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pseudotuberculosis. live recombinant Corynebacterium lymphadenitis by use of a single oral dose of Protection of sheep against caseous
Brucella abortus strain 19 was recovered at necropsy from cows vaccinated with reduced doses of strain 19, three to six months previously. Strain 19 was isolated from four of nine cows vaccinated with 5.8 x 10(9) viable cells during... more
Brucella abortus strain 19 was recovered at necropsy from cows vaccinated with reduced doses of strain 19, three to six months previously. Strain 19 was isolated from four of nine cows vaccinated with 5.8 x 10(9) viable cells during pregnancy. In two of these nine cows vaccination had led to colonisation of the uterus with erythritol-utilising mutants of strain 19 and consequent abortion or premature calving. In one of another group of nine pregnant vaccinated with 2.8 x 10(8) viable cells, the vaccine strain was recovered from the colostrum. There was no interference with the course of pregnancy in over 700 cattle vaccinated with 3 x 10(8) viable cells. Thirteen of these cows, serologically positive approximately three months after vaccination, were necropsied and strain 19 was isolated from six. This lower dose has already been shown to produce immunity equal to that of higher doses and is recommended as suitable for whole herd vaccination.
Three groups, each of 14 mature Jersey heifers, were vaccinated. They were mated about 2 months later and those that became pregnant were challenged at about 6.5 months of pregnancy by the conjunctival application of virulent Brucella... more
Three groups, each of 14 mature Jersey heifers, were vaccinated. They were mated about 2 months later and those that became pregnant were challenged at about 6.5 months of pregnancy by the conjunctival application of virulent Brucella abortus. Group 1 heifers received 2 doses of B. abortus 45/20 vaccine 2 months apart. Only 5 of the 14 heifers became pregnant, and of these 5 only one resisted challenge. Group 2 heifers received only one dose of 45/20 vaccine, 5 of the 10 challenged resisted infection. Group 3 heifers received 3 x 10(8) cfu of strain 19. Six of the 10 heifers challenged resisted infection. All of 5 non-vaccinated control cattle became infected. It appeared advantageous to give only one dose of 45/20 rather than 2 as presently recommended. A single dose of 45/20 vaccine induced resistance to virulent B. abortus approximately equal to that given by the reduced dose of strain 19. One dose of 45/20 vaccine stimulated transient serological positivity in 2 of 28 heifers wh...
Four experiments are summarized. Initially an attempt was made to modify strain 19 calfhood vaccination so as to eliminate the persistent serological reactions which interfere with eradication programmes. Later the project broadened into... more
Four experiments are summarized. Initially an attempt was made to modify strain 19 calfhood vaccination so as to eliminate the persistent serological reactions which interfere with eradication programmes. Later the project broadened into a search for an effective method of vaccination that could be applied when required, to all ages of cattle, thus allowing calfhood vaccination to be safely stopped. In the first experiment, reducing the age at vaccination with strain 19 to 1 month practically eliminated the serological response to vaccination, but the resulting immunity was not satisfactory. However, vaccination at 1 month followed by a booster consisting of a reduced dose of strain 19, given conjunctivally 1 year later, stimulated an immunity at least equal to that given by conventional calfhood vaccination. The vaccination of pregnant cows with either of 2 reduced dose levels of strain 19 gave better immunity than calfhood vaccination with the full dose. Uterine strain 19 infectio...
... in the Republic of Ireland in 1974 (Noonan et al., 1975) and since then have been found in all of the 26 counties (Dolan, 1993 ... badgers led to a dramatic and significant reduction in the prevalence of infection in associated cattle... more
... in the Republic of Ireland in 1974 (Noonan et al., 1975) and since then have been found in all of the 26 counties (Dolan, 1993 ... badgers led to a dramatic and significant reduction in the prevalence of infection in associated cattle populations ( [O'Mairtin et al., 1998] and [Griffin et ...
