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identification by spoligotyping of a caprine genotype in mycobacterium bovis strains causing human tuberculosis.we have used spoligotyping to characterize 18 mycobacterium bovis strains isolated from cattle and 23 m. bovis strains isolated from goats. the spoligotypes revealed that caprine strains form a separate and well-differentiated group that we refer to hereafter in this abstract as the caprine genotype. to evaluate the importance of this genotype as a cause of tuberculosis in other animal species, including humans, we applied the spoligotyping method to 112 strains, including to all isolates identi ...19979399549
vaccination of the badger (meles meles) against mycobacterium bovis.this review summarizes current knowledge relating to the potential development of a vaccine against mycobacterium bovis (m. bovis) for use in badgers. the serious economic loss incurred from bovine tuberculosis in ireland is highlighted. the problems presented to national bovine tuberculosis eradication programmes by wildlife reservoirs of m bovis, in particular the badger and the brushtail possum, are outlined. badger and possum ecology and the epidemiology of m. bovis infection in these specie ...19968870197
achievements and difficulties in maintaining the tuberculosis-free status of hungarian cattle herds.the tuberculosis-free status of hungarian cattle herds in the period between 1988 and the end of 1993 is evaluated. an epidemiological analysis of tuberculin tests, laboratory assays and allergic tests yielding positive results, summarized in three tables, is given with respect to the cattle population of hungary. the origin of positive reactions obtained in the tuberculin tests was traced in different farms of a total of 323 communities. on those farms, the diagnostic slaughter and examination ...19958882736
distribution of lesions in cattle infected with mycobacterium bovis.detailed postmortem examinations were conducted on 30 cattle from a dairy herd with bovine tuberculosis to determine the distribution of lesions in mycobacterium bovis-infected cattle. twenty-four different tissue specimens from each animal were examined for gross lesions and collected for bacteriologic culturing and histologic examination. tuberculosis was confirmed in 15 cattle with evidence of infection in 1 or more of the following tissues: medial retropharyngeal, parotid, tracheobronchial, ...19968844579
effect of dexamethasone treatment of tuberculous cattle on results of the gamma-interferon test for mycobacterium bovis.the purpose of this project was to determine the effect of dexamethasone (dex) treatment of tuberculous cows on antigen-stimulated gamma-interferon (gamma-ifn) production in a commercial mycobacterium bovis gamma-interferon test (gamma-ifn test) developed for diagnosis of bovine tuberculosis. in the gamma-ifn test an enzyme-linked immunosorbent assay is used to detect bovine gamma-ifn in the plasma from whole blood samples cultured with m. bovis and mycobacterium avium tuberculin purified protei ...19968941967
rapid diagnosis and strain differentiation of mycobacterium bovis in radiometric culture by spoligotyping.an assessment of spoligotyping for rapid detection and strain typing of mycobacterium bovis isolates in radiometric culture was made. spoligotyping was applied to bactec 12b broth cultures of 54 lesioned bovine lymph node specimens from 44 herds in northern ireland. a nucleic acid sequence capture technique was performed on bactec cultures at growth index points of approximately (approximately) 60, approximately 200, and 999. definitive spoligotype patterns were obtained for 90.4% and 94.2% of a ...19989646467
detection of mycobacterium avium subsp. paratuberculosis in infected tissues by new species-specific immunohistological procedures.we have previously described the cloning and sequencing of a gene portion coding for the terminal part of a 34-kda protein of mycobacterium avium subsp. paratuberculosis, the etiological agent of johne's disease (p. gilot, m. de kesel, l. machtelinckx, m. coene, and c. cocito, j. bacteriol. 175:4930-4935, 1993). the recombinant polypeptide (a362) carries species-specific b-cell epitopes which do not cross-react with other mycobacterial pathogens (m. de kesel, p. gilot, m.-c. misonne, m. coene, a ...19989665946
specific differentiation between mycobacterium bovis bcg and virulent strains of the mycobacterium tuberculosis complex.a pcr procedure based on the intergenic region (ir) separating two genes encoding a recently identified mycobacterial two-component system, named senx3-regx3, was developed and was shown to be suitable for identifying mycobacterium bovis bcg. the senx3-regx3 ir contains a novel type of repetitive sequence, called mycobacterial interspersed repetitive units (mirus). all tested bcg strains exclusively contained 77-bp mirus within the senx3-regx3 ir, whereas all non-bcg m. tuberculosis complex stra ...19989705376
brucella abortus strain rb51 as a vector for heterologous protein expression and induction of specific th1 type immune responses.brucella abortus strain rb51 is a stable, rough, attenuated mutant widely used as a live vaccine for bovine brucellosis. our ultimate goal is to develop strain rb51 as a preferential vector for the delivery of protective antigens of other intracellular pathogens to which the induction of a strong th1 type of immune response is needed for effective protection. as a first step in that direction, we studied the expression of a foreign reporter protein, beta-galactosidase of escherichia coli, and th ...200010816476
[tuberculous lymphadenitis caused by mycobacterium bovis].according to the literature, bovine tuberculosis in man has been an illness of minor importance in norway, unlike in sweden and denmark. this situation cannot be explained, since in former days infected cattle were a problem in the southeastern part of the country in particular. no case of bovine tuberculosis in humans has been reported in norway since 1940. recently we have observed a 29-year-old female immigrant from india with cervical lymph node tuberculosis caused by m. bovis. a second case ...19968650657
dna fingerprinting of mycobacterium bovis strains by restriction fragment analysis and hybridization with insertion elements is1081 and is6110.strains of mycobacterium bovis, the causative organism of bovine tuberculosis, can be clearly distinguished from each other by restriction fragment analysis. this method of dna fingerprinting has been used for many epidemiological studies in new zealand, but the technique presents practical difficulties that hinder its widespread use. the insertion element is6110 is being widely used as a dna probe for distinguishing restriction fragment polymorphisms among strains of mycobacterium tuberculosis. ...19938099083
molecular epidemiological studies of mycobacterium bovis infections in humans and animals in sweden.forty-nine isolates of mycobacterium bovis from humans and animals in sweden were analyzed by restriction fragment length polymorphism (rflp) patterns probed by the insertion element is6110. most isolates had patterns indicating the presence of only one or two genomic copies of the is6110 insertion element. this simple type of pattern was found in all human isolates. in contrast, isolates from m. bovis infections in five herds of farmed deer in sweden showed a specific rflp pattern with seven ba ...19958586698
