The distribution of gross lesions of tuberculosis caused by Mycobacterium bovis in feral ferrets (Mustela furo) from Otago, New Zealand

The distribution of gross lesions of Mycobacterium bovis was examined in 94 tuberculous feral ferrets (Mustela furo) collected from 1992 to 1995 from areas of Otago endemic for bovine tuberculosis. Overall, 56.4% of tuberculous ferrets had single-site lesions, 24.5% had multiple infections and 19.1% had generalised infections. The mesenteric lymph node was the most common site of infection (34.5% of all lesions), with the retropharyngeal (17%) and the prescapular lymph nodes (16.4%) also frequently infected. Only 2.9% of lesions involved the respiratory tract. Of single-site lesions, 60.4% were in the mesenteric lymph node. The high proportion of lesions in the alimentary tract suggests that the ingestion of infectious material, possibly carrion or prey, is an important source of infection. Peripheral lymph nodes contributed to 24.5% of all infections, suggesting that within species transmission by social contact such as fighting and mating also occurs. Open and respiratory lesions were found in 11.7% of tuberculous ferrets, which suggests that ferrets are potentially infectious and therefore may be involved in the transmission of bovine tuberculosis to domestic stock and other mammals. The distribution of gross M. bovis lesions in ferrets is compared to those observed in possums (Trichosurus vulpecula) and badgers (Meles meles).     

Isolation of Mycobacterium bovis from the respiratory tracts of skin test-negative cattle.

Mycobacterium bovis was isolated from the respiratory tracts of three cattle which registered negative to tuberculin testing; no tuberculous lesions were found and the culture of lymph nodes and other tissues proved negative. One animal was from a group of five calves which had been inoculated intranasally with M bovis, and the organism was recovered once only from nasal mucus sampled 100 days after inoculation. The second animal had had contact with experimentally infected cattle which were excreting M bovis and the third was from a commercial farm. The results of ELISAS for antimycobacterial antibodies and interferon-gamma, and of lymphocyte transformation assays are presented. The animals' immune responses provided evidence that each of them had been challenged.     

Mycobacterium bovis in the anterior respiratory tracts in the heads of tuberculin-reacting cattle

Twenty-five of 50 randomly selected tuberculin-reacting cattle were confirmed as tuberculous in the laboratory. All 25 cattle had macroscopic lesions in lymph nodes associated with the respiratory tracts but only one had lung lesions. M bovis was isolated from the anterior respiratory tracts in the heads of four of the 25 tuberculous animals and from a nostril lesion found in a fifth. For at least three of these five animals, the intervals between the final tuberculin test and their previously negative tests indicated that infection had established relatively rapidly. Four of them had been tuberculin tested solely because they were animals in contiguous 'at risk' herds. It would appear that although M bovis can be isolated from the anterior respiratory tracts in the heads of tuberculin-reacting cattle, it is unlikely that primary foci of infection exist in regions other than the lungs or associated tissues. The study demonstrates the potential for reactors with lesions to excrete M bovis and the continued importance of infected cattle in the epidemiology and eradication of the disease.     

Naturally occurring tuberculosis in white-tailed deer

OBJECTIVE: To determine the distribution of lesions and extent of tissues infected with Mycobacterium bovis in a captive population of white-tailed deer. DESIGN: Cross-sectional study. ANIMALS: 116 captive white-tailed deer. PROCEDURE: Deer were euthanatized, and postmortem examinations were performed. Tissues with gross lesions suggestive of tuberculosis were collected for microscopic analysis and bacteriologic culture. Tissues from the head, thorax, and abdomen of deer with no gross lesions were pooled for bacteriologic culture. Tonsillar, nasal, oral, and rectal swab specimens, fecal samples, and samples of hay and pelleted feed, soil around feeding sites, and water from 2 natural ponds were collected for bacteriologic culture. RESULTS: Mycobacterium bovis was isolated from 14 of 116 (12%) deer; however, only 9 of 14 had lesions consistent with tuberculosis. Most commonly affected tissues included the medial retropharyngeal lymph node and lung. Five of 14 tuberculous deer had no gross lesions; however, M bovis was isolated from pooled tissue specimens from the heads of each of these deer. Bacteriologic culture of tonsillar swab specimens from 2 of the infected deer yielded M bovis. Mean (+/- SEM) age of tuberculous deer was 2.5 +/- 0.3 years (range, 0.5 to 6 years). Mycobacterium bovis was not isolated from feed, soil, water, or fecal samples. CONCLUSIONS AND CLINICAL RELEVANCE: Examination of hunter-killed white-tailed deer for tuberculosis commonly includes only the lymph nodes of the head. Results of such examinations may underestimate disease prevalence by as much as 57%. Such discrepancy should be considered when estimating disease prevalence.     