In studies of Mycobacterium bovis infection in animals a definitive diagnosis requires the isolation of the organism. However, the optimum conditions for the primary isolation of M. bovis have not been determined. The aim of this study... more
In studies of Mycobacterium bovis infection in animals a definitive diagnosis requires the isolation of the organism. However, the optimum conditions for the primary isolation of M. bovis have not been determined. The aim of this study was to determine for primary isolation of M. bovis, (a) the incubation time required to achieve maximum sensitivity (i.e., the number of positive samples identified), (b) the effect of decontaminants on bacterial growth rates, and (c) the influence of media and the number of slopes of media on the number of positive samples detected. Two agar-based media, modified Middlebrook 7H11 (7H11) and tuberculosis blood agar (B83), and an egg-based medium, Stonebrink's (SB) were compared. Three decontaminants, 2% (w/v) sodium hydroxide (NaOH), 0.75% (w/v) and 0.075% (w/v) cetylpyridinium chloride (CPC, also called hexadecylpyridinium chloride, HPC) and 0.5% (w/v) benzalkonium chloride (BC) were evaluated against treatment with sterile distilled water. The inoculated media slopes were incubated for up to 15 weeks. Colonies first appeared after 2 weeks on all media types and 75% of positive slopes were identified by 8 weeks. An incubation time of 15 weeks was required to identify all positive samples. The slowest growth was associated with inocula that contained the fewest viable bacilli. The time to the appearance of colonies was influenced by medium type: the median time to detection of colonies was 28 days on 7H11 and B83, and 36 days on SB. However, SB returned the greatest number of positive samples. Decontamination procedures increased the minimum incubation time required to detect positive cultures, probably due to the toxic effect of the decontaminants. Increasing the number of inoculated slopes resulted in an increased number of positive samples and a decreased time to the detection of colonies. Overall, the detection of M. bovis was significantly influenced by the choice of media, the decontaminant and the duration of incubation of cultures.
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... Financial & competing interests disclosure Both Eamonn Gormley and Leigh Corner are principal investigators leading an oral BCG vaccine trial on badgers in Ireland. ... Vaccine 13, 1123–1130 (1995). 12 Griffin JFT, Mackintosh CG,... more
... Financial & competing interests disclosure Both Eamonn Gormley and Leigh Corner are principal investigators leading an oral BCG vaccine trial on badgers in Ireland. ... Vaccine 13, 1123–1130 (1995). 12 Griffin JFT, Mackintosh CG, Slobbe, L, Thomson AJ Buchan GS. ...
As part of ongoing culling operations, European badgers (Meles meles) were captured using stopped restraints in winter (October to December 2005) and summer (May to June 2006) in the Republic of Ireland. A subset of these badgers, those... more
As part of ongoing culling operations, European badgers (Meles meles) were captured using stopped restraints in winter (October to December 2005) and summer (May to June 2006) in the Republic of Ireland. A subset of these badgers, those caught during four consecutive nights, was examined postmortem to determine the frequency and severity of physical injuries resulting from capture in the restraints. The skin and the tissues underlying the restraint of 343 badgers were assessed for injury by visual examination. There was an absence of skin damage or only minor skin abrasions in 88% of badgers; an absence of subcutaneous tissue injury or only localized subcutaneous tissue injury in 69%; and an absence of muscle injury or only slight muscle bruising in 99% of badgers. Only 2% of badgers had cuts to the skin and 5.5% had extensive subcutaneous edema, whereas 1.2% had areas of hemorrhage and tearing of the underlying muscle. Our results show that the majority of badgers examined sustained minimal injuries attributable to capture in stopped restraints.
