A one health-focused literature review on bovine and zoonotic tuberculosis in Pakistan from the past two decades: challenges and way forward for control.

Bovine tuberculosis (bTB), caused by Mycobacterium bovis (M. bovis), is a globally prevalent zoonotic infectious disease. World Organization for Animal Health (WOAH) estimates indicate that up to 10% of the total human TB cases in developing countries are attributed to M. bovis. Pakistan ranks 4th in global milk production with a livestock population of over 212 million animals. Over 8 million families are involved in raising these animals as a means of livelihood. To date, there is an absence of national-level data on the prevalence of bTB and an effective control program is still lacking. The multifaceted impacts and substantial economic losses render addressing bTB a daunting, but highly important challenge. In this review, we summarise all the freely available literature on M. bovis infection from Pakistan using Google scholar and PubMed databases. A total of 40 animal studies were identified using search terms: "bovine tuberculosis in Pakistan, bTB, Pakistan, Mycobacterium bovis in Pakistan, M. bovis in Pakistan"; while seven human studies were identified using the terms: zoonotic tuberculosis in Pakistan', 'M. bovis in humans Pakistan', 'zTB in TB patients in Pakistan". We have summarized all these studies to identify critical risk factors involved in transmission of bTB among animals and humans. Despite lack of comprehensive and geographically representative studies, the literature suggests a varying prevalence of bTB in animals, ranging from as low as 2% to as high as 19%. Regarding zTB prevalence in humans, estimates range from 1.5% to 13% in high-risk group of farm and abattoir workers, with notably higher percentages in extra-pulmonary TB cases. The review also addresses the challenges that Pakistan faces in formulating an effective policy for the control and eradication of bTB. We conclude with one-health based recommendations as a way forward for controlling TB caused by M. bovis in cattle and humans.

The Establishment of a Novel γ-Interferon In Vitro Release Assay for the Differentiation of Mycobacterial Bovis-Infected and BCG-Vaccinated Cattle.

BCG vaccination is increasingly reconsidered in the effective prevention of bovine tuberculosis (bTB). However, the primary challenge in BCG vaccination for cattle is the lack of a technique for differentiating between infected and vaccinated animals (DIVA). This study aimed to establish a novel DIVA diagnostic test based on an interferon-gamma in vitro release assay (IGRA). The plasmid encoding three differential antigens (Rv3872, CFP-10, and ESAT-6) absent in BCG genes but present in virulent M. bovis was previously constructed. Thus, a recombinant protein called RCE (Rv3872, CFP-10, and ESAT-6) was expressed, and an RCE-based DIVA IGRA (RCE-IGRA) was established. The RCE concentration was optimized at 4 µg/mL by evaluating 97 cattle (74 of which were bTB-positive, and 23 were negative) using a commercial IGRA bTB diagnostic kit. Further, 84 cattle were tested in parallel with the RCE-IGRA and commercial PPD-based IGRA (PPD-IGRA), and the results showed a high correlation with a kappa value of 0.83. The study included BCG-vaccinated calves (n = 6), bTB-positive cattle (n = 6), and bTB-negative non-vaccinated calves (n = 6). After 3 months post-vaccination, PPD-IGRA generated positive results in both vaccinated and infected calves. However, RCE-IGRA developed positive results in infected calves but negative results in vaccinated calves. In conclusion, this DIVA method has broad prospects in differentiating BCG vaccination from natural infection to prevent bTB.

Very low prevalence of bovine tuberculosis in cattle in Sylhet district of Bangladesh.

Bovine tuberculosis (bTB), a chronic zoonotic disease in cattle, has a substantial socio-economic and public health impact. This study was conducted to estimate the prevalence and geographical distribution of bTB in the Sylhet district of Bangladesh. A cross-sectional study was conducted at all 12 upazilas of Sylhet district, which included 512 randomly selected cattle from 48 farms. Selected animals were tested with the Caudal Fold Tuberculin (CFT) test to identify bTB-positive cattle. Out of 512 cattle, only one animal was identified as a reactor, providing an estimated prevalence of 0.19% (95% Confidence Interval; 0-0.58%). The only positive reactor was found in Zakiganj upazila. As the prevalence of bTB in cattle in Sylhet appears to be low, it indicates that most of the upazilas of Sylhet district are free of the bTB infection. This prevalence is lower than the reported prevalence in other parts of Bangladesh. Thus, attempts should be made to maintain the current situation of bTB infection in cattle of Sylhet district.

