Introduction Strains of Leptospira interrogans belonging to two very closely related serovars, Ic... more Introduction Strains of Leptospira interrogans belonging to two very closely related serovars, Icterohaemorrhagiae and Copenhageni, have been associated with disease in mammalian species and are the most frequently reported agents of human leptospirosis. They are considered the most pathogenic serovars and represent more than half of the leptospires encountered in severe human infections. Material and Methods Nineteen such isolates from the United Kingdom – human, domestic and wildlife species – were typed using three monoclonal antibodies (F12 C3, F70 C14 and F70 C24) in an attempt to elucidate their epidemiology. They were further examined by restriction endonuclease analysis (REA), multiple-locus variable-number tandem repeat analysis (MLVA) and lic12008 gene sequence analysis. Results Monoclonal antibody F12 C3, which is highly specific for Icterohaemorrhagiae and Copenhageni, confirmed that all the strains belonged to these two serovars. Sixteen strains were identified as Copen...
<p>Phylogenetic reconstruction based on maximum likelihood estimation. Branch lengths are p... more <p>Phylogenetic reconstruction based on maximum likelihood estimation. Branch lengths are proportional to the number of substitutions per site and branch values are the bootstrap values assigned to the edges (i.e. the branch support values).</p
<p>MAT titres of reference serogroup antigens with antisera specific for each GWTS Isolates... more <p>MAT titres of reference serogroup antigens with antisera specific for each GWTS Isolates 1, 2 & 3.</p
<p>Genomic DNA from GWTS isolates #1 (1), #2 (2) and #3 (3) were compared by REA to that of... more <p>Genomic DNA from GWTS isolates #1 (1), #2 (2) and #3 (3) were compared by REA to that of <i>L</i>. <i>alstonii</i> isolates of serovar Pingchang (4), serovar Sichuan (5), MS 267 (6), MS 311 (7) and MS 316 (8). L = DNA Marker.</p
The greater white-toothed shrew (Crocidura russula) is an invasive mammalian species that was fir... more The greater white-toothed shrew (Crocidura russula) is an invasive mammalian species that was first recorded in Ireland in 2007. It currently occupies an area of approximately 7,600 km2 on the island. C. russula is normally distributed in Northern Africa and Western Europe, and was previously absent from the British Isles. Whilst invasive species can have dramatic and rapid impacts on faunal and floral communities, they may also be carriers of pathogens facilitating disease transmission in potentially naive populations. Pathogenic leptospires are endemic in Ireland and a significant cause of human and animal disease. From 18 trapped C. russula, 3 isolates of Leptospira were cultured. However, typing of these isolates by standard serological reference methods was negative, and suggested an, as yet, unidentified serovar. Sequence analysis of 16S ribosomal RNA and secY indicated that these novel isolates belong to Leptospira alstonii, a unique pathogenic species of which only 7 isolates have been described to date. Earlier isolations were limited geographically to China, Japan and Malaysia, and this leptospiral species had not previously been cultured from mammals. Restriction enzyme analysis (REA) further confirms the novelty of these strains since no similar patterns were observed with a reference database of leptospires. As with other pathogenic Leptospira species, these isolates contain lipL32 and do not grow in the presence of 8-azagunaine; however no evidence of disease was apparent after experimental infection of hamsters. These isolates are genetically related to L. alstonii but have a novel REA pattern; they represent a new serovar which we designate as serovar Room22. This study demonstrates that invasive mammalian species act as bridge vectors of novel zoonotic pathogens such as Leptospira.
