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Mycobacterium bovis (M. bovis) is the primary agent of bovine tuberculosis (TB) in Mediterranean buffalo, which has a negative economic impact on buffalo herds. Improving TB diagnostic performance in this species represents a key step to eradicate efficiently this disease. We have recently shown the utility of the IFN-γ assay in the diagnosis of M. bovis infection in Mediterranean buffaloes (Bubalus bubalis), but other cytokines might be useful immunological biomarkers of this infection. We therefore investigated the utility of key immune cytokines as diagnostic biomarkers of M. bovis infection in this species. Thirty-six Italian Mediterranean buffaloes were used in this study: healthy animals (N = 11), infected (IFN-γ test positive, no post-mortem lesions, no M. bovis detection; N = 14), and affected (IFN-γ test positive, visible post-mortem lesions; N = 11). Heparin blood samples were stimulated with bovine purified protein derivative (PPD-B), alongside controls, and 18–24 h later plasma were collected. Levels of 14 key cytokines were measured: IFN-γ, IL-17, IL-4, IL-10, TNF, IL-1α, IL-1β, IL-6, IP-10, MIP-1α, MIP-1β, MCP-1, IL-36Ra, and VEGF-A. We observed that both infected and affected animals released higher levels of IFN-γ, IL-17, IL-10, TNF, IL-1α, IL-6, MIP-1β, in response to PPD-B compared to healthy subjects. Mycobacterium bovis infected animals released also higher levels of IL-1β and IP-10 in response to PPD-B compared to healthy subjects, whereas only tendencies were detected in affected animals. Affected animals only released MIP-1α in response to PPD-B compared to healthy subjects and in this group higher values of PPD-B specific TNF was also observed. Finally, canonical discriminant analysis (CDA) was used to generate predictive cytokine profiles by groups. Our data suggest that IL-10, TNF, IL-1α, IL-6, MIP-1β could be useful biomarkers of TB in Mediterranean Buffalo and can improve the TB diagnostic performance in this specie. [ABSTRACT FROM AUTHOR]

Sustainable improvement of production in most tropical dairy systems is a significant challenge, because the cattle breeds with the most genetic potential for milk output relative to native tropical breeds have not been selected for these types of environments. Multiplex genome editing provides a potential solution to introduce beneficial sequence variants (SVs) into elite animals for tropical adaptation in a single generation. Bovine sequence variants for heat tolerance, stature, milk yield, and disease-tolerance traits were identified and genotyped across indigenous African, dairy Zebu, and dairy Taurine breeds to validate those targets best suited for introgression by genome editing. In vitro fertilized embryos from a series of matings were used to produce embryonic stem cells (ESCs) and were subsequently multiplexed edited prior to cloning by somatic cell nuclear transfer. A set of best target SVs for genome editing was established for the Holstein and Gir breeds. ESCs were produced and cloned following treatment and validation screening for multiplex alterations of up to four target genes. Currently, 12 animals have been born, and all the mature males have produced viable semen that will be submitted for regulatory review in a series of countries in Sub-Saharan Africa and South America. Multiplex genome editing based on homology-directed repair combined with cloning of bovine ESCs provides an opportunity to initiate genetic improvement of polygenic traits in cattle. Combining genomics and genome editing provides new opportunities to breed more resilient dairy animals for the tropics that should improve animal and farmer livelihoods. Current dairy breeding systems in tropical zones rely on crossbreeding between native, adapted breeds, and foreign high-performance genetics to increase milk yield. When the proportion of foreign-derived genetics becomes too high in subsequent generations of breeding, the animals typically fail to fit in the production environment due to unmanageable heat stress and susceptibility to endemic diseases. To overcome these types of genetic by environmental challenges, we bred animals with traits delivered through a multiplex gene editing platform that will contribute to significant and sustainable production gains for tropical dairy production systems. Photographs by Haley Jo. This article belongs to the Collection: Proceedings of the Annual Conference of the International Embryo Technology Society, Fort Worth, TX, USA, 18–22 January 2025. [ABSTRACT FROM AUTHOR]

