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Irish soil and land-use classifications as predictors of numbers of badgers and badger setts.

This study investigated possible associations between soil types, land use and badger numbers in an area of the Irish Midlands where badger removal had been conducted during 1989-1994. For this purpose, the area was divided into approximately 2500 geo-referenced square grids of 0.5 degrees km per side. For the outcomes (setts per grid, badgers per grid and tuberculous badgers per grid), Poisson models of land use, of soil type, and a combination of these two were developed. Influential grids were removed and the models adjusted for over-dispersion in the badger outcomes. Mineral-based soils, dry and very-dry peat soils supported increased numbers of setts and badgers. High-quality pasture was the major land use (pastures often are found on mineral-based soils) and supported increased numbers of setts, badgers, and tuberculous badgers. "Natural" areas also supported more setts and broad-leaf forested areas were associated with increased tuberculous badger numbers. Discontinuous urban areas tended to decrease sett numbers per grid. Hedgerow length was not an important predictor given the information on soil type and land use. Spatial correlations existed for badger setts in 1 degrees km grids, for badger numbers in 1.5 degrees km grids, and for tuberculous badgers in 2 degrees km grids. The latter two grids have approximately the same area as the territory size used by a social group of badgers. There were no spatial correlations at the smallest (0.5 km) grid size.

Agriculture↗

Cloning and sequencing of badger (Meles meles) interferon gamma and its detection in badger lymphocytes.

The European badger (Meles meles) has been identified as a reservoir for Mycobacterium bovis and is implicated in the maintenance and transmission of tuberculosis in cattle. There is a need for a sensitive test of M. bovis infection in badgers and the current serodiagnostic test used for this purpose has low sensitivity. As observed for other species, assay of interferon-gamma (IFNgamma) produced in response to M. bovis antigens is a more sensitive test of tuberculosis. With this objective in sight, we report the first step in the development of an ELISA for badger IFNgamma. The badger IFNgamma gene was cloned and sequenced and used to generate a specific polyclonal antibody to the cytokine. The gene sequence demonstrated regions that were conserved within the IFNgamma genes of other mammals. The badger sequence was most similar to the canine, showing similar structural organisation of the gene and 88% amino acid identity. Rabbits were immunised with DNA encoding badger IFNgamma and the resulting polyclonal antiserum demonstrated specificity for canine IFNgamma by immunoblot of a commercial recombinant canine IFNgamma. The antiserum was used to detect intracellular badger IFNgamma by flow cytometry analysis of badger lymphocytes stimulated with mitogen.

Amino Acid Sequence↗

Production and characterization of a monoclonal badger anti-immunoglobulin G and its use in defining the specificity of Mycobacterium bovis infection in badgers by western blot.

A mouse monoclonal anti-badger IgG antibody was produced to investigate the specificity of the antibody response of badgers infected with Mycobacterium bovis. The monoclonal antibody generated was directed against badger IgG heavy chain and appeared to be species restricted, reacting only with badger and dog IgGs but not cat, rabbit, mouse, guinea pig, bovine or ferret IgGs. This monoclonal antibody detection system functioned well in both ELISA and Western blot analyses and was successfully used to investigate the humoral response of the badger to M. bovis infection. Sera from infected badgers detected a 25 kDa antigen which was not detected by sera from M. bovis culture-negative animals. This antigen was conserved in all field strains of M. bovis tested and seroconversion to it was detected during experimental infection. The immunodominance of this antigen in the badger during infection with M. bovis suggests that this 25 kDa polypeptide is a suitable candidate on which to base an antibody detection test for M. bovis infection.

Animals↗

Tuberculosis in East Sussex. II. Aspects of badger ecology and surveillance for tuberculosis in badger populations (1976-1984).

Following the disclosure of Mycobacterium bovis infection in badgers in East Sussex in 1976, badgers have been examined from and around farms on which cattle have become infected, but with no other attributable source of infection. These farms are confined to the downland of the south-west of the county and M. bovis has been confirmed in badger populations utilising their land. The available evidence indicates that M. bovis infection in badgers is also confined to this area. A detailed study in one area on the South Downs suggested that M. bovis is endemic in the badger population and therefore presents a continued risk for cattle occupying the area.

Animals↗

Mycobacterium bovis in the European badger (Meles meles): epidemiological findings in tuberculous badgers from a naturally infected population.