Infection, genetics and evolution : journal of molecular epidemiology and evolutionary genetics in infectious diseases, 2015
Strains of Leptospira interrogans belonging to two very closely related serovars - Bratislava and... more Strains of Leptospira interrogans belonging to two very closely related serovars - Bratislava and Muenchen - are known to cause widespread infection of the horse population in many parts of the world. Conventional serological typing of isolates has been unable to differentiate between wildlife, pig, dog and possibly horse maintained isolates and therefore has been unable to provide further insight into their diversity and the relationship between them. Twenty-one such European isolates of serovar Bratislava and Muenchen were examined by restriction endonuclease analysis and multiple-locus variable-number tandem repeat analysis in an attempt to elucidate their epidemiology. The restriction pattern types were identified and fell into one of four REA designed pattern types, B1, B2a, M1, M2a. Nine strains from Northern Ireland and two from Germany belonged to B2a, which is a ubiquitous strain being originally isolated from a large number of wild and domestic animal species in the UK. Fi...
In order to manufacture many kinds of fiax fabric, it is necessary for the fihre in the yam to lo... more In order to manufacture many kinds of fiax fabric, it is necessary for the fihre in the yam to lose some of its non-ceUulosic components (ie, hemicelluloses, pectin, etc.) hefore it is made into cloth. An accurate determination of the remaining noncellulosic constituents left ...
A total of 855 sera from dogs in Greece were tested for antibodies to strains belonging to the Po... more A total of 855 sera from dogs in Greece were tested for antibodies to strains belonging to the Pomona, Grippotyphosa and Australis serogroups of Leptospira to assess exposure levels to these serogroups, possible associations with clinical disease and to evaluate whether these findings support the inclusion of additional serovars in dog vaccines. Antibodies were detected in 110 (12·9%) dogs. The highest seroprevalence (4·9%) was to the proposed novel serovar Altodouro belonging to the Pomona serogroup. This serovar also showed a statistically significant association with clinical disease. Serovar Bratislava antibodies were found in 3·4% of sera. Consideration should be given to the inclusion of serovars belonging to the Pomona serogroup and serovar Bratislava in future dog vaccines for the Greek market.
A Leptospira strain (designated RIM 139) was isolated from the kidney of a house-mouse, Mus muscu... more A Leptospira strain (designated RIM 139) was isolated from the kidney of a house-mouse, Mus musculus, in Trás-os-Montes e Alto Douro region of northern Portugal. The isolate showed typical leptospiral motility and morphology under dark-field microscopy and was pathogenic for hamsters. Species determination was carried out on basis of PCR products generated by species-specific primers and by sequencing of the secY gene. Putative serogroup typing was performed using the microscopic agglutination test (MAT) with a panel of rabbit anti-Leptospira sera representative of the major pathogenic serogroups. Serovar identification was carried out by a combination of monoclonal antibodies and cross-agglutinin absorption test (CAAT). The novel nature of the strain was confirmed by restriction endonuclease analysis (REA). Results showed that RIM 139 represents a new serovar. The name Altodouro is proposed for this new serovar. Speciation findings – PCR analysis of the ompL1 gene and sequencing of the secY gene – indicated that it belonged to Leptospira kirschneri as did amplification with G1/G2 and B64-I/B64-II the primer sets. Highlights ► A new virulent Leptospira strain was isolated from the house-mouse in Portugal. ► The strain was typed as Leptospira kirschneri, serogroup Pomona. ► MAbs, CAAT and REA analysis showed that the strain is a new serovar.
Introduction Strains of Leptospira interrogans belonging to two very closely related serovars, Ic... more Introduction Strains of Leptospira interrogans belonging to two very closely related serovars, Icterohaemorrhagiae and Copenhageni, have been associated with disease in mammalian species and are the most frequently reported agents of human leptospirosis. They are considered the most pathogenic serovars and represent more than half of the leptospires encountered in severe human infections. Material and Methods Nineteen such isolates from the United Kingdom – human, domestic and wildlife species – were typed using three monoclonal antibodies (F12 C3, F70 C14 and F70 C24) in an attempt to elucidate their epidemiology. They were further examined by restriction endonuclease analysis (REA), multiple-locus variable-number tandem repeat analysis (MLVA) and lic12008 gene sequence analysis. Results Monoclonal antibody F12 C3, which is highly specific for Icterohaemorrhagiae and Copenhageni, confirmed that all the strains belonged to these two serovars. Sixteen strains were identified as Copen...