Background: The growing use of real-time PCR (qPCR) as a diagnostic method for bovine TB (bTB) requires rapid and effective DNA extraction methods, which are crucial for its success. Automated DNA extraction methods based on magnetic beads are a promising alternative to conventional silica column-based protocols (COL protocol) due to their high throughput capacity and reduced hands-on time. This study aimed to assess the performance of the MagMax CORE Nucleic Acid Purification kit and the KingFisher Flex instrument (KF protocol) as an alternative for scaling up the use of qPCR in bTB diagnosis. Methodology: Performance was evaluated with two different real-time PCR (qPCR) protocols, based on the IS6110 element and the QuantiFast and VetMAX™ (QF and VM protocols) kits, on 145 frozen tissue homogenates confirmed as either bTB-positive or negative through a composite reference standard based on microbiological culture, column-based extraction, and qPCR, as well as on negative tissue samples spiked with 106 to 103 CFU/ml of M. bovis BCG. Results: The performance of both qPCR protocols was very high on samples extracted using the KF protocol, with positive percent agreement (PPA) values of 89.04% [95% Confidence Interval (CI): 79.54–95.15%] and 93.15% [95% CI: 84.74–97.74%] for the QF and VM protocols, respectively, and negative percent agreement (NPA) values of 100% [95% CI: 95.01–100.00%]. A higher variability was identified in samples analysed with the same qPCR protocol but different extraction methods. Higher Ct values were identified for samples extracted using the KF protocol in both routine and spiked samples, likely due to using the same amount of starting material for both extraction methods, which was lower than recommended by the manufacturer for the KF protocol. Discussion: The results of this study indicate that the MagMAX CORE Nucleic Acid Purification kit coupled with a KingFisher Flex instrument is a valuable alternative for the extraction of MTBC DNA from bovine tissues. However, the increased variability and Ct values suggest that a larger amount of starting material is recommended for this methodology, warranting further studies. [ABSTRACT FROM AUTHOR]

Simple Summary: To investigate the effects of air pollutants and bioclimatic variables on tuberculosis in domestic ruminants, we simulated the risk distribution of tuberculosis in cattle, buffaloes, sheep, and goats in China using the maximum entropy ecological niche model, and evaluated the effects of environmental factors. The risk factors that most significantly influenced the prevalence of tuberculosis were the nitrogen dioxide (NO2) level, mean temperature of the coldest quarter, cattle distribution density, sheep distribution density, ozone (O3) level, and precipitation of the driest month. The predicted map of tuberculosis risk in cattle, buffaloes, sheep, and goats indicated that the high-risk regions were mainly distributed in South, North, East, and Northwest China. The findings of this study provide a valuable reference for the prevention and control of tuberculosis in cattle, buffaloes, sheep, and goats in China. Tuberculosis is a zoonotic chronic respiratory infectious disease caused by the Mycobacterium tuberculosis complex. The outbreak and epidemic of tuberculosis can seriously threaten human and veterinary health. To investigate the effects of environmental factors on tuberculosis in domestic ruminants, we collected data regarding the prevalence of tuberculosis in cattle, buffaloes, sheep, and goats in China (1956–2024) from publicly published literature and available databases. We identified the key risk factors among six major air pollutants and 19 bioclimatic variables; simulated the risk distribution of tuberculosis in cattle, buffaloes, sheep, and goats in China using the maximum entropy ecological niche model; and evaluated the effects of environmental factors. The area under the curve of the model was 0.873 (95% confidence interval, 0.851–0.895). The risk factors that most significantly influenced the prevalence of tuberculosis were the nitrogen dioxide (NO2) level, mean temperature of the coldest quarter, cattle distribution density, sheep distribution density, ozone (O3) level, and precipitation of the driest month. The predicted map of tuberculosis risk in cattle, buffaloes, sheep, and goats indicated that the high-risk regions were mainly distributed in South, North, East, and Northwest China. Improved surveillance is needed in these high-risk areas, and early preventive measures must be implemented based on the risk factors identified to reduce the future prevalence of tuberculosis in cattle, buffaloes, sheep, and goats. [ABSTRACT FROM AUTHOR]