This study investigates the course of tuberculosis in a naturally infected badger population, its impact on the population and the risk of spread to other species in the light of capture data and post-mortem findings from 47 tuberculous badgers, stratified by age group and sex, accrued since 1975. The findings are compared with those for 260 badgers from the same population in whom no evidence of infection was detected. Detailed estimates of seasonal variations in bodyweight for uninfected male and female cub, yearling and adult badgers are presented and compared to the weights at post-mortem examination of the tuberculous badgers, in whom poor condition and weight loss were the principal presenting signs. Lesions were seen especially in the lungs and associated lymph nodes, and in the kidneys. Organisms were detected intermittently in faeces, urine, sputum and discharging bite wounds. Infected animals could survive for nearly 2 years and produce cubs successfully.

Animals↗

Role of infected, non-diseased badgers in the pathogenesis of tuberculosis in the badger.

The lungs and kidneys of 15 badgers which had no visible lesions of tuberculosis but from which Mycobacterium bovis was isolated from pooled collections of lymph nodes were serially sectioned. Lesions of tuberculosis were detected by histopathology in the the lungs of 13 and in the kidneys of one of them. The lesions were mostly typical early stage granuloma-lesions which were considered to be the primary foci of infection. These lesions suggest an early containment phase of arrested development previously not observed and provide further evidence on which to propose a hypothesis for the pathogenesis of tuberculosis in the badger.

Animals↗

Value of existing serological tests for identifying badgers that shed Mycobacterium bovis.

In the UK there has been a sharp rise in the incidence of bovine tuberculosis since the early 1990s and the badger has been identified as an important wildlife reservoir for this infection. Infected badgers can excrete Mycobacterium bovis, putting other badgers and cattle at risk of becoming infected. Vaccination has been proposed as an approach to reducing the excretion of M. bovis by tuberculous badgers. In order to evaluate the efficacy of a badger vaccine it will be necessary to accurately determine the number of badgers excreting M. bovis without removing them for post-mortem evaluation. The existing live tests for tuberculosis in the badger (culture, indirect ELISA, Western blot) have not been assessed for their ability to detect badgers excreting M. bovis. Over the past 18 years, badgers from 31 social groups have been trapped and sampled in a study area of the Cotswold escarpment. We have examined the serological responses of 128 badgers trapped between 1985 and 1998 from social groups where M. bovis infection was endemic. These responses were compared with culture from faeces, urine, tracheal aspirates and bite wound swabs taken from these animals while alive. ELISA was found to be more sensitive than Western blot in detecting badgers excreting M. bovis. The majority of culture-positive badgers excreted M. bovis intermittently over the period of study. As a result, there was only a 27.5% chance of sampling a badger for culture when it was excreting M. bovis. In contrast, a positive ELISA result correctly predicted 68.2% of badgers with a history of excreting M. bovis. In the absence of alternative live tests for the badger, the Brock Test indirect ELISA appears to be more valuable than culture for measuring the effect of vaccination on reducing the number of badgers at risk of transmitting tuberculosis.

Animals↗

Interactions between cattle and badgers at pasture with reference to bovine tuberculosis transmission.

The normal behaviour of badgers, as described here, would not result in direct transmission of tuberculosis from badgers to cattle via air expired by badgers or via bodily contact. All activities of wild badgers in cattle fields at three different sites were observed at night for a total of 359 h. In addition, all activities of pairs of wild caught badgers were observed in a 0.5 ha enclosure with an artificial sett on 20 nights when cattle were present. Badger foraging in cattle fields was infrequent during dry conditions and variable at other times. Clover fields were preferred to grass pastures in two autumnal studies and under dry conditions the badgers did not prefer to forage on short pasture. Badgers consistently avoided close contact with cattle by changing routes from sett to foraging site and by foraging much less in areas of fields occupied by cattle. When foraging they preferred to remain at least 10-15 m from cattle and they avoided compact groups of cattle more than individuals. Some cattle would move towards badgers, especially if they were carrying out unusual behaviour, but badgers fled rapidly from every approach. In all potential encounters badgers were able to keep at least 2-3 m from approaching cattle. Badgers in the artificial sett delayed entering the enclosure if cattle were within 15 m of the entrance. When cattle were managed on a strip-grazing (rotational) system the whole area which they occupied was avoided by the badgers. However, badgers came closer to individual cattle and foraged in areas grazed by the cattle more if the cattle were set-stocked. If cattle are managed so that they are concentrated in a small area (rotational) system the risk of disease transmission is minimized because they are less likely to encounter badgers or their fresh products than are cattle managed on a large area (set-stocked) system.

Animals↗

Bovine tuberculosis in badgers in four areas in Ireland: does tuberculosis cluster?