<p>Phylogenetic reconstruction based on maximum likelihood estimation. Branch lengths are p... more <p>Phylogenetic reconstruction based on maximum likelihood estimation. Branch lengths are proportional to the number of substitutions per site and branch values are the bootstrap values assigned to the edges (i.e. the branch support values).</p
<p>MAT titres of reference serogroup antigens with antisera specific for each GWTS Isolates... more <p>MAT titres of reference serogroup antigens with antisera specific for each GWTS Isolates 1, 2 & 3.</p
<p>Genomic DNA from GWTS isolates #1 (1), #2 (2) and #3 (3) were compared by REA to that of... more <p>Genomic DNA from GWTS isolates #1 (1), #2 (2) and #3 (3) were compared by REA to that of <i>L</i>. <i>alstonii</i> isolates of serovar Pingchang (4), serovar Sichuan (5), MS 267 (6), MS 311 (7) and MS 316 (8). L = DNA Marker.</p
The greater white-toothed shrew (Crocidura russula) is an invasive mammalian species that was fir... more The greater white-toothed shrew (Crocidura russula) is an invasive mammalian species that was first recorded in Ireland in 2007. It currently occupies an area of approximately 7,600 km2 on the island. C. russula is normally distributed in Northern Africa and Western Europe, and was previously absent from the British Isles. Whilst invasive species can have dramatic and rapid impacts on faunal and floral communities, they may also be carriers of pathogens facilitating disease transmission in potentially naive populations. Pathogenic leptospires are endemic in Ireland and a significant cause of human and animal disease. From 18 trapped C. russula, 3 isolates of Leptospira were cultured. However, typing of these isolates by standard serological reference methods was negative, and suggested an, as yet, unidentified serovar. Sequence analysis of 16S ribosomal RNA and secY indicated that these novel isolates belong to Leptospira alstonii, a unique pathogenic species of which only 7 isolates have been described to date. Earlier isolations were limited geographically to China, Japan and Malaysia, and this leptospiral species had not previously been cultured from mammals. Restriction enzyme analysis (REA) further confirms the novelty of these strains since no similar patterns were observed with a reference database of leptospires. As with other pathogenic Leptospira species, these isolates contain lipL32 and do not grow in the presence of 8-azagunaine; however no evidence of disease was apparent after experimental infection of hamsters. These isolates are genetically related to L. alstonii but have a novel REA pattern; they represent a new serovar which we designate as serovar Room22. This study demonstrates that invasive mammalian species act as bridge vectors of novel zoonotic pathogens such as Leptospira.
Infection, genetics and evolution : journal of molecular epidemiology and evolutionary genetics in infectious diseases, 2015
Strains of Leptospira interrogans belonging to two very closely related serovars - Bratislava and... more Strains of Leptospira interrogans belonging to two very closely related serovars - Bratislava and Muenchen - are known to cause widespread infection of the horse population in many parts of the world. Conventional serological typing of isolates has been unable to differentiate between wildlife, pig, dog and possibly horse maintained isolates and therefore has been unable to provide further insight into their diversity and the relationship between them. Twenty-one such European isolates of serovar Bratislava and Muenchen were examined by restriction endonuclease analysis and multiple-locus variable-number tandem repeat analysis in an attempt to elucidate their epidemiology. The restriction pattern types were identified and fell into one of four REA designed pattern types, B1, B2a, M1, M2a. Nine strains from Northern Ireland and two from Germany belonged to B2a, which is a ubiquitous strain being originally isolated from a large number of wild and domestic animal species in the UK. Fi...