mycobacterium paratuberculosis: a potential food-borne pathogen?mycobacterium paratuberculosis commonly infects dairy cattle, leading to johne's disease, which is also known as paratuberculosis. the infection is chronic progressive, and incurable. as the infection progresses, excretion of m. paratuberculosis in feces and milk occurs, and the bacterium spreads through the blood to multiple internal organs. consequently, raw products originating from cattle may harbor m. paratuberculosis. thermal treatments, such as pasteurization, are commonly relied on to ki ...19979436127
characterization of a specific mycobacterium paratuberculosis recombinant clone expressing 35,000-molecular-weight antigen and reactivity with sera from animals with clinical and subclinical johne's disease.johne's disease is a chronic enteritis of ruminants associated with enormous worldwide economic losses for the dairy cow- and goat-rearing industries. management limitations and eradication programs for this disease have been hampered by the lack of a simple and specific diagnostic test for the detection of subclinical cases. we used a recombinant clone expressing a 35,000-molecular-weight mycobacterium paratuberculosis antigen (p35 antigen) from a previously constructed expression library of m. ...19979196196
a multiplex approach to molecular detection of brucella abortus and/or mycobacterium bovis infection in cattle.a multiplex amplification and detection platform for the diagnosis of mycobacterium bovis and brucella abortus infection simultaneously in bovine milk and nasal secretions was developed. this system (designated the bovine pathogen detection assay [bpda]-pcr) consists of duplex amplification of species-specific targets (a region of the bcsp31k gene of b. abortus and a repeat-sequence region in the hsp65 gene of m. bovis, respectively). this is followed by a solid-phase probe capture hybridization ...200010878051
dynamic changes in circulating and antigen-responsive t-cell subpopulations post-mycobacterium bovis infection in cattle.bovine tuberculosis is a threat to animal and human health in several countries. greater understanding of the immunology of the disease is required to develop improved tests and vaccines. this study has used a model of bovine tuberculosis, established in the natural host, to investigate the dynamic changes that occur in the circulating t-cell subpopulations after infection. when the phenotypic composition of the peripheral blood lymphocytes was determined pre- and post-experimental infection, th ...19968698385
glycolipid antigen for use in diagnostic assays for bovine tuberculosis.a glycolipid antigen, was isolated, purified and characterized from mycobacterium bovis an5. chemical analysis (thin-layer chromatography, nuclear magnetic resonance and infrared spectra) showed that this glycolipid was a 2,3-di-o-acyl trehalose (dat), similar to the dat of m. tuberculosis. this antigen was used to establish elisa-based serodiagnostic tests for m. bovis-infected cattle. the sensitivity and specificity of the assay were investigated using sera of cattle from tuberculosis-free her ...19979765827
role of cd8+ and wc-1+ gamma/delta t cells in resistance to mycobacterium bovis infection in the scid-bo mouse.the role of various effector t cell populations in the bovine immune response to mycobacterium bovis infection is poorly understood. this is largely due to the difficulties associated with performing in vivo challenge studies in the natural host species. in this report, we utilized a fetal bovine-severe combined immunodeficient (scid-bo) xenochimeric mouse model to study the protective role of two putative effector cell types, cd8+ t cells and a subpopulation of gamma/delta t cells that express ...19999886243
an mpb-64 flanking sequence specific for mycobacterium bovis.a clone carrying a plasmid with the mpb-64 gene and 3' flanking sequences (plasmid pmba122) was detected during the screening of a mycobacterium bovis genomic library with sera from infected cattle. when the pmba122 insert was used as a probe in southern blots against pvuii-digested mycobacterial dna, it distinguished the different m. tuberculosis complex species. this probe hybridized with a 7-kb band in m. tuberculosis, a 5-kb band in m. bovis and a 3-kb band in m. tuberculosis complex strains ...19979765821
evaluation of a five-antigen elisa for diagnosis of tuberculosis in cattle and cervidae.to determine the validity of a 5-antigen elisa for detection of tuberculosis in cattle and cervidae.19968790550
effect of tuberculosis on milk production in dairy cows.to evaluate differences in milk production between cows with positive and negative caudal fold tuberculin test results in a mycobacterium bovis-infected dairy herd.19989743727
protection against aerosol mycobacterium tuberculosis infection using mycobacterium bovis bacillus calmette guérin-infected dendritic cells.in the lung, dendritic cells (dc) are key antigen-presenting cells capable of triggering specific cellular responses to inhaled pathogens, and thus, they may be important in the initiation of an early response to mycobacterial infections. the ability of dc to enhance antigen presentation to naive t cells within the lungs was characterized with respect to mycobacterium bovis bacillus calmette guérin (bcg) vaccination against m. tuberculosis infection. in vitro derived dc were infected with bcg, w ...199910382760
detection of mycobacterium bovis lymphocyte stimulating antigens in culture filtrates of a recombinant mycobacterium smegmatis cosmid library.culture filtrates derived from a mycobacterium bovis cosmid library in mycobacterium smegmatis were screened for bovine lymphocyte stimulatory antigens using peripheral blood mononuclear cells (pbmc) from cattle vaccinated with a low dose of mycobacterium bovis bcg. lymphocyte proliferation and interferon-gamma (ifn-gamma) production were used as cellular response markers for antigen recognition. in the primary screen, approximately 28% of all culture filtrates (cf) stimulated responses by pbmc ...199910438049
usefulness of spoligotyping in molecular epidemiology of mycobacterium bovis-related infections in south america.two hundred twenty-four mycobacterium bovis isolates, mainly from south american countries, were typed by spoligotyping, and 41 different spoligotypes were identified. a total of 202 m. bovis isolates (90%) were grouped into 19 different clusters. the largest cluster contained 96 isolates (42.8%) on the basis of the most frequently observed spoligotype, spoligotype 34. nineteen m. bovis isolates from humans in argentina had spoligotypes and polymorphic gc-rich repetitive sequence (pgrs) types th ...19999889207
identification of mycobacterium bovis in bovine clinical samples by pcr species-specific primers.tuberculosis, caused by mycobacterium bovis is emerging as the most important disease affecting cattle. furthermore, it results in a major public health problem when transmitted to humans. due to its difficult and non-specific diagnosis, m. bovis has been declared to be one of the etiologic agents causing significant economic loss in the cattle industry. our group evaluated a more rapid and specific method, based on a new polymerase chain reaction species-specific primers, which amplifies a 470- ...199910369566
