es el ahorro de recursos obviando la reiteracin anual de costosos censos o la implementacin de registros de baja aceptacin y cumplimiento por la poblacin. A partir de una encuesta por muestreo aleatorio de poblacin canina urbana, se presenta un modelo prospectivo para la estimacin del tamao poblacional. Su construccin se bas en el modelo terico utilizado por el Instituto Nacional de Estadsticas y Censos (INDEC) para la prospeccin de poblaciones humanas. Los indicadores utilizados fueron: natalidad, mortalidad postneonatal, mortalidad general en mayores de 10 das, proporcin de hembras en la poblacin, porcentaje de hembras frtiles, porcentaje de hembras sin cuidado que evite la concepcin, proporcin de hembras que paren y cachorros nacidos en una camada anual. El modelo implica la decisin de incluir en la encuesta las variables necesarias para continuar con la prospeccin de la poblacin a travs de un modelo matemtico que slo demanda la observacin de su vigencia por muestreos poblacionales peridicos y de bajo costo.

136

SEVENTH SESSION

RESULTADOS DEL PROYECTO MASIVO DE ESTERILIZACIN QUMICA CON GLUCONATO DE ZINC NEUTRALIZADO CON ARGININA EN PERROS MACHOS RESIDENTES DE LOS ESTADOS DE: HIDALGO, MXICO Y PUEBLA
S. Vernica Gutirrez Cedillo 1, Fernando Vargas Pino 2, Oscar Velzquez Monroy 3, Juan Manuel Balderas Torres 4, Salvador Alanis Valdez 5, Erick Canales Vargas 1, 2, 3, 5
1) Centro Nacional de Vigilancia Epidemiolgica y Control de Enfermedades, Mexico 2) Secretaria de Salud del Mxico, Mexico 3) SESA Puebla, Mexico 4) SESA Edo. Mxico, 5) SESA Hidalgo, Mexico

Dr. Fernando Vargas Pino - zoonosis@podernet.com.mx

Mxico

Introduccin: El vnculo del perro con el hombre, y por extensin con su familia es ancestral, de ah lo complejo que resulta intentar disociarlo con actividades que terminen con su exterminio. En la actualidad, la Secretara de Salud de Mxico aplica ms de 16 millones de dosis de vacuna antirrbica canina, lo cual es mayor al universo de nios protegidos en el Programa Nacional de Inmunizaciones. La abundancia de perros en la calle representa un reto para la salud pblica y para la poblacin en su conjunto, ya no es posible convivir con este exceso, se reportan 106,233 agresiones por perro y de stas 31,844 inician tratamiento antirrbico, con un gasto slo de biolgicos antirrbicos humanos de $2,856,067 US Dlls. En el ao 2001 inicia la estrategia estabilizacin de la poblacin canina que considera la esterilizacin de los perros y gatos y la donacin voluntaria de animales no deseados. El Proyecto de esterilizacin masiva y gratuita emplea tcnicas quirrgicas simplificadas y el uso de anestsicos de ultra corta duracin. Los rendimientos acumulados de animales esterilizados del 2001 al 2005 fueron de 518 mil cirugas, por sexo corresponde al 85% de hembras (440.3 mil) y de machos el 15% (77.7 mil). Conforme al Modelo de estimacin de cachorros que ya no se incorporan a la poblacin canina como resultado de la esterilizacin, tenemos que por cada hembra esterilizada dejan de nacer menos cachorros, que por cada macho esterilizado; por lo que se busc con la poblacin propietaria de perros fomentar la esterilizacin de los machos, mediante mtodos qumicos que no extirparan las gnadas, de fcil aplicacin y de manera masiva, lo cual se hizo posible con el uso del Gluconato de zinc, neutralizado con Arginina (GZA) de manara intratesticular. Objetivos: La importancia de este proyecto fue el corroborar que el GZA produce esterilidad en perros de diversas edades y establecer una metodologa para asegurar la adecuada aplicacin. El universo de estudio fue de 9 mil perros (3 mil por cada estado), de 3 a 36 meses de edad. Los criterios principales de exclusin utilizados: criptorqudeos, testculos menor a 10 mm o mayor a 27 mm, dermatitis y ulceras testicular, y Tumor Venreo Transmisible. Resultados. Para corroborar el porcentaje de fertilidad en los perros machos, se selecciono una muestra de 217,

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aplicando una prueba de espermatobioscopia con resultados de 34.6% (75 perros) de fertilidad. Se han esterilizado a 7,713 perros: Puebla, 3,000; Mxico 3,663 e Hidalgo 1,060. La distribucin total por edad fue: 60.4% de 2 a 3 aos, 24.5% de 1 a 2 aos, 10.3% de 6 a 12 meses y el 4.8% de 3 a 6 meses. Observamos que el 98.8% estaban vacunados contra la rabia. En cuanto a las reacciones indeseables (fstula o lcera) presentadas pos aplicacin, slo en el 3.1% (239 perros) de lo animales ocurri, ligeramente arriba de lo esperado (3%), de stos se castr a 90 perros (37.7%); la distribucin de los grupos de edad ms afectados fueron el de mayores a 24 meses con el 75.9% y de acuerdo al tiempo en presentarse esa reaccin, la mayora se concentr en las 48 hrs. con el 77.7% (179 perros). Se realiz un seguimiento durante 6 meses en 50 perros, evalundolos por pruebas de espermatobioscopia, encontrndose: 42 perros presentaron azoospermia, 10 aspermia y 1 oligospermia; y mediacin del dimetro testicular obteniendo en promedio una disminucin del 75.7% . Conclusin: El GZA, produce esterilidad en los machos sin modificar su comportamiento y se requiere una adecuada aplicacin por personal veterinario capacitado.

138

SEVENTH SESSION

THE IMPACT OF THE PROGRAM OF AMBIENT HEALTH IN THE CONTROL OF THE URBAN RABIES IN THE CITY OF RECIFE
Sandra Souto Arajo 1, Geraldo Andrade Filho 1, Joo A. Nascimento Jnior 1
1) Secretaria Municipal de Sade, Recife, PE, Brasil

Dr. Sandra Souto de Arajo - soutoaraujo@gmail.com

Brasil

The City of Recife is located on the coast of Pernambuco, in the Northeast region of Brazil. It has an extension of 209 Km and an estimated population of 1.546.502 inhabitants. The epidemiological situation of the animal rabies in Recife is historically endemic. Before the implantation of the Environmental Health Program (EHP), the confrontation of this zoonose consisted basically of the capture of dogs and cats, annual vaccination of the canine and feline population and sending of harvested samples to laboratories. The most significant reduction on the cases of rabies occurred between the years 2002 and 2003, first year of the of implantation of the EHP , whose purpose was to control the typical diseases of developing metropolis, such as animal rabies, filariose, leptospirose, dengue fever, etc., trying, through integrated actions between the City Department of Health and the organized Civil Society, to identify problems and point out solutions to improve the quality of life and health conditions of the population of Recife. The aim of this study is to point out the strategies used in the EHP for the confrontation of this important zoonose. With the implantation of the program, the actions previously taken to control the rabies were decentralized, and now the six Sanitary Districts - SD are in charge of monitoring and directing these actions, with the technical support of the Center of Environmental Monitoring. The animal vaccination campaign started to happen every six months, there was an increasing on the fleet of cars to take the animals from the streets, going from 3 cars to 7, covering the central area and the six SDs, more qualification for Environmental Health Agents, increasing and standardizing of the sending of samples to laboratories, valuation of the ownership by the Environmental Health Agents in the community. After the implantation of the EHP , an increasing on the vaccination coverage was observed with the introduction of the after-campaign carried by EHA, the increment of the sending of samples, made it possible the absence of sub-notification of the cases, as it was happening in previous years, considerable reduction of nomadic dogs in the streets of Recife and, consequently, the reduction of the cases that showed the following evolution in the years: 10 cases in 2000; 25 cases in 2001; 43 cases in 2002; 12 cases in 2003; 2 cases in 2004; and no cases of animal rabies since May of 2004 in Recife. We conclude that the strategy used by the EHP , besides efficient, diminished significantly the period of the current stage of controlling of this zoonose, showing a correct strategy of action in areas of increased incidence of this problem.

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POSTER I

POSTER I

142

POSTER I

ESTUDIO EPIDEMIOLGICO DE LA ENCEFALITIS RBICA HUMANA EN EL ESTADO DE MXICO, 1979- 2003


S.L. Prez 1, M.C. Gutirrez 2, G.G. Soto 2
1) Instituto de Salud del Estado de Mxico, 2) Facultad de Medicina Veterinaria y Zootecnia de la Universidad Autnoma del Estado de Mxico

Dr. Luis S. Prez Sotelo - luis_msp07@yahoo.com.mx

Mxico

Introduccin: Mxico, ha ocupado uno de los principales lugares en rabia en las Amricas. Objetivo: Analizar retrospectivamente el comportamiento de la enfermedad en casos presentados de 1979 a 2003 en el Estado de Mxico. Material y Mtodo: estudio epidemiolgico retrospectivo, exploratorio y descriptivo, considerando: estudio epidemiolgico del agredido y del animal agresor, se dividi la informacin en cuatro perodos. Resultados: en el primero perodo, se detectaron 40 casos, 26 hombres y 14 mujeres, tasa de mortalidad de 0. 13 25 por 100, 000 habitantes, tendencia de 0. 23 a 0.0 29, el segundo registr 4 2, 28 hombres y 14 mujeres, tasa de mortalidad de 0.08 y tendencia de 0.078 a 0.08 2, el tercero present 48 casos, 39 hombres y 9 mujeres, tasa de mortalidad de 0.047, tendencia de 0.08 1 a 0.0 1 2, y el cuarto registr 3 casos, todos hombres, tasa de mortalidad de 0.005 por 100, 000 y tendencia de 0.0 1 a 0.00. De 133 casos referidos, la mayora varones que habitaban en medio rural y semirural, del grupo etreo escolar y clase econmica 1. Respecto al animal agresor 87.97% fueron caninos, los cuales 45.86% tenan dueo, trasmitiendo la rabia por mordedura en 85.7 1% de los casos, generalmente en miembros superiores 27.07%. La signologa humana fue: principalmente nerviosa, presentndose la muerte en menos de cinco das. Los municipios ms afectados rodean al Distrito Federal. Conclusiones: La clase ms afectada es de bajos recursos, en los primeros perodos las personas murieron por ignorancia, y en los ltimos por desconocimiento mdico de la normatividad vigente. Palabras clave: Encefalitis Rabica Humana

143

POSTER I

RELATO DE CASO: AVALIACIN DE LOS TRATAMIENTOS ANTIRBICOS EN PERSONAS QUE TUBIERON CONTACTO CON BOVINO POSITIVO PARA RABIA
S.P .H. Rivero 1, E.P . Caldas 2, N. Sanberg, A. Rocha, E. Boff, E. Cemin, R. Cavagnoli
1) Secretaria Estadual de Sade do Rio Grande do Sul, RS, Brasil, 2) Secretaria Estadual de Sade, RS, Brasil

Dr. Patricio Rivero - segundo-hoyos@saude.rs.gov.br / riverovet@terra.com.br

Brasil

Introducin: La rabia es una enfermedad infecciosa aguda causada por un virus del genero lyssavirus, familia Rhabdoviridae, que ataca mamiferos, incluyendo al hombre, con 100% de letalidad.En el Brasil los principales reservatorios son el perro y los murcielagos hematofagos y no hematofagos. Otras especies atacadas son: bovinos, equinos, suideos, monos.Entre los aos de 1986 a 2005 son noticiados 755 casos de rabia humana en el Brasil, principalmente causado por ataques de murcielagos hematofagos e perros.-OBJETIVOS:Informar nuestra experiencia frente a la necesidad de aplicar tratamientos antirbicos y su evaluacion de personas que tubieron contacto con bovino con rabia, confirmado laboratorialmente.Informar las acciones interdisciplinares y multiinstitucionales desarrolladas por las Secretaria Estadual de la Salud del RS(5CRS/SES/Caxias do Sul), Secretaria Municipal de la Salud (SMS)y Educacin de Flores da Cunha(RS) y la Inspectoria Veterinaria y Zootecnia (IVZ/SSA/RS)local. METODOS: Colectar el material encefalico de bovidos con suspecha de Rabia (0 2 amostas) por medico veterinario de IVZ/SSA/RS y enviado al Laboratorio de Referencia IPVDF/FEPAGRO/SCT, para a realizacion de la prueba de Imunoflorescencia Directa.Los resultados obtenidos fueron informados a los tecnicos de la 5CRS, SMS y IVZ/ SSA/RS.A seguir fue realizado un levantamiento del numero de personas expuestas, la aplicacin de la vacuna antirbica humana, inicialmente FUENZALIDA&PALACIOS(F&P) y posteriormente vacuna de cultivo en clula diploide humana y suero antirbico heterlogo, seguinda la Norma Tcnica Operacional/Rabia/ 1997. Despues fue colectado muestras de suero de 27 personas sometidas a tratamiento para evaluar la respuesta imunologica satisfactoria (titulo adecuado> 1: 25) en teste de sueroneutralizacion em raton mus musculus(SN-cdg).La aplicacion del suero antirbico fue realizado en ambiente hospitalar bajo supervision mdica (Hospital Geral de Caxias do Sul). RESULTADOS: 27 personas recibieron tratamiento de vacunas y/o suero antirabico heterologo.Fueron utilizadas 19 2 dosis de vacunas F&P y 2 1 dosis de vacuna diploide humana, mas 19 pacientes recibieron suero antirbico heterlogo.Ocurrio un caso de reaccin adversa blanda (edema, eritema, prurido y rubor)a la vacuna F&P .La vacuna diploide humana fue empleada como refuerzo vacunal de las personas con respuestas imunologicas bajas(titulo adecuado< 1: 25) frente a la vacuna F&P y para complementar en caso de reaccion adversa a la misma.Inicialmente en el 1teste 18 personas, de las 27, fueron realizadas a evaluacion de la respuesta imunologica

144

POSTER I

(SN-cdg), de las cuales 09 no respondieron adecuadamente(titulo adecuado< 1: 25).De estas 09 personas con inadecuada respuesta imunologica, 06 tubieron titulos de anticuerpos adecuados en un 2teste.De las 09 personas con respuestas adecuadas(titulo adecuado> 1: 25) 03 personas tubieron, en un 2 teste, titulos de anticuerpos protectores bajos, talves debido a la interferencia de las vacunas y/o suero antirabico aplicados. Y de las 09 personas restantes tubieron sus titulos de anticuerpos protectores adecuados atraves de SN-cdg.CONCLUSIONES: La pesquisa de los titulos de anticuerpos protectores contra la rabia, nos ofrece parametros y garantia de la correcta supervision de las personas que tubieron contacto con animales con confirmacion laboratorial de rabia.Esta antropozoonosis debe ser trabajada sobre el punto de vista de diversas profesionales de salud y integracion de reparticiones municipales y estaduales de salud publica/animal, de educacion, entre otras, considerando las medidas preventivas y de intervencion.

145

POSTER I

EPIDEMIOLOGICS ASPECTS ABOUT HUMAN RABIES IN PERNAMBUCO


F.D.F. Bezerra 1, M.L.A.N. Ribeiro 1, A.P . Lobo 1, D.M. Ferreira 1
1) Secretaria de Estado da Sade de Pernambuco, PE, Brasil

Dr. Francisco Duarte - dfbezerra@hotmail.com

Brasil

The Pernambuco State is located on east center of Brazil North-east region, with a population of 8.50 2.606 inhabitant, presents a territorial extension of 98.3 1 1.6 16 m 2, the predominate vegetation is the buster and the relief is formed by coast plain, central high plain beyond of the Pernambuco depression is divided in five mesoregions: Metropolitan, buster zone, rural, Pernambuco backwoods and San Francisco backwoods. Characterizing the epidemiologic aspects of human rabies in Pernambuco, in the period from 1975 to 2005, to help on for the proposition of central measure. Characterizing the cases of human rabies by demographics aspects, physiographic, clinical and epidemiologic on Pernambuco state. This project consists in a descriptive epidemiologic study of human rabies in Pernambuco state, in the period from 1975 to 2005, from the 193 confirmed cases and reported to Sanitary Monitoring of Pernambuco Diretory. The Pernambuco rural region presented the most number of cases (69 cases) in the period studied. The cases age changed from 1 to 80 years old. In relation to origin of cases, registered predominance in urban area with 5 2, 6% of cases. As for the sex noticed that 74, 5% of cases happened in the male sex. The dog was responsible for 84, 4% of cases. The period incubation varied from 7 to 5 1 1 days. The Pernambuco state presented a rediction in cases of human rabies on study period, this done could be impute to increase of vaccination covering on the last years. The rabies cases happened with more predominance on urban areas and the dog was the principal responsible for the aggressions. To keep the population informed through the Health Continuous System, emphating the necessity to seek the Health Service for starting the actions and utilizing the Family Health Program to follow the vaccination diagram, to avoid this way, the treatment abandonment.

146

POSTER I

EPIDEMIOLOGIC CONTROL PROGRAM OF RABIES IN PERNAMBUCO: EPIDEMIOLOGIC AND OPERATIONAL ASPECTS


M.L.A.N. Ribeiro 1, F.D.F. Bezerra 1, D.M. Ferreira 1
1) Secretaria de Estado da Sade de Pernambuco, PE, Brasil

Dr. Francisco Duarte - dfbezerra@hotmail.com

Brasil

Rabies is a zoonose which etiologic agent is a Rabdovirus, it is neurotropic, and it is 100% lethal. Although it may affect any animal species, it attacks specially domestic carnivores. Dogs are the most affected group. Even though the occurrence of rabies in dogs (main transmitters of rabies to human beings) has been reduced, the disease still represents a problem to public health. The objective of this project is to study the status of the Canine Rabies Control Program in Pernambuco from 2000 to 2005, as well as identify the occurrence of the rabies virus in the 184 towns that compose the State. The project also intends to identify the towns that have met the requirement set by the Programao Pactuada Integrada (PPI) of providing samples to the diagnostic lab as well as evaluate through the Regional Health Management (GERES) the percentage of treated people and the rate of abandonment of the treatment for the disease. It is a descriptive study, with theoretical investigation and analysis. The data came from the Investigation Files on epidemiological monitoring of the National Control Program Against Rabies, Guide of Epidemiological Monitoring, Monthly report of positive and negative samples from the National Cattle breeding Agrarian Lab of Pernambuco (LANAGRO) and Goals of the Programao Pactuada Integrada - patent 1 17 2 (PPI). In Pernambuco, there was a notable reduction of the prevalence of positive cases as a result of the creation of strategies during the implementation of an epidemiological monitoring for this disease. The prophylactic treatment is used as a tool to avoid death of those infected by rabid animals. It is important to maintain a large supply of vaccine in order to avoid the occurrence of new cases

147

POSTER I

DIFFICULTIES FACED FOR PROFESSIONALS TO PERSUADE PERSONS TO REALIZE PROPHYLAXIS FOR RABIES
Vnia Maria Alves de Carvalho 1, Romualdo Oliveira Spndola 1, Rosngela Cavalcante de Carvalho 1, Joo Pereira da Silva 1, Romildo Francisco dos Santos 1, Marcelo Yoshito Wada 2
1) Centro de Controle de Zoonoses, Teresina, PI, Brasil, 2) Secretaria de Vigilncia em Sade, Ministrio da Sade, DF, Brasil

Dr. Vnia Carvalho - vaniacarvalho58@yahoo.com.br

Brasil

Introduction: Rabies is a viral zoonosis with a mortality rate 100%, but it could preventable through prophylaxis. The last human rabies case of Teresina municipality, Piau State was identified in 1989. This study was performed to report difficulties faced for professionals to persuade persons to realize pos-exposure-prophylaxis (PEP). Methods: Data were collected from a standard form and reports of health professionals from May to June, 2006. Regularly active search was performed to guarantee complete treatment prophylaxis. Results: During an investigation of rabies in a dog transmitted by a non-vampire-bat occurred in urban area, four months of age, domiciliated and non vaccinated, there were identified sixteen persons that had any kind of contact with the positive animal, which two had pre-exposure-prophylaxis. All of them were oriented to look for assistance and to realize PEP . From sixteen patients, ten had been taken to realize prophylaxis on the hospital for the Health Assistance, saying that they had difficulties with transport, three persons looked for assistance alone and three refused prophylaxis, saying that they hadnt any kind of contact with the animal. Conclusions: There is only one way to prevent rabies pos exposure. Persons need to be oriented for immediately PEP . It is important to decentralize vaccine for rabies, facilitating access for population. Active search and education health is essential in cases of refuses and abandonment of prophylaxis and an organized system of surveillance is needed involving animal and human health. This investigation shows the importance to vaccine domestic animal for rabies and cares that are necessary to avoid contact with others animals, mainly wild animals as bats.