We described the distribution of badger populations in four different areas in the Republic of Ireland. The data came from periodic targeted badger-removal and subsequent post-mortem examinations conducted between 1989 and September 1997, and from a formal badger-removal project in the same areas from 1997 through 1999. Records were complete for 2292 badgers regarding the date of capture, tuberculosis status, geographical area and specific sett from where the badgers were snared. Of 3187 setts, 2290 had no badgers recorded against them (i.e. were inactive). The badger-level prevalence of tuberculosis differed among areas (range 13-29%). Badger populations were highly clustered by sett, and this result was similar over the four study areas. The median number of badgers per active sett was 2. Tuberculous badgers also clustered within a sett. The third quartile of tuberculous badgers was 1 per active sett. The prevalence of tuberculous badgers within a sett was not related to the total number of badgers. There was little evidence of spatial clustering with only one local cluster of tuberculous setts in each of three areas, and none in the fourth area. After adjusting for the number of badgers per sett, only one area had spatial clusters identified.

Animals↗

The association between the bovine tuberculosis status of herds in the East Offaly Project Area, and the distance to badger setts, 1988-1993.

The proximity of farms to badger setts was compared between farms that had experienced a tuberculosis breakdown and those that had not, over the 6 year period from 1988 to 1993. The data were derived from a badger removal study conducted in East Offaly County in the Republic of Ireland. Badger removal began in 1989 and continued through 1993; by the end of 1990, approximately 80% of all badgers caught in the 6 year period had been removed. All badgers were examined, grossly, for evidence of tuberculosis. Tuberculosis status of the approximately 900 study herds was based on the results of the single intradermal comparative skin test and/or lesions of bovine tuberculosis. All herds were tested at least once annually. The number of herds experiencing bovine tuberculosis declined over the period, particularly in the years 1992 and 1993. The data on farm and badger sett location were stored and analysed, initially, in a geographical information system. Owing to the badger removal programme, the distance between the barn yard of a typical farm and the nearest occupied badger sett increased, by about 300 m year-1, and by about 600 m year-1 to the closest infected sett. In bivariate analyses, in the years 1988 and 1989, the risk of tuberculosis declined with increasing distance to a badger sett containing one or more tuberculous badgers. In multivariable logistic regression analyses, year and the average number of cattle tested per farm per year were controlled. A second identical analysis was conducted to control for the repeated observations on the same herds using generalised estimating equations. In both analyses, the risk of a multiple reactor tuberculosis breakdown decreased for herds at least 1000 m away from an infected badger sett, and increased as the number of infected badgers per infected sett increased. Despite the significantly reduced risk of a breakdown with increasing distance to infected badger setts, the relationship was not strong (sensitivity and specificity of the model in the low 70% range) and explained only 9-19% of tuberculosis breakdowns.

Animals↗

Laboratory study of Mycobacterium bovis infection in badgers and calves.

Two experiments with badgers infected with Mycobacterium bovis are described. In the first, badgers were infected by intravenous inoculation of a bovine isolate of M bovis. The course of the disease in these and its spread to healthy badgers and calves was monitored by clinical, immunological and bacteriological means. In the second experiment a group of naturally infected badgers were observed for a period of up to four years. They were found to excrete M bovis in their faeces for periods of between 165 and 1305 days before they died of tuberculosis or were killed. M bovis was also shed in the urine. The badgers in both experiments were examined regularly and blood samples were taken for complement fixation tests. Faeces, urine, pus and sputum were also collected for cultural and biological tests and the badgers were skin tested using Weybridge bovine and avian tuberculin. The skin tests were uniformly negative while the complement fixation test were positive in some infected badgers but gave very variable results. Only the isolation of M bovis gave a definite diagnosis of tuberculosis in the living badger but a number of badgers which were found to have tuberculosis at post mortem were not detected while alive by this method. Environmental samples from the yards, including badger faeces, soil, hay, scrapings from feeding bowls and water were regularly examined for the presence of M bovis but apart from faeces only one water sample was positive, indicating that the organism did not persist for long in the environment. In both experiments calves developed sensitivity to bovine tuberculin after six months' exposure to infected badgers. The experiments further demonstrate the potential of a badger population to become endemically infected with M bovis and to act as a source of infection for cattle.

Animals↗

Spatial relationship between Mycobacterium bovis strains in cattle and badgers in four areas in Ireland.