Mycobacterium bovis , the causative agent of bovine tuberculosis (bTB), is a zoonotic pathogen that contributes to economic losses in the cattle industry and poses a public health risk worldwide. Bacillus Calmette-Guerin, or BCG, is a live attenuated strain of M. bovis that is used for human vaccination against tuberculosis and is considered a potential vaccine candidate against bTB. However, BCG affords widely variable levels of protection against challenge and interferes with current diagnostic methods, and as such, it is not currently approved for use as a livestock or wildlife vaccine in the United States. Many efforts have been made to develop bTB vaccines that are reliable and do not interfere with diagnostic testing, but BCG continues to be the most effective option. Previous work has shown that a T helper 1 immune response is essential for protection against virulent M. bovis infection, characterized by CD4+ central and effector memory T cells. In an effort to identify an efficacious bTB intervention strategy, the study presented here used an in vitro recall response assay and concurrent evaluation of CD4+ T cell proliferation and cytokine production to characterize the surface and functional phenotypes of memory responses to BCG vaccination in cattle. Our findings enhance understanding of the bovine immune response to BCG and provide insights into the development of improved vaccines for the control of bTB. [ABSTRACT FROM AUTHOR]

Paratuberculosis is a chronic granulomatous enteritis, caused by Mycobacterium avium subspecies paratuberculosis (Map), that affects ruminants worldwide. Vaccination has been considered the most cost-effective method for the control of this disease in infected dairy herds. However, currently available vaccines do not provide complete protection and interfere with the diagnosis of both paratuberculosis and bovine tuberculosis, limiting its use. Because of that, efforts are being made for the development of new vaccines. The primary objective of this study was to evaluate the efficacy of two whole-cell inactivated experimental vaccines against paratuberculosis in goats, administered through the oral (OV) and intradermal (IDV) routes, and compare them with that of the commercial subcutaneous vaccine Gudair® (SCV). Over an 11-month period, the effect of vaccination and a subsequent Map challenge on the specific peripheral immune responses and Map- DNA fecal shedding were recorded. At the end of the experiment, tissue bacterial load and lesion severity were assessed. The experimental vaccines did not induce specific humoral immune responses and only elicited mild and delayed cellular immune responses. Although the OV reduced lesion severity, neither this vaccine nor the IDV prototype was able to reduce fecal shedding or tissue bacterial load. Moreover, although the SCV did not confer sterile immunity, it outperformed both experimental vaccines in all these parameters. [ABSTRACT FROM AUTHOR]

Introduction: Bovine tuberculosis is one of the primary infectious diseases affecting cattle. Although several countries have managed to eradicate this zoonosis it remains endemic and uncontrolled across many countries in Africa, Asia, Latin America, and the Middle East. Brazil launched its national control and eradication program in 2001, and since then, epidemiological studies have been carried out to define optimal control strategies and to enable the management of the process in each region. Methods: This study covered the state of Pará, which was divided into three regions, in each of which a pre-established number of properties were randomly selected, and within each property, a minimum number of animals were drawn to be tested by the tuberculin test to classify the farm as infected or free of the disease. A questionnaire was administered to the selected properties to identify the risk factors for the disease. Results: A total of 976 properties comprising 17,151 animals were tested. The prevalence of infected properties in the regions ranged from 3.1% [1.3; 4.9] to 18.6% [14.3; 22.9], while tuberculin-positive animals ranged from 0.24% [0.09; 0.40] to 4.8% [2.4; 7.3]. The introduction of cattle untested for bTB and the renting of pastures have been identified as risk factors for the disease in this state. Discussion: Pará has one of the highest prevalences of bovine tuberculosis in Brazil, and the implications of these results for public health were discussed. We therefore proposed that the state's Official Veterinary Service, together with the private dairy and meat sectors, pursue the objective of controlling or eradicating the disease, along with mechanisms to verify its effectiveness. The differences between the two objectives were discussed, but in both cases, an education program is necessary to inform cattle and buffalo breeders that they need to test animals for tuberculosis before introducing them to their farms, and also to avoid renting pastures for cattle to graze and rest while moving on foot, as these practices have contributed the most to the spread of bTB in the state. [ABSTRACT FROM AUTHOR]