IS6110 Copy Number in Multi-Host Mycobacterium bovis Strains Circulating in Bovine Tuberculosis Endemic French Regions.

IS6110 is an insertion sequence found in the Mycobacterium tuberculosis complex, to which Mycobacterium bovis belongs, which can play a role in genome plasticity and in bacterial evolution. In this study, the abundance and location of IS6110 on M. bovis genomic data of French animal field strains were studied. A first analysis was performed on a panel of 81 strains that reflect the national M. bovis population's genetic diversity. The results show that more than one-third of them are IS6110 multicopy and that 10% have IS6110 in a high copy number (more than 6 copies). Multicopy strains are those circulating in the regions where prevalence was above the national average. Further study of 93 such strains, with an IS6110 copy number of 10-12, showed stability of IS6110 copy number and genome location over time and between host species. The correlation between M. bovis multicopy strains and high bovine tuberculosis (bTB) prevalence leads us to consider whether their epidemiological success could be partly due to genetic changes originated by IS6110 transposition.

Defined Antigen Skin Test for Bovine Tuberculosis Retains Specificity on Revaccination With Bacillus Calmette-Guérin.

The Bacillus Calmette-Guérin (BCG) vaccination provides partial protection against, and reduces severity of pathological lesions associated with bovine tuberculosis (bTB) in cattle. Accumulating evidence also suggests that revaccination with BCG may be needed to enhance the duration of immune protection. Since BCG vaccine cross-reacts with traditional tuberculin-based diagnostic tests, a peptide-based defined antigen skin test (DST) comprising of ESAT-6, CFP-10, and Rv3615c to detect the infected among the BCG-vaccinated animals (DIVA) was recently described. The DST reliably identifies bTB-infected animals in experimental challenge models and in natural infection settings, and differentiated these from animals immunized with a single dose of BCG in both skin tests and interferon-gamma release assay (IGRA). The current investigation sought to assess the diagnostic specificity of DST in calves (Bos taurus ssp. taurus × B. t. ssp. indicus; n = 15) revaccinated with BCG 6 months after primary immunization. The results show that none of the 15 BCG-revaccinated calves exhibited a delayed hypersensitivity response when skin tested with DST 61 days post-revaccination, suggesting 100% diagnostic specificity (one-tailed lower 95% CI: 82). In contrast, 8 of 15 (diagnostic specificity = 47%; 95% CI: 21, 73) BCG-revaccinated calves were positive per the single cervical tuberculin (SCT) test using bovine tuberculin. Together, these results show that the DST retains its specificity even after revaccination with BCG and confirms the potential for implementation of BCG-based interventions in settings where test-and-slaughter are not economically or culturally feasible.

Engaging With Farmers to Explore Correlates of Bovine Tuberculosis Risk in an Internationally Important Heritage Landscape: The Burren, in the West of Ireland.

Bovith recene tuberculosis (bTB) continues to be a pathogen of concern in several countries globally. Analysis of areas that have higher incidences of bTB outbreaks has demonstrated how risk is not equally distributed, and local data collection, analysis and participatory engagement is required to develop tailored approaches. The Burren, an internationally important heritage landscape, has been an area of higher bTB incidence for many years in Ireland, and owing to its unique geology and farming heritage a survey was developed to engage with local farmers to gain greater insight into farming practices and bTB control to inform tailored approaches. The survey gathered data on the farm and animal management approaches being used within the Burren, including local farming techniques like the use of "winterage" (grazing exposed limestone dominated uplands). Thematic analysis of free text responses was undertaken. Quantitative data were then explored using statistical models to assess associations with recent (<3 years) self-reported bTB breakdown risk. There was a high number of responses demonstrating a high degree of willingness to engage on the issue. Thematic analysis suggested that wildlife and its management (culling and vaccination), testing quality, and its impact on the bTB scheme, and pessimism around eradication were important themes. Statistical analysis suggested that increasing bTB risk was primarily related to increasing herd-size and the percentage of herd owner's land inaccessible to those attempting to locate badger setts. There was less evidence for associations relating to the amount of time, or which season (i.e., summer), farmers utilized "winterage". The results of the study will feed back directly to local bTB management plans and further stakeholder engagement and is an exemplar for local tailoring of national control measures in situations of high incidences of bTB outbreaks in particular areas.