Pulmonary lesions and Mycobacterium bovis excretion from the respiratory tract of tuberculin reacting cattle

Although it is generally recognised that tuberculous lesions are present in lymph nodes associated with the respiratory tract in approximately 90 per cent of reactors with confirmed infection, lung lesions are found in only 1 to 2 per cent of such cases during abattoir examination. When lung lesions are not detected, it has been claimed that such cattle are non-excretors and thus unimportant in the epidemiology of the disease. In this study the lungs of 55 reactor cattle were sliced into sections approximately 0.5 cm thick. Tuberculous lesions were evident in over 70 per cent of lungs from reactors with concurrent lesions in lymph nodes of the respiratory system. Further, M bovis was isolated from single samples of nasal and, or, tracheal mucus taken at slaughter in 19 per cent of confirmed cases. Several of these reactors had a clear tuberculin test less than six months previously indicating recent infection. This study confirms the continued importance of the infected bovine in the epidemiology and current eradication of bovine tuberculosis. It is suggested that all tuberculous cattle with lesions in respiratory lymph nodes, rather than being regarded as non-excretors, should be considered as possible excretors and thus important sources of infection for other cattle both within and between herds.     

Epidemiology of Mycobacterium bovis infection in feral ferrets (Mustela furo) in New Zealand: II. Routes of infection and excretion

Detailed necropsies of 228 ferrets captured from eight areas in the North and South Islands provided material for an investigation into the epidemiology of tuberculosis in wild ferrets. Seventy-three of the 228 (32%) animals examined were diagnosed as tuberculous, by culture of pooled lymph nodes and detailed histopathological examination. The prevalence of bovine tuberculosis was 96% in 24 ferrets taken from areas in which tuberculous possums were common. None of 35 animals under 4 months of age were found to be infected, and the prevalence of infection was shown to rise with age, such that for each 6 month age increment there was a 2.8 times greater risk of becoming infected. The most common route of infection appeared to be via the alimentary tract, as 79% of 38 animals, in which the initial lesions could be reasonably determined, had these lesions associated with the digestive tract. Samples from potential sites of excretion from infected ferrets were submitted for culturing. The most common route of excretion was via the oral cavity, with M. bovis recovered from 15 of 64 (23%) oral swabs. Mycobacterium bovis was also isolated from four of 64 (6%) tracheobronchial lavage samples, ten of 63 (16%) faecal samples, two of 29 (7%) urine samples and one of 8 (12.5%) mammary glands. The disease in ferrets appears to be principally maintained by ingestion of tuberculous carrion. Although a moderate number of ferrets excrete M. bovis orally, there appears to be only minor intraspecific transmission by bite wounding. The findings provided no evidence to support the occurrence of pseudo-vertical transmission.     

Evaluation of the interferon-gamma (IFN-γ) assay to diagnose Mycobacterium bovis infection in pigs

Mycobacterium bovis recognizes as hosts a wide spectrum of animal species. In particular epidemiological situations, high prevalence of infection is found also in pigs. In the present study, we evaluated the capability of the interferon-gamma (IFN-γ) assay to identify pigs infected with M. bovis. The results of the immune-diagnosis were correlated to the findings of the post mortem inspection and the bacterial culture of lymph nodes. Blood samples of 146 pigs, belonging to a local breed of Sicily reared in free or semi-free roaming conditions, were collected to assess the specificity and the sensibility of the IFN-γ assay. Thirty-one pigs, from M. bovis free herds, did not react to the IFN-γ assay, yielding a specificity of 100%. The IFN-γ assay identified 15 out of 19 animals positive to the bacterial culture and 22 out of 26 animals with tuberculous lesions, with a sensibility of 78.9-84.6%, respectively. Out of 26 reactors to the test, 15 pigs (57.7%) confirmed to be infected after the bacterial culture and 22 (84.6%) had tuberculous lesions. The IFN-γ assay was able to reveal 4 animals with no visible lesions (NVL). Together, these findings support the feasible use of the IFN-γ assay as an intra vitam tool for the surveillance and management of M. bovis infection in swine populations.     