a versatile model of disease transmission applied to forecasting bovine tuberculosis dynamics in white-tailed deer populations.a model was derived for disease transmission in dynamic host populations and its application was demonstrated in forecasting possible outcomes of a bovine tuberculosis (mycobacterium bovis) epidemic in a white-tailed deer (odocoileus virginianus) population. the approach was mechanistic, based disease transmission on the probability of each susceptible individual becoming infected per unit time, and afforded the flexibility necessary to model epidemics in dynamic wildlife populations. this appro ...19989813841
the public health risks of cervid production in the united states of america.cervid species represent a growing livestock enterprise in the united states of america (usa). the zoonotic threat of bovine tuberculosis (mycobacterium bovis) is the only significant public health risk posed by this alternative livestock industry. this paper examines the potential sources of tuberculosis exposure as related to public health and compares and contrasts the status of tuberculosis in cervidae with the situation in the cattle industry in the usa. based on the existing prevalence of ...19979501369
immunochemical characterization of the mpb70/80 and mpb83 proteins of mycobacterium bovis.mpb70 and mpb80 (mpb70/80) and mpb83 are closely related antigens which are highly expressed in mycobacterium bovis. mpb70/80 are soluble secreted antigens, while mpb83 is an exported lipoprotein associated with the bacterial surface. in the present study, these antigens had different mobilities in sodium dodecyl sulfate-polyacrylamide gel electrophoresis under reducing and nonreducing conditions. these differences may be explained by the fact that mpb70 and mpb83 both have two internal cysteine ...19989529066
detection of mycobacterium bovis in milk by polymerase chain reaction.a simple method was developed for dna extraction of mycobacterium bovis in milk and further detection of the bacterium by polymerase chain reaction (pcr). milk previously seeded with m. bovis was used as the starting material. the procedure involved overnight digestion of a milk sample with proteinase k at 56 degrees c and phenol extraction, followed by ethanol precipitation and pcr. the amplification pattern obtained was analyzed with primers bw8-bw9 which amplify a 248 bp in strains of m. bovi ...19989820115
investigations on the prevalence of bovine tuberculosis and brucellosis in dairy cattle in dar es salaam region and in zebu cattle in lugoba area, tanzania.a study between august 1995 and december 1997 included 343 dairy cattle on 20 farms in the dar es salaam region and 2289 zebu cattle on 39 bomas in the lugoba area (coast region). the aim was to establish the prevalence of bovine tuberculosis (mycobacterium bovis) and bovine brucellosis (brucella abortus). in the single intradermal tuberculin test (sit), 0.9% (3/343) of the animals in dar es salaam tested positive and 1.2% (4/343) were doubtful. positive reactors were found in 10% (2/20) of the ...200010907285
identification of a novel dna probe for strain typing mycobacterium bovis by restriction fragment length polymorphism analysis.bovine tuberculosis caused by mycobacterium bovis remains a significant disease of farmed cattle in many countries despite ongoing tuberculosis eradication programs. molecular typing methods such as restriction fragment length polymorphism (rflp) analysis and spoligotyping have been used to identify related herd breakdowns in an attempt to identify more precisely the route of infection into cattle herds and to trace the transmission of bovine tuberculosis. a recent geographical survey of irish m ...200010790088
characterization of exochelins of the mycobacterium bovis type strain and bcg substrains.pathogenic mycobacteria must acquire iron in the host in order to multiply and cause disease. to do so, they release abundant quantities of siderophores called exochelins, which have the capacity to scavenge iron from host iron-binding proteins and deliver it to the mycobacteria. in this study, we have characterized the exochelins of mycobacterium bovis, the causative agent of bovine and occasionally of human tuberculosis, and the highly attenuated descendant of m. bovis, bacillus calmette-guéri ...199910085056
failure to identify non-bovine reservoirs of mycobacterium bovis in a region with a history of infected dairy-cattle herds.the state of texas had the most (cumulative) tuberculous cattle herds of any state in the united states during the decade ending in 1997. of the cumulative 18 infected herds in texas, 12 herds were concentrated in el paso county (designated the 'el paso milkshed'). to identify whether non-bovine reservoirs were a source of mycobacterium bovis infection of cattle in this region, an investigation was conducted on the premises of 14 dairy herds (12 tuberculous and 2 non-affected herds) between may ...200010665951
the effect of a badger removal programme on the incidence of tuberculosis in an irish cattle population.the risk of a confirmed tuberculous herd restriction was examined using a logistic model for herds involved in the east offaly badger research project, ireland, from 1988-1995. cattle herds present in the badger-removal area had a significantly lower proportion of new confirmed tuberculous herd restrictions compared with cattle from an area where no systematic badger removal was attempted.19989541950
prevalence of tuberculosis in cattle in the dangme-west district of ghana, public health implications.bovine tuberculosis due to mycobacterium bovis is a major cause of human gastrointestinal tuberculosis in developing countries where bovine milk is often not pasteurised before use. little information is available on the prevalence of the disease in african cattle and its zoonotic impact. the ghanaian government, through its peri-urban dairy cattle development project, is promoting the use of milk and dairy products from local cows in selected districts of the country, including the dangme-west ...200010913759
dendritic cells induce cd4+ and cd8+ t-cell responses to mycobacterium bovis and m. avium antigens in bacille calmette guérin vaccinated and nonvaccinated cattle.few data are available regarding the induction of memory t-lymphocyte responses in cattle following bacille calmette guérin (bcg) vaccination. studies of the immune response induced by bcg vaccination provide an insight into the basis of antimycobacterial immunity that could be exploited for the development of more effective vaccination strategies. we used autologous dendritic cells (dc) infected with mycobacterium bovis bacille calmette guérin (bcg) or pulsed with purified protein derivative fr ...200010972905
vaccination of cattle with mycobacterium bovis culture filtrate proteins and interleukin-2 for protection against bovine tuberculosis.in this study vaccines prepared from culture filtrate proteins (cfp) of mycobacterium bovis and interleukin-2 (il-2) were tested in cattle for their capacity to stimulate immune responses and to protect against an intratracheal challenge with virulent m. bovis. nine groups of cattle were vaccinated with combinations of different doses of cfp and bovine il-2 mixed with a monophosphoryl lipid a (mpl) adjuvant. an additional group was vaccinated with m. bovis bcg. immune responses in b1p-il-2-vacci ...200010992489
