Spatial analysis of Mycobacterium bovis infection in white-tailed deer (Odocoileus virginianus) in Michigan, USA

The wild white-tailed deer (Odocoileus virginianus) population in Michigan, USA, has endemic Mycobacterium bovis. We determined whether there were spatial clusters of retrospective TB cases in white-tailed deer in northeastern Michigan and identified specific factors associated with the spatial clusters. Data from hunter-harvested deer (age, gender, TB status, and geographic section) were collected by the Michigan Department of Natural Resources (MDNR) during TB surveillance from 1995 to 2002. Land cover (vegetation, land-use) and land type (soil types and drainage characteristics, landforms) described potential deer habitats. Specific locations of large-scale supplemental feeding sites were collected from the MDNR aerial surveillance program from 1997 to 2002. Analyses were conducted using principal components derived from environmental data (and other risk factors) on spatial clusters of disease (identified by the spatial scan statistic). Spatial effects were incorporated into the multivariable analyses by using a neighborhood approach. A total of 420 deer with M. bovis infection were identified from 1995 to 2002, out of 39,451 harvested deer from 3216 TRS units, and spatial clusters of cases were identified. A total of seven principal components of environmental data were generated. Clusters were associated with the presence of large expanses of deciduous forests on moraine ridges separated by low areas of forested wetlands, and the presence of many small lakes. Factors that promoted congregation of deer for extended periods of time (natural cover, access to water, and less human contact) appeared to be associated with increased odds of TB positivity. This suggests that there are specific areas where interventions can be implemented to reduce congregation of animals and disrupt the cycle of infection transmission.     

Spatial-temporal model of haemorrhagic disease in white-tailed deer in south-east USA, 1983 to 2000

The present study constructed a spatial-temporal statistical model to identify the risk and protective factors for haemorrhagic disease (HD) in white-tailed deer in the five states of Alabama, Georgia, South Carolina, North Carolina and Tennessee. The response variable was binary, indicating the presence or absence of HD in an individual county, measured annually from 1983 to 2000. Predictor variables included climatic factors of temperature, rainfall, wind speed and dew point, remotely sensed data of normalised difference vegetation index (NDVI) and land surface temperature derived from archived remotely sensed advanced very-high-resolution radiometer (AVHRR) satellite data, elevation, a spatial autocorrelation (SA) term and a temporal autocorrelation term. This study first applied principal component factor analysis to reduce the volume of climatic data and remotely sensed data. Then, a generalised linear mixed model framework (GLMM) was used to develop a spatial-temporal statistical model. The results showed that the area under receiver operating characteristic curve (ROC) was 0.728, indicating a good overall fit of the model. The total prediction accuracy over the 18 year period with optimal cut-off probability was 67 per cent. The prediction accuracy for individual years ranged from 48 to 75 per cent.     

Managing the wildlife reservoir of Mycobacterium bovis: the Michigan, USA, experience

Historical, social and economic factors combined to provide a focus where bovine tuberculosis has become established in free-ranging wildlife in northeastern lower Michigan. White-tailed deer, the primary reservoir and maintenance host of tuberculosis, are highly valued by the public, and particularly hunters, for cultural and economic reasons. Since 1995, significant progress has been made in defining and reducing the reservoir of tuberculosis in deer. As yet, no other wildlife species has been shown to play an epidemiologically important role in the disease cycle. The importance of deer and deer hunting to Michigan has uniquely shaped tuberculosis control policies, and poses ongoing challenges as wildlife managers strive to maintain momentum for broad control strategies, and develop focused strategies that are publicly acceptable. Even if momentum and funding can be maintained, tuberculosis will likely continue to be present for a decade or longer. Thus, cattle producers waiting for tuberculosis to be eradicated from wildlife to eliminate risks to their herds and markets face disappointment for the foreseeable future. Such unrealistic expectations also place Michigan's federal tuberculosis accreditation status at perpetual risk. Accredited free status is unlikely to be regained without accompanying changes in cattle management. In Michigan, management of tuberculosis has clearly demonstrated that social issues and public approval are likely to be the critical limiting factors in control.     