148

POSTER I

IMMUNOPATHOLOGIC IN RABIES INFECTION IN LINES OF MICE SELECTED FOR HIGH OR LOW ACUTE INFLAMMATORY REACTION
Samira M. Achkar 1, Idrcio L. Sinhorini 2, Orlando G. Ribeiro 3, Maria Luiza Carrieri 1, Rodrigo S. Ceretta 1, Cleide A. Consales 1
1) Instituto Pasteur, So Paulo, SP , Brasil, 2) Faculdade de Medicina Veterinria e Zootecnia, USP , So Paulo, SP , Brasil, 3) Instituto Butantan, So Paulo, SP , Brasil

Dr. Samira Achkar - samira_achkar@yahoo.com.br

Brasil

Rabies infection is a severe and lethal disease that presents a slight inflammatory response during the infection process. We analyzed the immunopathological mechanisms that occur in the CNS (Central Nervous System) using two lines of mice genetically selected for maximal (AIRmax) or minimal (AIRmin) acute inflammatory reactivity. As viral samples, we adopted the antigenic variant 3 (AgV3) rabies virus, from hematophagous bats and a fixed virus strain (PV 1 43/3). Titration of specific antibodies was performed by Enzyme Immunosorbent Assay. We observed a slight increase in IgG and IgG 1 isotype in the infected AIRmax mice. The incubation period, determined by intracerebral inoculation with 100LD50 was 6-7 days for PV 1 43/4 and 9- 10 days for AgV3. No difference in viral replication was found between AIRmax and AIRmin mice. Mortality was 100% with both viral strains. Histopathological analysis of the brain and spinal cord tissues showed inflammatory foci in all CNS, however, no differences were observed in the number of neutrophils. Negri bodies were also observed in practically all sites analyzed. Taken together, our results suggest that the inflammatory reaction is not a determining factor in susceptibility to rabies infection.

149

POSTER I

ASPECTS OF RABIES PROPHYLAXIS, TERESINA MUNICIPALITY, PIAU STATE, 2005


Fernando Luiz Lima de Oliveira 1, Carla Edlane Bezerra Sales, Jos Luciano F . H. Acioli Lins
1) Gerncia de Controle de Zoonoses, Teresina, PI, Brasil

Dr. Fernando Oliveira - fernandovet@gmail.com

Brasil

This study describes some aspects of rabies prophylaxis, frequently found in Teresina municipality, from October to December, 2005, that it is not present in the form of the National Reporting System of the Brazilian Ministry of Health and the goal was to subsidize control of the rabies in Teresina. There were applied standard questionnaire for all persons who looked for rabies prophylaxis on the reference hospital. A total of 653 cases were identified, being 65.6% adults and 54.7% males. Dog was responsible for 545 accidents (83.5%) and 87% were male. The residence of the owner was the place where occurred 64% of the accidents. Spatial distribution of the cases occurred in three regions of Teresina, 3 1.7% in north, 33, 7% in southeastern and 34.6% in south. In conclusion, this study demonstrate the importance to have health education, mainly for the ages with more occurrence, alerting about the behavior of the animals and how to manipulate them including all regions of the city.

150

POSTER I

IMPORTANCE OF PREEXPOSITION PROPHYLAXIS OF RABIES IN HIGH-RISK GROUPS


Vinicius Oliveira Drummond 1, Larissa Campos Aquino
1) Universidade de Braslia, Braslia, DF, Brasil

Dr. Vinicius Drummond - vinaforadecasa@yahoo.com.br

Brasil

Rabies is a zoonosis caused by a virus from genus Lyssavirus, family Rhabdoviridae that may be prevented by vaccination. The transmission is caused by the infected saliva of the illness animal. According to World Health Organization (WHO), 55, 000 people die annually, all around the world, from rabies. Rabies high-risk group includes veterinarians, veterinary students and staff, rabies laboratory technicians and wildlife officers. These professionals are more exposed to rabies virus, being able to get infected, if they dont have an adequate preexposure prophylaxis. The WHO recommends three doses of human anti-rabies vaccine, at day 0, 7 and 28, having a serum sample tested for antibodies every 6 or 1 2 months, depending on the virus exposure level, receiving a booster vaccine when the titer is less than 0.5 I.U. /ml. The preexposure prophylaxis doesnt eliminate the postexposure treatment after exposure to the virus, but simplify the therapy eliminating the need of rabies serum, and diminishing the number of vaccine doses, minimizing adverse reactions to multiple doses. Moreover, enhances immunity in cases of delay on postexposure treatment, and promotes protection to people with innaparent exposure to rabies. Human rabies still is a zoonosis with great epidemiologic importance because of high lethality (100%). Recently, Ministry of Healt reported a case of a Brazilian veterinary that doesnt had the preexposure prophilaxis, had contact with a diagnosticated sick animal on his duty, and even so, do not receive the treatment postexposure. This veterinary died. Based on that, the veterinarian, as public health professional, must have the preexposure prophylaxis and give orientation to his employees to do the same, remembering that many of them dont have any knowledge about the disease, and its implications on public health.

151

POSTER I

DOGS BITE IN A VETERINARY STAFF ON VETERINARY HOSPITAL OF UNIVERSITY OF BRASILIA (UnB) - CASE REPORT
Vinicius Oliveira Drummond 1, Larissa Campos Aquino
1) Universidade de Braslia, Braslia, DF, Brasil

Dr. Vinicius Drummond - vinaforadecasa@yahoo.com.br

Brasil

Rabies is an antropozoonosis caused by a RNA-virus, whose transmission is almost nearly caused by the bite of an infected mammal, who will inject virus by the saliva, causing encephalitis, and other clinical signs that havent cure, leading to death. According to literature, children less than 15 years old are the most bitten, but professionals like veterinarians and veterinary staff, are susceptible to be attacked too, having contact with the rabies virus. The objective of this study is to report an aggression case to veterinary staff in Veterinary Hospital of University of Brasilia (UnB). On 2006, July 5th, a dog, Rottweiller, came to Veterinary Hospital of University of Brasilia to radiographic exams. Then, it was taken to his owners car. The staff helped to carry the dog, holding his head, when the dog get scared and bite the staffs face. He was led immediately to Regional Hospital of Asa Norte (HRAN), where the wound was cleaned. Then, was noted that the left side of the nose was ripped and scratched. Following instructions from Ministry of Health, based on the localization of the bite and the fact that the functionary doesnt had the rabies preexposure prophylaxis, he received one dose of anti-rabies vaccine on day 0, and one dose on day 2. Even the animal wasnt suspicious to have rabies, it was placed in observation for 10 days, and was clinically health. Then the postexposure treatment was suspended. The staff remained in the HRAN until the plastic surgery. After seven days of rest and antibiotic therapy, he returned to his duty, normally. This report demonstrate the importance of the preexposure prophilaxy to veterinarians and staff, to avoid serious complications about the welfare of those who lead daily with animals that can transmit diseases to men, even fatal ones.

152

POSTER I

ECOLOGY AND MANAGEMENT OF VAMPIRE BAT ATTACKS UPON HUMAN BEINGS IN THE POOR VILLAGES FROM THE AMAZONAS STATE, NORTHERN BRAZIL
J.C.L. Vitor 1, W. Uieda 2, A.C.R. Campos 1, L.M. Oliveira 1, S.N. Martins 1
1) Gerncia de Controle de Zoonoses, Secretaria Estadual de Sade do Amazonas, AM, Brasil, 2) Universidade Estadual Paulista, Botucatu, SP , Brasil

Dr. Wilson Uieda - wilson.uieda@funasa.gov.br

Brasil

Desmodus rotundus is a common vampire bat in most regions of Brazil, where the cattle is its main food source. On the other hand, in the north region this bat species is not frequent, specially in Amazonas state, where the cattle raising is still incipient. In the Amazonian region D. rotundus has been observed feeding mainly on chickens, dogs and humans. In 200 2, the Gerncia de Controle de Zoonoses of the Amazonas state started a field study and control of aggressions by D. rotundus on humans. Untill 2006, 20 municipalities were visited, of which 17 had cases of bled people (Autazes, Barcelos, Codajs, Juta, Tapau, Santa Isabel do Rio Negro, Manaus, Novo Airo, Novo Aripuan, Presidente Figueiredo, Apu, Envira, Humait, Manaquiri, Rio Preto da Eva, So Gabriel da Cachoeira and Boca do Acre) and only three did not (Manicor, Eurinep and Careiro do Castanho). In three municipalities (Tapau, Barcelos and Santa Isabel do Rio Negro) the vampire victims have been found in several villages located at the edges of Purus and Negro rivers. In 44 villages, we recorded an average of 1 1, 3 (N=495) victims, mainly children. The vampire bat bites on human body were observed most frequently on toes, but also on scalp, fronthead, nose, hands and legs. Wooden houses with many free spaces and gaps, could favor the incoming of vampires but oil-lamps and mosquito nets have protected the sleeping victims against vampire bat attacks. The geographic isolation and hard access have been the great obstacles to vampire bat control and efficient anti-rabic immunization of human populations in the most municipalities of Amazonas State. Between 2004 and 2005, 174 bats of 15 species (1 1 Phyllostomidae, 3 Molossidae and 1 Emballonuridae) were submitted to rabies exams at Lacen (Manaus, AM) and Instituto Evandro Chagas (Belm, PA) and all were negative.

153

POSTER I

EPIDEMIOLOGIC CONTROL PROGRAM OF RABIES IN PERNAMBUCO: EPIDEMIOLOGIC AND OPERATIONAL ASPECTS


M.L.A.N. Ribeiro 1, F.D.F. Bezerra 1, D.M. Ferreira 1
1) Secretaria de Estado da Sade de Pernambuco, PE, Brasil

Dr. Maria de Lourdes de Almeida Nunes Ribeiro - lourdinha.ribeiro@hotmail.com

Brasil

Rabies is a zoonose which etiologic agent is a Rabdovirus, it is neurotropic, and it is 100% lethal. Although it may affect any animal species, it attacks specially domestic carnivores. Dogs are the most affected group. Even though the occurrence of rabies in dogs (main transmitters of rabies to human beings) has been reduced, the disease still represents a problem to public health. The objective of this project is to study the status of the Canine Rabies Control Program in Pernambuco from 2000 to 2005, as well as identify the occurrence of the rabies virus in the 184 towns that compose the State. The project also intends to identify the towns that have met the requirement set by the Programao Pactuada Integrada (PPI) of providing samples to the diagnostic lab as well as evaluate through the Regional Health Management (GERES) the percentage of treated people and the rate of abandonment of the treatment for the disease. It is a descriptive study, with theoretical investigation and analysis. The data came from the Investigation Files on epidemiological monitoring of the National Control Program Against Rabies, Guide of Epidemiological Monitoring, Monthly report of positive and negative samples from the National Cattle breeding Agrarian Lab of Pernambuco (LANAGRO) and Goals of the Programao Pactuada Integrada - patent 1 17 2 (PPI). In Pernambuco, there was a notable reduction of the prevalence of positive cases as a result of the creation of strategies during the implementation of an epidemiological monitoring for this disease. The prophylactic treatment is used as a tool to avoid death of those infected by rabid animals. It is important to maintain a large supply of vaccine in order to avoid the occurrence of new cases.

154

POSTER I

ANLISIS COMPARATIVA DE LOS PROGRAMAS DE CONTROL Y PREVENCIN DE LA RABIA EN AMRICA LATINA


Gustavo Deslandes de Carvalho 1
1) Universidade Estcio de S, RJ, Brasil

Dr. Jos Carlos Souza - souzajcp@agricultura.gov.br

Brasil

Se realiz un anlisis de la evolucin epidemiolgica de la rabia en los pases de Amrica Latina en el perodo de 1996 al 2005, para verificar si hubo progreso en el control de la transmisin de la rabia canina y de la rabia silvestre para el hombre. A partir de los datos obtenidos en la Organizacin Pan-Americana de la Salud acerca de los procedimientos profilcticos de la rabia canina y humana, junto con los del control preventivo de la agresin de los murcilagos obtenidos en la Organizacin Mundial de la Salud, Secretaria de Vigilancia Epidemiolgica del Ministerio de Salud de Brasil y de la Secretaria de Salud de Mxico, fue posible identificar los puntos de riesgo para la ocurrencia de la rabia humana: tratamiento profilctico pos-exposicin, tasa de vacunacin canina, nmero de puestos de salud, nmero de dosis de vacunas, cantidad de sueros hiperinmunes disponibles para el tratamiento anti-rbico y nmero de muestras de tejidos cerebrales caninos enviadas para diagnstico laboratorial. De acuerdo con las informaciones sobre el nmero de casos de rabia transmitida por murcilagos hematfagos y perros, se ha observado pases que registraron bajo porcentaje en el tratamiento profilctico pos-exposicin y baja tasa de vacunacin canina. Cinco pases presentaron bajo nmero de puestos de salud y en nueve el nmero de estas unidades de tratamiento fueron muy expresivas. Se observ adems que algunas naciones fueron clasificadas en los padrones muy baja, baja, media y alta, en relacin al nmero de dosis de vacunas y mililitros de sueros hiperinmunes disponibles para tratamiento antirrbico completo. Tambin fue detectado que pases enviaron un bajo nmero de muestras de sistema nervioso de perros callejeros para diagnstico y monitoreo de la circulacin viral. El anlisis de los datos permiti concluir que hubo una reduccin significativa en los casos de rabia humana transmitida por perros y gatos en los pases latinos americanos en los ltimos diez aos aunque haya habido fallas en los programas nacionales de control de la rabia, recomendndose mejoras necesarias en relacin a los puntos de riesgo identificados con el objetivo de la erradicacin de la rabia. Palabras-clave: Rabia, epidemiologa, profilaxis, control.

155

POSTER I

DEVELOPMENT OF AN IMUNOPEROXIDASE INHIBITION ASSAY (IIA) FOR RABIES ANTIBODY DETECTION


H.B.C.R. Batista 1, E. Schmidt 1, J. Reis, F .K. Robin, T.F. Teixeira, D. Maletich, A.C. Franco, J.C.A. Rosa, P .M. Roehe 1
1) Universidade Federal do Rio Grande do Sul, RS, Brasil

Dr. Helena Batista - hruthner@yahoo.com.br

Brasil

In this study an imunoperoxidase inhibition assay (IIA) for detection of rabies antibodies in sera from different species antibodies to rabies viruses reported. The test does not require live animals or UV microscopy, what makes it cheap and easy to introduce in any diagnostic laboratory. Chick embryo related (CER) cell monolayers prepared in 96 well plates and infected Pasteur virus (PV). Seventy two hours post-infection the monolayers were frozen and fixed in 4 % paraformaldehide . Fixed monolayers were rinsed and incubated for 1 hour at 37 oC with test sera diluted (1: 2, 1:4, 1, 8, 1: 16, 1:3 2, 1:64 and 1: 1 28) . Next, plates were rinsed again in wash fluid (PBS-T80) and incubated under the same conditions with an appropriate dilution of anti-rabies nitially serum (prepared in rabbits). The last incubation was performed with an anti-rabbit/peroxidase conjugate. The IIA was nitially evaluated with 15 human sera, of which 6 were positive and 9 were negative for rabies antibodies by the mouse neutralization test. In adition, sera from 24 hematophagous bats of unknown rabies antibody status were also tested. The 6 antibody-positive and 9 antibody-negative has its results confirmed by the IIA. From the hematophagous bat sera, 2 1were negative and 3 were positive for antibodies by IIA. These results demonstrate that the IIA was able to detect anti-rabies antibodies in sera from humans and bats. Future steps in the standardization of the assay will require testing with a larger number of samples, including a known reference serum, as well as the determination of its correlation coefficient against standard serological tests.

156

POSTER I

RABIES VIRUS STABILITY AFTER TWENTY PASSAGES IN MICE


H.B.C.R. Batista 1, E. Schmidt, T.F . Teixeira, A.C. Franco, P .M. Roehe 1
1) Universidade Federal do Rio Grande do Sul, RS, Brasil

Dr. Helena Batista - hruthner@yahoo.com.br

Brasil

This work was carried out to examine the extent of the antigenic stability of three rabies virus (RV) isolates originated from distinct natural hosts after serial passage in laboratory mice. The antigenic profiles of the viruses were determined by monoclonal antibody (Mab) analysis against a panel of anti-lyssavirus antigens. Isolate RV 1 is a typical street dog virus, RV 2 was isolated from an insectivorous bat though it displayed an antigenic and genomic profile commonly found in hematophagous bats, RV3 was also isolated from an insectivorous bat and presented an antigenic profile usually found in viruses having insectivorous bats as natural host species. The three viruses were submitted to twenty intracerebral passages in 3-4 weeks old swiss albino mice. After each passage the isolates were again examined with the Mab panel in indirect immunofluorescence tests. RV 1 and RV 2 maintained the same antigenic profile throughout the twenty passages. However, RV3 revealed a modification in its antigenic profile at the eigth passage. These results indicate that the antigenic composition of RV is rather stable and is usually not affected by a number of passages in a host different from the natural host of origin. The limited antigenic modifications observed on one of the isolates (i.. RV3) after the eigth passage in mice might provide clues on what changes are necessary to lead to the adaptation of virus to a particular host species. Further studies are being carried out to examine the genomic profile of the passaged viruses.

157

POSTER I

A COLLABORATIVE STUDY ON THE TESTING OF EQUINE ANTIRABIES SERUM BY A VIRUS NEUTRALIZATION TEST IN BHK- 2 1 CELLS
Wlamir Corra de Moura 1, Neuza Maria Frazzatti Gallina 2, Regina Maria Mouro Fuches 2
1) INCQS-FIOCRUZ, RJ, Brasil, 2) Instituto Butantan, SP , Brasil

Dr. Wlamir Moura - wlamir.moura@incqs.fiocruz.br

Brasil

A collaborative study was realized to validate the virus neutralization test in cell culture to determine the potency of equine antirabies serum, by the Instituto Nacional de Controle da Qualidade em Sade (INCQS) of Brazil and the Instituto Butantan. The test utilized to determine the potency of the serum was the virus neutralization in BHK- 2 1 cells cultivated on 96 wells microplates. It was based on the recommendations of the fifth edition of the European Pharmacopoeia and the tests described by Trimarchi and Smith in Laboratory Techniques in Rabies. The medium effective dose (ED50) and the potency (IU/mL) were calculated by the probit method. The accuracy, intra-assay precision, inter-assay precision and reproducibility (inter-laboratory precision) were evaluated in two batches produced at Instituto Butantan and in the 2nd International Standard for Rabies Immunoglobulin (IR). The geometric mean of ED50 obtained with the IR at both laboratories (1/39, 39) was similar to that obtained by Trimarchi (ED 100= 1/3 2), demonstrating the accuracy of the test. The intra-assay precision was observed when one batch of equine antirabies serum was tested in 10 replicates in a single assay and it was found a coefficient of variation (%CV) of 8, 8%. The %CV of the ED50 of the batches tested presented variations of 1 2, 4- 19, 8% in the INCQS and of 13, 2- 2 2, 0% in the Instituto Butantan, showing the inter-assay precision. The %CV obtained with the two batches of equine antirabies serum and the IR tested at the two laboratories, between 1 2, 8% to 16, 3%, demonstrated the reproducibility. The study showed that the virus neutralization test in BHK- 2 1 cell is suitable as a potency test for equine antirabies serum, presenting accuracy, repeatability, inter-assay precision and reproducibility, validating the test in these laboratories. This method is an alternative to the mice virus neutralization potency test for equine antirabies serum realized actually in Brazil.

158

POSTER I

ANTIGENIC AND GENETIC CHARACTERIZATION OF FIRST RABIES VIRUS ISOLATED FROM EUMOPS PEROTIS BAT OF BRAZIL
J.G. Castilho 1, F.M. Canello 2, K.C. Scheffer 1, S.M. Achkar 1, M.L. Carrieri 1, I. Kotait 1
1) Instituto Pasteur, SP , Brasil, 2) Centro de Controle de Zoonoses de Ribeiro Preto, SP , Brasil

Dr. Juliana Galera Castilho - juliana.castilho@uol.com.br

Brasil

In Brazil, although the main transmitters of rabies are dogs and vampire bats, the role of others species, such as insectivorous and frugivorous bats, deserves special attention, once rabies virus has been isolated from 36 bats species. This study describes the first isolation of rabies virus from an insectivorous bat Eumops perotis, belonging to Molossidae family and Eumops genus, from which two species have already been found rabies positive: Eumops auripendulus and Eumops glaucinus. The specimen was found in an urban area in the city of Ribeiro Preto and identified according to morphological and morphometrical characteristics in Pasteur Institute of So Paulo. The isolation was carried out in N 2A cells culture and in adult mice, which only one animal died, on seventeenth day post inoculation. With this brain, a second passage was made and all mice inoculated died, presented an incubation period of 9 to 14 days. The sample was submitted to antigenic typing, using a panel of eight monoclonal antibodies (CDC/Atlanta/USA) and presenting a non-compatible result. The sequence analysis of DNA to the nucleoprotein gene located between nucleotides 10 2 and 1 284 was aligned with homologous sequences present in Genbank using the CLUSTAL/W method and Bioedit and Mega 2. 1 softwares. The phylogenetic analysis showed that the virus isolate segregated in the same cluster related with other virus isolated from insectivorous bat, genus Nyctinomops ssp, isolates from Brazil, showing high degree of nucleotide similarity (98, 8%) each other. This result shows the first isolation of rabies virus in this bat species.