Evaluation of Abattoir Inspection for the Diagnosis of Mycobacterium bovis Infection in Cattle at Addis Ababa Abattoir

Detailed postmortem examinations were conducted to evaluate the efficiency of meat inspection procedures and to determine the distribution of lesions in Mycobacterium bovis-infected cattle. The study involved routine inspection at slaughter, collection of tissues for detailed examination in the laboratory, and bacteriological examination to identify M. bovis. Additionally, a 10-year (1992--2001) meat inspection record was analysed to determine tuberculosis trends in the past decade. chi2-Test and simple regression were used to analyse the data. Out of 1350 cattle examined, 1.5% were found with tuberculous lesions. Routine abattoir inspection detected only 55% of cattle with confirmed lesions. Fifty-four per cent of tuberculous lesions were found in the lungs and thoracic lymph nodes, 23% in the lymph nodes of the head, and the remaining 23% in the mesenteric and other lymph nodes of the carcase. M. bovis was additionally isolated from an animal that had no gross lesions of tuberculosis. On average, the annual rate of whole-carcase condemnation due to generalized tuberculosis was 0.024% and it has increased annually by 0.34% over the past decade. The rate of whole-carcase condemnation indicates a high degree of TB transmission and requires immediate attention from both the economic and public health points of view. The lower sensitivity of routine abattoir inspection confirms the importance of improving necropsy procedures.     

Granuloma development in cattle after intratonsilar inoculation with Mycobacterium bovis

OBJECTIVE: To examine the temporal development of tuberculous lesions in cattle inoculated with Mycobacterium bovis. ANIMALS: 15 mature crossbred cows obtained from a herd with no history of M bovis infection. PROCEDURE: Inoculation of cattle was done by intratonsilar instillation of 1.48 X 10(5) to 5.4 X 10(7) colony-forming units of M bovis strain 2045T. At 3 to 4 hours, 4 weeks, 6 weeks, and 8 weeks after inoculation, tissues were examined for gross and microscopic lesions and processed for isolation of M bovis. RESULTS: Retropharyngeal lymph nodes from cattle examined 4 weeks after inoculation contained microgranulomas consisting of aggregates of macrophages with few neutrophils. Retropharyngeal lymph nodes from all cattle examined 6 and 8 weeks after inoculation contained multiple, large, coalescing granulomas consisting of central areas of necrosis with mild fibrosis, numerous macrophages, lymphocytes, plasma cells, multinucleated giant cells, and neutrophils. Three of 8 cattle examined 6 or 8 weeks after inoculation had lesions in nonretropharyngeal sites with morphologic characteristics similar to that seen in retropharyngeal lymph node granulomas from cattle examined 4 weeks after inoculation. CONCLUSION: Granulomas can develop in draining lymph nodes of cattle in as little as 4 weeks after inoculation via intratonsilar instillation of M bovis. Intralesional morphologic changes between 4 and 6 weeks after inoculation indicate an increase in cellular chemotaxis and differentiation. Dissemination of bacteria to distant sites most likely was by lymphatic and hematogenous routes after establishment of the primary infection in retropharyngeal lymph nodes.     

Surveillance of wildlife for Mycobacterium bovis infection using culture of pooled tissue samples from ferrets (Mustela furo)