risk factors for the between-herd spread of mycobacterium bovis in canadian cattle and cervids between 1985 and 1994.microorganisms of the genus mycobacterium cause tuberculosis in many animal species including humans. generally, mycobacterium bovis (m. bovis) infects cattle and cervids, but it has the potential to infect virtually all species of mammals. this study examined and analysed the data from the nine outbreaks of tuberculosis in canadian cattle and cervids from 1985 to 1994. for the purposes of this study, a positive herd was one with at least one culture-positive animal. a reactor herd had at least ...199910448941
a lymphocyte transformation assay for the detection of mycobacterium bovis infection in the eurasian badger (meles meles).the eurasian badger (meles meles) is a significant wildlife reservoir of mycobacterium bovis in great britain. improved control strategies against the disease in badgers require the development of diagnostic tests and vaccines. here, we report the development of a comparative lymphocyte transformation assay (lta) using bovine and avian tuberculin as antigen to detect cell-mediated responses in m. bovis-infected badgers. in a pilot study, the performance of this assay was compared with the existi ...199910507289
identification of a mycobacterium bovis bcg auxotrophic mutant that protects guinea pigs against m. bovis and hematogenous spread of mycobacterium tuberculosis without sensitization to tuberculin.tuberculosis remains one of the most significant diseases of humans and animals. the only currently available vaccine against this disease is a live, attenuated vaccine, bacillus calmette-guérin (bcg), which was originally derived from mycobacterium bovis and despite its variable efficacy is the most widely administered vaccine in the world. with the advent of the human immunodeficiency virus-aids pandemic concern has been raised over the safety of bcg. moreover, since bcg sensitizes vaccinated ...200011083835
tuberculosis in adult beef cattle of mexican origin shipped direct-to-slaughter into texas.to evaluate differences in prevalence of tuberculosis (tb) in adult beef cattle that originated from different states in mexico and were shipped direct-to-slaughter into texas in 1995.19989491166
molecular typing of mycobacterium bovis isolates from cameroon.in order to gain a better understanding of the molecular epidemiology of mycobacterium bovis isolates in cameroon, 75 isolates of m. bovis collected in three provinces of northern cameroon were studied by spoligotyping. for 65 of these isolates, typing was also carried out by pulsed-field gel electrophoresis (pfge) with drai, and 18 of the isolates were also typed by restriction fragment length polymorphism (rflp) analysis with probe is6110-rhs. molecular typing of the isolates by these techniqu ...200111136775
vaccination of mice and cattle with plasmid dna encoding the mycobacterium bovis antigen mpb83.a scientific review of bovine tuberculosis in great britain has concluded that the development of a cattle vaccine holds the best prospect for long-term disease control. recent reports of successful dna vaccination against mycobacterium tuberculosis in small animal models have raised the possibility of using a similar strategy to produce vaccines against mycobacterium bovis infection in cattle. to test this possibility, balb/c mice were immunized with dna encoding the m. bovis antigen mpb83. the ...200010875801
characterization of the early antibody response in bovine tuberculosis: mpb83 is an early target with diagnostic potential.a 26-kda antigen has been shown to be a dominant antibody target in mycobacterium bovis-infected cattle. in this study, that antigen was used as an immunogen to raise a panel of mouse monoclonal antibodies. the majority of those bound to native protein with a molecular mass of 26 kda and to recombinant mpb83, strongly suggesting that mpb83 is an important b-cell antigenic target in bovine tuberculosis. in order to provide assessment of the potential of measuring antibody responses to the native ...200111285116
use of innovative methods in the eradication of bovine tuberculosis.during final phases of eradication programmes, strains of mycobacterium sp. not belonging to the tuberculosis complex increase their relative frequency and are responsible for positive skin test reactions. moreover, the specificity of any indirect diagnostic test, such as the skin test, is never completely accurate, therefore even when tuberculosis infection is completely eradicated, a number of false positive reactions are to be expected. the aim of this paper is to evaluate the performances of ...200010900822
estimates of within-herd incidence rates of mycobacterium bovis in canadian cattle and cervids between 1985 and 1994.we analysed the individual-animal data from six of the nine outbreaks of tuberculosis in canadian cattle and cervids from 1985 to 1994. a "positive/reactor" animal was one which had either a positive culture or a positive or suspicious reaction on a mid-cervical, comparative cervical, or gross or histopathological test for tuberculosis. individual-animal data were collected only for herds which had one or more positive/reactor animals. data were collected from the outbreak records in the regiona ...200010821964
mycobacterium bovis infection and control in domestic livestock.bovine tuberculosis, caused by mycobacterium bovis, is a well-known zoonotic disease which affects cattle world-wide. the public health risk has been alleviated in many countries by the introduction of pasteurisation, but the disease continues to cause production losses when poorly controlled. the office international des epizooties classifies bovine tuberculosis as a list b disease, a disease which is considered to be of socio-economic or public health importance within countries and of signifi ...200111288521
dna amplification assay for rapid detection of bovine tubercle bacilli in semen.tubercle bacilli shed in the semen can be a potential hazard for unlimited number of cows through artificial insemination. we have evaluated the efficacy of a dna amplification technique by polymerase chain reaction (pcr) for the detection of tubercle bacilli in fresh and frozen semen using spiked samples. the test was based on insertion sequence is 1081 and could detect as low as 10-100 bacterial cells per ml of spiked semen. the specificity of the test was 100%. the method was applied to semen ...199910565436
differentiation among members of the mycobacterium tuberculosis complex by molecular and biochemical features: evidence for two pyrazinamide-susceptible subtypes of m. bovis.the variations in biochemical as well as molecular characteristics among several members of the mycobacterium tuberculosis complex that are not m. tuberculosis have been assessed to facilitate an unambiguous species identification. altogether, 96 m. tuberculosis complex strains including 52 m. bovis isolates and 44 m. africanum isolates were analyzed by spoligotyping. the strains could be clustered into five spoligotype groups. all m. bovis isolates showed the typical absence of the spacers 39 t ...200010618079
lymphocyte subset proliferative responses of mycobacterium bovis-infected cattle to purified protein derivative.despite highly successful eradication efforts in several countries, mycobacterium bovis infection of cattle remains a significant health concern worldwide. immune mechanisms of resistance to and/or clearance of m. bovis infection of cattle, however, are unclear. recent studies have provided evidence supporting a role for cd4(+), cd8(+), and gammadelta tcr(+) t cells in the response of cattle to m. bovis. in the present study, we utilized a flow cytometric-based proliferation assay to determine t ...200011137124