159

POSTER I

RABIES VIRUS NEUTRALIZING ANTIBODIES: COMPARISON OF TWO EVALUATION TEST IN CELL CULTURE
L.B. Chaves 1, A.L.C. Mazutti 1, G.M.M. Caporale 1, K.C. Scheffer 1, A.C.R. Silva 1
1) Instituto Pasteur, So Paulo, SP , Brasil

Dr. Luciana Botelho Chaves - lbchaves@pasteur.saude.sp.gov.br

Brasil

The World Health Organization (WHO) recommends that anyone at increased risk of exposure to rabies virus should be submitted to pre-exposure immunization schedule and the evaluation of virus neutralizing antibodies (VNA) performed periodically using the rapid fluorescent focus inhibition test (RFFIT). People with serum titer e 0.5 IU/mL are considered immunized. Since 199 2, the laboratory of Instituto Pasteur - So Paulo has used the simplified fluorescent inhibition microtest (SFIMT) that was standardized and implanted there in substitution of indirect immunofluorescence for titration of rabies VNA. Similar to RFFIT, the SFIMT is a cell culture virus neutralizing test performed in microplates with the advantage of rapid reading. The aim of this study was compare these two tests and the use of Pasteur Virus (PV) and Challenge Virus Standard (CVS) on the evaluation of VNA in serum samples. Fifty human serum samples collected from persons that performed periodical evaluation of VNA titres were tested twice in different days by SFIMT and RFFIT using CVS and PV strains for both techniques. Considering the titres < 0.5 IU/mL as negative and e 0.5 IU/mL as positive, 3 2 samples (64%) were positive and 18 (36%) were negatives by RFFIT with both CVS and PV strain. By SFIMT, 33 (66%) samples were positive and 17 (34%) was negative for CVS and 34 positive (68%) and 16 negative (3 2%) for PV. This qualitative analysis showed a significant agreement among the methods with kappa > 0.9 (p< 0.05). By intra-class correlation test the quantitative analysis showed more than 80% of agreement (p < 0.0 1) between the methods. In conclusion, the results obtained by SFIMT are comparable those obtained with the RFFIT and the use of CVS or PV strain doesnt affect those results.

160

POSTER I

COMPARISON OF RFFIT PERFORMED IN LAB-TEK AND IN 96-WELL MICROTITRE PLATES


L.B. Chaves 1, A.L.C. Mazutti 1, G.M.M. Caporale 1, K.C. Scheffer 1, A.C.R. Silva 1
1) Instituto Pasteur, So Paulo, SP , Brasil

Dr. Luciana Botelho Chaves - lbchaves@pasteur.saude.sp.gov.br

Brasil

The rapid fluorescent focus inhibition test (RFFIT) became the test of choice in most modern laboratories for measuring virus-neutralizing antibodies to determine the potency of rabies serum and immunoglobulins for postexposure treatment, and to evaluate the immunogenicity of human or animal rabies vaccines. Although RFFIT has good specificity, sensitivity and reproducibility it has the disadvantage of no automated manipulation of microtitre plates. The use of 8-well Lab-Tek chamber slides limited the amount of serum samples tested. The aim of this study was verify if the adaptation of the classic RFFIT in Lab-Tek slides to the 96 well microtitre plates affected the specificity, sensibility and reprodutibility of the test. Twenty-four human serum samples were titrated twice by classic RFFIT in Lab-Tek and by RFFIT in 96 well microplate. For each serum dilution 50 or 18 microscopic fields, for Lab-tek and 96 well microplate respectively, in a single well were evaluated for the presence of infected cells at x 200 magnification. As results, in a qualitative analysis, considering titres < 0.50 IU/mL as negative and 0.50 IU/mL as positive, 15 samples (6 2%) were positive and 9 (38%) samples were negative in both tests. The measure of the kappa = 1.0 prove almost perfect agreement between the tests (p < 0.00 1). In the quantitative analysis, the value of the intra-class correlation is C > 0.98 (p < 0.00 1) that demonstrate a high agreement among the tests. In conclusion, the adaptation of the classic RFFIT to 96 well microplates is perfect satisfactory to performed the evaluation of virus neutralizing antibodies in human serum samples.

161

POSTER I

THE SO PAULO STATE/BRASIL RABIES CONTROL PROGRAM, UNDER THE VISION OF ITS COORDINATION (1996 - 2005)
Neide Yumie Takaoka 1, Maria de Lourdes A. B. Reichmann 1, Tereza Mitiko Omoto 1, Maria Luiza Carrieri 1, Ivanete Kotait 1
1) Instituto Pasteur, So Paulo, SP , Brasil

Dr. Neide Yumie Takaoka - pasteur@pasteur.saude.sp.gov.br

Brasil

The State Rabies Control Program Coordination has its head control at Instituto Pasteur/SP since 1996, analyzed the epidemiological disease situation as well as the other activities which compose the Program. The evidence was that the number of human cases during 1985 - 1997 maintained 0 - 2 cases/year, with the transmitting the canine specie. In relation to the Canine Vaccination Campaign, it was verified that many counties presented low vaccination covering and that the Program was not adequately developed. A wide training program was started in all state regional, aiming the canine and feline rabies control as well as the eradication of human rabies transmitted by dog, with the collaboration of health education personnel. In 1996, it was registered around 100 cases in dogs and cats, in 1997 it was reduced to in 90%. The last human rabies case transmitted by dog was in 1997. The last rabies case of companion animal (dog) by the canine variant occurred in 1998. Both cases occurred at the same region. The period of 1990 - 1997 had the register of more than 1000 lab diagnoses in companion species (94% dogs and 6% cats), in 5 2 counties, being, probably in its totality by canine variant. From 1998 to 2005, however, the number of labs diagnoses were around 35 cases, being its occurrence dispersed and caused by bat variants. Besides the features before mentioned, the program improvement contributed the implantation of laboratory tests to identify infeccion source and the change to in Vero cells culture vaccine. In conclusion, the State Rabies Control Program Coordination reached its goal: de cem para sem - zero cases of canine transmitted rabies in So Paulo, until now.

162

POSTER I

POLYMERASE CHAIN REACTION (RT-PCR) AND HEMINESTED-RT PCR (Hn RTPCR) EVALUATION IN FRESH AND DECOMPOSED CENTRAL NERVOUS SYSTEM SAMPLES FOR RABIES DIAGNOSIS
D.B. Arajo 1, J. Megid 1, R.C. Silva 1, M.F.A. Almeida 2, H. Langoni 1
1) Faculdade de Medicina Veterinria e Zootecnia , Botucatu, SP , Brasil, 2) Centro de Controle de Zoonoses , So Paulo, SP , Brasil

Dr. Daniele Bastos Arajo - daniellebastos@yahoo.com.br

Brasil

The use of sensitive and specific methods for rabies diagnosis is an important tool for the control and prophylaxis of the disease. Reverse-Transcriptase Polymerase Chain Reaction (RT-PCR) has been used in rabies diagnosis with good results, even in decomposed materials. In this study, we evaluated RT-PCR and Heminested RT-PCR (hnRTPCR) in 100 suspected brain samples from different animal species. The materials were evaluated in fresh and decomposed conditions (left at room temperature for 7 2 hours for decomposition). The RT-PCR and hnRT-PCR results were compared with previous results from Direct Fluorescent Antibody Test (FAT) and Mouse Inoculation Test (MIT). Were evaluated 100 samples, being 50 previously positive to FAT and MIT. Out of these positive samples 45 (90%) resulted positive to molecular techniques applied to fresh material, 18 (36%) to RT-PCR and 27 only to hnRT-PCR (54%). Only 98 samples were evaluated in decomposed conditions, 48 previously positive to FAT and MIT. When these decomposed materials were tested for the molecular techniques 29 (60, 4%) were positive, 13 (27%) to RT-PCR and another 16 (33, 3%) only to hnRT-PCR. No false positive results in molecular techniques were observed in fresh and decomposed materials from negative previously diagnosed samples. The results demonstrated higher sensitivity of hnRT-PCR compared with RT-PCR for fresh and decomposed materials but low sensitivity of both techniques in decomposed samples which could be explained by RNA viral denaturation due to tissue decomposition.

163

POSTER I

EVALUATION OF THE HUMAN ANTIRABIC SERVICE, MINAS GERAIS, BRAZIL, 1999 TO 2004
Valria de Melo Rodrigues e Oliveira 1, Cristiana Ferreira Jardim de Miranda 1, lvio Carlos Moreira 2, Pedro Lcio Litgh Pereira 2, Jos Ailton da Silva 2
1) Secretaria de Estado da Sade, Minas Gerais, MG, Brasil, 2) Universidade Federal de Minas Gerais, MG, Brasil

Dr. Cristiana Jardim - cristiana.jardim@saude.mg.gov.br

Brasil

The decrease of the cases of human rage in Brazil by canine transmission, shows that the evaluation of the human antirabic service becomes a need of the service, before the correlation lack among aggression, indication of the treatment and local epidemic situation. The objective of this work was to describe and to evaluate some aspects of the human antirabic service in Minas Gerais, from 1999 to 2004. Its took place a study retrospective descriptive observacional, correlating the risk areas for human rage transmitted by dog and the information registered in the several systems of information of SUS-MG. They were appraised 13 2.45 2 registrations of the National System of Offences of Notification - SINAN, and 339.0 1 2 registrations done by the service of Zoonoses of the Superintendency of Epidemiology of the General office of Health of Minas Gerais. It was verified that the treatment indication is excessive in the bass areas and medium risk, since most of the aggressions is made by healthy dogs, unlike the areas of high risk, where a decrease of the indication of the treatment was observed. The systems of information involved possess isolated bases, not acting as net. There is still so much for silent municipal district as for bad quality of the information. The surveillance of the event possesses aspect, however the interfaces dont communicate, generating insecurity to the responsible doctor for the service to just indicate the animal aggressors observation. Key words: information, rage, risk, human antirabic service.

164

POSTER I

HEALTH EDUCATION PROGRAM FOR RABIES, MARANHO STATE, BRAZIL, 2006


Sonivalde Santana Monte 1, Aldenir Paiva de Carvalho 1, Alan Arajo Nascimento 1, Margarida Paula Correa S1, Teresinha Lisieux C. dos Santos 1, Edilene Santos Silva1, Roberto Carlos Negreiros de Arruda 2, Marcelo Yoshito Wada 3
1) Agncia Estadual de Defesa Agropecuria do Maranho, MA, Brasil, 2) Superintendncia Federal de Agricultura do Maranho, MA, Brasil, 3) Secretaria de Vigilncia em Sade, Ministrio da Sade, DF, Brasil

Dr. Sonivalde Santana Monte - sonivalde@yahoo.com.br

Brasil

Introduction: Rabies is a zoonotic disease with a mortality rate of 100%. Health education is one of the most important activities to prevent diseases. The Agriculture Department of Maranho State(AGED-MA) have implemented a specific program for rabies to avoid occurrence of human and animal cases of rabies. Methods: This project is divided in two activities. The first one is realized in public schools, where theater parts are done, lectures are ministered and folders and magazines are distributed. The second one is named Field Day, where a specific locality is chosen, and activities of education are realized for farmers and professionals that care animals, after this, a practice class is done, including demonstrations of animal contention and vaccination of herbivorous and human prophylaxis. Drawings, certificates and homage are distributes at the end of the sessions. Results: Three municipalities: Morros(MO), So Joo dos Patos(SJP) and Rosrio(RO) were chosen to realize this program. A total of 39 1 children were trained, MO(33%), SJP(36%) and RO(3 1%). A total of 680 farmers and professionals realized the Field Day, MO(29%), SJP(43%) and RO(28%). In these actions are discussed rabies in herbivorous, dogs, cats, bats and wild animals and prophylaxis for human rabies. Conclusions: This program has increased tax vaccination in animals and demonstrated to be an important action to clarify farmers about rabies and to alert the necessity to vaccine animals and to realize prophylaxis in case of exposure to suspected rabid animal or bite, mainly about pre-exposure prophylaxis for professionals who care animals and farmers. This program has contributed a lot, and it is recommended to continue it in all municipalities. It has demonstrated the necessity to stand out an integrated action between Agriculture and Health Departments.

165

POSTER I

INFLUENCE OF DIFFERENT SERUM-FREE MEDIA IN THE RABIES VIRUS PRODUCTION


Sandra Harue Fujimura 1, Vanessa Harumi Takinami 1, Claudia Regina Menezes Botelho 1, Neuza Maria Frazatti Gallina 1
1) Instituto Butantan, So Paulo, SP , Brasil

Dr. Sandra H. Fujimura - sanhfujimura@yahoo.com.br

Brasil

Vero cell culture is an important substrate to rabies vaccine production. The fetal calf serum or human albumin utilized in culture media for cell growth can contain a prion contamination. The use of serum-free medium in this culture is a new strategy to obtain a high purity product. In this study the influence of three different serum-free medium in the rabies virus production were evaluated: VP-SFM AGT (GIBCO), Ex-Cell (Sigma) and HyQSFM4MegaVir (HyClone). Vero cells (1.6 x 106 cells/flask) were infected with PV rabies virus (MOI= 0.008) and maintained in serum-free media at 34C during 1 1 days. Samples of the supernatants of the cultures were taken each 24h to determine de rabies virus titers in BHK- 2 1 cells. The results obtained in the kinetic studies in rabies virus culture infected, maintained in VP-SFM AGT, Ex-Cell and HyQ SFM4MegaVir were 103.8_ 104.8, 10 2.0_ 104.6 and 10 2.6_ 103.7 FFD50/0, 05ml, respectively. The yield index (virus production x cells growth) obtained during all the experiment were similar in the cultures with VP-SFM AGT (6.08) and Ex-Cell (7.08) media. The values encountered with the HyQ SFM4MegaVir medium were lower than the others (0.48). The results showed that the VP-SFM AGT and Ex-Cell media are indicated to rabies virus production.

166

POSTER I

ANALYSIS OF THE LAST 16 YEARS (1990 - 2005) - POST EXPOSURE PROPHYLAXIS SO PAULO (BRASIL) STATE
T.M. Omoto 1, N.Y. Takaoka 1
1) Instituto Pasteur, So Paulo, SP , Brasil

Dr. Tereza Omoto - terezaomoto@hotmail.com

Brasil

After the analysis of the last 16 years which was divided into two periods (1990- 1997 and 1998- 2005) it was verified an increase of the number of patients who came to health services after a mammal injury. In the first period, the average number of patients was 60.000/year. During the second period it reached the double (1 20.000). Such increase was greater than the populational, possibly it occurred by the increase of consciousness of the population and as result of better awareness regarding the notification by the county services. Until 199 1, the files copies were sent to the central (state). By 199 2 it was developed a monthly bulletin, in which each county informed the number of patient seen, treated and other data related to Rabies Prophylaxis. The greatest number of register occurred between 1996 and 1998, after the rabies Control Program in the State of So Paulo came under the coordination of Instituto Pasteur of So Paulo (1996). At that period its register had rate 35.000/ 10.000 inhabitants. In 1999 occurred the implanting of SINAN (National System of Injury Notification).This is an informatized system containing individual data in which the files are available from local level to the federal level. The humans, after injury caused by mammal, represent the greatest volume of files to be digitized. Maybe by this reason, the numbers decreased. It might be caused by the lack of files registration, besides a worse information quality, with their reflexes on the increase of the percentage of cases where it does not appear the animal specie involved. In the present time, the number of registers is about 1 10- 1 20 thousand persons/year (30 persons / 10.000 inhabitants). Before 2000, the year when occurred the substitution of the anti-rabies human vaccine (Fuenzalida & Palcios to VERO Cell Culture) the proportion of patients treated was 50%, and it became 35%. In the beginning of 1990, the injuries caused by dogs represented 90% and in nowadays 85%, by cats it didnt change (7, 5 - 9%), by herbivorous it was 1% and reached 4, 5% in years of bovine rabies epidemic. Injuries caused by bats, from 0, 1% to 0, 45%. It is the understanding that the analysis of these data should be on charge of the State level. It started paying more attention to the counties with rate under 10 patients / 10.000 inhabitants / year what indicate lack of consciousness and /or difficulties of access to information or to local health services. The high percentage of treatments could mean that patients without prophylaxis indication were not registered and / or the necessity of professional training. After this, the State Program developed training program on data collection, data registration and local team training.

167

POSTER I

LABORATORIAL PROFILE OF THE RABIES DIAGNOSIS IN EVANDRO CHAGAS INSTITUTE, 2003 - 2005
E.S. Travassos da Rosa 1, T.F . Barbosa 1, L.M.N. Casseb 1, A.S. Pereira 1, D.B.A. Barbosa 1, H.C. Peixoto 2, A.L. Begot 3, R.J.S. Lima 3, P .F.C. Vasconcelos 1
1) Instituto Evandro Chagas / SVS / MS, 2) Universidade Federal de Agricultura da Amaznia, AM, Brasil, 3) Ncleo de Doenas Endmicas do Estado do Par, PA, Brasil

Dr. Elizabeth Salb T. da Rosa - elizabethsalbe@superig.com.br

Brasil

Introduction: Rabies is an important disease occurring in the northern Brazil that includes the States of the Acre, Amap, Amazonas, Maranho, Par, Rondnia, and Roraima. Objective: To evaluate the rabies based on the laboratorial diagnosis situation in the northern region in 2003- 2005 period. Method: 8, 934 central nervous system (CNS) samples were analyzed as follows: human beings (43), dogs (5, 998), cats (276), bovine (8 1), equine (2 2), bats (2, 097), and (403) of other animal species. Direct Immunofluorescency Assay and attempts of viral isolation by the cranial inoculation in newborn mice were used. Results: A total of 166 CNS specimens were positives, 2 2 (5 1. 16%) from humans, 83 (1.38%) canine, 7 (2.53%) feline, 40 (49.38%) bovine, 7 (3 1.8 1%) equine, 6 (0. 28%) bats and 1 (100%) swine. All human cases occurred in Par State due Desmodus rotundus battransmitted outbreaks of human rabies, in 2004 (10), cities of Portel (region of Marajo) and Viseu (Northeastern of Para) and 2005 (1 2), in Augusto Corra, Northeastern of Para. With regard the canine rabies, of the 83 confirmed cases, 48 (57.83%) occurred in 2003 (Acre, Amazonas, Par and Rondnia), 18 (2 1.68%) in 2004 (Acre, Par and Rondnia) and 17 (20.48%) in 2005 (Par and Rondnia), while among feline rabies, 7 cases were diagnosised 2 (28.57%) in 2003 (Acre and Par), 3 (4 2.85%) in 2004 and 2 (28.57%) in 2005, all in Par state. Regarding to the 40 confirmed cases in bovines, 8 (20%) occurred in 2003 (Acre, Amazonas, Par and Rondnia), 9 (2 2.5%) in 2004 (Acre and Par) and 23 (57.5%) in 2005, being 1 in Amazonas and 2 2 in Par. From the 7 equine positive 4 (57. 14%) occurred in 2004 (Acre and Par) and 3 (4 2.85%) in 2005 in Par. All positive bats were from Par State, 3 of them were non hematophagous bats, and the single positive swine was from Augusto Corra, Par state. Conclusions: In the Northern region two different cycles are present, urban cycle occurring among domestic animals without human cases, the aerial cycle affecting humans, bovines and swine. The surveillance of bat attacks should be improved to prevent the occurrence of bat-transmitted rabies. Financial support: IEC

168

POSTER I

THE PROPHYLAXIS PROFILE OF THE HUMAN RABIES IN THE STATE OF CEAR


Nlio Batista de Morais 1, David Caldas Vasconcelos 1, Lcia Maria de Silva Alencar, Benedito Nelson Rolim 1, Jarier de Oliveira Moreno 1
1) Secretaria de Estado da Sade do Cear, CE, Brasil

Dr. Nlio Morais - neliom@ig.com.br / neliomo@ig.com.br

Brasil

INTRODUCTION: The state of Cear is located in the Northeast area of Brazil, it possesses an area of 146.8 17km 2 and an approximate population of 7. 106.605 inhabitants. The Rabies along all its implantation period of the program since 1973 always came as a problem of health public face to its epidemic compounds in the urban areas, wild and rural. OBJECTIVE: It intends to analyze the structure of services of health, the demand and the profile of the attendance and peoples surveillance assisted in the units of health with aggression complaint for animals. MATERIAL AND METHODS: The data were collected through the VE-7 records, reports of the Secretary of Health of Cear and of the National Program of Control of the Rabies. RESULTS: Starting in 2003 the state started to adopt the human antirabic vaccine of cellular cultivation. In the period from 2003 to 2005 the state assisted 8 2.67 1 people for aggression of animals, of these 64.99 2 (78, 6 1%) were directed for treatment. It was applied 169.0 20 doses of vaccines that it corresponds to 2, 8 doses per person treated. From 2003 to 2005 were registered 2.5 20 abandonments that corresponds to 3, 9% of the treated people. Of the 8 2.67 1 assisted people were identified 7 1.558 attacked by dogs and cats that corresponds to 86, 5% of the aggressions. The state possesses 184 municipal districts with units of health for attendance with antirabic vaccine and serum and it is applied in 100 municipal districts, all in hospitalares units. CONCLUSION: The treatment indications are considered high in the state, because most of the aggressions happens for dogs and cats, many susceptible to observation. The professionals rotatividade in the attendance area as well as the qualification of the same ones is a gap that propitiates this situation. The index of abandonment treatment in the state is one of the smallest ones in Brazil, face the performance of the Health Program of the Family acting in way decentralized in the rural areas of the state. In the last three years the state moved forward in a significant way in the control of the urban Rabies, parallel to this began in 2005 a training of medical professionals and nurses on the technical norm of the Rabies that aims in 2007 to reach a profile of larger quality in the area of human prophylaxis.