AIM: To compare culture results of homogenates of pooled lymph nodes from individual ferrets with and without macroscopic lesions of bovine tuberculosis for the presence of Mycobacterium bovis, and to determine whether homogenates from 10-30 ferrets could be combined and cultured without loss of sensitivity as a possible method for improving cost-effectiveness of surveillance for M. bovis infection in wildlife populations. METHODS: Numbers of colony forming units (cfu) of M. bovis present in cultures of homogenates of pooled lymph nodes from individual ferrets known to be infected and having no visible lesions (NVL) or macroscopic lesions consistent with bovine tuberculosis were determined. Prevalences of M. bovis infection in populations of ferrets in the Marlborough region of the South Island of New Zealand were determined by culturing homogenates of pooled lymph nodes from individual animals. Samples from homogenates from North Canterbury were combined to form pools representing 10, 20 and 30 animals and also cultured for M. bovis. RESULTS: Fewer M. bovis cfu were isolated from ferrets with NVL (mean=0.77 log10) compared with ferrets with macroscopic lesions (mean=3.22 log10; p<0.05). The mean prevalence of infection in eight different surveys involving 427 ferrets from the Marlborough region was 18% (range 8-44%), which included a small number of animals with macroscopic lesions of tuberculosis. Pooling of samples from up to 30 different ferrets with NVL did not reduce the sensitivity of detecting M. bovis infected populations. CONCLUSION: Culturing of pools of lymph node samples detected a significant proportion of M. bovis-infected ferrets that would otherwise have gone unnoticed based on samples that had only macroscopic lesions. Culturing of samples pooled from up to 30 different ferrets could provide significant cost savings in surveys of wildlife for the presence of M. bovis infection without any apparent loss of sensitivity.     

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.     

The rise and fall of tuberculosis in a free-ranging chacma baboon troop in the Kruger National Park

A single troop of free-ranging chacma baboons (Papio ursinus) was found to be infected with tuberculosis caused by Mycobacterium bovis. It is assumed that some members of the troop originally became infected when feeding on a tuberculous carcass in the veld or on tuberculous material scavenged at a nearby post mortem facility. Subsequently, apparent aerosol transmission took place while sleeping in an unused room. Oral transmission probably also occurred due to continuous contamination of the floor of this room and the common, narrow access (a train bridge crossing the Sabi River) to it with faeces and urine. A macroscopic prevalence of 50 % was found and the disease was noted to progress rapidly in infected baboons. A variety of organs had typical tuberculous lesions, of which the spleen, lungs and mesenteric lymph nodes were consistently, grossly affected. Using Restriction Fragment Length Polymorphism analysis, all but one of the baboon isolates were found to be identical to the most common African buffalo (Syncerus caffer) isolate (genotype 1) in this Park. The opportunistic sleeping facility was made inaccessible to the troop, which was forced to revert to sleeping in trees. A follow-up survey six months after closure, demonstrated that the disease had disappeared from the troop, and that no spillover infection had occurred into neighbouring troops.     

Experimental Mycobacterium bovis infection of cattle: effect of dose of M. bovis and pregnancy on immune responses and distribution of lesions

Groups of 18-month-old cattle were inoculated intratracheally with 5 x 10(5) colony forming units (high dose) or 500 colony forming units (low dose) of Mycobacterium bovis to determine an appropriate dose to induce lesions similar to those seen in the natural disease. An additional group of 21-28 weeks pregnant cattle were inoculated with the high dose of M. bovis to determine if pregnancy increased the susceptibility of cattle to M. bovis infection. By 23-24 weeks after challenge, the high dose of M. bovis had induced extensive lung lesions, and tuberculous lesions were observed in the lymph nodes of the head, neck, and thoracic and abdominal cavities. In contrast, the low dose of M. bovis induced predominantly small lesions (< 1 cm diameter) which were localised to the lungs and pulmonary lymph nodes. The lesions induced by the low dose were similar to those seen in the natural disease in cattle. The majority of the high dose group cattle produced strong antibody responses to M. bovis culture filtrate, while only one low dose animal produced a detectable response. All of the M. bovis-inoculated cattle produced strong cellular immune responses to bovine PPD (skin test and interferon-gamma responses). Pregnancy did not appear to affect the susceptibility to M. bovis infection, and immune responses of the cattle in this group at the end of the study were similar to those in the high dose non-pregnant group. However, from the first test after calving, the interferon-gamma responses of peripheral blood cultures to bovine PPD were low compared with the responses prior to calving.     

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.     