[immunological methods for the diagnosis of bovine tuberculosis--state of the art].an overview is given about the most common immunological methods for the diagnosis of bovine tuberculosis. these methods are based on cell mediated immune reactions on the one hand (intradermal skin test, interferon-gamma-test, lymphocyte transformation test) or on antibody production against mycobacterium bovis on the other hand (elisa). the advantages and disadvantages of the methods as well as their specificity and sensitivity are discussed.200011084757
tuberculosis in deer: perceptions, problems and progress.since the emergence of deer farming as an alternative farming enterprise over the past 30 years, there has been an increasing awareness of the potential threat posed by tuberculosis (tb) to domesticated deer. tb, caused by mycobacterium bovis, has been found in deer in every country involved with deer farming. different types of tb control policies, which vary from whole-herd depopulation to selective testing and slaughter of reactor animals, have been implemented. extensive research has been ca ...200011061957
a one-tube nested polymerase chain reaction for the detection of mycobacterium bovis in spiked milk samples: an evaluation of concentration and lytic techniques.the objective of this study was to evaluate the use of a one-tube nested polymerase chain reaction (otn pcr) with 5 concentration and lytic treatments for the detection of mycobacterium bovis in experimentally inoculated milk samples (spiked samples). otn pcr and the following treatments were tested in inoculated samples: 1) centrifugation; 2) c18-carboxypropylbetaine + capture resin 1 + proteinase k (cb18-ch-pk); 3) centrifugation + capture resin 1 + proteinase k; 4) centrifugation + capture re ...200111289205
an esat6 knockout mutant of mycobacterium bovis produced by homologous recombination will contribute to the development of a live tuberculosis vaccine.strains of the mycobacterium tuberculosis complex are being rationally attenuated in order to develop better tuberculosis vaccines than bcg, and it would be helpful if new vaccines lacked an immunogenic protein which could be used as a skin test reagent for determining infection status.200011052907
infection of red deer, cattle, and humans with mycobacterium bovis subsp. caprae in western austria.twelve cases of mycobacterium bovis subsp. caprae infection have occurred in four humans, three cattle, and five red deer in western austria since 1994. dna-fingerprinting of the isolates suggested transmission in and between these species over several years. contact with cattle, but not with goats, was found to be associated with three of four human cases.200212037107
optimal models to evaluate the protective efficacy of tuberculosis vaccines.bcg has been used widely as a vaccine to prevent tuberculosis (tb) for 80 years, yet there is still considerable controversy about its efficacy. many experimental variables have obscured the true efficacy of bcg. the absence of appropriate animal models for the study of protective efficacy and the lack of in vitro correlates of protective immunity have impeded progress. laboratory animal studies, which have contributed to understanding the pathogenesis, heritability of resistance and immunology ...200111463234
towards more accurate diagnosis of bovine tuberculosis using defined antigens.diagnostic accuracy is of paramount importance in test-and-slaughter programmes for the eradication of bovine tuberculosis (tb). currently applied methods, such as in vivo skin testing and in vitro interferon-gamma (ifn- gamma) testing, utilize purified protein derivatives (ppds), which are poorly-defined mixtures containing many individual antigenic components. it is known that false-positive responses to these reagents can occur in cattle which are not infected with tb, largely because of that ...200111463225
wildlife disease reservoirs: the epidemiology of mycobacterium bovis infection in the european badger (meles meles) and other british mammals.mycobacterium bovis infection has been confirmed in a wide range of mammals hosts throughout the world. the european badger (meles meles) and the brushtail possum (trichosurus vulpecula) are implicated as significant sources of infection for domestic cattle in the uk and new zealand respectively. the risk of transmission of infection between a wildlife population and domestic animals will be determined by both the epidemiology of the disease and the ecology of the host. in the uk, surveys by the ...200111463223
molecular fingerprinting confirms extensive cow-to-cow intra-herd transmission of a single mycobacterium bovis strain.in this study we have characterized m. bovis isolates from a herd of cattle in uvalde, texas in which 52 of the 193 animals selected at random in 1994 from a herd of 331 were caudal fold skin-test positive. thirty-two of 52 skin-test positive cattle had gross lesions at slaughter, and isolations of m. bovis were made from 29 animals. the herd was comprised of red devon cattle purchased between 1978 and 1980 (n = 26) and breeding bulls (n = 3) introduced at later times, and all were tuberculosis ...199910596810
polymerase chain reaction detection of mycobacterium tuberculosis complex and mycobacterium avium organisms in formalin-fixed tissues from culture-negative ruminants.in the us eradication program for bovine tuberculosis, a definitive diagnosis depends on the isolation of mycobacterium bovis. however, in some cases bacterial culture is unsuccessful, even though the tissue is considered suspicious by histopathology because granulomatous lesions and acid-fast organisms are present. the purpose of this study was to determine if polymerase chain reaction (pcr) tests on formalin-fixed tissue would successfully identify the organisms observed in suspect lesions fro ...200212079743
immunological responses of eurasian badgers (meles meles) vaccinated with mycobacterium bovis bcg (bacillus calmette guerin).wildlife species, such as the badger (meles meles), may act as maintenance hosts for mycobacterium bovis and contribute to the spread and persistence of tuberculosis in associated cattle populations. targeted vaccination of badgers against tuberculosis is an option that, if successfully employed, could directly facilitate the advancement of bovine tuberculosis eradication in affected areas. in this study, the immunological responses of a group of badgers vaccinated subcutaneously with low doses ...200111389955
the epidemiology of tuberculosis in free-ranging african buffalo (syncerus caffer) in the kruger national park, south africa.the presence of bovine tuberculosis (mycobacterium bovis) in the kruger national park (knp) was determined for the first time in 1990. it was diagnosed in an african buffalo (syncerus caffer) bull, which was found recumbent and in an emaciated and moribund state near the south-western boundary fence. this prompted an investigation into the bovine tuberculosis (btb) status of the knp, with emphasis on its epidemiological determinants and risk factors. this report documents the findings of surveys ...200111585089
spoligotype diversity of mycobacterium bovis strains isolated in france from 1979 to 2000.the molecular fingerprints of 1,349 isolates of mycobacterium bovis received between 1979 and august 2000 at agence française de sécurité sanitaire des aliments (afssa) have been obtained by spoligotyping. the majority of the isolates (1,266) were obtained from cattle living in france. an apparently high level of heterogeneity was observed between isolates. one hundred sixty-one spoligotypes were observed in total, of which 153 were from french isolates. the two predominant spoligotypes, designa ...200111574583