169

POSTER I

EPIDEMIOLOGY OF HUMAN RABIES POST-EXPOSURE PROPHYLAXIS AND RELATIONSHIP BETWEEN RABIES CONTROL STRATEGIES IN CENTRO SUL AREA OF RIO DE JANEIRO, BRAZIL
Ana Luisa Poerner 1, Maria Julia Salim Pereira 1
1) Universidade Federal Rural do Rio de Janeiro, RJ, Brasil

Dr. Ana Lusa Poerner - anaperrone@ufrrj.br

Brasil

INTRODUCTION Dog still remains as the most important reservoir and vector of human urban rabies and is responsible for most of the aggressions that require post-exposure treatment (PEP). Rabies is considered a serious public health problem and the main control activities developed in Brazil are preventive human rabies treatment and canine mass vaccination. Intervening in the animal reservoir can control canine rabies and hence human exposure. OBJECTIVES Evaluation of canine vaccination and human post-exposure treatment (PEP) trends and epidemiological profile of animal aggression. METHOD Records from animal agression and canine vaccination in centro-sul area of Rio de Janeiro state between years 2000 and 2005 were obtained from State Health Office and were entered into an Epi Info version 200 2. Data from animal vaccination and human post-exposure treatment were submitted to simple linear regression analysis to study trends and the levels of relation among them. RESULTS Of 4.3 10 exposures, dogs were responsible for 86, 2%, and most of them were healthy (44, 8%). Among those exposed, 65% received PEP treatment. Most of the patients were male (57, 4%) and had less than 14 years old (35%). Bite exposures accounted for 8 1, 5% of the wounds and most were superficial (60, 8%), unique (59, 7%) and located in hands/feet (36, 6%) or lower limbs (35, 1%). The aggression was not provoked in most of the cases (60, 1%) and often happened in peoples spare time (50, 8%). The incidence of rabies PEP increased from 5, 2 cases per 10.000 persons in 2000 to 23, 9 cases per 10.000 in 2005. Canine vaccination ranged from 44.340 animals in year 2000 to 53.88 1 in 2005. There is an ascended trend in human post-exposure treatment, ignoring the decrease in canine rabies and high levels of vaccination coverage. CONCLUSION There is no integration between the actions developed in animal and human rabies prevention and control.

170

POSTER I

IMPLEMENTACIN DE UNA RT-PCR ANIDADA COMO ALTERNATIVA PARA LA DETECCIN DEL VIRUS DE LA RABIA EN MUESTRAS INFECTADAS
V. Fuentes-Gmez, E. Rojas-Anaya, B. Cortz-Garca, S. Nadin Davis, E. Loza Rubio 2, 1
1) Canadian Food Inspection Agency, Neapan, Ontario, Canada, 2) Centro Nacionalde Investigaciones en Microbiologa Animal, INIFAP , Mexico

Dr. Elizabeth Loza Rubio - loza.elizabeth@inifap.gob.mx

Mxico

La rabia es una enfermedad cosmopolita que afecta animales domsticos y silvestres, esto constituye la mayor fuente de la enfermedad a humanos. Por este motivo es necesario un diagnstico oportuno y certero. La prueba estndar es la inmunofluorescencia directa, sin embargo los resultados negativos se tienen que confirmar por la prueba de inoculacin del ratn, esta requiere ms de quince das para arrojar resultados. Por lo anterior el objetivo de este trabajo es el desarrollo de una RT-PCR anidada para aumentar la sensibilidad del diagnstico del virus en muestras sospechosas, en comparacin con los resultados obtenidos en otro estudio Loza-Rubio E, Rojas AE, Banda RVM, Nadin-Davis S, Cortez GB. Detection of multiple strains of rabies virus RNA using primers designed to target Mexican vampire bat variants. Epidem Infect. 2005, 133:9 27-934. Se colectaron 50 muestras de cerebro de diferentes especies y regiones de Mxico. Se consideraron muestras comprendidas entre 2000- 2006. Se extrajo el RNA total, como control positivo se utiliz RNA de cerebros de ratn infectado con el virus de la cepa CVS, y como control negativo RNA de un ratn sin infectar. A partir de ste se sintetizo el cDNA y con este se realizo la RT-PCR utilizando los iniciadores SuEliI y SuEliII, los cuales amplifican una regin de 75 1pb del gen N. Para aumentar la sensibilidad del mtodo se realiz una PCR anidada empleando iniciadores que amplifican 16 2 pb. El resultado fue que el 90% de las muestras resultaron positivas con el RT-PCR utilizando los iniciadores SuEliI y SuEliII, mientras que el 94% fueron positivas por la RT- PCR anidado. Estos resultados demuestran que la PCR anidada aumenta las posibilidades de deteccin del virus de la rabia. Para demostrar que dicha prueba aumenta la sensibilidad, es necesario realizar otros ensayos as como aumentar el nmero de muestras

171

POSTER I

THE PROFILE OF THE PROPHYLAXIS OF HUMAN RABIES IN THE REGIONAL CELL OF HEALTH OF CAUCAIA - CE
Jarier de Oliveira Moreno 1, Nlio Batista de Morais 1, David Caldas Vasconcelos 1
1) Secretaria de Estado da Sade do Cear, CE, Brasil

Dr. Jarier de Oliveira Moreno - cetrav@saude.ce.gov.br

Brasil

INTRODUCTION: The Regional Cell of Health of Caucaia is located in the Northeast area of the state of Cear, composed of 1 1 municipal districts with total population of approximately 5 28.3 15 inhabitants. Since 1973, with the implantation of the National Program of Control of Rabies, that zoonose comes as a problem of health public, due its interfaces with the urban atmosphere, rural and wild. OBJECTIVE: Analyzing the actions and services of health, in what concerns to the peoples attendance attacked potentially by animals transmitters of the Rabies virus in the period from 2003 to 2005. MATERIAL AND METHODS: The data were collected through the records VE-7, reports of the Secretary of Health of Cear and of the National Program of Control of the Rabies. RESULTS: In agreement with the analyzed period 6.6 2 2 people were assisted, in which 5.333 (80, 5%) were directed for treatment. 18. 2 16 vaccine doses were applied, totaling 3, 4 doses per person applied. The treatment abandonments were registered in 3, 9% of the treated people. Dogs and cats were responsible for 95, 8% of the total of aggressions. The high percentile of treatment indication is related to the great number of aggressions for dogs and cats, associated to the epidemic conditions of the municipal districts, what came to reinforce the control of the human rabies in the last three years. The professionals of health rotatividade which accomplish the attendances did the Secretary of the Health of the State of Cear accomplished trainings on the technical norm of the Rabies. The low number of treatment abandonments is due the performance of the Program of Health of the Family that acts in way decentralized in the drones rural of those municipal districts. CONCLUSION: The control increment of urban rabies associate to the professionals of health qualification favors in the improvement of quality pattern of prophylaxis actions of the human rabies.

172

POSTER I

EVALUATION OF THE ANTI-RABIES REEXPOSURE TREATMENTS IN THE CLINIC OF THE PASTEUR INSTITUTE OF SO PAULO, BRAZIL
Vera Lucia Bolzan 1, Ricardo Siqueira Cunha 1, Andrea de Cssia Rodrigues Silva 1, Graciane Maria Medeiros Caporale 1
1) Instituto Pasteur, So Paulo, SP , Brasil

Dr. Vera Bolzan - vlbolzan@ig.com.br

Brasil

In cases of risk exposition to the rabies virus, immune patients will have to receive doses from reinforcement of the rabies vaccine. The objective was to evaluate the treatments of re exposed in Pasteur Institute of So Paulo. The cases reports of clinic attendance among January of 2003 until December of 2005 had been evaluated. In this period 3 1 2 patients had received doses from rabies vaccine of cellular culture (VERORABIES, Aventis-Pasteur) and the searched neutralizing antibodies in 1 10 patients. Of the treated patients, 53.73% were female. The average age was of 44years (varying of 4 to 83 years). The animals involved in the accidents were: 63.78% dogs, 18.9 1% cats, 6.09% monkeys, 6.09% bats, 2, 24% other wild animals, 2, 89% others. The expositions was light in 36, 54% and serious in 63, 46%. Patients received 5 doses in 10, 58%, and 2 doses of reinforcement in 89, 4 2%. The number of previous treatments was 1 (85, 26%), 2 (8, 09%), 3 or more (1, 9%) The interval of time between the treatments was of 17, 7 years (varying of 4 months to 63 years). The average of doses of vaccine was of 1 1, 3 (varying of 3 to 40 doses). After 10 days of treatment 98% of the patients had titles of neutralizing antibodies higher than 0, 5 UI/ml, with geometric average of 2, 0 UI/ml (varying of 0, 3 to 4, 0 UI/ml). Two patients had not answered (a HIV patient and another patient without known cause). The treatment can stimulate the immune response, independent of the time out, the number of doses of vaccine. The immune memory is enough since that the initial treatment has been complete and other diseases do not exist that compromise immune reply of the patient.

173

POSTER I

ANALYSIS OF INFORMATION RELATED TO PROPHYLACTIC TREATMENT AGAINST RABIES GIVEN TO RESIDENTS OF JABOTICABAL, SO PAULO, BRAZIL
D.F.R. Frias 1, A.A.B.Carvalho 1, M.G. Buzinaro 1, S.L.S. Lages 1
1) Faculdade de Cincias Agrrias e Veterinrias, Universidade Estadual Paulista, Jaboticabal, SP , Brasil

Dr. Adolorata Aparecida Bianco Carvalho - adbianco@fcav.unesp.br

Brasil

The stablishment of an appropriate prophylactic treatment is fundamental to the control of rabies. Efforts made by the Public Health services have significantly reduced the number of rabies cases in both humans and animals at most of Brazilians States. However, the number of persons treated with post-exposure prophylaxis (PEP) has not proportionally reduced. The main goal of this study was to analyze information about PEP given to residents of Jaboticabal - SP , Southern Brazil, between the years 2000 and 2005. A total of 2, 740 official records of patients involved in accidents with animals were analyzed. From these records, PEP was not indicated for 508 (18.5%) patients and the aggressive animals were kept under observation for ten days, and 2, 23 2 (8 1.5%) patients received at least one dose of vaccine. For these 2, 23 2 patients, 88.8% received three doses of vaccine, 9.4% received five doses, 1.4% received one dose, 0. 2% received thirteen doses, 0. 1% received two doses and 0. 1% received four doses. After the beginning of PEP , 2 2.3% of patients quit treatment after the animals were observed and claimed to be non-rabid. Concerning to the condition of the animals during the accident, 75.4% of the animals were not rabid, 5.0% were ignored, 2.6% were suspected to be rabid, 0.8% was observable and 0. 1% was rabid. Moreover, 16. 1% of the records did not give this information. As to the period of animal observation, 3 1.9% of observed animals were healthy, 1.8% disappeared, 1.8% died and 0.7% was submitted to euthanasia, however, 63.8% of the records did not give this information. The data analysis can identify failures in the Public Health services and the results can be used to ameliorate the rabies surveillance system and services offered to the citizens of this town.

174

POSTER I

MEASUREMENT OF THE IMMUNE RESPONSE TO RABIES VACCINATION USING A NOVEL AND TRADITIONAL ASSAYS
S.M. Moore 1, M.W. Wilkerson 1, C.R. Wyatt 1, R.D. Davis 1, D.J. Briggs 1
1) Kansas State University, Manhattan, KS, USA

Dr. Susan Moore - smoore@vet.ksu.edu

USA

Detection of neutralizing antibodies to rabies virus is the basis for determination of adequate immunity while specific lymphocyte responses to rabies vaccination have not been fully investigated. The objective of this study was to develop a nonradioactive multi-parameter flow cytometry assay to identify antigen specific memory lymphocytes in individuals previously vaccinated against rabies virus and compare it against the tritiated thymidine method. A cell tracking dye (carboxyfluorescein succinimidyl ester) was used in combination with surface staining for CD4 and CD8 molecules to determine the response of memory lymphocytes to killed rabies virus in an antigen recall assay. Rabies virus-specific lymphocyte responses were compared to antibody responses in the same individuals. Lymphocytes and serum samples from 10 healthy subjects previously vaccinated with cell culture rabies vaccine and from five non-vaccinated subjects were analyzed. Memory T lymphocyte responses to rabies virus were identified in all vaccinated subjects, some noted as early as 3 days after stimulation and others not until 7 days of stimulation. The proliferation index of the CFSE assay correlated well with the simulation index of the [3H] thymidine assay (kappa statistic = 0.73). An inverse relationship between antibody and lymphocyte responses to rabies virus was detected in the vaccinated subjects. Two vaccinated individuals with high lymphocyte and low antibody responses produced Th 1 type cytokines (IFN- gamma and TNF-alpha) to rabies virus stimulation, whereas two vaccinated individuals with low lymphocyte and high antibody responses did not produce these cytokines. Currently this CFSE assay is being adapted to veterinary species for the detection of cellular immunity memory response to various recall vaccine antigens.

175

POSTER I

PRINCIPAL RISK FACTORS FOR HUMAN RABIES TRANSMITTED BY VAMPIRE BATS, MARANHO STATE, BRAZIL, 2005
Jakeline Maria Trinta Rios 1, Jos Orlando de S. Martins, Salim Jorge Waquin Neto, Daniel S. Saraiva, Henrique Jorge dos Santos, Mildes Mendes Pereira, Graa Lrio Leite, Marcelo Yoshito Wada 2
1) Secretaria de Sade do Maranho, MA, Brasil, 2) Secretaria de Vigilncia em Sade, Ministrio da Sade, DF, Brasil

Dr. Jakeline Rios - jakeline.rios@gmail.com

Brasil

Introduction: Rabies is a zoonotic disease with a mortality rate of 100%. During 2004 and 2005, outbreaks of human rabies transmitted by vampire-bats occurred in two Brazilian states: Par and Maranho. Methods: A descriptive study was performed to describe risk factors and actions taken based on observations in four municipalities (Godofredo Viana(GV), Carutapera(CA), Cndido Mendes(CM) and Turia(TU)) where there were identified human rabies cases in 2005. Results: There were reported 24 human rabies cases during the period study: GV3, CA 2, CM 2 and TU(17). Among identified cases, 2 1(87.5%) were confirmed by laboratorial criteria and three (1 2.5%) by clinical and epidemiological criteria. All municipalities were located in edges of rivers with a lot of trees (Avicennia germinans) with characteristics of fen. In GV, it was found cracks caused by pan. Other important factors included deforestation, cultural habits, lack of information, isolated localities, open residences and poverty. Conclusions: This investigation demonstrated that the mainly risk factor found was information lack about necessity of prophylaxis for rabies in case of exposure to vampire-bats. The resistance to realize prophylaxis and access to some areas prejudiced actions as pos-exposure prophylaxis, including application of vaccine and immunoglobulin. Most of the human rabies cases were children and fisherman and were attacked in their foot, whose habits were to sleep in beaches without any kind of protection. Actions taken included capture and control of vampire-bats, pre and pos-exposure prophylaxis, orientation for communities and professionals about how to protect their residences, training about control and prevention measures, education in radio, schools, churches and other mechanisms.

176

POSTER I

DESCRIPT OF TWO HUMAN RABIES CASES THROUGH CANINE TRANSMITION IN THE STATE OF MARANHO IN 2006 AND THE IMPORTANCE OF REVACCINATE THE DOG WHICH WERE ALREADY VACCINATED
Jakeline Maria Trinta Rios 1, Jos Orlando de Souza Martins 1,2, Salim Jorge Waquim Neto 1,2, Daniel S. Saraiva 2, Gleusa Maria Barreto Vieira 3, Natal Jata de Camargo 3, Marcelo Yoshito Wada 3
1) Secretaria de Sade do Maranho, MA, Brasil, 2) Secretaria Municipal de Sade de So Lus, MA, Brasil, 3) Secretaria de Vigilncia em Sade, Ministrio da Sade, DF, Brasil

Dr. Jakeline Rios - jakeline.rios@gmail.com

Brasil

Introduction: The Rabies is an acute viral encephalitis, which represents 100% of death and the dog is its main source of transmition in urban areas. The disease keeps on being a serious pubic-health problem despite the increasing advances on the control program against it. In Maranho there is a historical sequence of various cases of the disease involving young does which had only one or even no anti-rabies vaccination, that fact was discovered during the investigation. Goal: Describe two cases of human rabies registered in 2006 in which transmition happened through three-to-six-month-old dogs and which there wasnt a explanation to the aggression, therefore there are reports on the epidemiological investigation which demonstrate contact with animals (dogs). In one of the cases there was a contact with a dog, which died with symptoms similar to the rabies ones. On the second case it was involved a young dog which died by trampling. Method: Describing through the reports and epidemiological investigation in loco, searching subsides for the detecting the possible rabies transmitter on both cases since theres no aggression report by dogs or cats. Results: Since there are no manners of identifying the transmitter species of rabies in both cases. It was made, after the death of patients, the gathering of the brain tissue and it was diagnoses human rabies, and it was also made the antigenic typology and was discovered that it was the variant 2 (dog). Conclusion: Through the occurrence of the cases already mentioned, we can say that even without the aggression certainty by dogs or cats, the rabies can be transmitted through direct contact with mucous or simple scratch by young animals through play, which cant be even noticed, mainly with these animals. Both of the investigated of the dog vaccination campaign on the cities dont know the importance of making the vaccination to reinforce the animal already vaccinated, increasing in the way the transmition cycle of urban rabies.

177

POSTER I

BUDGETARY AND FINANCIAL RESOURCES VERSUS HUMAN AND CANINE RABIES CONTROL IN THE MUNICIPAL DISTRICT OF RIO BRANCO - ACRE BRAZIL, FROM 1997 TO 2002
Jos da Conceio do Nascimento Guimares 1, Francisco Glauco de Arajo Santos, Henrique Jorge de Freitas
1) Secretaria Estadual de Sade do Acre, AC, Brasil

Dr. Jos da Conceio do Nascimento Guimares - joconasgui@ibest.com.br

Brasil

The rabies or encephalomyelitis acute and incurable is an viral anthropozoonosis. It is most one studied diseases in the world in view of your lethality occur in 100% of the cases, as well as to introduce problems tied to public health, besides demanding big sums in cash and investments for your eradication and/or control. The number of cases notified of human rabies comes decreasing, in Brazil, since decade of 80. Municipal district of Rio Branco Acre/Brazil has been investing in infrastructure, specialized personnel contract, their agents training, free-roaming dogs capture, laboratories diagnosis of the illness, as well as in vaccination campaigns, with seen to the control and/or eradication of this disease. The aim of this work is to evaluate if the budgetary and financial resources allocated for the municipal district of Rio Branco - Acre/Brazil, in the period from 1997 to 200 2 were enough for the human and canine rabies control. It observed are the budgetary and financial insufficient resources for the human and canine rabies effective control, that the capture, the sampless collection for laboratories diagnosis (identification of the areas with positive cases), free-roaming dogs vaccination and sacrifice have been contributing for the incidence decrease of human and canine rabies cases, as well as the canine antirabies vaccination campaigns house to house, the clearness campaigns to the population, regarding the risk of the free-roaming dogs presence in public ways and the danger of contracting the canine rabies, by the communication means in mass, as well as the lectures at fundamental and intermediate teaching schools have been improving enough the free-roaming dogs capture teams work in the districts, but they have been being insufficient for the human and canine rabies control and eradication in the municipal district of Rio Branco - Acre/Brazil.