The aim was to develop an endobronchial infection procedure for the study of Mycobacterium bovis infection in badgers. The badgers were anaesthetised and a cannula was passed per os to the tracheal bifurcation. When in place 1 ml of M.... more
The aim was to develop an endobronchial infection procedure for the study of Mycobacterium bovis infection in badgers. The badgers were anaesthetised and a cannula was passed per os to the tracheal bifurcation. When in place 1 ml of M. bovis suspension was inoculated. Three concentrations of M. bovis suspension were used; <10 colony forming units (cfu), approximately 10(2) cfu and approximately 3 x 10(3) cfu. The badgers were examined at three weekly intervals for clinical signs of disease and a tracheal aspirate was collected at each examination. The badgers were euthanased 17 weeks post infection (pi) and at the post mortem examination a wide range of tissues were examined for gross and histopathological lesions of tuberculosis and cultured for M. bovis. A sample of bronchial alveolar lavage (BAL) fluid was collected at post mortem for culture. At post mortem examination 17 weeks after infection, gross and histopathological lesions of tuberculosis were observed in all badgers inoculated with the high and medium dose and 1/3 inoculated with the low dose. M. bovis was recovered from all inoculated badgers. Infection in the high dose group was more widely disseminated than in the other groups. The number of sites with gross and histopathological lesions increased with increasing dose of M. bovis. All tracheal aspirates were negative on culture and only one BAL, collected from a badger of the high dose group, was positive on culture. No clinical signs due to the experimental infection were observed. The endobronchial route of inoculation is an effective route for establishing experimental infection, and could be used for studies of tuberculosis pathogenesis, immunology of M. bovis infection in badgers and for challenging badgers in vaccine protection studies. Badgers appeared to be very susceptible to infection by this procedure even with a dose of < 10 cfu but appear to control and limit the resulting infection.
The eradication of tuberculosis (Mycobacterium bovis infection) from cattle herds may be compromised if infected wildlife species, such as European badgers (Meles meles), share the same environment and contribute to transfer of infection.... more
The eradication of tuberculosis (Mycobacterium bovis infection) from cattle herds may be compromised if infected wildlife species, such as European badgers (Meles meles), share the same environment and contribute to transfer of infection. Options for dealing with tuberculosis in this wild reservoir host are limited by conservation and social concerns, despite a clear implication that infected badgers are involved with the initiation of tuberculosis in cattle herds. Vaccination of badgers against M. bovis, if successfully employed, would directly facilitate the completion of bovine tuberculosis eradication in affected areas. Vaccine trials in captive badgers have established that the M. bovis bacille Calmette-Guérin (BCG) vaccine can induce a protective response that limits the distribution and severity of tuberculosis disease following experimental challenge. The protective effect of the vaccine has been demonstrated when the vaccine was delivered by subcutaneous injection, deposited on mucous membranes, and given orally in a lipid formulation. A large-scale field trial of oral BCG vaccine has been designed to measure the protection generated in wild badgers subjected to natural transmission of infection and to estimate vaccine efficacy. These parameters will be estimated by comparing the prevalence of M. bovis infection in vaccinated and nonvaccinated badgers. The results will provide a framework for the development and implementation of a national strategy to eliminate the disease in badger populations and if successful will remove this major impediment to bovine tuberculosis eradication.
Mycobacterium bovis is endemic in badger (Meles meles) populations of Ireland and the United Kingdom and infected badgers are a potential source of infection for cattle. In domestic livestock tuberculosis causes economic losses from lost... more
Mycobacterium bovis is endemic in badger (Meles meles) populations of Ireland and the United Kingdom and infected badgers are a potential source of infection for cattle. In domestic livestock tuberculosis causes economic losses from lost production and the costs associated with eradication programmes, and in addition there is a risk of zoonotic infection. Whereas culling is currently used to control tuberculous badger populations in Ireland, vaccination, if it were available, would be preferred. A study was undertaken to examine the protective responses of badgers vaccinated either by the subcutaneous or mucosal (intranasal and conjunctival) routes with bacille Calmette-Guérin (BCG), when challenged with M. bovis by the endobronchial route. Three groups of badgers were used. The first group (n=4) was vaccinated with approximately 5 x 10(5) colony forming units (cfu) of BCG by subcutaneous injection. In the second group (n=5) badgers were vaccinated via the mucosal route by instilling 1.0 x 10(5)cfu into each conjunctival sac and spraying 1.0 x 10(5)cfu into each nostril (final vaccine dose of 4 x 10(5)cfu). The control (n=5) badgers served as a non-vaccinated group. Twelve weeks post-vaccination all badgers in the three groups were challenged with approximately 10(4)cfu of M. bovis by endobronchial inoculation. At 12 weeks post-infection all badgers were examined post-mortem to assess the pathological and bacteriological responses to challenge. Gross and histological lesions of tuberculosis were seen in all challenged badgers and M. bovis was recovered from all challenged badgers. However, across six of the eight parameters used to measure disease severity, the infection in the vaccinated badgers was significantly less severe than in the control group. The BCG vaccine induced a significant protective effect in the badgers and the protective immunity was generated by subcutaneous and mucosal vaccination.