Background: Bovine paratuberculosis is a chronic infectious disease of ruminants primarily caused by Mycobacterium avium subsp. paratuberculosis (MAP). It is essentially a chronic granulomatous enteritis characterized by intractable diarrhea, progressive lethargy, and thickening of the intestinal mucosa with the formation of crumpled pouches. Bovine paratuberculosis not only adversely affects milk production and the quality of dairy products but also poses a significant threat to the economic development of dairy farming and human food security. This systematic review and meta-analysis was conducted to assess the prevalence of MAP infection among cattle herds in mainland China Results: A total of 62 studies with data from 102,340 cattle in 24 provinces in China were selected after matching the assessment criteria. In China, the overall estimated prevalence of MAP infection in cattle was 8%(7727/102340). Interestingly, the MAP infection rate in cattle in southern China was estimated to be 2% (6/281), which was significantly lower compared with other regions of China, and the highest infection rate was 12% (1914/16008) in eastern China. MAP infection rates were related to age, average herd size, type of use, season, detection method, and sample type. Moreover, the MAP infection rate in cattle did not correlate with the publication date of the studies. Conclusion: The analysis identified age, average herd size, type of use, and season as significant potential risk factors associated with PTB pool positivity. In addition, the detection method and sample type can also potentially affect the incidence of detected PTB. [ABSTRACT FROM AUTHOR]

The development of vaccines and effective diagnostic methods for bovine tuberculosis requires an understanding of the immune response against its causative agent, Mycobacterium bovis. Although this disease is primarily investigated and diagnosed through the assessment of cell-mediated immunity, the role of B cells and antibodies in bovine tuberculosis has been relatively undervalued and understudied. Current evidence indicates that circulating M. bovis-specific antibodies are not effective in controlling the disease. However, local humoral immune responses may contribute to either defence or pathology. Recent studies in animal models and cattle vaccine trials suggest a potential beneficial role of B cells in tuberculosis control. This review discusses the role of B cells and antibodies in bovine tuberculosis and explores antibody-based diagnostics for the disease, including traditional techniques, such as different ELISA, new platforms based on multiple antigens and point-of-care technologies. The high specificity and sensitivity values achieved by numerous antibody-based tests support their use as complementary tests for the diagnosis of bovine tuberculosis, especially for identifying infected animals that may be missed by the official tests. [ABSTRACT FROM AUTHOR]

Cholangiocarcinoma is a malignant tumour of bile duct epithelial cells, an uncommon primary hepatic neoplasm in cattle. A 9‐year‐old Simmental cow presented with watery diarrhoea, crusty skin lesions, loss of appetite and hepatomegaly. Suspecting severe liver disease, the cow was euthanased due to poor prognosis. During gross postmortem examination, large numbers of multifocal to coalescing nodules often with soft necrotic centres ranging from miliary to 7 cm in size were present in the liver, lung, mesentery, peritoneum and lymph nodes. These features were suggestive of bovine tuberculosis. However, direct tissue smears stained by Ziehl Neelsen stain were negative for the presence of acid‐fast bacteria, and subsequent histopathological and immunohistochemical investigations confirmed the presence of metastasizing cholangiocarcinoma. This case reminds clinicians and pathologists to keep an open mind during the diagnostic process and serves to highlight the value of histopathology in diagnosing seemingly characteristic gross lesions. [ABSTRACT FROM AUTHOR]

European badgers (Meles meles) play an important role in the epidemiology of bovine tuberculosis (caused by Mycobacterium bovis) in England, but little is known about the prevalence of atypical mycobacteriosis. Badgers are also known to be infected by other infectious agents, and the relationship between mycobacteriosis and concomitant infections needs further investigation. Overall, 88 badger carcasses from the Midlands of England collected between July 2016-August 2017 were selected for histopathological examination based on the degree of autolysis (mild), mycobacterial culture results and a balanced sex ratio. Mycobacteria were cultured from 44 badgers, of which 31 were mycobacteria belonging to the M. tuberculosis complex (MTBC) (based on IS6110 PCR and Hsp64 and/or rRNA PCR and sequencing) and 13 were non-tuberculous atypical mycobacteria (NTM). Mycobacteria were not cultured from the remaining 44 animals. Histologically, the most common findings were silica-laden macrophages (85%), granulomas (53%), sarcocystosis (47%), nephritis (31%), portal/periportal hepatitis (26%), ulcerative dermatitis (18%). Culturable mycobacteriosis was associated with higher prevalence of granulomas (p < 0.001) and lower prevalence of hepatitis (p = 0.003). NTM (M. nonchromogenicum, M. avium complex, M. hassiacum, M. malmoense, M. vaccae.) infections were associated with granulomatous pneumonia, and M. malmoense was associated with pyogranulomatous and ulcerative dermatitis. In conclusion, this study describes, for the first time, histological lesions associated with NTM in badgers, the histomorphology of which was similar to those caused by MTBC. In addition, the negative relationship between mycobacteriosis and periportal hepatitis may indicate a complex relationship between mycobacteriosis and other diseases, as previously observed with tuberculosis. [ABSTRACT FROM AUTHOR]