The prevalence and distribution of Mycobacterium bovis infection in European badgers (Meles meles) as determined by enhanced post mortem examination and bacteriological culture

The accurate diagnosis of Mycobacterium bovis infection in badgers is key to understanding the epidemiology of tuberculosis in this species and has significant implications for devising strategies to limit spread of the disease. In this study, badgers (n = 215) in the Republic of Ireland were examined at post mortem and tissues were collected from a range of anatomical locations and pooled into groups for bacterial culture of M. bovis. By assessing confirmed gross visible lesions (VL) alone, infection was detected in 12.1% of badgers. However, by including the results of all culture positive pooled samples, the overall infection prevalence increased significantly to 36.3%. Two-thirds (66.7%) of infected animals had no visible lesions (NVL). While the thoracic cavity (lungs and pulmonary lymph nodes) was found to be the most common site of infection, in a proportion of animals infection was absent from the lungs and draining lymph nodes and was confined to the lymph nodes of the carcase or the head. This may indicate an early extrapulmonary dissemination of infection or alternatively, in the case of the head lymph nodes, a secondary pathogenic pathway involving the lymphoid tissues of the upper respiratory tract (URT).     

Natural infection of red deer with bovine tuberculosis

Six bovine tuberculosis-free red deer hinds were introduced in October 1993 to a 1.8 ha enclosure, within a larger field study site known to contain tuberculous possums, and kept there for 9 months. A Mycobacterium bovis-infected possum was found in the vicinity of the deer enclosure 3 weeks after the introduction. Subsequently, a further eleven infected possums were found in the area. The deer were monitored by repeated composite antibody detection ELISA and lymphocyte transformation assays for tuberculosis, interpreted in parallel, by skin testing and by routine culturing of samples collected from potential excretion sites. Lymphocyte transformation assay evidence of M. bovis infection in four hinds was first observed 4 months after introduction. One other hind became bovine tuberculin lymphocyte transformation assay positive in the 5th month. Positive or equivocal bovine reactivity remained evident at most test episodes. A comparative cervical skin test performed in July 1994, shortly before slaughter, was positive in these five hinds. Mycobacterium bovis was recovered off swabs from the oropharyngeal tonsils of two hinds during routine sampling. Detailed necropsy of the six deer revealed a single typical tuberculous lesion in only one, but culturing of various tissue specimens ascertained that the five blood test and comparative cervical skin test-positive animals were all infected. Mycobacterium bovis was cultured from the oropharyngeal tonsils of four and medial retropharyngeal lymph nodes of two of the deer with no typical gross lesions. Six additional tuberculosis-free hinds were introduced to the enclosure in April 1994 and kept there for 12 months. Four of these animals showed a positive lymphocyte transformation assay response to M. bovis after 9 weeks, but no significant reactivity thereafter. Concurrent observational studies suggest that five of the first six deer probably became infected through close inspection and investigation of the tuberculous possums, although the possibility of deer-to-deer transmission cannot be totally excluded. The likely deer-possum contact, and thus exposure to M. bovis, was related to the curiosity and social ranking of the hinds. The second group appear to have had transient exposure to M. bovis, possibly caused by direct contact with the infected hinds introduced earlier. This group never showed any curiosity toward, or interaction with, possums during the periods of observation.     

Tuberculosis in kudus (Tragelaphus strepsiceros) in the Kruger National Park.

Five kudus (Tragelaphus strepsiceros), three bulls and two cows, within the Greater Kruger National Park complex, were diagnosed with generalized tuberculosis caused by Mycobacterium bovis. The lesions seen in these animals were similar to those previously reported in kudus and included severe tuberculous lymphadenitis of the nodes of the head and neck (that resulted in noticeable uni- or bilateral swelling beneath the ear), thorax, and the mesentery. All the animals also suffered from severe granulomatous pneumonia. The lesions in the lungs were more severe cranially and had a miliary distribution elsewhere in the lungs. Based on the DNA patterns of the M. bovis isolates, at least some of these kudus were infected with strains commonly present in tuberculous buffaloes, lions, cheetahs, and baboons in the Park whereas other strains from these kudus were quite different and may reflect another source of infection. The presence of tuberculous kudus in the Park is expected to complicate control measures that may be instituted to contain or eradicate the disease in the Park.     

Mycobacterium bovis infection in North American elk (Cervus elaphus).