Performance and Agreement Between WGS Variant Calling Pipelines Used for Bovine Tuberculosis Control: Toward International Standardization.

Whole genome sequencing (WGS) and allied variant calling pipelines are a valuable tool for the control and eradication of infectious diseases, since they allow the assessment of the genetic relatedness of strains of animal pathogens. In the context of the control of tuberculosis (TB) in livestock, mainly caused by Mycobacterium bovis, these tools offer a high-resolution alternative to traditional molecular methods in the study of herd breakdown events. However, despite the increased use and efforts in the standardization of WGS methods in human tuberculosis around the world, the application of these WGS-enabled approaches to control TB in livestock is still in early development. Our study pursued an initial evaluation of the performance and agreement of four publicly available pipelines for the analysis of M. bovis WGS data (vSNP, SNiPgenie, BovTB, and MTBseq) on a set of simulated Illumina reads generated from a real-world setting with high TB prevalence in cattle and wildlife in the Republic of Ireland. The overall performance of the evaluated pipelines was high, with recall and precision rates above 99% once repeat-rich and problematic regions were removed from the analyses. In addition, when the same filters were applied, distances between inferred phylogenetic trees were similar and pairwise comparison revealed that most of the differences were due to the positioning of polytomies. Hence, under the studied conditions, all pipelines offer similar performance for variant calling to underpin real-world studies of M. bovis transmission dynamics.

Prevalence of Bovine Tuberculosis in Yaks Between 1982 and 2020 in Mainland China: A Systematic Review and Meta-Analysis.

Bovine tuberculosis (bTB) is a major chronic bacterial disease in cattle and is the major economic and animal welfare issue in the world. Although the economic costs and public health safety risks associated with the disease are considerable, the overall epidemiology of the Chinese yak (Bos grunniens) bTB is unclear. To fully reveal the basic prevalence of yak bTB in different regions of China, we searched five databases including PubMed, Science Direct, CNKI (China National Knowledge Infrastructure), Wanfang and Chongqing VIP. Based on the incidence and prevalence of yak tuberculosis in China from 1982 to 2020, a meta-analysis of yak bTB in China was established for the first time. By formulating the search formula, 97 studies were searched in five databases. According to the established exclusion criteria and excluded comments and repeated and irrelevance research, we finally selected 19 cross-sectional studies, which showed the prevalence of bTB in Chinese yaks. Random-effect meta-regression model analysis showed that the estimated prevalence of 122,729 yaks in China was 1.0 (95% confidence interval [CI]: 0.0-1.0). The regional prevalence varies greatly, northwest China prevalence rate 0. 39% (95% CI: 0.2-0. 64) and southwest China prevalence rate 2.59% (95% CI: 1.94-3.34); in terms of province level, the prevalence was highest in Tibet 2.59% (95% CI: 1.94-3.34), followed by Xinjiang 2.36% (95% CI: 0.86-4.58), and Shanxi has the lowest 0.00% (95% CI: 0.00-0.98). This systematic review and meta-analysis identified the estimated prevalence of bTB in Chinese yaks and estimated the underlying factors associated with bTB, including geographic location, sampling year, age, and TB detection method. Provide evidence to plan corresponding disease control strategies for policymakers and to assess future economic risks accurately.

Driving Adoption and Commercialization of Subunit Vaccines for Bovine Tuberculosis and Johne's Disease: Policy Choices and Implications for Food Security.

Infectious animal diseases, such as Johne's disease (JD) caused by Mycobacterium avium paratuberculosis (MAP) and bovine tuberculosis (bTB) caused by Mycobacterium bovis, have been a challenge to the livestock industry globally, impacting negatively on animal, human and environmental health, and overall food security. Despite several industry-led and government initiatives and programs aimed at preventing and reducing losses associated with JD and bTB outbreaks, JD has remained endemic in many parts of the world while there have been incidental outbreaks of bTB. While several studies focus on sustainable intensification of food (crop) production as a critical solution to food insecurity, following the existential interconnection between animals, humans and the environment recognized by one health, we frame food security through the lens of animal disease prevention and control, given the importance of livestock products to human health and livelihood. Vaccination has been a popular strategy successfully used in controlling other infectious diseases. The paper focuses on an alternate strategy of two subunit vaccines with companion diagnostics targeted at individual pathogens to attain satisfactory immunological responses for JD and bTB. We examine gaps in vaccine policies, commercialization, and potential strategies that would strengthen animal disease prevention and enhance food security. The potential of public-private partnership in strengthening private sector participation in effective animal disease control and health delivery and the implications for global food security are discussed.