Bovine tuberculosis (bTB) has significant socio-economic and welfare impacts on the cattle industry in parts of the world. In the United Kingdom and Ireland, disease control is complicated by the presence of infection in wildlife, principally the European badger. Control strategies tend to be applied to whole populations, but better identification of key sources of transmission, whether individuals or groups, could help inform more efficient approaches. Mechanistic transmission models can be used to better understand key epidemiological drivers of disease spread and identify high-risk individuals and groups if they can be adequately fitted to observed data. However, this is a significant challenge, especially within wildlife populations, because monitoring relies on imperfect diagnostic test information, and even under systematic surveillance efforts (such as capture-mark-recapture sampling) epidemiological events are only partially observed. To this end we develop a stochastic compartmental model of bTB transmission, and fit this to individual-level data from a unique > 40-year longitudinal study of 2,391 badgers using a recently developed individual forward filtering backward sampling algorithm. Modelling challenges are further compounded by spatio-temporal meta-population structures and age-dependent mortality. We develop a novel estimator for the individual effective reproduction number that provides quantitative evidence for the presence of superspreader badgers, despite the population-level effective reproduction number being less than one. We also infer measures of the hidden burden of infection in the host population through time; the relative likelihoods of competing routes of transmission; effective and realised infectious periods; and longitudinal measures of diagnostic test performance. This modelling framework provides an efficient and generalisable way to fit state-space models to individual-level data in wildlife populations, which allows identification of high-risk individuals and exploration of important epidemiological questions about bTB and other wildlife diseases. Author summary: Wild animals commonly harbour infectious diseases with risk of spillover to humans and livestock. We fitted an individual-level stochastic spatial meta-population model of bovine tuberculosis (bTB) transmission to data from a long-term longitudinal study of the European badger (Meles meles). Our framework provides an efficient and generalisable means of fitting state-space models to individual-level data, to identify high-risk individuals and explore important epidemiological questions. We develop a novel estimator for the individual effective reproduction number, providing quantitative evidence for the presence of superspreader badgers (those individuals most responsible for onward transmission of infection), despite the population-level effective reproduction number being less than one. Predicting the hidden burden of infection in individuals and social groups is critical for disease management but challenging in practice, since monitoring relies on imperfect surveillance and diagnostic testing. Our results suggest that control of bTB in badgers might be substantially increased by targeting interventions at high-risk groups. [ABSTRACT FROM AUTHOR]

Background: Bovine tuberculosis (bTB) is the most important animal health concern in Wales. Annual testing across all cattle herds in Wales commenced in 2010. In 2017, a new geographic division of Wales was conceived, with bespoke cattle controls and eradication milestones reflecting the geographical heterogeneity of bTB distribution. Methods: This observational study uses descriptive analysis and Poisson regression modelling to analyse bTB surveillance data from all herds in Wales. Results: Since 2010, there has been a significant decrease (p < 0.0001) in bTB incidence (8.6%‒6.3%) and plateauing prevalence across Wales. Conversely, there has been an increase in bTB incidence and prevalence in discrete areas. Recurrence and persistence remain important drivers of bTB infection. One of the sharpest declines in bTB incidence was observed in an intensive action area where enhanced cattle control and wildlife vaccination were implemented in an area of high bTB prevalence. Increased herd size, dairy herd type and herd location are important risk factors affecting the rate of bTB incidents in Wales. Limitations: This study includes data from Wales only. Conclusions: Improvements in trends of bTB occurred from 2010 to 2021, but the spatial variations described in this paper support the continued need for regionally adapted surveillance and control measures. [ABSTRACT FROM AUTHOR]