178

POSTER I

HUMAN AND CANINE RABIES CONTROL PROGRAM EVALUATION IN THE ACRE STATE, BRAZIL, FROM 1996 TO 2002
Maria do Carmo Portela 1, Francisco Glauco de Arajo Santos
1) Secretaria de Estado da Sade do Acre, AC, Brasil

Dr. Maria do Carmo Portela - mariadocarmo.portela@ac.gov.br

Brasil

The rabies is a fatal viral zoonosis and a public health serious problem. This work analyzes the activities developed by the Human and Canine Rabies Control Program in the Acre State, Brazil, from 1996 to 200 2, from the activities defined by the Health State Department. The data were going obtained from the analyses accomplished in the epidemiological filing card sending monthly from Zoonosis Control Program State Coordination, by the 2 2 Acre municipal districts, incorporating the next aspects: municipal districts that accomplish free-roaming dogs systematic capture, municipal districts with canine antirabies vaccination campaign, human and canine rabies cases notified for municipal district, human and canine samples sending to the rabies laboratories diagnosis for the municipal district, people that began the antirabies treatment human regarding the people attended in the health units, municipal districts that notify the Rabies Control actions through the notification filing card VE-7. The data were going tabulated and analyzed statistically using the Program SPSS. From 2 2 municipal districts that investigated the canine rabies, only 0 2 accomplished her systematically. Even so these municipal districts obtained success, regarding not human rabies occurrence. However the municipal districts dont be with the structured rabies Epidemic Vigilance, and although activities accomplished in the canine rabies control are deficient, the data point that in this period occurred an evolution in the actions in the Acre State. It observes that there isnt an expressed preoccupation by the municipal managers concerning the risk elimination of the human and canine rabies in the regional of Alto Juru, Tarauac and Envira and Sena Madureira. That being the case, the research suggests to the municipal managers owed her attention to the control measures of this disease, in the epidemic chain breakage, having in mind the possibility of occurrence of the new in human and in animal cases.

179

POSTER I

HUMAN RABIES AND THEIR ANIMAL EXPOSURES: DESCRIPTIVE ANALYSIS FROM TRANSMISSION CYCLES IN TWENTY YEARS
Marcelo Rezende de Oliveira 1; Mauro da Rosa Elkhoury 2; Marlia Lavocat Nunes 2; Marcelo Yoshito Wada 2
1) Estudante de Medicina Veterinria, 9 perodo, Faculdades Integradas do Planalto Central FIPLAC; 2) Secretaria de Vigilncia em Sade, Ministrio da Sade, Brasil.

Dr. Marcelo Rezende de Oliveira- medvetmarcelo@yahoo.com.br

Brasil

Rabies is a zoonoses transmitted by a RNA virus, genus Lyssavrus. It can be classified in four transmission cycles that occurs in terrestrial and aerial mammals. Usually present two clinical manifestation: furious or paralitical and progress to death. A retrospective and descriptive study of the human rabies cases reported to Brazilian Ministry of Health was performed between 1986-2005. Cycles of transmission were classified according animal exposure: urban cycle when transmission occurred by dog or cat; aerial cycle when it occurred by bat; wild cycle when it occurred by wild mammals and rural cycle when it occurred by herbivorous. Temporal anaylsis was performed for each fifteen years.There were reported 756 human rabies cases in period study. Urban cycle was responsible for 71.3% of the cases; aerial cycle for 17.6%; wild cycle for 4.4% and rural cycle for 0.3%. Among urban cycle, 94.4% had dogs as animal exposure, while in wild cycle the mainly animals exposures were primate (42.4%) and fox (39.4%). Temporal analysis showed that urban cycle was steady between 1986-2000 and a decline trend from 2001 was observed; while aerial cycle demonstrated an abnormal trend. Wild cycle and ignored cycle showed a continuous decline trend. In last twenty years, human rabies was endemic in Brazil, but has demonstrated clearly different cycles of transmission. Investigations about exposure animals are improving. It is recommended local analysis in states and municipalities have realized to contribute among control and prevention measures.

180

POSTER I

POSTER II

181

POSTER II

182

POSTER II

DOG RABIES PREVENTION PROPOSAL BY THE STATE HEALTH SECRETARY FOR THE COUNTIES OF RIO DE JANEIRO
P .R. Barbosa 1, F .R.M. Azevedo 1, D.R. Cruz 1, P .C. Romijn 1
1) Coordenao de Vigilncia Epidemiolgica Ambiental, Secretaria de Estado de Sade do Rio de Janeiro, RJ - Brasil

Dr. Paulo Roberto de Barbosa - vigiambiental@saude.rj.gov.br

Brasil

SES-RJ developed a project in which the traditional actions are questioned for their fundaments. Knowledge on viral immunology and cytology has evolved, and concepts should be revised up on different comprehension of phenomena due to scientific progress. The final goal, avoiding human and pet rabies, is worked on through the development of a technical view of the preventive actions that could/should be taken when risk factors of the disease occurrence are present. In a state with no dog rabies, educational presentations incentives the alertness on evidence of rabies infected bats, such as those found with abnormal behavior, as well as the presence of bites by haematophagous bats on animal of economical interest and dogs. The project includes a detailed methodology, starting with dog and cat population statistics counting, up to integration with agricultural organisms/institutions for bat monitoring and the presence of bat caves, and the elaboration of herbivore rabies maps in each county and its neighbors. The monitoring and population control of Desmodus rotundus bats is carried out by Health, Agricultural and Environment organizations. The recollection of street dogs is incentivated with the sending of samples for rabies laboratory diagnosis. The results permitted the human population involved to have knowledge on the subject and to collaborate, permitting a reduction in fatalities due to zoonoses in general, the knowledge of risk regions and factors, a better vaccination campaign planning, immediate identification of possible risk situations, evidence of the necessity of immunological competence checking of vaccinated animals and the occurrence of sub-clinical infection in wild mammals. It is concluded that a situation diagnosis is made possible by simply monitoring in a specific county, and the risk factors to be considered for the next years. It is suggested to introduce a discussion about this zoonosis, as well as others, as a pedagogical component.

183

POSTER II

ACCIONES DE VIGILANCIA EPIDEMIOLOGICA QUE PERMITEN AL ESTADO DE BAJA CALIFORNIA (MEXICO) MANTENERSE POR 24 AOS SIN CASOS DE RABIA URBANA 2001-2006
Francisco Vera Gonzlez, Rolando Ortiz Quinez, Jos Alfredo Cubillas Araiza, Carlos Alberto Delgado Snchez, Daniel Luna Rubio 1
1) Instituto de Servicios de Salud Publica del Estado de Baja - California - Mexico

Dr. Daniel Luna Rubio - zoonosisbc@yahoo.com.mx

Mxico

En el periodo que se informa se atendieron con oportunidad 7,450 personas agredidas de las cuales nicamente se les aplico Vacuna Antirrbica Humana a 235 cabe sealar que en este Estado de Baja California es donde menos biolgico se aplica con un 3.15% el ms bajo en todo el pas. VACUNACIN ANTIRRBICA CANINA cada ao se incrementa la aplicacin de mas dosis de este biolgico en el ao del 2001 se aplicaron 207,534 en el Estado de Baja California y comparando con el 2006 se administraron 234,565 dosis faltando una campaa de reforzamiento en el mes de septiembre, esperando llegar a 250,000 dosis aplicadas al termino de este ao. CONTROL DEL PERRO CALLEJERO la captura del perro callejero es indispensable para controlar la poblacin de los mismo y disminuir las agresiones a personas, en el la Entidad operan 8 Centros Antirrbicos en las ciudades de Mexicali con 2, en Tecate, en Tijuana 2, Rosarito y Ensenada con 2, Administrados por los Municipios e ISESALUD y estos han capturado un total de 114,900 en este periodo, de los cuales aproximadamente el 92% de ellos son sacrificados compasivamente al permanecer por 72 horas y no ser reclamados por sus dueos. MUESTRAS ENVIADAS AL LABORATORIO el diagnostico de laboratorio representa una actividad muy importante para el manejo correcto y oportuno de las personas agredidas el total de muestras enviadas a los laboratorios fue de 3,467 por supuesto todas negativas en el mismo periodo. ESTERILIZACION DE MASCOTAS. La Secretaria de Salud del Estado de Baja California y en coordinacin con el Colegio de Mdicos Veterinarios Especialistas en Pequeas Especies as como con el apoyo de la Universidad Autnoma del mismo Estado y asociaciones protectoras de animales se ha realizado 8,989 esterilizaciones de mascotas en el periodo que se informa. El Estado de Baja California ha obtenido por tercera ocasin, el primer lugar Nacional en las actividades de prevencin y control de la rabia (2001, 2003 y 2005).

184

POSTER II

CONTRIBUTION OF ZOONOSIS DIAGNOSIS SERVICE, UNESP-BOTUCATU/SPBRAZIL, TO RABIES DIAGNOSIS IN THE PERIOD OF 1991 TO 2005
Helio Langoni 1, Veruska Maia da Costa 1, Fabio Hiroto Shimabukuro 1, Rodrigo Costa da Silva 1, Benedito Donizete Menozzi 1
1) Ncleo de Pesquisas em Zoonoses, UNESP-Botucatu, SP , Brasil

Dr. Veruska Costa - verusket@yahoo.com.br

Brasil

The Zoonosis Diagnosis Service, of UNESP-Botucatu/SP-Brazil, realizes diagnostic activities in rabies since 1991, as credential laboratory to Health State Secretary of So Paulo State - Pasteur Institute/SP . A survey referent to the period of 1991 to 2005 was realized, where 3728 samples were received to the analysis, involving 986 (26.95%) samples of the canine specie, 296 (8.09%) feline, 621 (16.97%) bovine, 1590 (43.46%) chiroptera, 56 (1.53%) equidea, 32 (0.87%) ovine and caprine, 17 (0.46%) swine, 2 (0.05%) bubaline, 21 (0.57%) rodents and 37 (1.01%) wild animals. Amongst the received samples, 70 (1.87%) were considered improper to the diagnosis, being analyzed 3658 samples. For the direct fluorescent antibody test (DFAT) and biological test (mice inoculation), 92 (2.51%) samples were positive, 9/986 (0.91%) to canine specie, 2/296 (0.67%) to feline, 68/621 (10.95%) to bovine, 6/1590 (0.38%) to chiroptera, 7/56 (12.5%) to equidea and 2/17 (11.76%) to swine. Between 1991 and 1997, oscillations had occurred in the percentage of positivity to canine and feline species, that diminished gradually, have not been registered anyone positive diagnosis of the disease to both species after this period. With the control of rabies in dogs and cats in the beginning of the decade of 1990, in many cities of So Paulo state, an intensification of the vigilance occurred in other animal species, being observed an increase of positivity in chiropteras and bovines. Independent of the animal specie involved among the 92 positive samples, there was a concordance of 99.69% between the used tests. However, 11 (11.95%) were negative to DFAT and positive to biological test, demonstrating a sensitivity of 88.04% and specificity of 100%.

185

POSTER II

RABID BATS AND PETS


Maria Isabel R. Bofill 1, Angelika Bredt 1, Rodrigo M. B. Rodrigues 1, Klaus M. Paranayba 1, Pricles N. T. Massunaga 1
1) Secretaria de Estado de Sade do Distrito Federal, Braslia, DF, Brasil

Dr. Maria Isabel Rao Bofill - bofillrao@ig.com.br

Brasil

Since 1988 the Health Secretary of Distrito Federal (DF), Mid-Western Brazil, recorded 55 cases of interaction between pet animals and bats in urban environments. They involved 11 bat species, being seven insectivorous (Molossus molossus, Molossops planirostris, Nyctinomops laticaudatus, Eumops glaucinus, Eptesicus brasiliensis, Lasiurus sp. and Lasiurus blossevillii), three phytophagous (Glossophaga soricina, Platyrrhinus lineatus and Artibeus lituratus) and one omnivorous bat (Phyllostomus discolor). From those 46 bats were submitted to rabies exams, only three were rabid: N. laticaudatus, L. blossevillii and A. lituratus. In two of these cases, the encounter of a siamese cat and an individual of N. laticaudatus (24/09/2003) and of a mixed race female dog and an individual of L. blossevillii (17/02/2004) was occurred in a 6th floor apartment of two residential buildings. Another encounter of a female dog Pinscher and a bat A. lituratus was recorded on a residential house garden (14/06/2004). Although pet animals had no updated rabie vaccination, their soroneutralization exams showed a good level of rabies antibodies. After collecting a blood sample from the pets one dose of anti-rabic vaccine was applied in the days 0 and 30. These pet animals are still living until now. In the urban environments, dogs and cats are considered important bat killers and the rabies transmission could carry out if theses encounters involve a rabid bat and a deficiently rabies vaccined pet. This interaction could take to a life risk to pet animals and their owners.

186

POSTER II

FIRE, BATS AND THE RISK OF RABIES TRANSMISSON


Maria Isabel Rao Bofill 1, Angelika Bredt 1, Wilson Uieda 2
1) Secretaria de Estado de Sade do Distrito Federal, Braslia, DF, Brasil, 2) UNESP Botucatu, SP , Brasil

Dr. Maria Isabel Rao Bofill - bofillrao@ig.com.br

Brasil

Nyctinomops laticaudatus (Molossidae) is a common insectivorous bat in urban areas of Distrito Federal (DF), MidWestern Brazil. Colonies with hundred to thousand bats can be found harboring in expansion joints and other building gaps. This colonial species caused several constraints to humans such as incoming movements, feces accumulation and bad odor. Its management could take the incoming movements to other buildings and consequently to promote agonistic interactions between intruders and resident bats. This interaction is one the rabies transmission route among bats. Near 07:00h of December 27th 2005 a big fire started on a ten-story building in Braslia, DF . After two hours a massive running of N. laticaudatus was observed on the burning building and part of them flew into the neighboring building and other part fallen down to the public ways where 37 burned bats were caught and submitted to rabies exams. No bats positive to rabies were found. After two weeks, the intruders were not observed harboring in neighboring building. After 22 days, a bat with fire burned pelage was found on the ground distant six km from the fire to the northeast, suggesting that some bats are still looking for the definitive roosts. This roost change caused by fire event and the long search for a good harbor could cause physical stress in some bats. The body contact among stranger bats increases the chances of rabies occurrence. We suspect that there is a relationship between the fire and a rabid N. laticaudatus found after 72 days, distant four km from the fired building to the southeast. Between 2000 and 2005, only four other rabid N. laticaudatus were found in Braslia. The possible consequences of the fire on that bat colony from Braslia must be considered in the eventual management of colonial bat species.

187

POSTER II

MACROSCOPIC AND MICROSCOPIC FINDING IN A BOVINE RABIES CASE AT DISTRITO FEDERAL (BRAZIL)
Santos Jnior, H. L.1; Ecco, R. 2; Elias, F.1; Barros, R. M.1; Correia Neves, D. L1; e Mondadori R. G.1.
1)Unio Pioneira de Integrao Social, 2)Universidade Federal de Minas Gerais

Dr. Helvcio Leal - helvecio17@ig.com.br

Brasil

A crossbred calf, four month-old, was submitted to a necropsy procedure. Macroscopically, cranioventral region of the lungs anterior lobes, showed an intense red and firm cut surface. A white and viscous substance was observed in bronchi and bronchioles. The meninges showed diffuse hyperemia. Three different fragments were sent to a specialyzed laboratory: 1 a 0,5 cm slice from the cerebellum, 2 a 0,5 cm slice from the bulbus and, 3 the rostral end of one telencephalic hemispheres. Lung and nervous system (telencephalic hemispheres, cerebellum, brainstem and trigeminal (Gasserian) ganglion) fragments were collected, fixed in 10% formalin, routinely processed and stained with H&E. Microscopically, the lung showed remarkably suppurative diffuse bronchopneumonia associated with vegetal fibers at bronchioles lumen. The aspiration pneumonia, associated with rabies, is related to the ganglioneuritis and consequently deglutition difficult. Central nervous system microscopical analysis revealed leptomeningitis, diffuse moderate lymphocytes perivascular cuffing, focal gliosis, diffuse nonsuppurative inflammation of trigeminal ganglion and discrete nonsuppurative myelitis. In the cytoplasm of Purkinje cerebellum cells it was observed egg-shape eosinophilic corpuscles, single or multiple, from 2 to 6 micrometer in diameter. The Negri bodies are visualized in approximately 50% of confirmed rabies cases, it depends on the stage and course of the disease, and the virus concentration and strain. The utilizing fluorescent antibody labeling confirmed the rabies diagnostic. Rabies differential diagnostics includes: bovine herpesvirus-5 meningoencephalitis, polioencephalomalacia of ruminants, listeriosis, botulism, tetanus and hepatotoxic plants as senecio spp.

188

POSTER II

BODY CONTACT ON BAT SPECIES HARBORING IN THE SAME DAY ROOST: A POSSIBLE WAY TO INTERSPECIFIC RABIES TRANSMISSION AMONG BATS
W. Uieda 1, P .C. Romijn 2, C.A.M. Cattaneo 3, E.C.N. Rubio 4
1) Universidade Estadual Paulista, Botucatu, SP , Brasil, 2) Centro de Vigilncia Epidemiolgica Ambiental, Secretaria Estadual de Sade do Rio de Janeiro, RJ, Brasil, 3) Secretaria Estadual de Agricultura do Rio de Janeiro, RJ, Brasil, 4) Fundao Educacional Serra dos rgos, Brasil

Dr. Wilson Uieda - wilson.uieda@funasa.gov.br

Brasil

Rabies transmission among individuals of different species has not been easily explained because it depends on the species involved, on the types of relationship among them and on the place where they are harboring. Our goal is to report the interaction among bat species that were harboring in a farm house of the Municipality of Caucaia during August of 2003, 2004 and 2005. This study is part of the field works of a Course of Wildlife Rabies Management organized by the Health Secretariat of Cear, Northeastern Brazil. Seven bat species were observed in the farm house: Molossus molossus, M. rufus (Molossidae), Phyllostomus hastatus, Carollia perspicillata, Anoura caudifera, Trachops cirrhosus (Phyllostomidae) and Noctilio albiventris (Noctilionidae). Body contact was observed during day time in two situations: 1) a frequent body contact, but not constant, was observed among M. molossus, M. rufus and P . hastatus while they were harboring between tile and wood of the roof of a room, 2) a constant body contact (back of one on belly of another) between C. perspicillata and A. caudifera while they were hanging together in the kitchens roof. We suspect that body contact could have occured between M. molossus and N. albiventris because they were observed flying out from the roof by the same openings, though not observed resting together in another roof of the room. During the body contacts, agonistic behavior, licks and bites could have happened among bats of these species and favored the intra and interspecific transmission of rabies while they were harboring together. Just one individual of the carnivorous bat T. cirrhosus was observed in the farm house of Caucaia and it could have eventually participated on the rabies transmission during its predatory behavior on other bat species. The observation of animal behavior in wild is important to understand rabies transmission.

189

POSTER II

PRESENCE OF RABIES VIRUS IN FRUITFUL BAT OF THE SPECIES ARTIBEUS LITURATUS IN VITRIA-ES
A. M. Galimberti, M. A. Guerra, A. L. D. S. Capezzuto, L. C. B. Tavares 1, A. M. Zago
1) Instituto de Defesa Agropecuria e Florestal do Estado do Esprito Santo, ES, Brasil

Dr. Luiz C.B.Tavares - lcbtav@yahoo.com.br

Brasil

Presence of rabies virus in fruitful bat of the species Artibeus lituratus in Vitria-ES A. M. Galimberti1, M. A. Guerra2, A. L. D. S. Capezzuto3, L. C. B. Tavares4, A. M. Zago5 After the capture of four (04) bats in the neighborhood of Consolao, in Vitria, capital of Esprito Santo State, identified as fruitful of the species Artibeus lituratus, two (02) of then were introduce to the urban fauna, and the other two (02), submitted to exam of epidemic investigation in the Laboratrio de Diagnstico de Raiva do Instituto de Defesa Agropecuria e Florestal do Estado do Esprito Santo- IDAF-ES. The samples were negative for the technique of direct immunifluoresce, on March 22, 2006, however, one of the samples was positive in the exam of cerebral inoculation in mice, six days after the inoculation, therefore on March 28, 2006. With the determination of the presence of the rabies virus in the sample, the following actions were accomplished: a) intensified the capture of dogs and cats in the area of Lourdes neighborhoods and Consolao, in an area of approximately 2 km around the positive focus (bat), b) it was priorized and intensified the sending of samples (encephalon) of the removed animals of the neighborhoods of the focus, for diagnosis of rabies, c) the professionals of the Centro de Controle de Zoonoses, qualified thirty (30) community agents of health and six 6 agents of environmental health, that work in the area of the focus, and in this training it was approached, the procedures be taken in the case of a the citizen find a bat in your residence or suffer an aggression, d) concluding, it was accomplished the supply of vaccination for the animals of the area that were not vaccinated in the last campaign of anti rabies vaccination, which reached 83% of covering, and for those that borns after the campaign, totaling two hundred and forty six (246) vaccinated animals.