Bovine Tuberculosis Case Intervention Using the T.SPOT.TB Assay to Screen Dairy Workers in Bailey County, Texas.

Background: One potential exposure on a dairy farm is Mycobacterium bovis or bovine tuberculosis (bTB)-an infectious zoonotic pathogen. The prevalence of tuberculosis among dairy workers in the U.S. is unknown largely due to insufficient surveillance and testing practices. Our objective was to determine the prevalence and risk factors of LTBI among dairy workers potentially exposed to cattle infected with bTB in two Bailey County, Texas dairy farms in 2016. Methods: This study involved a secondary analysis of data that were collected by Texas Department of State Health Services (DSHS) Public Health Region 1 (PHR 1). A total of 140 dairy workers were tested using the T.SPOT.TB test assay. As a proxy for occupational exposures, we used three categories of cattle exposure groups based on work task, duration, and conditions of exposure to cattle-high, medium, low. Results: Positive LTBI was found among 14/140 (10.0%) of the dairy workers tested with 12/87 (13.8%) in Dairy A and 2/53 (3.8%) in Dairy B. All LTBI cases were determined to be from Hispanic workers with 71.4% indicating having been vaccinated with the BCG vaccine in their country of birth and none indicated previously known exposure to TB. The high category of cattle exposure group experienced the highest prevalence of LTBI (64.3%), followed by the medium cattle exposure group (28.6%), and the low cattle exposure group (7.1%). Conclusion: Our findings suggest that the prevalence of LTBI among dairy workers in Bailey County, Texas is higher than demographically comparable workforces. Future efforts should focus on the development, delivery, and evaluation of a tuberculosis-and other zoonotic diseases-health and safety training which can become a part of a more comprehensive safety management and training program on dairy farms.

Global Distribution and Evolution of Mycobacterium bovis Lineages.

Mycobacterium bovis is the main causative agent of zoonotic tuberculosis in humans and frequently devastates livestock and wildlife worldwide. Previous studies suggested the existence of genetic groups of M. bovis strains based on limited DNA markers (a.k.a. clonal complexes), and the evolution and ecology of this pathogen has been only marginally explored at the global level. We have screened over 2,600 publicly available M. bovis genomes and newly sequenced four wildlife M. bovis strains, gathering 1,969 genomes from 23 countries and at least 24 host species, including humans, to complete a phylogenomic analyses. We propose the existence of four distinct global lineages of M. bovis (Lb1, Lb2, Lb3, and Lb4) underlying the current disease distribution. These lineages are not fully represented by clonal complexes and are dispersed based on geographic location rather than host species. Our data divergence analysis agreed with previous studies reporting independent archeological data of ancient M. bovis (South Siberian infected skeletons at ∼2,000 years before present) and indicates that extant M. bovis originated between 715 and 3,556 years BP, with later emergence in the New World and Oceania, likely influenced by trades among countries.

Is moving from targeted culling to BCG-vaccination of badgers (Meles meles) associated with an unacceptable increased incidence of cattle herd tuberculosis in the Republic of Ireland? A practical non-inferiority wildlife intervention study in the Republic of Ireland (2011-2017).