A naturally occurring outbreak of Mycobacterium bovis infection in captive wild elk (wapiti) in Montana was confirmed by mycobacteriologic examination. Twenty-eight of 143 elk responded to M. bovis purified protein derivative (PPD) tuberculin injected intradermally in the cervical region (SCT). The results of comparative cervical tuberculin skin tests conducted within 9 days of SCT revealed greater responses to M. bovis PPD tuberculin than to M. avium PPD tuberculin in 23 of 28 elk responding. At necropsy, several grossly visible tuberculous lesions were observed in the parenchyma of the lung, thoracic lymph nodes, and submandibular lymph nodes. Microscopic examination of appropriately stained tissue sections revealed the presence of granulomatous lesions containing acid-fast bacilli. An enzyme-linked immunosorbent assay (ELISA) was developed using a sarkosyl extract of M. bovis (antigen) and peroxidase-labeled protein G (conjugate); reactions were detected in the sera of 8 of 9 elk responding to M. bovis PPD tuberculin. Lymphocyte blastogenic assay responses were detected using M. bovis antigens in 7 of 9 elk positive on skin tests using M. bovis PPD.     

Geographical association between the genotype of bovine tuberculosis in found dead badgers and in cattle herds

In a survey, 457 badgers that had been found dead in Wales were postmortem-examined, and samples were examined by histology and by extended culture (for up to 12 weeks). Mycobacterium bovis was cultured from 55 badgers (12.0 per cent), and the histology typical of M bovis infection was seen in a further six (1.3 per cent). The prevalence in badgers in each of 10 geographical areas varied between 0 and 26 per cent (P<0.001), and was associated with the incidence of confirmed M bovis infection in cattle herds in the same areas (P<0.01). In northern Wales, bTB was rare in both hosts. An infected badger was 12.3 times more likely to be within 5 km of a confirmed cattle bTB breakdown than an uninfected badger. The M bovis isolates from badgers belonged to one of four genotypes defined by spoligotype and variable number tandem repeat type. These genotypes were also found in 290 concurrent confirmed herd breakdowns, and tended to be similar to the genotypes in badgers in the same geographical areas. When badgers and cattle no more than 30 km apart were compared, the genotype diversity was greater in cattle than in badgers (P=0.016), suggesting that the movement of cattle plays a greater part in the spatial distribution of M bovis than the movement of badgers.     

Pathologic findings and association of Mycobacterium bovis infection with the bovine NRAMP1 gene in cattle from herds with naturally occurring tuberculosis

OBJECTIVE: To determine necropsy and Mycobacterium bovis culture results in cattle from herds with tuberculosis, the role of the bovine NRAMP1 gene in resistance and susceptibility to infection with M bovis, and the association between magnitude of the tuberculous lesions and various types of M bovis isolates. ANIMALS: 61 cattle from herds with tuberculosis in Texas and Mexico. PROCEDURE: 61 cattle were evaluated by necropsy; 59 had positive and 2 had negative caudal fold tuberculin intradermal test (CFT) results. Thirty-three cattle with positive CFT results were genotyped to evaluate polymorphism of the 3' untranslated region of the bovine NRAMP1 gene, using single-stranded conformational analysis, 9 were resistant to M bovis with no tuberculous lesions and negative M bovis culture results, and 24 were susceptible with tuberculous lesions and positive M bovis culture results. Isolates of M bovis were analyzed by restriction fragment length polymorphism (RFLP) on the basis of IS6110 sequences and direct-repeat fingerprinting patterns. RESULTS: 21 (35.6%; 21/59) cattle with positive CFT results had tuberculous lesions or positive culture results; in addition, 1 of 2 cattle with negative CFT results had tuberculous lesions and positive culture results. Tuberculous lesions were most common in the thorax (35/63; 55.5%) and lymphoid tissues of the head (10/63; 15.9%). Tuberculous lesions varied from 1 to 11/animal; 8 of 21 (38.1%) had solitary lesions. Associations were not found between resistance or susceptibility to infection with M bovis and polymorphism in the NRAMP1 gene or between the magnitude of the lesions and various RFLP types of M bovis isolates. CONCLUSIONS AND CLINICAL RELEVANCE: The NRAMP1 gene does not determine resistance and susceptibility to infection with M bovis in cattle.