vaccination of the brushtail possum (trichosurus vulpecula) against mycobacterium bovis infection with bacille calmette-guérin: the response to multiple doses.in new zealand, the brushtail possum (trichosurus vulpecula) is the principal wildlife vector of bovine tuberculosis. control of infected possum populations contributes to the control of tuberculosis in domestic livestock. vaccination is potentially a complementary strategy to population control, but to be cost-effective, administration of the vaccine to possums would need to be from an appropriately designed automatic vaccinator. possums themselves would activate the vaccinator so that it would ...200211750141
modulation of mycobacterium bovis-specific responses of bovine peripheral blood mononuclear cells by 1,25-dihydroxyvitamin d(3).historically, administration of vitamin d has been considered beneficial in the treatment of tuberculosis. the interaction of this vitamin [i.e., 1,25-dihdroxyvitamin d(3) [1,25(oh)(2)d(3)]] with the antitubercular immune response, however, is not clear. in the present study, in vitro recall responses of peripheral blood mononuclear cells (pbmc) from cattle infected with mycobacterium bovis were used to study the immune-modulatory effects of 1,25(oh)(2)d(3) on m. bovis-specific responses in vitr ...200111687464
a guinea pig model of low-dose mycobacterium bovis aerogenic infection.in order to develop a model of mycobacterium bovis infection with pathogenetical relevance, a modified version of the henderson apparatus was used to deliver infectious aerosols directly to the snouts of guinea pigs. aerosols generated from 10(6), 10(7), 10(8)cfu/ml m. bovis suspensions established disease in every animal, with estimated retained doses of 10, 100, 1000 cfu, respectively. for comparison, other guinea pigs were inoculated with 100 cfu m. bovis intramuscularly (i.m.). pathology and ...200111337137
identification of tuberculosis in cattle slaughtered in mexico.to determine epidemiologic factors associated with tuberculosis (tb) in dairy cattle slaughtered in 6 important regions for milk production in mexico.200010630785
risk factors associated with the occurrence of bovine tuberculosis in cattle in the southern highlands of tanzania.a study was conducted in the southern highlands of tanzania to determine the prevalence of bovine tuberculosis and the risk factors associated with the occurrence of the disease in cattle of different categories and in different climatic zones. the overall prevalence of the disease was 13.2%, and 51% of the herds tested contained reactor cattle. assessment of risk factors was based on comparisons of the reactivity of the cattle in the single comparative intradermal tuberculin test (scitt). older ...200111767006
[specific features of mycobacteria isolated from patients and animals with tuberculosis and from environmental objects].two hundred samples from environmental objects, such as soil, manure, water, feeds, et.), 129 isolates from tuberculosis patients' sputum, and 222 samples from tuberculin-responsive animals were studied. mycobacteria of atypical forms were found to be widely distributed in the environment. the causative agents of bovine-type tuberculosis could be isolated from both tuberculin-responsive animals and environmental objects of tuberculosis-burdened firms. the cultural morphological and biochemical p ...200111588963
[application of rapid detection for mycobacterium tuberculosis with phage splitting assay].to study the significance of rapid identification for mycobacterium tuberculosis with phage splitting assay.200111770425
modulation of immune responses to mycobacterium bovis in cattle depleted of wc1(+) gamma delta t cells.it is accepted that cell-mediated immune responses predominate in mycobacterial infections. many studies have shown that cd4(+) t cells produce th1 cytokines, such as gamma interferon (ifn-gamma), in response to mycobacterial antigens and that the cytolytic activity of cd8(+) cells toward infected macrophages is important. however, the extent and manner in which gamma delta t cells participate in this response remain unclear. in ruminants, gamma delta t cells comprise a major proportion of the p ...200211854237
identification of novel mycobacterium tuberculosis antigens with potential as diagnostic reagents or subunit vaccine candidates by comparative genomics.an independent review for the british government has concluded that the development of a cattle vaccine against mycobacterium bovis holds the best long-term prospects for tuberculosis control in british herds. the development of complementary diagnostic tests to differentiate between vaccinated and infected animals is necessary to allow the continuation of test-and-slaughter-based control policies alongside vaccination. vaccination with m. bovis bacillus calmette-guérin (bcg), the only available ...200212438379
evaluation of the abbott lcx mycobacterium tuberculosis assay for direct detection of mycobacterium bovis in bovine tissue samples.the commercial lcx amplification assay, usually employed to detect the mycobacterium tuberculosis complex in respiratory specimens, was evaluated by comparing the results it gave with those obtained using lowenstein-jensen solid medium and pathological findings in 55 lymph nodes from cattle with positive and 10 lymph nodes from cattle with negative skin tests for tuberculosis. fifty-three cultures (51 and 2, respectively) were positive for m. bovis, while the results for the lcx assay and the hi ...200211860084
masking of two in vitro immunological assays for mycobacterium bovis (bcg) in calves acutely infected with non-cytopathic bovine viral diarrhoea virus.acute infection of calves, previously vaccinated with bacille calmette-guerin (bcg), with non-cytopathic viral diarrhoea virus (bvdv) resulted in the temporary suppression of two in vitro assays used to monitor mycobacterium bovis infection. lymphocyte proliferation and interferon-gamma production by whole blood cultures containing purified protein derivatives prepared from mycobacterium avium (ppd-a) and m bovis (ppd-b) were markedly suppressed. the implication is that acute infections of cattl ...200111700926
generalized tuberculosis in a 45 day-old calf.the objectives of the present study were to describe pathomorphological and immunohistochemical features of generalized natural tuberculosis in a 45 day-old female calf. the characteristic lesion of tuberculosis was productive type which was located in the lung, liver, and especially in the lymph nodes (mediastinal, bronchial, mesenterial, portal, prescapular, renal and caudal sternal lymph nodes). mycobacterium bovis antigens were seen generally in the cytoplasma and around the macrophages, rar ...200111765603
mycobacterium bovis: polymerase chain reaction identification in bovine lymphonode biopsies and genotyping in isolates from southeast brazil by spolygotyping and restriction fragment length polymorphism.diagnosis of the mycobacterium tuberculosis complex by direct pcr of mediastinal lymphnode dna and microbiological tests were compared in cattle suspicious of bearing tuberculous-like lesions detected during slaughter. the pcr procedure applied on dna samples (n=54) obtained by adding alpha -casein into the thiocyanate extraction mix was positive in 70% of the samples. pcr confirmed the identification of 23 samples (100%) that grew in culture, 9 samples (60%) that failed to grow in culture, plus ...200111562707