Lesion development in white-tailed deer (Odocoileus virginianus) experimentally infected with Mycobacterium bovis

The recent discovery of tuberculosis in free-living white-tailed deer in northeastern Michigan underscores the need for increased understanding of the pathogenesis of tuberculosis in wildlife species. To investigate lesion development in white-tailed deer, 32 deer were experimentally infected by intratonsilar instillation of 300 colony-forming units of Mycobacterium bovis. Three deer each were euthanatized and examined at days 15, 28, 42, and 56 after inoculation, and five deer each were euthanatized and examined at days 89, 180, 262, and 328 after inoculation. Microscopic lesions first were seen in the medial retropharyngeal lymph node and lung 28 and 42 days after inoculation, respectively. Lung lesions were present in 12 (38%) of 32 deer, involving 23 lung lobes. Left caudal and right middle and caudal lobes were involved in 17 (74%) of the 23 affected lung lobes. Lesions in the medial retropharyngeal lymph node first appeared as granulomas composed of aggregates of macrophages and Langhans-type giant cells. Some early granulomas contained centrally located neutrophils. As granulomas developed, neutrophils were replaced with a central zone of caseous necrosis that first showed signs of mineralization 42 days after inoculation. Granulomas increased in size as the zone of caseous necrosis expanded. Peripheral fibrosis, first seen at 56 days after inoculation, progressed to only a thin fibrous capsule by 328 days after inoculation. By the termination of the study, the central necrotic core of the granuloma contained abundant liquefied necrotic material and grossly resembled an abscess. Although tuberculous lesions in white-tailed deer follow a developmental pattern similar to that in cattle, fibrosis is less pronounced and the advanced lesions may liquefy, a change seldom reported in cattle. An understanding of lesion development will aid in the identification of the spectrum of disease that may be seen in this important wildlife reservoir of tuberculosis.     

Cell mediated and humoral immune responses of white-tailed deer experimentally infected with Mycobacterium bovis

The objective of this study was to improve the understanding of immune responses of whitetailed deer (Odocoileus virginianus) infected with Mycobacterium bovis. Ten mature, female, white-tailed deer were inoculated by intratonsilar instillation of 2 x 10(3)or 2 x 10(5)colony-forming units of M. bovis. Lymphocyte proliferation and humoral response to M. bovis PPD and the M. bovis protein, MPB70 were measured. Deer were tested for exposure to M. bovis by the comparative cervical skin test. Biopsy specimens of skin test sites were examined microscopically and immunohistochemically. The comparative cervical skin test correctly identified all M. bovis -inoculated deer as exposed to M. bovis. Lymphocyte proliferative responses to MPB70 were more consistent than responses to M. bovisPPD in M. bovis -inoculated deer. Antibody responses were more prominent in deer with disseminated disease than in deer with localised disease. The cellular components of delayed-type hypersensitivity reactions at skin test sites were similar to tuberculin reactions in other species. T lymphocytes of the gamma/delta phenotype were seen in increased numbers in M. bovisPPD injection sites.     

Comparison of postmortem techniques for the detection of Mycobacterium bovis in white-tailed deer (Odocoileus virginianus).

A retrospective study of various diagnostic postmortem techniques used in a 4-year surveillance program for detection of Mycobacterium bovis infection in wild white-tailed deer (Odocoileus virginianus) was conducted. The tests evaluated were routine histopathology, acid-fast staining, detection of acid-fast bacilli in culture, and an M. tuberculosis group-specific genetic probe applied to pure cultures. Each of these techniques were compared with a reference or "gold standard" of mycobacterial culture and identification. Histopathology, the most rapid form of testing for M. bovis infection in white-tailed deer samples, had a sensitivity of 98% and a specificity of 87%, resulting in a positive predictive value of 94%. The detection of acid-fast bacilli by staining was less sensitive than histopathology (90%), but its higher specificity (97%) resulted in a positive predictive value of 99%. The detection of acid-fast bacilli on culture was both highly specific (93%) and sensitive (100%). The group-specific genetic probe had the highest sensitivity and specificity and produced results in complete agreement with those of mycobacterial culture, suggesting that this technique could be used as the new "gold standard" for this particular wildlife tuberculosis surveillance program.     

Epidemiology and pathogenesis of Mycobacterium bovis infection of red deer (Cervus elaphus) in New Zealand