The conclusion from the randomised badger culling trial was that localised badger culling not only fails to control but can actually increase the incidence of bovine tuberculosis in cattle. Professor Simon More and colleagues from... more
The conclusion from the randomised badger culling trial was that localised badger culling not only fails to control but can actually increase the incidence of bovine tuberculosis in cattle. Professor Simon More and colleagues from University College Dublin question that conclusion, arguing that the data do not provide sufficient evidence to rule out alternative hypotheses.
Bronchoalveolar lavage (BAL) fluid is evaluated for the diagnosis and study of lung disease and airway inflammation. Cytologic profiles for BAL fluid have not been reported for badgers and may be useful in understanding the pathogenesis... more
Bronchoalveolar lavage (BAL) fluid is evaluated for the diagnosis and study of lung disease and airway inflammation. Cytologic profiles for BAL fluid have not been reported for badgers and may be useful in understanding the pathogenesis of pulmonary diseases such as Mycobacterium bovis. The aim of this study was to evaluate cytologic and microbial findings in BAL fluid from captive European badgers (Meles meles) and identify correlates with the results of concurrently collected blood and fecal samples. BAL fluid (by a nonbronchoscopic method) and jugular venous blood samples (for routine CBC) were obtained from 23 captive tuberculosis-free anesthetized badgers on 2 occasions 4 weeks apart. Fecal samples were collected for routine parasitology. Morphologic evaluation and 100-cell differentials were done on cytocentrifuged BAL specimens. Pellets from centrifuged BAL were aerobically cultured for bacteria. With the 2 BAL samples from each of the 23 badgers combined, the median (range) cell percentages were 73.0% (5-95%) neutrophils, 7.5% (2-16%) macrophages, 8.0% (0-27%) lymphocytes, and 9.5% (0-92%) eosinophils. Macrophages frequently contained silica-like crystals. Other findings included ciliated epithelial cells, goblet cells, mucus, and Aelurostrongylus sp. larvae. A light growth of Streptococcus, Pasteurella, or Escherichia coli was cultured in 6 badgers. Trypanosoma pestanai were identified in blood from 10 badgers and fecal parasites (mainly coccidia) were found in 20 badgers. No correlation was found between BAL and CBC results and the presence of parasites. The predominance of neutrophils in BAL fluid from badgers differs from the predominance of macrophages found in BAL from other species. This difference may reflect the burrowing lifestyle or the unique immune response of badgers.
The Mycobacterium bovis strain, bacille Calmette-Guérin (BCG) is one of the most widely used human vaccines and remains one of the safest vaccines available. It has been used in human populations for over 80 years and 100 million children... more
The Mycobacterium bovis strain, bacille Calmette-Guérin (BCG) is one of the most widely used human vaccines and remains one of the safest vaccines available. It has been used in human populations for over 80 years and 100 million children receive the vaccine annually. It has also been employed extensively for vaccine studies in laboratory animal hosts and is currently being developed for use in a variety of livestock and wild animals. Despite the large number of doses delivered since is first usage in 1921, reports of adverse reactions arising from the use of the BCG vaccine are relatively uncommon and where serious reactions do occur they are often the result of vaccination of immuno-compromised individuals. Factors that may influence the development of adverse reactions to BCG include the potency and dose of the vaccine strain, the route of delivery, the age and immune status of the host, and the skill levels of the operator administering the vaccine. Circumstances affecting the notification of adverse reactions include the lack of clear case definitions of abnormal vaccine reactions, and a scarcity of systematic surveillance and functioning reporting systems. With continued use of the BCG and the development of a new generation of prophylactic and therapeutic vaccines against tuberculosis in different host species, the risk factors associated with adverse reactions may need to be reappraised.