tuberculosis in badgers; a review of the disease and its significance for other animals.this review examines the current state of knowledge of aspects of tuberculosis in the badger. the gross pathology and pathogenesis are elaborated as well as the immune mechanism, diagnosis of infection and excretion and viability of infected products. the epidemiology in badgers is considered, as is the significance of infection in this species for other wildlife species as well as domestic animals sharing the same habitat. trials of the effects of the removal of badgers on the occurrence of tub ...200011124091
control of mycobacterium bovis infections and the risk to human populations.conventional control methods based on test-and-slaughter policies have, in several countries, led to the successful eradication of bovine tuberculosis in cattle. however, new approaches for control of bovine tuberculosis are required in developing countries and those with a wildlife reservoir of infection. recent developments include improved diagnostics and evaluation of new vaccination strategies.200211932198
the hook lake wood bison recovery project: can a disease-free captive wood bison herd be recovered from a wild population infected with bovine tuberculosis and brucellosis?the hook lake wood bison recovery project (hlwbrp) is a wildlife conservation project aimed at recovering a captive, disease-free herd of wood bison (bison bison athabascae) from a wild herd infected with bovine tuberculosis (mycobacterium bovis) and brucellosis (brucella abortus). the disease eradication protocol that we have used involves a combination of techniques, including (1) orphaning of newborn wild-caught calves to minimize exposure to b. abortus and m. bovis, (2) testing calves for ma ...200212381597
experimental inoculation of north american opossums (didelphis virginiana) with mycobacterium bovis.eight north american opossums (didelphis virginiana) were inoculated with 1 x 10(5) colony forming units of mycobacterium bovis to investigate their potential as reservoir hosts for bovine tuberculosis in michigan. four animals received this dose orally and four were inoculated intramuscularly (i.m.). in each group, two animals were euthanized 1 mo postinoculation (pi) and two at 2 mo pi. four control animals were housed separately and sacrificed in the same manner as those inoculated. one of fo ...200212038125
the p2x7 purinergic receptor on bovine macrophages mediates mycobacterial death.p2x7 is an atp gated purinoceptor that has been linked to various immune responses. p2x7 appears to be expressed ubiquitously in the immune system and thus may be important as an effector pathway or play significant roles in cell activation/death. 2',3'-(4-benzoyl)benzoyl atp is the most potent agonist of this receptor and atp in its fully dissociated form (atp(4-)) also activates the receptor. high concentrations of atp can cause the p2x7 receptor to induce pore formation on the surface of the ...200111292527
kinetics of interferon-gamma (ifn-gamma) release in the peripheral blood of calves vaccinated with bcg.a study was conducted on 13 friesian-zebu crossbred calves, the ages of which lie between 6 and 18 months, to investigate the kinetics of ifn-gamma release in the peripheral blood following bacille calmete guerine (bcg) vaccination. after being screened for bovine tuberculosis (btb), the calves were vaccinated with 1 ml inoculums containing 6 x 10(6) cfu of bcg. the level of ifn-gamma in the peripheral blood was measured two times before vaccination and seven times after vaccination. using a san ...200212033646
mycobacterium tuberculosis mammalian cell entry operon (mce) homologs in mycobacterium other than tuberculosis (mott).the cloned mammalian cell entry gene mce1a from mycobacterium tuberculosis confers to non-pathogenic escherichia coli the ability to invade and survive inside macrophages and hela cells. the aim of this work was to search for and characterize homologs of the four m. tuberculosis mammalian cell entry operons (mce1, mce2, mce3 and mce4) in mycobacteria other than tuberculosis (mott). the dot-blot and polymerase chain reaction (pcr) experiments performed on 24 clinical isolates representing 20 diff ...200212052567
antigen recognition and immunomodulation by gamma delta t cells in bovine tuberculosis.this report describes the in vitro proliferative responses of peripheral blood gammadelta t cells to defined mycobacterial protein ags and the immunomodulatory effect of gammadelta t cells in cattle infected with mycobacterium bovis. gammadelta t cell responses were specific to m. bovis infection because they were detected in cattle either experimentally or naturally infected with m. bovis, but were not present in uninfected controls. proliferating gammadelta t cell cultures produced enhanced le ...200111313400
tuberculosis, caused by mycobacterium bovis, in a kudu (tragelaphus strepsiceros) from a commercial game farm in the malelane area of the mpumalanga province, south africa.tuberculosis, caused by mycobacterium bovis, was diagnosed for the first time, in a kudu cow from a commercial game ranch in the malelane area of the mpumalanga province close to the kruger national park. this diagnosis has important implications for the eradication of the disease in commercial and communal livestock in the area. kudus are considered to be a potential maintenance host and, because of discharging fistulae in the parotid area where the lymph nodes are commonly infected, they have ...200111769357
use of synthetic peptides derived from the antigens esat-6 and cfp-10 for differential diagnosis of bovine tuberculosis in cattle.in great britain an independent scientific review for the government has concluded that the development of a cattle vaccine against mycobacterium bovis infection holds the best long-term prospect for tuberculosis control in british herds. a precondition for vaccination is the development of a complementary diagnostic test to differentiate between vaccinated animals and those infected with m. bovis so that testing and slaughter-based control strategies can continue alongside vaccination. to date ...200111329460
immunogenicity of mycobacterium tuberculosis rd1 region gene products in infected cattle.current immuno-diagnostic tests for the detection of mycobacterium bovis infection in cattle rely on the use of tuberculin ppd as antigens. however, the use of a cattle vaccine is effectively prohibited because bcg, the only potentially available cattle tb vaccine, compromises the current tuberculin test. the main objective of this study was to identify specific antigens, which could increase the test sensitivity to levels achieved with tuberculin. our approach utilized the availability of the g ...200212296851
responses of bovine wc1(+) gammadelta t cells to protein and nonprotein antigens of mycobacterium bovis.wc1(+) gammadelta t cells of mycobacterium bovis-infected cattle are highly responsive to m. bovis sonic extract (mbse). in mycobacterial infections of other species, gammadelta t cells have been shown to respond to protein and nonprotein antigens, but the bovine wc1(+) gammadelta t-cell antigenic targets within mbse require further definition in terms of the dominance of protein versus nonprotein components. the present study sought to characterize the wc1(+) gammadelta t-cell antigenic targets ...200212379688