Bovine tuberculosis (BTB) remains as a costly disease of cattle-herds in the Republic of Ireland (ROI). This persistence is partially attributable to the presence of M. bovis infection in a wildlife reservoir, the European badger (Meles meles). Thus, both area-wide and limited-area targeted-badger-culling have been part of the ROI-BTB control/eradication program to help reduce the future incidence of a cattle-herd BTB breakdown (i.e. a "new herd-level occurrence of BTB"). However, neither badger-culling practice can be sustained as a major component in the ongoing BTB eradication program in the ROI. Vaccination of badgers with Bacille Calmette-Guérin (BCG) has been proposed as an alternative to badger culling. Thus, in 2011, a five-year non-inferiority study was implemented in seven counties in the ROI. This study was designed to compare and contrast the cattle-herd-BTB-incidence in areas where intramuscular badger vaccination would be implemented versus the cattle-herd-BTB-incidence in the remaining area of the same county where targeted-badger-culling was maintained as the standard treatment response to probable badger-sourced BTB breakdowns. Our outcome of interest was a new cattle-herd-BTB-episode (breakdown) with a total of >2 standard skin-test (SICTT) reactors detected during the episode. Treatments (badger vaccination or targeted badger culling) were cluster allocated based on where the majority of the herd owner's land was located. To assess the impact of the two treatments, we compared the incidence-risk, of our defined outcome, for cattle herds in the area under vaccination to the outcome incidence-risk for cattle herds in the remainder of the same county after 4 and 5 years of having implemented badger vaccination. A random-effects logit model with adjustment for clustering by treatment, and statistical control of herd-type, herd-size and five-year prior-BTB-episode history was used for our analyses. Although not included in the logistic model, a relative badger density metric based on the annual number of badgers captured-per-sett-night of capturing effort was developed for each treatment area; this metric indicated that relative badger density was approximately 40 % higher in vaccination areas than in the targeted badger-culling areas during our study. Overall, our study results indicated that vaccination was not inferior to targeted badger-culling in four counties and badger vaccination was deemed to produce ambivalent results in one (County Cork North) of the seven study sites in the ROI. A post-study investigation, in County Galway, where vaccination was deemed inferior to target culling, revealed that widespread purchases of cattle from a nearby cattle mart, by herd owners in the vaccination-area, was associated with the increased herd and vaccination-area risk of BTB. No single "biasing hypothesis" was evident for the apparent vaccine inferiority in the second study site (County Monaghan) where vaccination was deemed inferior to targeted culling; hence no further investigations were conducted.

Tracing cross species transmission of Mycobacterium bovis at the wildlife/livestock interface in South Africa.

BACKGROUND: Bovine tuberculosis (bTB) affects cattle and wildlife in South Africa with the African buffalo (Syncerus caffer) as the principal maintenance host. The presence of a wildlife maintenance host at the wildlife/livestock interface acting as spill-over host makes it much more challenging to control and eradicate bTB in cattle. Spoligotyping and mycobacterial interspersed repetitive unit-variable number of tandem repeat (MIRU-VNTR) genotyping methods were performed to investigate the genetic diversity of Mycobacterium bovis (M. bovis) isolates from cattle and wildlife, their distribution and transmission at the wildlife/livestock interface in northern Kwa-Zulu Natal (KZN), South Africa. RESULTS: SB0130 was identified as the dominant spoligotype pattern at this wildlife/livestock interface, while VNTR typing revealed a total of 29 VNTR profiles (strains) in the KZN province signifying high genetic variability. The detection of 5 VNTR profiles shared between cattle and buffalo suggests M. bovis transmission between species. MIRU-VNTR confirmed co-infection in one cow with three strains of M. bovis that differed at a single locus, with 2 being shared with buffalo, implying pathogen introduction from most probably unrelated wildlife sources. CONCLUSION: Our findings highlight inter and intra species transmission of bTB at the wildlife/livestock interface and the need for the implementation of adequate bTB control measures to mitigate the spread of the pathogen responsible for economic losses and a public health threat.

Wildlife-cattle interactions emerge as drivers of bovine tuberculosis in traditionally farmed cattle.

Bovine tuberculosis (bTB) in cattle is recognized to be associated with several risk factors that include herd size, cattle movement, ownership of other domestic animals, confinement of cattle in enclosures at night, water sources, communal grazing area and proximity to wildlife, especially bTB maintenance hosts. A questionnaire survey was used to investigate the risk factors associated with Mycobacterium bovis (M. bovis) infection and transmission in traditionally farmed cattle at the wildlife/livestock interface in uMkhanyakude district, northern Kwa-Zulu Natal (KZN), South Africa. The questionnaire comprised of semi-structured questions that were used to gather data on livestock management practices and knowledge about bTB from 71 respondents from households that owned either bTB infected cattle herds or uninfected herds. Multiple correspondence analysis (MCA) was used to explore the association between the risk factors for M. bovis transmission and the bTB herd status. Bovine TB positive herds were associated with a herd size of n > 15, movement of cattle to areas adjacent to the game parks for grazing, cattle grazing inside the game parks as well as cattle sharing water and pasture with wildlife. The multivariable logistic regression model identified movement of animals to areas adjacent to the game parks and cattle sharing water with wildlife as highly significant risk factors for bTB infection in cattle. The findings of this study emphasized the need for the implementation of bTB control strategies in both cattle and wildlife populations for the successful control of the disease.