In order to manufacture many kinds of fiax fabric, it is necessary for the fihre in the yam to lo... more In order to manufacture many kinds of fiax fabric, it is necessary for the fihre in the yam to lose some of its non-ceUulosic components (ie, hemicelluloses, pectin, etc.) hefore it is made into cloth. An accurate determination of the remaining noncellulosic constituents left ...
A total of 855 sera from dogs in Greece were tested for antibodies to strains belonging to the Po... more A total of 855 sera from dogs in Greece were tested for antibodies to strains belonging to the Pomona, Grippotyphosa and Australis serogroups of Leptospira to assess exposure levels to these serogroups, possible associations with clinical disease and to evaluate whether these findings support the inclusion of additional serovars in dog vaccines. Antibodies were detected in 110 (12·9%) dogs. The highest seroprevalence (4·9%) was to the proposed novel serovar Altodouro belonging to the Pomona serogroup. This serovar also showed a statistically significant association with clinical disease. Serovar Bratislava antibodies were found in 3·4% of sera. Consideration should be given to the inclusion of serovars belonging to the Pomona serogroup and serovar Bratislava in future dog vaccines for the Greek market.
A Leptospira strain (designated RIM 139) was isolated from the kidney of a house-mouse, Mus muscu... more A Leptospira strain (designated RIM 139) was isolated from the kidney of a house-mouse, Mus musculus, in Trás-os-Montes e Alto Douro region of northern Portugal. The isolate showed typical leptospiral motility and morphology under dark-field microscopy and was pathogenic for hamsters. Species determination was carried out on basis of PCR products generated by species-specific primers and by sequencing of the secY gene. Putative serogroup typing was performed using the microscopic agglutination test (MAT) with a panel of rabbit anti-Leptospira sera representative of the major pathogenic serogroups. Serovar identification was carried out by a combination of monoclonal antibodies and cross-agglutinin absorption test (CAAT). The novel nature of the strain was confirmed by restriction endonuclease analysis (REA). Results showed that RIM 139 represents a new serovar. The name Altodouro is proposed for this new serovar. Speciation findings – PCR analysis of the ompL1 gene and sequencing of the secY gene – indicated that it belonged to Leptospira kirschneri as did amplification with G1/G2 and B64-I/B64-II the primer sets. Highlights ► A new virulent Leptospira strain was isolated from the house-mouse in Portugal. ► The strain was typed as Leptospira kirschneri, serogroup Pomona. ► MAbs, CAAT and REA analysis showed that the strain is a new serovar.
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Papers by Colm Gilmore
Highlights
► A new virulent Leptospira strain was isolated from the house-mouse in Portugal. ► The strain was typed as Leptospira kirschneri, serogroup Pomona. ► MAbs, CAAT and REA analysis showed that the strain is a new serovar.
Abbreviations
CAAT, cross-agglutinin absorption test; EMJH, Ellinghausen-McCullough-Johnson-Harris; MAT, microscopic agglutination test; MAb’s, monoclonal antibodies; PCR, polymerase chain reaction; REA, restriction endonuclease analysis
Keywords
Keywords
Leptospira; Leptospirosis; New serovar; House mouse
http://dx.doi.org/10.1016/j.meegid.2012.09.013
maria_neves_cardoso@msn.com
Highlights
► A new virulent Leptospira strain was isolated from the house-mouse in Portugal. ► The strain was typed as Leptospira kirschneri, serogroup Pomona. ► MAbs, CAAT and REA analysis showed that the strain is a new serovar.
Abbreviations
CAAT, cross-agglutinin absorption test; EMJH, Ellinghausen-McCullough-Johnson-Harris; MAT, microscopic agglutination test; MAb’s, monoclonal antibodies; PCR, polymerase chain reaction; REA, restriction endonuclease analysis
Keywords
Keywords
Leptospira; Leptospirosis; New serovar; House mouse
http://dx.doi.org/10.1016/j.meegid.2012.09.013
maria_neves_cardoso@msn.com