implications of tuberculosis in african wildlife and livestock.in most countries, tuberculosis caused by mycobacterium bovis is mainly a disease of domestic cattle and can be controlled successfully by means of a test-and-slaughter program. once the infection spills over into a wild animal species with maintenance host potential, conventional measures are no longer sufficient to provide effective control. in south africa, african buffaloes (syncerus caffer) represent the most important maintenance host for m. bovis. apart from transmitting the disease to pr ...200212381600
bovine tuberculosis in michigan wildlife and livestock.since 1994, the state of michigan has recognized a problem with bovine tuberculosis (tb), caused by mycobacterium bovis, in wild white-tailed deer from a 12-county area in northeastern lower michigan. a total of 65,000 free-ranging deer have been tested, and 340 have been found to be positive for m. bovis. the disease has been found in other wildlife species, and, in 1998, in domestic cattle, where to date 13 beef cattle and 2 dairy cattle herds have been diagnosed with bovine tb. unfortunately, ...200212381603
the wc1(+) gammadelta t-cell population in cattle: a possible role in resistance to intracellular infection.intracellular infections are important in veterinary medicine and detailed understanding of the associated immune responses is needed for optimal development of strategies based on diagnosis and vaccination. it is generally accepted that cell-mediated immune responses are of greatest importance in intracellular infections and recent studies from several bovine models of infection indicate that wc1(+) gammadelta t-cells have a number of possible levels of involvement, which remain incompletely de ...200212383642
association of a macrophage galactoside-binding protein with mycobacterium-containing phagosomes.mycobacteria reside intracellularly in a vacuole that allows it to circumvent the antimicrobial environment of the host macrophage. although the mycobacterial phagosome exhibits selective fusion with vesicles of the endosomal system, identification of host and bacterial factors associated with phagosome bio-genesis is limited. to identify these potential factors, mabs were generated to a membrane preparation of mycobacterial phagosomes isolated from m. tuberculosis-infected macrophages. a mab re ...200211906453
genotyping of mycobacterium bovis by geographic location within mexico.the spacer oligonucleotide typing (spoligotyping) method was used to differentiate 62 mycobacterium bovis isolates obtained from tissues with macroscopic lesions typical of tuberculosis in dairy cattle from different regions of mexico. our purpose was to see if a strain from one region was genetically different from those of other regions (with the long-term aim of doing molecular trace back of isolates obtained in the laboratory). results from the genetic analysis indicate that m. bovis isolate ...200212392876
mycobacterium bovis-infected white-tailed deer (odocoileus virginianus): detection of immunoglobulin specific to crude mycobacterial antigens by elisa.white-tailed deer (odocoileus virginianus) have recently emerged as a source of mycobacterium bovis infection for cattle within north america. the objective of this study was to evaluate the antibody response of m. bovis-infected deer to crude mycobacterial antigens. deer were experimentally inoculated with m. bovis strain 1315 either by intratonsilar instillation or by exposure to m. bovis-infected (i.e., in contact) deer. to determine the time course of the response, including the effects of a ...200212423028
a review of m. bovis bcg protection against tb in cattle and other animals species.bovine tuberculosis (tb) causes severe economic losses in livestock due to low production, animal deaths and condemnation of carcasses. it is also an important constraint in international trade of animals and animal products. a scientific committee in great britain in 1997 concluded that the development of a cattle vaccine would be the best option for long-term control of tb. however, vaccination of cattle currently is not accepted because the vaccine interferes with the skin reaction to the tub ...200312628767
oral delivery of mycobacterium bovis bcg in a lipid formulation induces resistance to pulmonary tuberculosis in mice.a lipid-based formulation has been developed for oral delivery of mycobacterium bovis bacille calmette-guérin (bcg) vaccine. the formulated m. bovis bcg was fed to balb/c mice to test for immune responses and protection against m. bovis infection. the immune responses included antigen-specific cytokine responses, spleen cell proliferation, and lymphocyte-mediated macrophage inhibition of m. bovis. oral delivery of formulated m. bovis bcg to mice induced strong splenic gamma interferon levels and ...200312496154
cultivation of mycobacterium bovis bcg in bioreactors.the mycobacterium bovis bcg vaccine for commercial use is classically produced as surface pellicles by culture on synthetic medium. under these conditions, reproducibility of the cultures and quality assessment are hampered by slow growth of the bacilli, the formation of bacterial aggregates and a high proportion of dead bacilli after processing and final formulation of the vaccine. here, we established dispersed cultures of m. bovis bcg in synthetic media in small-scale bioreactors. these cultu ...200212044554
vaccination of guinea pigs with dna encoding the mycobacterial antigen mpb83 influences pulmonary pathology but not hematogenous spread following aerogenic infection with mycobacterium bovis.protection of cattle against bovine tuberculosis by vaccination could be an important control strategy in countries where there is persistent mycobacterium bovis infection in wildlife and in developing countries where it is not economical to implement a tuberculin test and slaughter control program. the main aim of such a vaccination strategy would be to reduce transmission of infection by reducing the lung pathology caused by infection and preventing seeding of the organism to organs from which ...200211895982
the mpb83 antigen from mycobacterium bovis contains o-linked mannose and (1-->3)-mannobiose moieties.mycobacterium tuberculosis and mycobacterium bovis, the causative agents of human and bovine tuberculosis, have been reported to express a range of surface and secreted glycoproteins, although only one of these has been subjected to detailed structural analysis. we describe the use of a genetic system, in conjunction with lectin binding, to characterize the points of attachment of carbohydrate moieties to the polypeptide backbone of a second mycobacterial glycoprotein, antigen mpb83 from m. bovi ...200312517764
new generation vaccines and delivery systems for control of bovine tuberculosis in cattle and wildlife.advances in the understanding of protective immune responses to tuberculosis are providing opportunities for the rational development of improved vaccines for bovine tuberculosis. protection requires activation of macrophages through stimulation of a th 1 type immune response. ideally, a vaccine for cattle should induce protection without causing animals to react in a tuberculin test when exposed to mycobacterium bovis. a number of new tuberculosis vaccines including attenuated m. bovis strains, ...200212072232
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