Bovine tuberculosis is one of the most important common infectious diseases between humans and livestock. Cow's milk can be investigated as one of the transmission reservoirs of the disease. Our study was conducted to investigate the presence of Mycobacterium bovis (M. bovis) DNA in cow's milk from different regions of Lorestan province using Touch-down PCR (TD-PCR) method. So, 100 milk samples from industrial and traditional cattle farms were collected and evaluated according to the animal breed, the average age of the animal and the region where the animal was kept. Seven (26.9%) out of the 26 cow's milk samples contaminated with Mycobacterium spp., were diagnosed as M. bovis positive. The cattle with an average age of more than 5 years were most infected with Mycobacterium. Also, the crossbred cattle with an average age of 3 to 5 years, which were kept and raised in tropical areas, showed the highest rate of M. bovis infection. Based on our knowledge, this is the first study regarding the presence of Mycobacterium in cow's milk in Iran. [ABSTRACT FROM AUTHOR]

Epidemiologic research on zoonotic tuberculosis historically used Mycobacterium bovis as a surrogate measure; however, increased reports of human tuberculosis caused by other animal-associated Mycobacterium tuberculosis complex members like Mycobacterium orygis necessitates their inclusion. We performed a retrospective cohort study including persons infected with any animal-lineage M tuberculosis complex species in Alberta, Canada, from January 1995 to July 2021, identifying 42 patients (20  M bovis , 21  M orygis , 1 M caprae). Demographic, epidemiologic, and clinical characteristics were compared against persons with culture-confirmed M tuberculosis infection. The proportion of culture-positive infections caused by M orygis increased continuously from 2016 to 2020. Significantly more females at a higher median age were impacted by M orygis , with all patients originating from South Asia. Mycobacterium bovis caused significantly more extrapulmonary disease and disproportionately impacted young females, particularly those pregnant or postpartum. All infections were acquired abroad. These findings can aid in developing targeted public health interventions. [ABSTRACT FROM AUTHOR]

Persistent tuberculosis (TB) in cattle populations in England has been associated with an exchange of infection with badgers (Meles meles). A badger control policy (BCP) commenced in 2013. Its aim was to decrease TB incidence in cattle by reducing the badger population available to provide a wildlife reservoir for bovine TB. Monitoring data from 52 BCP intervention areas 200–1600 km2 in size, starting over several years, were used to estimate the change in TB incidence rate in cattle herds, which was associated with time since the start of the BCP in each area. A difference in differences analysis addressed the non-random selection and starting sequence of the areas. The herd incidence rate of TB reduced by 56% (95% Confidence Interval 41–69%) up to the fourth year of BCP interventions, with the largest drops in the second and third years. There was insufficient evidence to judge whether the incidence rate reduced further beyond 4 years. These estimates are the most precise for the timing of declines in cattle TB associated with interventions primarily targeting badgers. They are within the range of previous estimates from England and Ireland. This analysis indicates the importance of reducing transmission from badgers to reduce the incidence of TB in cattle, noting that vaccination of badgers, fertility control and on farm biosecurity may also achieve this effect. [ABSTRACT FROM AUTHOR]

Matrix metalloproteinases (MMPs) are involved in the breakdown of lung extracellular matrix and the consequent release of Mycobacterium tuberculosis into the airways. Recent studies indicate that kallikrein-related peptidase 12 (KLK12) regulate MMP-1 and MMP-9, suggesting that targeting the KLK12 gene could be a promising tuberculosis (TB) treatment. To maximise therapeutic potential, this strategy of silencing KLK12 needs to be delivered to the pathogenic cell population while preserving the immunoprotective and tissue homeostatic functions of other lung macrophages. Our research found that KLK12 is highly expressed in M2 macrophages, leading us to design mannose-based bovine serum albumin nanoparticles (MBNPs) for delivering siRNA to silence KLK12 in these cells. The results of in vitro experiments showed that MBNPs could accurately enter M2 macrophages and sustainably release KLK12-siRNA with the help of mannose and mannose receptor targeting. The results of the in vivo experiments showed that MBNPs could reach the lungs within 1 h after intraperitoneal injection and peaked at 6 h. MBNPs increased collagen fibre content in the lungs by decreasing the levels of KLK12/MMPs thereby limiting the progression of TB. Importantly, MBNPs provided greater alleviation of pulmonary TB symptoms and reduced bacterial load in both TB and drug-resistant TB models. These findings provide an alternative and effective option for the treatment of TB, especially when drug resistance occurs. RNA interference using small interfering RNA (siRNA) can target various genes and has potential for treating diseases such as tuberculosis (TB). However, siRNAs are unstable in the blood and within cells. This study presents bovine serum albumin nanoparticles encapsulating KLK12-siRNA (BNPs) synthesized via desolvation. A mannose layer was added (MBNPs) to target mannose receptors on M2 macrophages, facilitating endocytosis. The low pH-responsive MBNPs enhance lysosomal escape for siRNA delivery, downregulating the KLK12 pathway. Tests confirmed that MBNPs effectively inhibited Mycobacterium bovis proliferation, reduced granulomas, and decreased inflammation in a mouse model. This research aims to reduce antibiotic use, shorten treatment duration, and provide a novel TB treatment option. [Display omitted] [ABSTRACT FROM AUTHOR]