AIMS: This study was initiated to investigate aspects of the epidemiology, pathogenesis and transmission of tuberculosis in wild red deer, with the aim of determining whether this species may be considered a reservoir host of Mycobacterium bovis in New Zealand. METHOD: One hundred and six wild red deer (Cervus elaphus) carcasses from the Castlepoint and Hauhungaroa Range areas, which are endemic for bovine tuberculosis, were examined for the presence of M. bovis infection. Samples were also examined from 46 skin test-positive farmed deer killed at two deer slaughter premises. Where possible, a standard set of tissues and excretion site samples was collected for mycobacteriological examination. RESULTS: Fifty-eight infected deer were identified, and of these 28% showed no gross lesions. The prevalence of tuberculosis confirmed by culture in the wild deer was 32%. Only one of 18 deer younger than 1 year was infected. Mature deer (>2 years) were 12 times more likely to be infected than those under 1 year of age. Infected older deer were less likely to show typical gross lesions than younger animals. Mycobacterium bovis was isolated from the oropharyngeal tonsil of 34 of 56 (61%) of the infected deer, and this was the most commonly infected site. Gross lesions were found in 18 of the 34 infected tonsils and only one of these showed a purulent tonsillitis. Mycobacterium bovis was recovered from four of 53 nasopharyngeal tonsils, four of 53 oropharyngeal swabs, one of 53 tracheal and nasal swabs, and one of 46 faecal samples, but not from any urine specimens. CONCLUSION: These findings suggest that significant bacillary excretion from infected deer was uncommon, and is more likely to occur in severely affected animals. This study has confirmed the importance of mucosa-associated lymphoid tissues (MALT), particularly the oropharyngeal tonsil, in the pathogenesis of tuberculosis in deer. The findings justify investigation of the hypotheses that the prevalence of tuberculosis in wild deer in New Zealand is high due to transmission of infection from possums, and that in the absence of an infected possum population, the prevalence of tuberculosis in deer is likely to be low, and spatially patchy. CLINICAL RELEVANCE: The results suggest that about one quarter of infected deer show no detectable gross lesions. This implies that many infected carcasses may enter the food chain unrecognised and that the estimated sensitivity and specificity of diagnostic tests may be erroneous if there is a difference in test performance between those conducted on deer with or without gross lesions. Diagnostic sensitivity following slaughter may be improved by routine culture of oropharyngeal tonsils and careful examination of lungs for adhesions and small subpleural tubercles.     

The Epidemiology of Mycobacterium bovis in Buffalo in Iran

Mycobacterium bovis is the cause of bovine tuberculosis (bovine Tb) in animals and is considered to be zoonotic and accordingly it infects humans, although cattle are the main host. Buffalo can also be infected and develop bovine Tb. In Iran, almost half a million buffaloes are farmed, mainly in three provinces. In West Azerbaijan, which has the largest numbers of buffaloes, cattle and buffalo are often farmed together. According to the reports of the Iranian Veterinary Organization over the last 25 years, there have been no reports of bovine Tb in buffalo, although the disease is often reported in cattle in this province. Eighteen and 140 pathology specimens from cattle and buffalo, respectively, collected from West Azerbaijani abattoirs were cultured. From one buffalo specimen out of 140, M. bovis was recovered, whereas the pathogen was isolated from 13 cattle specimens. Spoligotyping showed a relatively higher polymorphism within these isolates compared with M. bovis isolated from other Iranian provinces.     

Molecular Epidemiology of Mycobacterium bovis in Humans and Cattle

Bovine tuberculosis (bTB), caused by Mycobacterium bovis (M. bovis), is a serious re‐emerging disease in both animals and humans. The evolution of the Multi‐ and Extensively drug‐resistant M. bovis strains (MDR‐TB and XDR‐TB) represents a global threat to public health. Worldwide, the disease is responsible for great economic losses in the veterinary field, serious threat to the ecosystem, and about 3.1% of human TB cases, up to 16% in Tanzania. Only thorough investigation to understand the pathogen's epidemiology can help in controlling the disease and minimizing its threat. For this purpose, various tools have been developed for use in advanced molecular epidemiological studies of bTB, either alone or in combination with standard conventional epidemiological approaches. These techniques enable the analysis of the intra‐ and inter‐species transmission dynamics of bTB. The delivered data can reveal detailed insights into the source of infection, correlations among human and bovine isolates, strain diversity and evolution, spread, geographical localization, host preference, tracing of certain virulence factors such as antibiotic resistance genes, and finally the risk factors for the maintenance and spread of M. bovis. They also allow for the determination of epidemic and endemic strains. This, in turn, has a significant diagnostic impact and helps in vaccine development for bTB eradication programs. The present review discusses many topics including the aetiology, epidemiology and importance of M. bovis, the prevalence of bTB in humans and animals in various countries, the molecular epidemiology of M. bovis, and finally applied molecular epidemiological techniques.     

Diagnostic detection methods for Mycobacterium avium subsp. paratuberculosis in white-tailed deer

This study compares the results and suitability of serological testing, microscopic examination, deoxyribonucleic acid (DNA) detection, and bacterial culture for detecting Mycobacterium avium subsp. paratuberculosis (Map) infection in asymptomatic farmed white-tailed deer (WTD) (Odocoileus virginianus). Deer were classified as infected if culture slants from their feces, lymph nodes, or ileum were positive, or if a polymerase chain reaction (PCR) assay detected Map DNA in any of its tissues. Deer identified as positive by agar gel immunodiffusion (AGID) testing or enzyme-linked immunosorbent assay (ELISA) but not by bacterial culture, Ziehl-Neelsen staining, or PCR assay were classified as suspect. Culture of tissues classified 10/16 (62.5%), histopathologic examination 1/16 (6.3%), tissue smears 4/16 (25%), culture slant (CS)-PCR on feces 12/15 (80%), CS-PCR on tissue 13/16 (81.3%), and direct PCR on uncultured tissues 5/16 (31.3%) deer as infected. The ELISA classified 2/15 (13.3%) deer as positive and therefore suspect. The AGID test was negative for all deer. Fifteen of 16 deer were positive by 1 or more tests; only 1 deer was negative on all 11 assays. The CS-PCR gave superior results on antemortem fecal testing as well as postmortem tissue testing and can be recommended for improving the detection of Map in WTD at every stage of infection.     