Global distribution and evolution of mycobacterium bovis lineages

Mycobacterium bovis is the main causative agent of zoonotic tuberculosis in humans and frequently devastates livestock and wildlife worldwide. Previous studies suggested the existence of genetic groups of M. bovis strains based on limited DNA markers (a.k.a. clonal complexes), and the evolution and ecology of this pathogen has been only marginally explored at the global level. We have screened over 2,600 publicly available M. bovis genomes and newly sequenced four wildlife M. bovis strains, gathering 1,969 genomes from 23 countries and at least 24 host species, including humans, to complete a phylogenomic analyses. We propose the existence of four distinct global lineages of M. bovis (Lb1, Lb2, Lb3, and Lb4) underlying the current disease distribution. These lineages are not fully represented by clonal complexes and are dispersed based on geographic location rather than host species. Our data divergence analysis agreed with previous studies reporting independent archeological data of ancient M. bovis (South Siberian infected skeletons at ~2,000 years before present) and indicates that extant M. bovis originated between 715 and 3,556 years BP, with later emergence in the New World and Oceania, likely influenced by trades among countries.

Risk Factors for Zoonotic Tuberculosis at the Wildlife-Livestock-Human Interface in South Africa.

A cross-sectional study was conducted to investigate the risk factors associated with zoonotic tuberculosis in humans and its transmission to people living at the wildlife-livestock-human interface. A questionnaire was administered to collect information on food consumption habits, food handling practices, and knowledge of zoonotic TB. Sputum samples were also collected from 150 individuals that belonged to households of cattle farmers with or without a bTB infected herd. In addition, 30 milk samples and 99 nasal swabs were randomly collected from cattle in bTB infected herds for isolation of Mycobacterium bovis (M. bovis). The sputum samples were screened for TB using the GeneXpert test and this was followed by mycobacterial culture and speciation using molecular techniques. No M. bovis was isolated from TB positive sputum samples and only one sample was confirmed as Mycobacterium tuberculosis (M. tuberculosis). M. bovis was isolated from 6.6% (n = 2/30) milk samples and 9% (n = 9/99) of nasal swabs. Ownership of a bTB infected herd and consumption of milk were recognized as highly significant risk factors associated with a history of TB in the household using multiple correspondence analysis (MCA) and logistic regression. The findings from this study have confirmed the potential for zoonotic TB transmission via both unpasteurized milk and aerosol thus, the role of M. bovis in human TB remains a concern for vulnerable communities.

Prevalence of Mycobacterium bovis infection in traditionally managed cattle at the wildlife-livestock interface in South Africa in the absence of control measures.

Cattle are the domestic animal reservoir for Mycobacterium bovis (M. bovis) which also affects other domestic animals, several wildlife species and humans leading to tuberculosis. The study area is in a resource-poor community that is surrounded by several game parks, where M. bovis infection has been previously diagnosed in wildlife. A cross-sectional study was carried out to determine the prevalence of M. bovis infection in 659 cattle from a total of 192 traditionally managed herds using the BOVIGAM® interferon gamma assay (IFN-γ). Infection was confirmed by post mortem examination and M. bovis isolation from three test-positive cattle. Genotyping of the M. bovis isolates was done using spoligotyping and VNTR (variable number of tandem repeats typing). The apparent M. bovis prevalence rate in cattle at animal level was 12% with a true population prevalence of 6% (95% Confidence interval (C.I) 3.8 to 8.1) and a herd prevalence of 28%. Spoligotyping analysis revealed that the M. bovis isolates belonged to spoligotype SB0130 and were shared with wildlife. Three VNTR profiles were identified among the SB0130 isolates from cattle, two of which had previously been detected in buffalo in a game reserve adjacent to the study area. The apparent widespread presence of M. bovis in the cattle population raises a serious public health concern and justifies further investigation into the risk factors for M. bovis transmission to cattle and humans. Moreover, there is an urgent need for effective bTB control measures to reduce infection in the communal cattle and prevent its spread to uninfected herds.