We investigated whether strains (restriction fragment length polymorphism, RFLP-types) of Mycobacterium bovis isolated from badgers and from cattle clustered among and within four areas in Ireland. The spatial scan test and nearest-neighbor analysis were used as the spatial cluster-detection techniques. In addition, for each of the major strains, associations between the distance to badger setts and the "centroid" of the cattle farm were assessed in a logistic model. Overall, between September 1997 and May 2000, 316 and 287 M. bovis samples, from badgers and cattle, respectively, were strain-typed. The distribution of strains in badgers, and separately in cattle, differed among areas. Within each of the four large areas, badgers and cattle tended to have similar strains; this is consistent with the sharing of M. bovis strains within an area. In more detailed within-area analyses, some spatial clusters of M. bovis strains were detected, separately, in both cattle and badgers. Almost half of the infected badger setts with a specific strain were located outside of the "detected" clusters. There was no association between the number of infected badgers with a specific M. bovis strain within 2 or 5 km distances to cattle herds, and the risk of the same strain in cattle. We speculate about the dynamic nature of badger movements, as an explanation for the absence of more clusters of most of the strains of M. bovis isolated from badgers, and its impact on trying to study transmission of M. bovis between cattle and badger.

Animals↗

Quantifying the risks of TB infection to cattle posed by badger excreta.

Despite strong circumstantial evidence to suggest that the main route of TB transmission from badgers to cattle is via contaminated badger excreta, it is unclear whether the associated risks are high enough to account for the prevalence of the disease in south-west England. To decide whether this was a viable route of transmission, cattle contact with badger excreta was investigated using a deterministic approach to quantify the risks to cattle posed by badger excreta. Levels of investigative and grazing contacts between cattle and badger urine and faeces could each account for the disease prevalence in south-west England. An infection probability of 3.7 x 10(-4) per bite from pasture contaminated with badger urine infected with Mycobacterium bovis could account for the prevalence of TB in cattle in south-west England. Infection probabilities of 6.9 x 10(-7) per investigation and 1.1 x 10(-7) per bite from badger latrines could each account for the prevalence of TB in cattle in the south-west. When considering only the high risk areas of south-west England these bounds fell by a factor of eight. However, badger excreta may still constitute a high level of risk to cattle. The levels of cattle contact with badger excreta are far higher than previously thought, suggesting that it is the probability of infection per given contact with infected badger excreta which has the greater influence on the probability of transmission and not the level of contact. The infection probability per cattle contact with infected badger excreta is in all likelihood extremely low.

Animals↗

Bovine tuberculosis in badger (Meles meles) populations in southwest England: an assessment of past, present and possible future control strategies using simulation modelling.

A spatial stochastic simulation model was used to compare the efficacy of different badger control policies and to determine the theoretical requirements for the control of endemic bovine tuberculosis in badger populations in southwest England. Culling-based strategies for controlling endemic disease were compared with strategies employing a yet-to-be-developed oral vaccine which would provide uninfected badgers with immunity to the infection. A comparative assessment was made of the efficacy of previous and proposed culling-based strategies employed by the Ministry of Agriculture, Fisheries and Food for the control of localized disease, and the potential for an oral vaccine-based strategy for the control of localized disease was examined. For endemic bovine tuberculosis, to achieve a reasonable probability (p > 0.70) of successful control with a strategy involving a single culling operation, a very high proportion of the badger population (> 90%) must be culled. Single vaccination would not be successful in combating endemic disease. However, strategies involving repeated annual vaccination would have a very high probability of eradicating endemic disease, even with a relatively low (40-50%) annual vaccination efficiency. The most successful culling-based strategies for the control of localized disease were the gassing and clean ring strategies. Compared with no control at all, the interim strategy only offered benefits of a lower probability of disease spread and persistence in populations with low disease-free equilibrium group sizes or low initial prevalences of infection. In all other instances the benefits were negligible. The live test strategy will offer an improvement over the interim strategy, but will not be as effective as either the gassing or clean-ring strategies. In addition, it is likely to necessitate the culling of approximately four times as many badgers each year as the interim strategy, and the proportion of those killed that are infected will be approximately half that under the interim strategy. The efficacy of a strategy involving annually repeated oral vaccination of the badgers within a similar area to that covered by the live test depended on the efficiency of vaccination. A vaccination efficiency of 20-60% represented an overall improvement in efficacy over the interim strategy, being equivalent to the live test strategy. However, only vaccination efficiencies of 60-80% or greater achieved similar results to the gassing strategy, and none were so successful as the clean-ring strategy. Recommendations for future management are provided. Reactive strategies based on culling or vaccination will not solve the problem of bovine tuberculosis in badgers. Proactive strategies directed in those areas with a recent history of bovine tuberculosis in badgers should be considered as an alternative short-term control measure. The only strategy likely to eradicate bovine tuberculosis from badger populations in the long term is the use of repeated vaccination in proactive control operations in areas with a history of bovine tuberculosis in the badger population. Analyses should be conducted to evaluate whether the economic benefits of the live-test strategy are likely to outweigh its economic and ecological costs and whether continued research into the development of a vaccine for badgers is likely to offer any significant long-term economic benefits.