Bovine tuberculosis (bTB) is a contagious disease that has a socio-economic impact on familial and industrial farms. Genetic selection can improve resistance against bTB. This study aimed to characterize the links between SLC11A1 and CARD15 gene polymorphisms and tuberculosis (TB) susceptibility. In total 200 animals were used (50 cases and 150 controls). Polymorphisms in the SLC11A1 and CARD15 genes were identified and analyzed with the restriction fragment length polymorphism (PCR-RFLP) method. Our results report the PstI PCR-RFLP marker for the SLC11A1-SNP1 site and the StyI PCR-RFLP marker for the CARD51-SNP1 site in Tunisian Holstein cows. Statistical analysis showed a significant association between SLC11A1-SNP1, CARD15-SNP1 and susceptibility/resistance to TB (P < 0.05). Two SLC11A1-SNP1 genotypes were susceptible to tuberculosis, i.e., the heterozygote CG and the homozygote CC, while one SLC11A1-SNP1 genotype, i.e., the GG mutated homozygote, was less susceptible to infection. Concerning the CARD15 gene, two genotypes are highly sensitive to the incidence of bovine tuberculosis, i.e., AA and AG, while the GG genotype is more beneficial and more tolerant of Mycobacterium bovis. SLC11A1 and CARD15 are two useful candidate genes associated with tuberculosis, and this information can be used to improve the health status of domestic cattle and humans. [ABSTRACT FROM AUTHOR]

Bovine tuberculosis (BTB) still represents a significant public health concern and economic issue for livestock breeders in Egypt. This research investigates the incidence of bovine tuberculosis in cattle slaughtered at the central abattoir in Aswan, Egypt, in 2023. A total of 720 cattle were checked antemortem and postmortem at the abattoir. The suspected lesions were analyzed with acid-fast staining, microscopy, histology, ELISA, and RT-PCR. Based on gross tubercle lesions, the overall occurrence of bovine tuberculosis in cattle slaughtered at Aswan abattoirs was 3.2% (23/720), with thoracic lymph nodes and lungs exhibiting the highest frequency of tubercle lesions compared to other organs. According to employed diagnostic tests, the prevalence of tuberculosis was estimated as 68.6, 61, 56.5, and 78.3% for microscopy, histopathology, ELISA, and RT-PCR, respectively. These findings highlight the incidence of bovine tuberculosis and the dissemination of tuberculous lesions among cattle slaughtered at Aswan abattoirs. Furthermore, such prevalence emphasizes the urgent need for a practical disease control approach and an organized surveillance system in the cattle population. One Health strategy is strongly advised to mitigate the zoonotic transmission to people and economic losses in the cattle industry. [ABSTRACT FROM AUTHOR]

This study determined the frequency of positive tests for bovine tuberculosis in animals and breeders/establishments in Rio Grande do Norte, Brazil. Data were provided by the Instituto de Defesa e Inspeção Agropecuária do Rio Grande do Norte (IDIARN), from its Unidade Local de Sanidade Animal e Vegetal (ULSAV'S), acquired in monthly reports issued by qualified Veterinarians under the National Program for the Control and Eradication of Brucellosis and Animal Tuberculosis (PNCEBT), from June 2012 to December 2021. For diagnosis, the comparative cervical test was used as a routine and confirmatory test. A total of 45,804 cattle were tested, 53 (0.1%) of which tested positive. Secondary data are essential in the evaluation of sanitary measures, allowing rapid generation of hypotheses about diseases and providing support for decision-making. Measures such as raising awareness among producers, sanitary control in the acquisition and sale of cows for reproduction, inspection of sanitary barriers, and conducting epidemiological surveys to understand the actual situation of this disease in the state of Rio Grande do Norte are crucial. [ABSTRACT FROM AUTHOR]