190

POSTER II

IMMUNE RESPONSE TO BHK-21 CELL RABIES VACCINE PRODUCED AT INSTITUTO BUTANTAN IN BOVINES
Neuza Maria Frazatti-Gallina 1, Regina Maria Mouro-Fuches 1, Daniella Ventini 1, Luiz Souza Reis 2, Sandro Eduardo Arenas2, Paulo Eduardo Pardo 2
1) Instituto Butantan, So Paulo, SP , 2) Universidade do Oeste Paulista - UNOESTE, Presidente Prudente, SP , Brasil

Dr. Neuza Maria Frazatti Gallina - galinajr@uol.com.br

Brasil

A new rabies vaccine, veterinary use, was developed at Instituto Butantan. This vaccine was obtained of BHK-21 cell cultures infected with PV virus strain. It was purified by chromatography, inactivated with -propiolactone and presented in freeze-dried vials. In this study the immunogenicity of this vaccine was evaluated in Nelore bovines. Thirty-six animals were vaccinated with one dose of the Butantans vaccine (Batch IB-01/03 with 1,7 IU/mL). Blood samples were taken on days 74 and 150 to determine the neutralizing antibody titers in BHK-21 cells (RFFIT). In these tests there were utilized the 2nd International Standard for Rabies Immunoglobulin as reference and CVS-11 virus strain for the neutralization. The mean neutralizing antibody titers obtained on days 74 and 150 after the immunization were 3,0 IU/mL and 1,6 IU/mL, respectively and only two of all animals presented antibody titers <0,5 IU/mL. The results obtained showed that the BHK-21 vaccine developed at Instituto Butantan induced a satisfactory immune response in 94% of the animals immunized with only one dose of vaccine.

191

POSTER II

SITUATION OF ANIMAL RABIES IN BRAZIL, 2005


Sabrina Lzaro Mendes 1, Rosely Cerqueira de Oliveira 1, Marcelo Yoshito Wada 1 Lucia Regina Montebello 1
1) Secretaria de Vigilncia em Sade, Ministrio da Sade, DF, Brasil

Dr. Sabrina Lzaro Mendes - mendes.sabrina@gmail.com

Brasil

Introduction: Rabies is a zoonotic disease transmitted for mammals with mortality rate of 100%. No specific treatment is available, but it can be preventable with a prophylaxis at an opportune time. The objective of this study is to describe animal rabies reported in time, place and species in Brazil during 2005. Methods: A descriptive study was performed with the animal-cases rabies which were reported using a standard form to Ministry of Health. State Secretariat of Health monthly report these cases to federal level. Animal cases were described in five different Brazilian Regions and 27 states. Results: A total of 1,638 cases were reported in Brazil. Region North had 121 animal cases: 18 dogs(19.8%), 3 cats(3%), 77 cattle(6,7%), 14 horses(10%), 9 bats(4,5%). Region Northeast had 261 animal cases: 67 dogs(73.6%), 7 cats(7%), 116 cattle(10,1%), 2 horses(44%), 13 bats(6,5%), 3 marmosets(100%), 53 foxes(100%). Region Southeast had 617 animal cases: 449 cattle(39.2%), 53 horses(38.4%), 115 bats(57,5%). Region South had 162 animal cases: 1 dog(1%), 129 cattle(11.2%), 6 horses(4.6%), 26 bats(13%). Region middle-west had 477 animal cases: 5 dogs(5.5%), 372 cattle(32.54%), 63 horses(45.65%), 37 bats(18.5%). Conclusions: Region North and Northeast had reported more rabid dogs and rabid cats, mainly in four states: Par, Maranho, Pernambuco and Bahia. For rabid cattle and rabid horses, two regions had reported more cases, mainly three states: Minas Gerais, Rio de Janeiro and Gois. For rabid bats southeast region and middle-west had more cases, mainly two states: So Paulo and Gois. For wild terrestrial mammals, region Northeast had more reported cases, mainly three states: Cear, Pernambuco and Piau. The results of this study demonstrated different distribution for animal rabies and each state need to implement surveillance according your situation. Animal surveillance for rabies demonstrated an importance to prevent human rabies cases and should be stimulated as routine.

192

POSTER II

CHARACTERIZATION OF DOG AND CAT BITES IN PORTO ALEGRE/BRAZIL IN THE YEAR OF 2005 AND ITS ROLE IN THE SURVEILLANCE OF RABIES
K.M. Dabdab 1, S.M.M. Duro, L.G. Daielo
1) Prefeitura Municipal de Porto Alegre, RS, Brasil

Dr. Karen Dabdab - kdabdab@yahoo.com

Brasil

Objectives: To characterize victims and aggressor animals (dogs and cats) involved in bite and/or scratch incidents in the city of Porto Alegre in the year of 2005 aiming the surveillance of Rabies. Methodology: 789 bite investigations reported in 2005 were analysed, 729 referring to dog bites and 60 referring to cat bites. The investigations were realized by domiciliary visits or by telephone aiming at the guarantee of observation of the bitting animal or remittance of sample for laboratorial diagnostic of Rabies. The analyse information about victims included sex, age, location of the bite wound. The information about the animals included sex, breed, with or without owner, the place where the animal observation was carried out. Results: Mostly of the dog bites had occurred in the legs of the victims (37.58%) and hands/feet (30.59%). The majority of dog bite victims were 0-19 years old (51.02%). Among the children of 0-5 years old, the higher incidence of dog bites was in the head/neck (33.69%) and hands/feet (26.08%). Regarding the bitting dogs, they were mostly males (75.66%), without defined breed (68.31%), and owned (86.28%). In the case of dog bite investigations, 88.34% had been observed and/or conducted to laboratorial examination. With relation to the cat bites, victims were predominantly women (75%) and the higher bite incidence location was hands/feet (66,66%). The aggressive cats were mostly undefined breed (83,33%), and owned (78,33%) and 68.08% of these belonged to the familys victim. Among the cats investigated, 81.66% had been observed and/or conducted to laboratorial examination. Conclusions: To know the profile of victims and the aggressive animals is fundamental to define strategies for bite prevention and education. Prevention of bites should be translated to reduction of the risks of transmision of Rabies.

193

POSTER II

CLINICAL ASPECTS OF RABIES IN BOVINE IN THE CENTRO-OESTE OF BRAZIL


J.R. Borges 1, A.R.C.Moscardini 2, L.M.C.C. Costa 3, A. Bredt 4, A.R. Neto Texeira 1, P .H.J. Cunha 2
1) Universidade de Braslia - UnB, 2) Universidade Federal de Gois - UFG, 3) Agncia Nacional de Vigilncia Sanitria, Braslia - ANVISA, 4) Centro de Controle de Zoonoses, Braslia - CCZ, Brasil

Dr. Augusto Ricardo Coelho Moscardini - augustomoscardini@hotmail.com

Brasil

Introduction Rabies is a disease that shows greater variations in clinical signs than other diseases with neurological manifestation. Vector and virus variant, are responsible for several ways of clinical manifestations, making it difficult to give the precise identification. Objective The goal of this study was to describe clinical aspects of rabies in bovines in the center region of Brazil. Method 12 bovines with rabies, confirmed in immunofluorescence test, were accompanied daily. These animals came from Distrito Federal region and were interned in Hospital Escola de Grandes Animais of UnB/SEAPA. The clinical data were logged in individual chip until the death of the animal. Results All animals showed ataxia, pelvic limbs paresis, sialorrhea, and tongue hypotony. Ataxia was, in all cases, the first sign related by owners. Paresis, sialorrhea and tongue hypotony were found when clinical signs developed to decubitus. Only one animal acquired sternal decubitus until death. Mean time of this disease was 5,3 days with a variation between 2 to 8 days. Other clinical signs were ruminal hypomotility and dehydration, as a consequence of decubitus or anorexia. Just one animal presented hyperthermia. Any animal showed behavioral changes. Conclusion Any disease with neurological clinic signs in bovine cattle must include rabies as a differential diagnosis, mainly, if the evolution time remain between two to eight days. Some clinical signs found in literature were not revealed in this work, as vocal alteration, bellow, aggressiveness, local itch, mutilation, seizures, blindness and head pressure against obstacles. Specific clinical signs for rabies in cattle, as all other disease that affects the nervous system, can not be described.

194

POSTER II

CEREBROSPINAL FLUID EVALUATION IN BOVINE WITH RABIES


A.R.C. Moscardini 1, G.R. Paludo 2, I.I. Ferreira 2, E.P . Novais 2, R.M.B. Rodrigues 3, J.R.J. Borges 2
1) Universidade Federal de Gois, GO, Brasil, 2) Universidade de Braslia, DF, Brasil, 3) Centro de Controle de Zoonoses, Braslia, DF, Brasil

Dr. Augusto Ricardo Coelho Moscardini - augustomoscardini@hotmail.com

Brasil

Introduction Nowadays, cerebrospinal fluid (CSF) analysis, help veterinarians to obtain and to use laboratorial results as central nervous system pathologies diagnosis. By the way, clinical diagnoses of animals through CSF analysis still require a lot of studies to become efficient. Objective To evaluate laboratorial parameters of CSF in bovines naturally infected for rabies virus Method Samples from the cerebelomedullary cistern of 10 bovines with rabies suspicion were collected, chilled on ice and sent for laboratorial analysis in 30 minutes. CSF density, pH, glucose and protein concentration and evaluation of cells were done. Results In physical examination of the CSF , all samples showed a clear aspect, without coagulation. Three animals revealed l and 70mincreased number of erythrocytes and leukocytes (310 blood cells/ l, 120 bloodml and 16 nucleated cells/ml, 86 blood cells/mnucleated cells/ l). Three samples showed normal proportion ofml and 37 nucleated cells/mcells/ lymphocytes (83%, 63%, and 64%). In four cases the percentage of monocytes was increased (> 50%) and in one sample, 30% of neutrophils was found. The protein concentration was increased just in one sample (50 mg/dl). Three CSF presented more than 105 mg/dl of glucose, but this value was considered normal. Only one animal showed lower values of glucose in CSF (28, 7 mg/dl). Conclusion The CSF analysis did not reveal a diagnostic tool for rabies, without any characteristic alterations. Therefore this exam showed an efficacy when showing differentiations of some encephalitis.

195

POSTER II

DIFFERENTIAL DIAGNOSTIC OF BOVINE RABIES IN THE WEST-CENTER REGION OF BRAZIL


A.R.C. Moscardini 1, G.M. Machado 2, M.I.R. Bossil 3, M.B. Castro 2, G.C. Reckziegel 2, J.R.J. Borges 2
1) Universidade Federal de Gois, GO, Brasil, 2) Universidade de Braslia, DF, Brasil, 3) Centro de Controle de Zoonoses, Braslia, DF, Brasil

Dr. Augusto Ricardo Coelho Moscardini - augustomoscardini@hotmail.com

Brasil

Introduction In spite of rabies have been studying so much, this illness in cattle can be easily misdiagnosed with others neurological diseases in these animals. The incidence and emergence of neurological disease in ruminants depend on geographic region. Clinical demonstration of some diseases like polioencephalomalacia, botulism, bacterial encephalitis, black leg and bovine herpesvirus type-5 meningoencephalitis can be confused with rabies, mainly, for clinical signs or for the condition of death. Objective The goal of the present research was to identify the main diseases with neurological signs in bovines of Distrito Federal and Gois that can be misdiagnosed with rabies. Method Through January 2003 to January 2005, 87 ruminants with neurological signs were assisted on Hospital Escola de Grandes Animais UnB/SEAPA. The clinical examination was done in all animals with rabies suspicion. All day, cardiac and respiratory frequency, behavior changes, ocular exams, sensibility test in limbs and anus and paralysis of tail, were evaluated. Rabies was confirmed with imunofluorescence and biological tests. Results 18 diseases with neurological signs were diagnosed. The research showed that rabies had significant importance, with 25% of 87 cases, black leg (8%) and polioencephalomalacia (7%). Leucosis (3%), lead poisoning (3%), botulism (7%), Palicourea marcgravii poisoning (3%), bovine herpesvirus type-5 meningoencephalitis (7%), bacterial encephalitis (3%), cerebral babesiosis (1%), malignant catarrhal fever (1%), pituitary abscess (2%) were also diagnosed. Conclusion At least, 18 bovine neurological diseases might be included as differential diagnosis when imunofluorescence and biological test to rabies indicate negative. Its very import to identify the neurological diseases that are more common in the region because it can help veterinary practcioners that work in field.

196

POSTER II

FIRST DETECTION OF EHV-4 IN AN EQUINE CASE OF SUSPECTED RABIES IN RIO DE JANEIRO STATE, BRAZIL
Carlos Henrique de Azeredo Lima 1, Paulo Eduardo Brando 2, Luz Alba M G Fornells 1, Waldyr Pessanha Junior 3, Carlos Henrique Campello Costa 1, Jorge Luis Teixeira Palmeira 3, Ivanete Kotait 4, Cludio de Moraes Andrade 1
1) PESAGRO, Rio de Janeiro, RJ, Brasil, 2) Universidade de So Paulo, SP , Brasil, 3) SEAAPI, Rio de Janeiro, RJ, Brasil, 4) Instituto Pasteur, So Paulo, SP , Brasil

Dr. Carlos Azeredo - carlos_azeredo@yahoo.com

Brasil

The rabies virus is the most prevalent agent of central nervous system (CNS) disease in herbivores in Brazil, causing encephalopathy and ultimately death. From 1996 to 2004, 29,969 cases of animal rabies were diagnosed all over the country (MAPA, 2004). In the Southeast Region, where Rio de Janeiro is located, 16,706 cases were confirmed in the same period of time (MAPA, 2004). Routinely, diagnosis is carried out aimed at finding only rabies virus. Due to the nature of the disease and its invariably fatal outcome, when rabies virus is not identified, no other agents are routinely searched for. On the other hand a negative result for rabies does not elucidate the causative agent of the encephalitis. EHV and some arboviruses are important and widespread nervous system pathogens in the horse population. This work reports a case of identification of equine herpesvirus-4 from a rabies-negative specimen submitted to the Rabies Reference Laboratory of Rio de Janeiro State. Routine laboratory testing for rabies was performed. The Direct Fluorescent Antibody (DFA) test in four fragments of the nervous system was carried out (marrow, hippocampus, cortex and cerebellum) aimed at finding rabies virus, and was negative. In the biological test, 50-62% of inoculated mice died after inoculation. The pattern of disease in animals in the biological test strongly suggested herpesvirus disease. EHV-1 and EHV-4 were tested for by a PCR assay described by Carvalho et al. (2000). The amplified sample resulted in a fragment of 333bp in the reaction corresponding to EHV-4. The result led to the detection and isolation of EHV-4 and was the first description of this virus in Rio de Janeiro State.

197

POSTER II

DESCRIPTIVE ANALYZE OF RABIES DISTRIBUTION, IN THE YEAR OF 2005, IN RIO DE JANEIRO STATE, BRAZIL
Carlos Henrique de Azeredo Lima 1, Luz Alba M. G. Fornells 1, Waldyr Pessanha Junior 2, Cludio de Moraes Andrade 1
1) PESAGRO, Rio de Janeiro, RJ, Brasil, 2) SEAAPI, Rio de Janeiro, RJ, Brasil

Dr. Carlos Azeredo - carlos_azeredo@yahoo.com

Brasil

Rio de Janeiro State is located at 22 degrees, 54 minutes south latitude, 43 degrees 14 minutes west longitude (22542 S 43142 W). The population is more than 14.391.282, occupying an area of 43.696,054 km. It is divided in six regions in agreement the physical and regional characteristics. The Laboratrio de Biologia Animal, PESAGRO-RIO, located in Niteri City, received 173 samples for the rabies diagnosis, in 2005. These samples were sent by 52 different cities A descriptive analyze were performed to determining distribution and spread of illness, portraying natural history of a health condition and estimate the impact of the rabies cases in the State. It was used samples distributed by city, regions, species and the sample delay time to get into the laboratory. The observed results were that: 32 cities had at list one positive results, the region IV and II together are responsible for more than 69% (35,62% and 34,25%) of the cases, followed by the regions III, V, I and VI (16,44%, 10,95%, 1,37% and 1,37%, respectively), the specie which had suffered more were the bovine with 51,78% (58/112), followed by the equine, 50% (11/22), ovine, 37,50% (3/9) and canine 7,69% (1/13), and 77,63% of the received samples took 6 days or less to reach the Rabies Reference Laboratory of Rio de Janeiro State to be submitted to the routine laboratory testing for rabies. These finds will help to evaluating prevention and control measures, facilitating planning, case detection and public health intervention when necessary.

198

POSTER II

EVALUATION OF RABIES VIRUS NEUTRALIZING ANTIBODIES TITRES OF PETS THAT MOVES TO THE EUROPE UNION FROM BRAZIL
K.C. Scheffer 1, A.M. Batista 1, A.C.R. Silva 1, P .S. Cruz, A.E.B. Costa 1, G.M.M. Caporale 1, L.B. Chaves 1
1) Instituto Pasteur - SP - Brasil

Dr. Andra de Cssia Rodrigues da Silva - acrsilva@yahoo.com.br

Brasil

Since July of 2004, the regulation of European Parliament and of the Council lays down the animal health requirements for both the movement of pets between Member States and from third countries. This regulation recommends a titration of rabies virus neutralizing antibodies (VNA) carried out by an approved laboratory by the Laboratoire dtudes et de recherches sur la rage et la pathologie des animaux sauvages of the Agence franaise de scurit sanitaire des aliments AFSSA. This titration has been performed since February of 2006 at Instituto Pasteur - So Paulo (IP-SP), the first approved laboratory in Brazil. According the World Health Organization, titres e 0,5 IU/mL of VNA protect in case of natural exposure to the rabies virus. The aim of this study was analyze the VNA titres obtained in serum samples of animals received at IP-SP laboratory from February to May of 2006, according the age group of animal. Titration of three hundred eighty eight serum samples of dogs and cats was performed by the rapid fluorescent focus inhibition test (RFFIT) and the data about age group, and specie of animal was analyzed from the file information. Three hundred forty seven animals (89,4%) showed satisfactory VNA titres and in forty one animals (10,6%) the titres were < 0,5 UI/mL, from those the majority was under one year old (63,4%) and had received only one dose of vaccine. The greater part of animals were adults, 195 (50,2%) were between 1 to 6 years old, 69 (17,8%) were above 6 years old, 105 (27,1%) were under one year old and from 18 there was no information about age. These results support the requirement of two doses of vaccine in pet animals at first vaccine.

199

POSTER II

EPIDEMIOLOGICAL PROFILE OF THE CONFIRMED CASES OF ANIMAL RABIES BETWEEN 2000 TO 2005
Ftima Cristina de Souza 1, Deborah Daniela M. T. Carneiro 1, Edgar Pinho Cerqueira 1
1) Secretaria de Sade do Estado da Bahia, BA, Brasil

Dr. Ftima Cristina - fcris@br.inter.net

Brasil

Rabies is an antropozoonosis transmited by mammals throught of virus rabic innoculation contained in the saliva of the animal infected. It is considered as of the serious problem of health public presented letality 100% and largest economic by high cost in the prevention assistence for people subjects at risk. The basics activities developed in the program of rabies include the preventive treatment against human rabies, animal vaccination, laboratorial diagnostic, epidemiological surveillance and health education. The objetive of this article present the epidemiological profile of rabies in the different species distribuided in Bahia State of 2000 to 2005. During this period were analyzed in the Laboratrio Central 8.902 samples of central nervous system of different animal species, coming from Bhatia State municipalities. There were confirmed for rabies 442 dogs, 41 cats, 329 bovines, 37 foxes, 9 equines, 9 sheeps, 5 bats, 4 buffalo, 3 goats and 2 asinine a total of 881 samples. In 2000 of 1.015 samples examined, 202 (19,90%) were positive for rabies, noting that of samples 104 dogs, 21 cats, 71 bovines, 1 bat and 5 others species. In the 2005 of 1.326 samples examined, 62 (4,67%) were positives for rabies with 18 dogs, 38 bovines, 5 foxes and 1 sheep registered. Cases of rabies in dogs and cats have been decreasing drastically while the number of cases in bovines, foxes, equines and bats have been a gradual increase and mark situation of rabies, attentive for system sending of samples, the increase of vaccination of dogs and cats, obligatory vaccination of cattle, horses, sheep and goats, control of the bats population and take precautions with the wild fauna.