Animals↗

Responses of dairy cows to badger urine and faeces on pasture with reference to bovine tuberculosis transmission.

Grazing cattle were observed when they encountered badger urine or faeces which, in all but the first study, came only from badgers which were not infected with bovine tuberculosis. The faeces were very strongly avoided and there was generally a strong avoidance of ingestion of badger urine. There was no evidence that cattle were attracted to badger latrines in an area where some infected badgers were present and cows actively avoided faeces up to 28 days old which was placed on grass turves or on pasture. 99.3% of cows took no bites from small grass plots contaminated with faeces and 88.7% of cows took no bites from urine-treated plots. There was generally avoidance of pasture treated with badger urine up to 14 days old. However, two cows out of 240 were willing to graze close to faeces and seven out of 240 were willing to graze near urine. Contaminated herbage was eaten most when attractive herbage became scarce. Wet weather did not reduce the strength of avoidance of urine. Some cows responded to badger urine, and to a lesser extent to faeces, by more sniffing, particularly when herbage was scarce. The odour of faeces, and sometimes that of urine, often resulted in the ejection of mouth contents. As a consequence of their avoidance of badger faeces and urine, the vast majority of cows are unlikely to contract tuberculosis from infected badgers by ingestion. Most cows totally avoid badger products so they are unlikely to be infected via inhalation. However, the small minority of unselective cows must be more at risk and this finding warrants further investigation.

Animals↗

Quantifying badger exposure and the risk of bovine tuberculosis for cattle herds in county Kilkenny, Ireland.

The objectives of the study were to quantify the levels of badger exposure for cattle and to test the hypothesis that increased badger exposure does not increase the risk of bovine tuberculosis (BTB) in a herd. Information that became available from the targeted removal of badgers over the study period, and from a badger-removal project in county Kilkenny, during 1996-1999 was used. The specific location of cattle within each farm, and the length of time that cattle spent in each farm field during the grazing season, and in the barnyard during winter, was used to build an exposure coefficient to quantify the amount of badger exposure that cattle encountered either on pasture or in the barn. The study design was a matched case-control study in which the control herds were selected using incidence density sampling. During the 4-year study period, 543 badgers were removed and of these 96 badgers were classified as tuberculosis positive; 96 BTB herd breakdowns occurred. There was a significant association between case herds and having a higher badger sett exposure coefficient during 1996-1998. No significant association between case herds and having a higher exposure coefficient based on the number of badgers, or the number of tuberculous badgers, during September 1997-December 1999 was found.

Animals↗

Experimental tuberculosis in the European badger (Meles meles) after endobronchial inoculation of Mycobacterium bovis: I. Pathology and bacteriology.

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.

Animal Diseases↗

Mitochondrial DNA reveals a strong phylogeographic structure in the badger across Eurasia.

The badger, Meles meles, is a widely distributed mustelid in Eurasia and shows large geographic variability in morphological characters whose evolutionary significance is unclear and needs to be contrasted with molecular data. We sequenced 512 bp of the mitochondrial DNA control region in 115 Eurasian badgers from 21 countries in order to test for the existence of structuring in their phylogeography, to describe the genetic relationships among their populations across its widespread geographic range, and to infer demographic and biogeographic processes. We found that the Eurasian badger is divided into four groups regarding their mitochondrial DNA: Europe, Southwest Asia, North and East Asia, and Japan. This result suggests that the separation of badgers into phylogeographic groups was influenced by cold Pleistocene glacial stages and permafrost boundaries in Eurasia, and by geographic barriers, such as mountains and deserts. Genetic variation within phylogeographic groups based on distances assuming the Tamura-Nei model with rate heterogeneity and invariable sites (d(T-N) range: 3.3-4.2) was much lower than among them (d(T-N) range: 10.7-38.0), and 80% of the variation could be attributed to differences among regions. Spatial analysis of molecular variance (samova), median-joining network, and Mantel test did not detect genetic structuring within any of the phylogeographic groups with the exception of Europe, where 50% of variation was explained by differences among groups of populations. Our data suggest that the European, Southwest Asian, and North and East Asian badgers evolved separately since the end of Pliocene, at the beginnings of glacial ages, whereas Japanese badgers separated from continental Asian badgers during the middle Pleistocene. Endangered badgers from Crete Island, classified as Meles meles arcalus subspecies, were closely related to badgers from Southwest Asia. We also detected sudden demographic growth in European and Southwest Asian badgers that occurred during the Middle Pleistocene.

Animal Migration↗