Five-year surveillance of human tuberculosis caused by Mycobacterium bovis in Bologna, Italy: an underestimated problem.

Human tuberculosis (TB) caused by Mycobacterium bovis surveillance is affected by a lack of data. The aims of the present study were: (i) to estimate the proportion of human TB caused by M. bovis over a period of 5 years in Bologna, Northern Italy, which, like most Western European countries, has been declared bovine TB-free; (ii) to compare the genetic profiles of M. bovis strains identified in humans with those circulating in cattle in the last 15 years in Italy. Among 511 TB patients, the proportion of human TB caused by M. bovis was 1·76%, significantly associated to extra-pulmonary localization (P = 0·004) and to being elderly (P < 0·001) and Italy-born (P = 0·036). The molecular epidemiology analysis by spoligotyping and Multilocus Variable Tandem Repeat Analysis confirmed that most M. bovis strains from Italy-born patients matched those circulating in cattle herds in Italy between 2001 and 2016. Two cases of Mycobacterium bovis BCG infection were also characterized. In conclusion, the rate of human TB caused by M. bovis was not negligible, highlighting the relevance of molecular typing in evaluating the effectiveness of programmes designed to eradicate TB in cattle in Italy.

Investigation of intra-herd spread of Mycobacterium caprae in cattle by generation and use of a whole-genome sequence.

Single nucleotide polymorphisms (SNPs) calculated from whole genome sequencing (WGS) are ideally suited to study evolutionary relationships of pathogens and their epidemiology. Mycobacterium caprae infections have been documented frequently in cattle and red deer along the Bavarian and Austrian Alps during the last decade. However, little is still known about the transmission within cattle holdings and possible alterations of the genomes of M. caprae during such events. The aim of this study was to study the molecular epidemiology of bovine tuberculosis (bTB) in selected herds based on isolate-specific genome-wide SNPs and to perform a phylogenetic network analysis. In total, 61 M. caprae isolates were collected originating from eight cattle farms over a period of twelve years between 2004 and 2015. Analysis of their sequence data revealed that the M. caprae isolates of an affected farm differ at all in a few SNPs. In contrast, many more SNPs were found when comparing the M. caprae genomes originating from different herds. The results demonstrated that the spread of bTB in the affected farms occurred by direct transmission between the members of each herd rather than between herds and a M. caprae introduction in farms after contact events e. g. on summer pastures can readily be traced by WGS analysis. Furthermore, we assembled a nearly complete whole genome sequence of M. caprae derived from several cattle isolates originating from bTB cases in the Bavarian Alpine region.

Prevalence and risk factors of bovine tuberculosis in dairy cattle in Eritrea.

BACKGROUND: The prevalence of bovine tuberculosis (BTB) in dairy cattle in the three major milk producing regions of Eritrea was assessed by subjecting 15,354 dairy cattle, 50 % of Eritrea's dairy cattle population, to the single intradermal comparative tuberculin test (SICTT). Skin test results were interpreted according to guidelines of the World Organization for Animal Health (OIE) with >4 mm as cutoff in skin thickness increase. In addition, we studied the relation between 'physiological' variables related to pregnancy and lactation, and the variable 'region' on the probability to be skin test positive. RESULTS: The BTB prevalences at animal and herd levels were: 21.5% and 40.9% in Maekel, 7.3% and 10% in Debub, and 0.2% and 1.6% in the Anseba region, respectively. Overall, in the regions included, prevalence was 11.3% (confidence interval (CI) 95% CI, 11.29 - 11.31%) and 17.3% (95% CI, 17.27-17.33%), at animal and herd level, respectively. Considering positive herds only, the animal BTB prevalence was 36.8%, 30.1%, and 1.8%, in Maekel, Debub and Anseba, respectively, and the overall animal prevalence within these herds was 32%. In adult dairy cattle the probability of positive reactivity in the SICTT test was highest in pregnant animals as compared to the other categories. CONCLUSION: This study reports persistent prevalence of BTB as defined by positive SICTT in the dairy sector of Eritrea, especially in the regions of Maekel and Debub that are located in the central highlands of the country. To our understanding this is the first report that has encompassed all the major dairy farms in Eritrea and it will be instrumental in advocating future BTB control programs in the dairy sector.