Mycobacterium bovis infection in a captive herd of Sika deer.

Infection with Mycobacterium bovis was diagnosed in a small privately owned herd of Sika deer. After postmortem examination of a deer with progressive pulmonary disease, diagnosis of infection with M bovis was confirmed by bacteriologic culture. The 2 remaining deer in this herd were euthanatized, necropsied, and confirmed to be infected with M bovis. Three cats in contact with the deer were also euthanatized and necropsied. One of these cats had lesions suggestive of mycobacterial infection in the colon and mesenteric lymph nodes. Infection of this cat with M bovis was not confirmed by bacterial culture. Mycobacteriosis, infrequently encountered in clinical veterinary practice, may be confused with disease caused by other infective agents or neoplasia. The zoonotic potential of these bacteria and a recent increase in human tuberculosis warrants continued surveillance of companion and food animal populations for mycobacterial infection.     

Spatial and spatial-temporal clustering analysis of hemorrhagic disease in white-tailed deer in the southeastern USA: 1980-2003

We used the space-time K function and Kulldorff's scan statistic to analyze the spatial and spatial-temporal clustering of hemorrhagic disease (HD) in white-tailed deer in Alabama, Georgia, South Carolina, North Carolina, and Tennessee. The HD occurrence data were binary presence/absence data acquired annually on a county basis from 1980 to 2003. Space-time K function was employed to globally examine the existence of spatial-temporal clustering in the HD data. Three approaches of Kulldorff's scan statistic, i.e., spatial clustering analysis for the entire period, spatial-temporal clustering analysis, and spatial clustering analysis by individual years, were applied to detect potential HD clusters. Statistically significant spatial clusters and spatial-temporal clusters were detected in the five southeastern states during the 24-year study period. Some clusters were observed in multiple years. Clusters were most evident in west Alabama, south Alabama, central South Carolina, and along the border between South Carolina and North Carolina. The identification of HD clusters may provide a means to better understand the causal factors related to the HD outbreaks. Results also have potential application in improving or designing effective surveillance programs for this disease.     

吉林省鹿源牛分枝杆菌MIRU分型研究

为了解鹿源牛分枝杆菌(M.bovis)基因多态性,初步建立适合吉林省鹿源M.bovis的基因分型方法,本研究利用12个分枝杆菌散在重复单位(MIRU)位点,对96株鹿源M.bovis吉林分离株进行MIRU基因分型研究.结果显示,12个位点中有8个位点(MIRU2、4、16、23、27、31、39及40)具有多态性,可以将96株菌株分为1 1个基因型,分辨力指数(H)为0.893,其中6个中度多态性位点的分辨力为0.877.其余4个位点(MIRU 10、20、24及26)未显示多态性.研究结果表明,鹿源M.bovis与其他宿主来源的M.bovis在基因分型上存在差异.MIRU位点对鹿源M.bovis具有良好的分辨力,该分型方法可以作为鹿结核病分子流行病学研究的有效方法.     

Molecular epidemiology of Mycobacterium bovis isolates from free-ranging wildlife in South African game reserves

Bovine tuberculosis is endemic in African buffalo and a number of other wildlife species in the Kruger National Park (KNP) and Hluhluwe-iMfolozi Park (HiP) in South Africa. It was thought that the infection had been introduced into the KNP ecosystem through direct contact between cattle and buffalo, a hypothesis which was confirmed in this study by IS6110 and PGRS restriction fragment length polymorphism (RFLP) typing. The molecular characterisation of 189 Mycobacterium bovis isolates from nine wildlife species in the HiP, including three smaller associated parks, and the Kruger National Park with adjacent areas showed that the respective epidemics were each caused by an infiltration of a single M. bovis genotype. The two M. bovis strains had different genetic profiles, as demonstrated by hybridisation with the IS6110 and PGRS RFLP probes, as well as with regard to evidence of evolutionary changes to the IS profile. While the M. bovis type in HiP was transmitted between buffaloes and to at least baboon, bushpig and lion without obvious genetic changes in the RFLP patterns, in the KNP a dominant strain was represented in 73% of the M. bovis isolates, whilst the remaining 27% were variants of this strain. No species-specific variants were observed, except for one IS6110 type which was found only in a group of five epidemiologically related greater kudu. This finding was attributed to species-specific behaviour patterns rather than an advanced host-pathogen interaction.