200

POSTER II

EVALUATION OF THE RABIES ACTIVITIES REALIZED IN CENTERS OF CONTROL OF ZOONOSIS - BRAZIL


Ricardo da Silva Teixeira Vianna 1, Daniel Andrade Simes Jorge 2, Francisco Anilton Alves Arajo 1, Marcelo Yoshito Wada 1
1) Secretaria de Vigilncia em Sade, Ministrio da Sade, DF, Brasil, 2) Faculdades Integradas do Planalto Central - FIPLAC, Valparaso de Gois, GO, Brasil

Dr. Marcelo Yoshito Wada - marcelo.wada@saude.gov.br

Brasil

Introduction. In Brazil, SVS have financed centers of control of zoonosis(CCZ). They are responsible for different activities, including actions to control rabies in animals. The objective of this study is to describe activities related with rabies control realized in different Brazilian CCZ. Methods. The National Secretariat of Health Surveillance(SVS) created a standard form about activities realized in Brazilian CCZ. This form was sent to CCZ, which fill the form and resent to SVS for evaluation. The data were analyzed in excel. Results. There were sent the form to 240 CCZ and 107(44%) answered the questions from 13 states: 64(60%) So Paulo, 11 (10%) Minas Gerais and 30% were from others states (Bahia, Amazonas, Acre, Rondnia, Roraima, Par, Mato Grosso, Maranho, Rio Grande do Norte, Cear and Tocantins). A total of 103(96%) have control program of rabies for dogs and cats implanted. Routine vaccine is realized in 69(65%), rabies campaigns for dogs and cats in 104(97%) and 69(65%) realized focus block. About samples for diagnosis of rabies, 101(94%) municipalities sent dogs, 93(875), 82(77%) bats, 20(19%) wild terrestrial animals and 37(35%) herbivorous. If we evaluate the praised percentage of 0.2% of samples that need to be sent for diagnosis of rabies, only 37(35%) municipalities reached this goal. Capture and control of bats is realized in 46(43%) CCZ and 65(61%) have an education health program about rabies in the municipality. Conclusions. Rabies program is implanted is almost all CCZ, otherwise some activities as routine vaccination and the praised percentage of 0.2%, capture and control of bats and education health program about rabies need to be implanted.

201

POSTER II

DOG RABIES TRANSMITTED BY A BAT IN URBAN AREA OF TERESINA MUNICIPALITYPIAU STATEBRAZIL2006. REPORTED CASE
Vnia Maria Alves de Carvalho 1, Romualdo Oliveira Spndola 1, Rosngela Cavalcante de Carvalho 1, Joo Pereira da Silva 1, Romildo Francisco dos Santos 1, Marcelo Yoshito Wada 2
1) Centro de Controle de Zoonoses, Teresina, PI, Brasil, 2) Secretaria de Vigilncia em Sade, Ministrio da Sade, DF, Brasil

Dr. Romualdo Spndola - romualdospindola@yahoo.com.br

Brasil

Zoonoses Control Service/ Municipal Secretariat of Health of Teresina municipality / Piau State 2 Zoonoses and Vectorborne Diseases Branch (COVEV). National Secretariat of Health Surveillance (SVS), Ministry of Health-Brazil (MoH) Introduction: Circulation of the rabies virus in Teresina municipality have been identified in different species of animals, causing outbreaks in herbivorous and wild animals. Teresina has demonstrated high vaccination covering in dogs and cats, however cases of rabies in dogs have been identified, mainly in rural area and usually have exposition history of wild animals. This study reports a rabies dog case transmitted by a bat in urban area. Methods: A descriptive study was performed. An investigation was conducted, after laboratorial confirmation. Dog rabies case was confirmed when brain tissue was reactive using direct immunofluorescent(IF) antibody testing, and virus was identified following intracerebral innoculation in suckling mice. Results: On 19, March, 2006, a dog, German Shepherd, four months, non vaccinated for rabies, domiciliated at urban area was attended in a particular veterinarian. The main complaint was poisoning. The animal was presenting vomiting, hypersalivation and prostration, evaluating with parasthesis, paralysis and death on 22, March. During the investigation, it was identified that this animal was bought on January from a hatch of nine puppies in a rural area. The parents were healthy and others puppies werent found. The rabies dog lived with two dogs and one cat and all had received more than one dosis of rabies vaccine, they didnt have access to the street or other animals. Residents reported presence of bats and eventually animals caught bats, which could be confirmed for the zoonoses control service which found a non-vampire-bat on the garden. The dog was confirmed rabies by laboratorial test and it was identified variant three, compatible with Desmodus rotundus. Concluso - This case shows the vulnerability to occur rabies transmitted by bats and the importance to monitor this cycle. It strengthens the necessity of vaccination for rabies in domesticated animals and rapid response of laboratorial test.

202

POSTER II

CHIROPTERA MONITORING IN A CAVE NORTHWEST OF MINAS GERAIS, 20042005


L. V. de Knegt 1, M.H. Marcos 2, M.H.S. Silva 2, C.F .J. Miranda 2, V.M.R. Oliveira 2
1) Secretaria de Vigilncia em Sade, Ministrio da Sade, DF, Brasil, 2) Secretaria de Estado de Sade - MG, Brasil

Dr. Marcelo Henrique Marcos - novo_mar@yahoo.com.br

Brasil

Introduction: Bats have occupied the first place in rabies transmission to humans in Brasil from 2004 to 2005. There are 168 species, of nine families in Brasil and 66, of 8 families, in Minas Gerais state. This richness is due the habitat variety on the state, which encompasses almost all the biomes in the countryside. The Tamboril cave is on the Cerrado area, in the municipality of Una. It is ten kilometers far from downtown and three kilometers far from the road BR-251. Objectives: The objective of this work was to identify the bat species that use the cave entrance and verify the presence of rabies virus on these animals. Methods: Two mist nets (2,5x7m) were set near ground, one night per month, between 6:00PM and 12:00PM, from October 2004 to November 2005. Results: There were captured 421 individuals, of 16 species in 3 families, included the 3 vampire species (Artibeus obscurus, Carollia perspicillata, Chrotopterus auritus, Desmodus rotundus, Diaemus youngi, Diphylla ecaudata, Eptesicus brasiliensis, Glossophaga soricina, Lonchorrhina aurita, Macrophyllum macrophylum, Peropteryx macrotis, Phylloderma stenops, Phyllostomus hastatus, Phyllostomus discolor, Tonatia sp, Trachops cirrhosus). From that, 7 were taken for rabies virus detection (1 Tonatia sp., 1 Trachops cirrosus, 2 Desmodus rotundus, 2 Diphylla ecaudata, 1 Glossophaga soricina). All results were negative. Conclusion: The lack of detection of the virus on the samples does not discard its presence in the bat community in the Tamboril cave, because the prevalence is often low in bat populations and size of sample, small. The high number of species that co-habit in the cave heighten the risk for transmission of rabies virus variants among species of bats. On the other hand, it seems that this species richness is unmatched on caves in Brasil, and the use of this cave in conservation policies should be considered.

203

POSTER II

THE EXPERIENCE OF ACTIVITIES OF BLOCKADE OF RABIES IN BATS IN BOTUCATU - SP , BRAZIL


Andr Peres Barbosa de Castro 1, Denise Aparecida Garcia 1, Jos Reinaldo Gasperini 1, Fernando Heglhammer de Oliveira 1, Valdinei Moraes C. da Silva 1, Jonas Lotufo Brant de Carvalho 2, Cassiano Victria 1
1) Vigilncia em Sade Ambiental, Botucatu, SP , Brasil, 2) Secretaria de Vigilncia em Sade, Ministrio da Sade, DF, Brasil

Dr. Andr P . B. de Castro - saudeambiental@botucatu.sp.gov.br / apbcastro@gmail.com

Brasil

Introduction: The bats are important in maintenance of wild cycle of rabies. In past decades the disordered expansion of cities and agricultural areas, has caused the presence of bats in urban areas. The consolidation of the Equipe de Vigilncia em Sade Ambiental - EVSA of the Botucatu-SP , has taken care to the population requests for resolution of doubts about these animals. Objectives: development and improvement of surveillance activities for rabies in bats and activities of blockade of primary focus for rabies prevention. Method: all dead and presenting behavior alterations bats are collected and send to laboratory for rabies diagnosis. When of notification of a positive case, the EVSA organizes activities of blockade through the re-vaccinated all dogs and cats in within of 500 meters from focus, and advised the inhabitants when of the meeting of bats in residential areas. Results: in the period of november of 2004 at july of 2006, the EVSA carried 11 blockade. In the year of 2004 eight blockade had been carried vaccinated or re-vaccinated 10,046 animals, in 2005 an blockade was made with a total of 2,015 animals, and in the 2006 already they had been re-vaccinated in the two activities 1,535 animals. Conclusions: Activities of blockade have been raised given important data on immunity of dogs and cats population, allowing improvement of annual vaccination program against rabies. In the year of 2005 there was a increment of 12,11% in cats vaccination due to priorization of these species in the animal immunization campaign. The localization of the cases has been typed or a geoprocessing software and has carried the search and surveillance of the colonies in this areas.

204

POSTER II

SURVELLAINCE OF RABIES IN BATS IN THE CITY OF BOTUCATU - SP


Cassiano Victria 1, Joo Edenilson Miranda 1, Jerian Edmundo Athanazio 1, Jonas Lotufo Brant de Carvalho 2, Benedito Donizete Menozzi 3, Helio Langoni 3, Andr Peres Barbosa de Castro 1
1) Vigilncia em Sade Ambiental - Prefeitura Municipal de Botucatu, SP , Brasil, 2) Secretaria de Vigilncia em Sade, Ministrio da Sade, DF, Brasil, 3) Faculdade de Medicina Veterinria e Zootecnia - UNESP Botucatu, SP , Brasil

Dr. Andr P . B. de Castro - saudeambiental@botucatu.sp.gov.br / apbcastro@gmail.com

Brasil

Introduction: Botucatu is located in center-west of So Paulo State, possess great part of your 1,428 km2 as agricultural exploration, however, inside of urban area finding of places of environment preservation, with abundance of trees that serve of shelter or food source, favoring the presence of bats. With consolidation of activities of Equipe de Vigilncia em Sade Ambiental - EVSA, Secretaria Municipal de Sade, for resolution of demands involving bats increased, taking to create bats surveillance program, integrating EVSA and Servio de Diagnstico de Zoonses - SDZ, of Faculdade de Medicina Veterinria e Zootecnia of UNESP , campus of Botucatu. Objectives: work aims at the improvement of bats surveillance program and blockade of primary focus when rabies positive notification, and to advise of population when the presence of these animals. Method: the intervention group of EVSA in cases of died or presenting behavior alterations bats, collects the specimen and send to SDZ for rabies diagnosis. The EVSA also development activities of localization the possible shelters and food sources of bats. This places are marked by GPS equipment for the geoprocessing, making possible the localization of colonies and accompaniment of dynamics these populations. None of colonies of bats is removed or captured, advised only in relation the shelters when these if find in residences. Results: in year of 2004, had been protocolled involving bats 247 demand, in 2005, 432, and in six first months of 2006, 165. In year of 2005 had been sent to ZDS a total of 262 samples, in six first months of 2006, 195 samples. Conclusions: bats program surveillance of colonies has produced of data and information about presence of bats in city, refining the control of rabies urban, a time that data have assisted in blockade of focus of bats rabies when of positive notification.

205

POSTER II

RABIA EM MURCILAGOS NO-HEMATFAGOS EN LA AREA URBANA DE CAAPAVA, SO PAULO, BRASIL


F.S. Galvo 1, M.C.A.M. Souza 1
1) Prefeitura Municipal de Caapava, SP , Brasil, 2) Secretaria de Agricultura e Abastecimento, SP , Brasil

Dr. Fanice Siqueira Galvo - fanfervet@yahoo.com.br

Brasil

La ciudad de Caapava, situada en la regin sudeste del Brasil, entre la Sierra del Mar e la Sierra de Mantiqueira, tiene caractersticas topogrficas que favorecen una amplia distribucin de murcilagos, principalmente el Desmodus roduntus, que es lo principal responsable por la transmisin de la rabia, en la regin. En el inicio de la dcada del 90 hubo un grande aumento del numero de casos positivos para rabia en animales de inters econmico, y despus la Prefectura Municipal pas a orientar intensamente la populacin con relacin a la presencia de murcilagos en las moradas, situadas en la rea urbana. Este trabajo result en el envo de diversos espcimen de murcilagos insectvoros y frugvoros para el laboratorio de diagnstico de la rabia. En mayo del 2004, uno espcimen de Artibeus lituratus fue positivo y en junio, del 2005, uno Myotis nigricans fue encontrado en el Parque Eldorado, periferia de la ciudad. En esta oportunidad fue realizado bloqueo con vacunacin, casa a casa, en perros y gatos, presentes en un rayo de 1000m, orientacin a la populacin, cuanto al riesgo de manipulacin de los murcilagos y pesquisada la presencia de estos animales y sus eventuales abrigos. Verificase que la degradacin ambiental tienen provocado el deslocamiento de los murcilagos para los centros urbanos, en busca de alimento e abrigo, o que, en algunas veces, permite la reintroduccin del virus de la rabia en las reas urbanas, colocando en riesgo los animales de estimacin de los humanos.

206

POSTER II

COURSE OF WILD RABIES - AN ALTERNATIVE TO IMPLEMENT ACTIONS


Nlio Batista de Morais 1, David Caldas Vasconcelos 1, Phyllis Catharina Romijn 2, Carlos Alberto Monteiro Cattaneo 3, Benedito Neilson Rolim 1
1) Secretaria de Estado da Sade do Cear, CE, Brasil, 2) Secretaria de Estado da Sade do Rio de Janeiro, RJ, Brasil, 3) Secretaria Estadual de Agricultura do Rio de Janeiro, RJ, Brasil

Dr. Nlio Morais - neliom@ig.com.br

Brasil

INTRODUCTION: Addressed for the control of urban Rabies, the current scenery appears for elimination of this zoonose. Since the beginning of 1990s that we attended the progress of wild species causing accidents and deaths to the man. OBJECTIVE: Analyzing viable alternatives that make possible the knowledge and decrease the risks of human Rabies cases of the wild atmosphere. MATERIAL AND METHODS: Reports of the National Program of Control of Rabies and of Health Secretary of Par, Maranho, Cear and Piau. RESULTS: In 2004, Brazil presented an inversion in the epidemic scenery, registering the bat (22 cases) as the largest transmitter of human cases front to the dog (05 cases), fact confirmed in 2005. In Brazil from 1990 to 1992 were registered 32 human cases for bat and marmmosets were being identified in Cear, for the transmission of four human cases in 1991. The foxes transmitted cases to the man in the period from 1986 to 1993. In the accomplished investigations, in its majority, victims didnt look for the units of health for ignorance of the risk. CONCLUSION: The absence of a national politics for the wild Rabies, addressed some states to implant control measures. Cear, with the project of control of the Wild Rabies, based in bases of education information, communication and the formation of human resources, for instance, a course of surveillance of the wild rabies. In its fourth edition in 2006 the course was decentralized for state of Piau, in 2007 it will be headquartered in the state of Maranho. Only of the gender in the country, the course has hourly load of 50 hours with theoretical and practical classes, qualifying technicians of Brazilian states. In Cear it is a surveillance tool, contributing significantly so that during the period from 1999 to 2005 the state just registers a human case for transmission of wild rabies in the period previous thirteen cases were confirmed.

207

POSTER II

CAUCAIA-CE: THE SCENERY OF THE RAGE IN THE WORLD


Jarier de Oliveira Moreno 1
1) Secretaria de Estado da Sade do Cear, CE, Brasil

Dr. Jarier de Oliveira Moreno - cetrav@saude.ce.gov.br

Brasil

INTRODUCTION: The municipal district of Caucaia is located in the Northeast area of the state of Cear, with population of approximately 303.970 inhabitants. The diversity of species susceptveis in the wild and rural atmosphere, allied the continuity of the occurrence of canine and feline cases in the urban atmosphere, associated the environmental factors, it establishes a link among those transmission cycles, placing the municipal district in prominence position in the epidemic picture of the rage. OBJECTIVE: to analyze the diversity of species and circulating variants of the area in the period from 1990 to 2005. MATERIAL AND METHODS: The data were collected through the records SEE-7, reports of the Regional Cell of Health of Caucaia, it Programs National of Control of the Rage, laboratories of diagnosis of Cear, Instituto Pasteur and USP-SP . RESULTS AND DISCUSSION: In the above-mentioned period, 83 canine cases were diagnosed, 10 felines, 25 bovine, 05 humans, 06 for fox, 01 for raccoon, 08 for bat and 05 for sagi. In the analyses antignicas, he/she identified variant type 2 in dogs, cats, as well as variant non compatible in sagis. In the year of 2002, he/she stands out an accentuated increase of canine cases in Caucaia (22 cases) what made possible 1 case humans occurrence in 2003. Before that, concrete measures were adopted, through the accomplishment of two annual campaigns of antirabic vaccination starting from 2003, and the implementao of the withdrawal of unrestricted dogs. The actions with relationship to the wild rage were centered mainly potentially in the notification and treatment of the aggressions by the species transmissoras, associated to the education process in health. CONCLUSION: The prophylaxis actions and control of the rage in the municipal district of Caucaia presented satisfactory results in the urban atmosphere. However, the occurrence of cases in several species of wild animals, as well as the narrowing of the connection link with the urban atmosphere suggests new strategies for the control of the rage in the municipal district

208

POSTER II

ANIMAL RABIES IN PERNAMBUCO FROM 2000 TO 2005


Camila Brederode Sihler 1, Francisco Duarte Farias Bezerra 2, Maria de Lourdes de Almeida Nunes 2, Danielle Mendona Ferreira 2, Vernica Isabel de Brito Alencar 1) Ministrio da Sade, Pernambuco, PE, Brasil, 2) Secretaria de Estado da Sade do Pernambuco, PE, Brasil Dr. Camila Brederode Sihler - camilasihler@gmail.com Brasil

The goal of eliminating Human Rabies as transmitted by dogs, established by the OPAS and approved by American countries, will require a great effort by the federal government. The epidemiologic vigilance of animal rabies, through vaccination, monitoring of the viral circulation through laboraty examinations and the identification of infected species, are some of the instruments of control. Objective: Mapping all cases of animal rabies in the State of Pernambuco, from 2000 to 2005. Method: After laboratory confirmation of animal rabies using the imunofluorescence test and a biological test carried out by LANAGRO (National Agricultural Laboratory of Pernambuco) the Environment Agency is informed using a standardized notification from the laboratory and a regulamentary VE-7 notification. These describe the species, time and place. Results: 612 cases were identified during the period (64% dogs, 11% cats, 12% cows, 1,5% horses, 11% wild animals and 0,5% sheep). The largest number of cases was in 2002, with 194 cases. Regarding cases in dogs, 50% of the cases were in metropolitan outskirts, especially in the municpalities of Recife, Jaboato and Cabo, whilst the interior represented the lowest number of cases (4,5%). Conclusion: The State of Pernambuco presented a reduction in the number of cases of animal rabies during the period, in the majority of the species, probably due to greater vigilance, such as the monitoring of viral circulation through laboratory examinations, the increase in vaccination, opportune blockades of outbreaks and the capture of stray animals. Notwithstanding, with the implementation of the monitoring of wild animals (an increase in material being sent to the laboratory) the number of cases increased, especially regarding bats. It is therefore recommended that the responsible ownership of animals, antirabies vaccination and education regarding the castration of pets be stimulated, stray dogs lose their protection and educative programs be developed.

209

POSTER II

ZOONOSIS CONTROL VERSUS SOCIAL PLEAS


M.L.A.B. Reichmann 1, R.G.F. Cordeiro 1
1) Instituto Pasteur, So Paulo, SP , Brasil

Dr. Maria de Lourdes Aguiar Bonadia Reichmann - mreichmann@saude.sp.gov.br

Brasil

The state of So Paulo has 645 municipalities and 40 million inhabitants. The pet population here has increased due to economic and social changes. Procedures for zoonosis control have been recommended by official bodies of knowledge, as WHO, the Health Ministry of Brazil and the State of So Paulo Health Office. It is necessary and fundamental that authorities develop a common understanding on the subject, including the executive, judiciary branches and the legislature to promote public health and preserve the natural environment. The executive branch should develop systematic programs with unequivocal, exact purposes that define zoonosis control, based on technical- scientific foundations. The legislature enacts laws regulating the interaction among humans and animals. The judiciary branch, to try concrete lawsuits proposed by the organized civil society, thus needs technical scientific grounds at its disposal, besides legal support for that purpose. The Attorney General submits conciliatory actions (Conduct Adjustment Term) to settle impasses generated by the conflict between the organized civil society and the zoonosis control services. The organized social civil society shall operate using definitions recognized by the different public branches. Health departments should offer the scientific grounds to respect such social plea. Only regular, continual meetings may establish consonance among the parties, furthering public health and environmental preservation, animal welfare and the respect to the legislation in force.