Vaccination of badgers by the subcutaneous, mucosal and oral routes with the Pasteur strain of Mycobacterium bovis bacille Calmette-Guérin (BCG) has resulted in significant protection against experimental infection with virulent M. bovis.... more
Vaccination of badgers by the subcutaneous, mucosal and oral routes with the Pasteur strain of Mycobacterium bovis bacille Calmette-Guérin (BCG) has resulted in significant protection against experimental infection with virulent M. bovis. However, as the BCG Danish strain is the only commercially licensed BCG vaccine for use in humans in the European Union it is the vaccine of choice for delivery to badger populations. As all oral vaccination studies in badgers were previously conducted using the BCG Pasteur strain, this study compared protection in badgers following oral vaccination with the Pasteur and the Danish strains. Groups of badgers were vaccinated orally with 10(8) colony forming units (CFU) BCG Danish 1331 (n = 7 badgers) or 10(8) CFU BCG Pasteur 1173P2 (n = 6). Another group (n = 8) served as non-vaccinated controls. At 12 weeks post-vaccination, the animals were challenged by the endobronchial route with 6 × 10(3) CFU M. bovis, and at 15 weeks post-infection, all of the badgers were euthanased. Vaccination with either BCG strain provided protection against challenge compared with controls. The vaccinated badgers had significantly fewer sites with gross pathology and significantly lower gross pathological severity scores, fewer sites with histological lesions and fewer sites of infection, significantly lower bacterial counts in the thoracic lymph node, and lower bacterial counts in the lungs than the control group. No differences were observed between either of the vaccine groups by any of the pathology and bacteriology measures. The ELISPOT analysis, measuring production of badger interferon - gamma (IFN-γ), was also similar across the vaccinated groups.
European badgers (Meles meles) are a reservoir host of Mycobacterium bovis and are implicated in the transmission of tuberculosis to cattle in Ireland and Great Britain. The development of a vaccine for use in badgers is considered a key... more
European badgers (Meles meles) are a reservoir host of Mycobacterium bovis and are implicated in the transmission of tuberculosis to cattle in Ireland and Great Britain. The development of a vaccine for use in badgers is considered a key element of any campaign to eradicate the disease in livestock in both countries. In this study we have vaccinated groups of badgers with approximately 5 x 10(5)cfu of the BCG vaccine delivered via two alternative routes, subcutaneous and mucosal (intranasal/conjunctival). Following experimental endobronchial infection with approximately 10(4)cfu of M. bovis, all badgers were euthanised at 12 weeks post-infection. At post-mortem examination both vaccinated groups had significantly reduced severity of disease compared with the non-vaccinated controls. The analysis of immune responses throughout the study showed that vaccination with BCG did not generate any detectable immunological responses as measured by IFN-gamma production in antigen-stimulated peripheral blood mononuclear cells (PBMC) and IgG serological responses. However, the levels of the responses increased following M. bovis infection, and the kinetic profiles corresponded to the severity of lesions recorded post-mortem. Significant differences were observed in the timing of development of the immune responses between vaccinates and controls. The results suggest that the immunological responses are associated with the levels of protective immunity and could be used as markers to monitor control of disease in badgers following vaccination.
European badgers (Meles meles) are a wildlife reservoir for Mycobacterium bovis infection (tuberculosis) in Ireland and the UK and are implicated in the transmission of infection to livestock. Vaccination of badgers with the human BCG... more
European badgers (Meles meles) are a wildlife reservoir for Mycobacterium bovis infection (tuberculosis) in Ireland and the UK and are implicated in the transmission of infection to livestock. Vaccination of badgers with the human BCG vaccine (Bacille Calmette Guerin) is considered as an important strategy to reduce the burden of disease in this species, and a pragmatic approach is likely to involve oral vaccination. In this study, we evaluated nine different flavours for use as attractants in a prototype oral vaccine bait for European badgers (M. meles): aniseed, apple, cocoa powder, carob powder, curry, fish, garlic, peanut and strawberry. The bait matrix was composed of a natural lipid formulation, developed as a vehicle for oral vaccination against tuberculosis in wildlife. A ‘food for work’ paradigm was employed during the trials to ensure the animals were actively seeking the baits. The trials showed carob and cocoa powders were equally attractive and more attractive than any of the other candidates. Carob and cocoa show potential as bait attractants for badgers and might form part of a novel vaccine delivery system.
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