210

POSTER II

RABIES IN THE NORTHWESTERN SAO PAULO STATE, BRAZIL: ANTIGENIC AND GENETIC CHARACTERIZATION OF VIRUS ISOLATES
L.H. Queiroz da Silva 1, S.R. Favoretto 2,5, E.M.S. Cunha 3, L.F . Martorelli 4, W.A. Pedro 1, K. Iamamoto, 1, M.C. Lopes 1, D.R. Sacramento 5, E.L. Durigon 5
1) Universidade Estadual Paulista, Araatuba, SP , Brasil, 2) Instituto Pasteur, So Paulo, SP , Brasil, 3) Instituto Biolgico, So Paulo, SP , Brasil, 4) Centro de Controle de Zoonoses, So Paulo, SP , Brasil , 5) ICB/USP , So Paulo, SP , Brasil

Dr. Silvana Regina Favoretto Lazarini - srfavoretto@usp.br

Brasil

During 1993 to 2003 the Rabies Laboratory at UNESP/Araatuba, in Northwestern Sao Paulo State, examined a total of 7,821 samples and 493 resulted positive, most of it (71 %) corresponding to dogs from an urban rabies epizootic registered in Araatuba area until 1997. In 1998 there was a change in the rabies epidemiological profile and from 1998 to 2003, in contrast to a single case of dog rabies, 31-bat rabies has been detected in that geographical area. Epidemiological studies were done by using antigenic and genetic characterization of those rabies virus (RABV) isolates obtained from different animal species. A total of 86 were tested by a panel of eight monoclonal antibodies (CDC origin) and 79 samples were genetically characterized by sequencing the nucleoprotein gene region from 1,157 to 1,476 positions. During 1993 to 1997 antigenic variant 2 (AgV-2), related to rabies virus maintained by dog populations was the responsible for rabies cases in dogs, cats, cattle and horses and antigenic variant 3 (AgV-3), related to Desmodus rotundus was detected in cattle from rural areas, with the presence of vampire bats attacks. After 1998 rabies was detected only in bats and four distinct antigenic variants were found: AgV3 in 50% (13/26) of the samples and in most of the cities (8/11), antigenic variant 4, related to Tadaria brasiliensis in 15% (4/26) of the samples in four cities and two different antigenic patterns not previously established by monoclonal panel (NC-1 in 31% of the cases and NC-2 in only one case). Nucleotide similarity analysis resulted in two main groups, representative of dog (A) and bats (B) genetic variants, this one divided in four sub-groups. The sub-groups was formed by frugivorous and insectivorous bats, cattle and one dog, showing different endemic cycles maintained by different animal species in this region.

211

POSTER II

IMPLEMENTACIN DE UN SISTEMA DE VIGILANCIA DE RABIA EN VAMPIROS EN LA COSTA DEL DEPARTAMENTO DE LIMA - PERU
J. Gmez-Benavides 1, A. Velasco-Villa 2, A. Escudero-Quintana 3, A. Cahuina 4, M. Chun 4, M. Salvador 4, I. Vargas 4, M. Villanueva 5, L. Suarez 1, C. E. Rupprecht 2
1) Direccin General de Epidemiologa del Ministerio de Salud, Lima, Peru, 2) Centers for Disease Control and Prevention, Atlanta, GA, USA, 3) Direccin de Salud II Lima Sur, Peru, 4) Direccin de Salud III Lima, Peru, 5) Centro Antirrbico de Lima - Direccin de Salud V Lima Ciudad, Peru

Dr. Jorge Luis Gomez Benavides - jgomez@oge.sld.pe

Peru

Introduccin En los valles costeros se ha identificado al D. rotudus, desde hace ms de 30 aos, y la primera epizootia de rabia silvestre fue notificada en julio del 2004, en el Valle de Mala, a 80 Km de Lima, cuando se detecta un mono (Cebus apella) y un vampiro, positivos a rabia por IFD, con Variante Antignica 3. Objetivo: Describir la implementacin y resultados de la vigilancia de rabia silvestre. Mtodo: Se revis los reportes elaborados por los servicios de salud, desde julio 2004 hasta julio 2006. Resultados: Dentro de las actividades ejecutadas se realiz la capacitacin a personal de salud sobre la rabia y tratamiento antirrbico, entrenamiento de 3 equipos de 8 personas en medidas de vigilancia y control de rabia silvestre en los 2 valles donde se identificaron epizootias, implementados con equipos de proteccin personal (incluyendo respiradores N95) y redes, con pre-exposicin antirrbica con VERO, identificacin y geo-referenciacin de 40 refugios de murcilagos, 6 tenan colonias positivas a rabia, se captur 1,526 murcilagos (>98% vampiros), 19/775 muestras fueron positivas a rabia (2.5%), eliminacin de 682 vampiros, bsqueda de signos neurolgicos en mamferos domsticos, ndice de mordeduras en el ganado 10%, sensibilizacin de grupos de riesgo y educacin sanitaria de la comunidad. Se identificaron 8 personas con mordeduras por vampiros, 7 de ellas en zonas urbanas, en 2/8 el vampiro era rabioso (IFD), del equipo de vigilancia, 9 fueron mordidas por murcilagos, todos recibieron vacuna antirrbica. Varios refugios tenan caractersticas comunes: ubicacin en tneles de canales de regado, longitud de 15 a 1500 m, oscuridad y humedad. Impacto: ausencia de casos humanos y de casos en ganado. Conclusin: La implementacin de la vigilancia y de medidas de control en forma oportuna reduce los riesgos de transmisin de la rabia de vampiros a los humanos y mamferos domsticos.

212

POSTER II

EFICACIA A CAMPO DEL VAMPIRICID GEL PARA EL CONTROL SELECTIVO INDIRECTO DE LOS MURCILAGOS HEMATFAGOS (Desmodus rotundus - E. Geoffroy, 1810) EN BRASIL
Neide Judith Faria Oliveira 1, Celso Akio Maruta, Edson Moura, Elbi Bernardes Queiroz Jnior, Emerson Marques Botelho
1) Valle S/A, Montes Claros, MG, Brasil

Dr. Neide Judith Faria de Oliveira - neide@vallee.com.br

Brasil

Desmodus rotundus es el principal transmissor de rabia de los herbivoros, as con el objetivo de evaluar la eficacia en la reduccin del ndice de ataque de los murcilagos (D. rotundus), una preparacin de varfarina de uso dorsal fue desarrolada. El experimento fue realizado en el estado de So Paulo, Brasil La poblacin inicial estimada de murcilagos fue 55 animales. Todas las propiedades rurales alrededor del rea fueran envolvidas, garantizando el control nun rayo de seis kilmetros del abrigo. Fueron localizadas diez propiedades que tenan bovinos y equinos, se busc el tratamiento de todos los animales atacados o no. El ndice de ataque, obtenido dividindo-se el nmero de mordeduras de murcilagos por el nmero de animales fue registrado antes y siete das despus del tratamiento. Despus de la contencin, los animales fueron inspeccionados, se hizo los registros de mordeduras y a preparacin de varfarina en gel (concentracin de 1%) fue empleada sobre la lnea dorsal de todos animales. La dosificacin utilizada fue la preconizada para el producto, 8,5g/100kg de peso corporal. De 1.258 animales, 1.212 bovinos y 46 equinos fueron tratados, totalizando 99%. El ndice de mordeduras fue del 20% antes del tratamiento. Siete das despus, el ndice de ataque registrado fue del 3%, significando reduccin del 85%. Adems, uma semana despus del tratamiento, en la colonia, se verific 18 murcilagos y de esos, 11 tenian alteraciones comportamentales, comprometiendo la viabilidad de esos en la naturaleza. Eso resultara en una reduccin del 87% en la poblacin en ese abrigo, esas eficacias fueron similar a otros mtodos eficaces de control de esos transmisores de rabia. Esos resultados indican que el Vampiricid Gel, deve ser usado en rebaos bovinos e equinos para control de la poblacin de murcilagos hematofagos y como una medida adicional al control de la rabia en herbvoros.

213

POSTER II

A CANINE KEPT IN OBSERVATION AFTER CONTACT WITH A RABIES POSITIVE TADARIDA BRASILIENSIS
E.P . Caldas 1, J. Predebon 1, A.A. Farinatti 1, N. Barreto 1, S.M. Pacheco 2, E. Zirbes 3, M.F .R. Machado 4, F.D. Andrade 4, H.B.C.R. Batista 5, J.C.A. Rosa 5, P .M. Roehe 5
1) Secretaria Estadual de Sade do Rio Grande do Sul, RS, Brasil, 2) Pontifcia Universidade Catlica do Rio Grande do Sul, RS, Brasil, 5) Universidade Federal do Rio Grande do Sul, RS, Brasil

Dr. Eduardo Caldas - eduardo.caldas@hotmail.com / eduardo-caldas@saude.rs.gov.br

Brasil

On february 10, 2006, in the urban area of Montenegro, RS, a seven months old Schnauzer bitch, non vaccinated against rabies, got in direct contact with an insectivorous bat Tadarida brasiliensis, by keeping it alive inside its mouth. Nervous tissue of the bat was collected and submitted to routine rabies laboratory testing, which resulted positive. Antigenic characterization of the isolate againts a panel of anti-lyssavirus monoclonal antibodies revealed an antigenic profile commonly found in viruses originated from haematophagous bats, despite recovered from an insectivorous bat. The bitch was maintained in observation for 180 days in an isolation facility constructed with two compartments, allowing alternate opening and facilitating cleaning and feeding without compromising the operators safety. The bitch was submitted to clinical exams weekly. At the end of observation period, the dog displayed no clinical signs compatible with rabies.

214

POSTER II

GRAY FOX RESEARCH TO SUPPORT ORAL RABIES VACCINATION PROGRAMS IN TEXAS: AN OVERVIEW
Jeff Root 1, Brian T. Mesenbrink 1, Bruce Leland 2, Mike R. Dunbar 1, Guy Moore 3 Randy W. DeYoung 4, Angeline Zamorano 4, Robert G. McLean 1
1) USDA-APHIS-WS-National Wildlife Research Center, Ft. Collins, USA - 2) USDA-APHIS-WS, San Antonio, USA - 3) Texas Department of State Health Services, Austin, USA - 4) Caesar Kleberg Wildlife Research Institute, Texas A&M University-Kingsville, Kingsville, USA

Dr. Jeff Root - jeff.root@aphis.usda.gov

USA

A study examining the ecology of the gray fox in relation to oral rabies vaccination programs in Texas was initiated in 2005 by NWRC scientists in cooperation with Texas Wildlife Services and Texas Department of State Health Services. A major objective of this study is to document the movements and the potential of long-distance movements of gray fox near a recent ORV zone break in west-central TX. To date, we have already documented several male gray fox which have moved many km in straight-line distance. An additional major objective of this project involves a landscape-genetics approach to assist gray fox oral rabies vaccination strategies in Texas. Specific objectives include 1) the identification of landscape features influencing dispersal and gene flow, 2) to estimate dispersal rates, and 3) to examine sex bias in dispersal. This study will be of great complimentary value to the telemetry study listed above. To date, in cooperation with the aforementioned collaborators, we have already collected over 400 DNA samples from gray fox. A third major objective, which compliments both genetic and telemetry studies, is the development of GIS habitat layers. These layers are presently being built and will be produced and maintained throughout the duration of this project.

215

POSTER II

POBLACIONES CON ALTA CONSANGUINIDAD COMO FACTOR DESENCADENANTE DE BROTES DE RABIA EN LAS POBLACIONES DE VAMPIROS
Ignacio Amezcua Osorio 1, Toni Piaggio 2, Elizabeth Prez Torres 1, Ral Clmaco Fernndez 1, Alejandro Jimnez Ramrez 3, Dennis Kohler 4, Luis Lecuona 5
1) Pecuary Promotion and Protection Committee in San Luis Potosi State, Mexico, 2) APHIS Science Fellow / Research Molecular Biologist Wildlife Genetics USDA / National Wildlife Research, 3) National Service of Agricultural and Food Health, Safety and Quality (SENASICA), 3) Ministry of Agriculture, Livestock, Rural Development, Fisheries and Nutrition (SAGARPA), Mexico, 4) Analytical Chemistry Project. National Wildlife Research Center, 5) USDA/APHIS/IS NAR, Mexico City, Mexico

Dr. Ignacio Amezcua Osorio - ignacio_murcio@hotmail.com

Mxico

Desde que se desarrollaron las tcnicas de control de las poblaciones de vampiros, se establecieron las bases para la prevencin y control de los brotes de rabia en el ganado en el Latinoamrica. Ahora es necesario la bsqueda de nuevas alternativas de control que estn basadas en el comportamiento del virus rbico en la poblaciones de vampiros, de esta forma combatiremos el origen del problema en estos quirpteros y por ende a la salud pblica y animal. El trabajo presentado en la reunin Internacional de Rabia en las Amricas, en Ottawa, Canad en el 2005, en el cual se mostr el hallazgo de un Desmodus rotundus con albinismo Tipo 1, como parte del control de la Rabia Paraltica Bovina en el estado de San Lus Potos, Mxico, por el Biol. Ignacio Amescua, nos dio las bases para pensar que en esas poblaciones de vampiros existe un alto grado de consanguinidad, factor que est siendo de mucha importancia para que el virus de la rabia se haya diseminado rpidamente en el estado, esto basado con los registros de focos de rabia en bovinos y las variantes antignicas identificadas, mismas que corresponden a los vampiros. En el ao 2001, las colonias de vampiros en el estado de San Luis Potos, fueron alcanzadas por el virus de la rabia, teniendo un comportamiento de distribucin que no corresponda al que se vena presentando en los estados de Veracruz e Hidalgo, y los cuales iban avanzando en promedio ao con ao alrededor de 30 a 40 kilmetros, fenmeno que no sucedi en San Luis Potos en el ao 2004 se pudo registrar el desplazamiento del virus en las poblaciones de vampiros de hasta 150 kilmetros en un solo ao, sorprendiendo a las autoridades para evitar su propagacin. Esto nos hizo pensar que algo estaba sucediendo en las poblaciones de vampiros, pero ignorbamos hasta entonces dicho factor o factores. Con el descubrimiento de un vampiro albino y al reporte del personal operativo donde se seala la identificacin de murcilagos hematfagos de pelaje ms claro del habitual negro o marrn oscuro, que puede ir desde el naranja al dorado o amarillo claro. Nos acerca ms a la teora de que existe un factor gentico en las poblaciones

216

POSTER II

de vampiros que predispone a la diseminacin del virus de la rabia en ellas y llevandonos a colaborar con la Dra. Toni J. Piaggio de la Universidad de Colorado con investigaciones cientficas a partir del material biolgico colectado, quedando pendiente an los resultados. Los resultados de estos estudios nos pueden brindar informacin valiosa sobre la biologa de los murcilagos hematfagos, as como el grado de susceptibilidad o resistencia que tienen este grupo de vampiros al virus rbico y continuar con la bsqueda de respuestas a nuevas incgnitas con el desarrollo de nuevas lneas de investigacin que amplen el conocimiento sobre la etologa del virus de la rabia en estos quirpteros y en un futuro tener la posibilidad de tener alternativas de prevencin de la transmisin hacia el ganado y el hombre.

217

POSTER II

VACCINATING CATTLE TO PROTECT AGANIST RABIES TRANSMISSION FROM RABID BAT POPULATIONS EN MEXICOS ENDEMIC ZONES: AN ANALYSIS USING ECONOMIC MODELS OF THE BENEFITS AND COST
S. Shwiff 1, R.A. Jimnez 2, O.L. Lecuona 3, K. Kirkpatrick 1, D. Kohler 1
1) United States Department of Agriculture / Wildlife Service, Colorado, USA, 2) National Campaign of Paralytic Rabies in Bovines, Mexico, 3) United States Department of Agriculture / Animal Plant Health Inspection Services, Mexico

Dr. Alejandro Jimenez - jdrbovin@senasica.sagarpa.gob.mx

Mxico

Rabies can be transmitted by vampires bats to domestic mammals through its unique feeding behavior. Vampire bats are a main vector of the disease and domestic cattle are the preferred prey, limiting the production capabilities of ranchers. Horses, sheep, pets and even humans may also be bitten and infected. Currently, multiple methods using anticoagulants for control of vampire bat populations exist and are being utilized. Additional problems arise when trying to understand the vampire bat vectored rabies effects in Mexico. Its has been estimated that only 40% of the rabid cattle cases are reported and recorded each year, grossly underestimating the economic loss due to the disease. Cattle interactions with vampires bats pose a potential economic loss to the Mexican economy for several reasons. Ranchers do not have the economic information necessary to determine if vaccination of their range beef cattle is cost effective. It is common for ranchers to under-report the incidence of rabidity in cattle for fear of increased economic impacts due to the reduction in the sale of their beef. Therefore, it is also unknown if vampire bat rabies is having an impact on the local economy due to the loss of cattle. A rabid cow must be destroyed and the rancher loses the asset value of the beef cow and all fixed costs of raising the cow. Additionally, this loss of cattle in an area may be have localized impacts on the economy. The purpose of this study is to develop a model of the economic issues surrounding the vampire bat rabies problem facing cattle ranchers in Mexico to correctly estimate the impact of the disease and identify which methods of preventions are most cost effective. Examine the benefits and cost over time of beef cattle vaccination. This may include an input-output analysis of localized and regional revenue and employment impacts due to vampire bat transmitted rabies.

218

POSTER II

POPULATION GENETIC STUDY OF DESMODUS ROTUNDUS IN AN AREA OF HIGH BAT RABIES INCIDENCE IN CATTLE, SAN LUIS POTOS STATE, MEXICO
Toni Piaggio 1, Ignacio Amezcua Osorio 2, Melissa Neubaum 1, Elizabeth Prez Torres 2, Ral Clmaco Fernndez 2, Alejandro Jimnez Ramrez 3, Ana Lilia Sandoval-Sanchez 4, Luis Lecuona 5, Dennis Kohler 1
1) United States Department of Agriculture/Wildlife Service Colorado, USA. 2) Pecuary Committee in San Luis Potosi State, Mexico, 3) National Campaign of Paralytic Rabies in Bovines, DF, Mexico, 4) Laboratory of Conservation Medicine ESM/IPN, Mexico, 5) United States Department of Agriculture / Animal Plant Health Inspection Services, Mexico

Dr. Alejandro Jimenez - jdrbovin@senasica.sagarpa.gob.mx

Mxico

Incidences of bat rabies in cattle have significantly increased in the state of San Luis Potos. There is still a need for alternative control methods based on knowledge about the behavior of these bats to control the spread and rate of occurrence in this region. Since D. rotundus transmits this disease to cattle we are taking a multi-disciplinary, multi-agency approach between SAGARPA, the State Committee of San Luis Potos, ESM/IPN, APHIS/IS, and APHIS/WS/NWRC to manage this disease. Based on data collected by SAGARPA, cases of bat rabies in cattle appear to have progressed from the southern regions of Mexico into more northern regions. Only 4 cases of rabies in cattle were reported from 1997-2001, it is believed that these cattle were initially transported from other states. Between 2002 and 2006, 277 cases were reported, thus it appears that rabies has increased in this area. In fact, San Luis Potos and neighboring states now account for the highest number of rabies cases in cattle in Mexico (60%). At the last International Meeting for Rabies in 2005, we presented a discovery of an albino Type 1 D. rotundus. There also have been D. rotundus pelage color variants identified from this area. The albino and other pelage variants could suggest low genetic diversity in these populations of D. rotundus, which could account for the expression of a recessive trait such as albinism and associated color variants. If this is true, this lower diversity may mean that these bats have a susceptibility to the rabies virus resulting in a higher prevalence in these populations. Alternatively, these bats may not have lower diversity. A fine-scale population level study of D. rotundus populations from San Luis Potos will help elucidate the relationship of population differentiation and population connectivity to the occurrence and prevalence of rabies and facilitate formulation of targeted management plans. This is a portion of an international multi-agency collaboration to support the Mexico/U.S. bi-national agreements of monitoring and surveillance for wildlife diseases.

219

POSTER II

PLATELIA RABIES II: A HELPFUL TOOL FOR MONITORING RACCOONS RABIES VACCINATION PROGRAMS BY ANTIBODIES TITRATION
M. Feyssaguet 2, I. Blanchard 2, S. Moore 1
1) Kansas State University, College of Veterinary Medicine, Kansas, USA - 2) Bio-Rad, Marnes-la-Coquette, France

Dr. Irene Blanchard - Irene.Blanchard@bio-rad.com

USA

Oral rabies vaccination (ORV) of wild Carnivores species takes an important place in the containment and eradication of rabies from one territory. To measure the success of ORV programs, serums of sampled animals are tested for anti-rabies antibody. A new microplate assay, PLATELIA RABIES II, for the detection and titration of antirabies antibodies in animal serum or plasma, has been developed by Bio-Rad Company and is validated for a use on dogs, cats and foxes. PLATELIA RABIES II is based on the ELISA technique with a microplate coated with an inactivated and purified Glycoprotein antigen and uses a Protein A-POD conjugate. Two protocols, qualitative or quantitative give the possibility of precise titration and the entire assay can be automated. The PLATELIA RABIES II assay has been evaluated at the Rabies Laboratory, Kansas State University, on 187 Raccoons (Procyon lotor) samples that were tested in parallel with the reference method RFFIT. Though each method has technical specificity, results using a 0.5 IU/ml threshold level show good correlation between ELISA and RFFIT and reveal that the PLATELIA RABIES II is able to detect anti-rabies glycoprotein antibodies with 100% specificity, 79% sensitivity and excellent reproducibility. The elimination of the need for high-level biohazard facilities, the ease of use, in addition to the rapid and reproducible results make the PLATELIA RABIES II kit a helpful tool in serological surveys following oral vaccination of raccoons.

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