Background/Objectives: Vaccines may improve the control and eradication of bovine tuberculosis. However, the evaluation of experimental candidates requires the assessment of the protection, excretion, transmission and biosafety. A natural transmission trial among likely infected animals was conducted. Methods: Seventy-four male heifers were randomly distributed (five groups) and vaccinated subcutaneously with attenuated strains (M. bovis Δmce2 or M. bovis Δmce2-phoP), a recombinant M. bovis BCG Pasteur (BCGr) or M. bovis BCG Pasteur. Then, they cohoused with a naturally infected bTB cohort under field conditions exposed to the infection. Results: A 23% of transmission of wild-type strains was confirmed (non-vaccinated group). Strikingly, first vaccination did not induce immune response (caudal fold test and IFN-gamma release assay). However, after 74 days of exposure to bTB, animals were re-vaccinated. Although their sensitization increased throughout the trial, the vaccines did not confer significant protection, when compared to the non-vaccinated group, as demonstrated by pathology progression of lesions and confirmatory tools. Besides, the likelihood of acquiring the infection was similar in all groups compared to the non-vaccinated group (p > 0.076). Respiratory and digestive excretion of viable vaccine candidates was undetectable. To note, the group vaccinated with M. bovis Δmce2-phoP exhibited the highest proportion of animals without macroscopic lesions, compared to the one vaccinated with BCG, although this was not statistically supported. Conclusions: This highlights that further evaluation of these vaccines would not guarantee better protection. The limitations detected during the trial are discussed regarding the transmission rate of M. bovis wild-type, the imperfect test for studying sensitization, the need for a DIVA diagnosis and management conditions of the trials performed under routine husbandry conditions. Re-vaccination of likely infected bovines did not highlight a conclusive result, even suggesting a detrimental effect on those vaccinated with M. bovis BCG. [ABSTRACT FROM AUTHOR]

Bovine tuberculosis (bTB) is a zoonotic disease with high economic impact and public health concern. However, the understanding of bTB in Indonesia is limited. Thus, this study aims to to estimate the pooled prevalence of bTB in cattle in Indonesia using a meta-analysis. Five online databases were searched for related articles. The articles were screened by two independent reviewers following the inclusion criteria. The included studies went through quality assessment using five pre-determined question. The necessary data were extracted and analysed using random effect model in R studio 4.3.0 with "meta" R package to estimate the pooled prevalence, heterogeneity level, and risk of publication bias. Subgroup meta-analysis was conducted for subgroup diagnostic method and Mycobacterium species. The results showed that six studies fit in inclusion criteria, representing 711 samples. The quality assessment showed the included studies were good quality. The estimated pooled prevalence of bTB in Indonesia was 9.71% (95% CI, 1.47-43.69%) with high heterogeneity. Based on subgroup meta-analysis, no significant different was found in between non-molecular and molecular diagnostic methods (10.55% [95% CI, 0.48-42.03] vs 6.43 [95% CI, 1.64-22.11], p = 0.92). Similarly, between M. bovis and M. tuberculosis, no significant different prevalence was found (2.04% [95% CI, 0.38-10.13] vs 6.55% [95% CI, 0.15-76.17], p= 0.57). The risk of bias assessment showed no indication of severe publication bias. This study indicates the necessity of bTB active surveillance and epidemiological study in Indonesia. Furthermore, a control and preventive strategy for zoonotic transmission mitigation should be developed. [ABSTRACT FROM AUTHOR]

In the United Kingdom, the management of bovine tuberculosis (bTB) challenges the coexistence of people and wildlife. Control of this cattle disease is hindered by transmission of its causative agent, Mycobacterium bovis, between cattle and badgers Meles meles.Badger culling has formed an element of bTB control policy for decades, but current government policy envisions expanding badger vaccination. Farming leaders are sceptical, citing concerns that badger vaccination would be impractical and potentially ineffective.We report on a 4‐year badger vaccination initiative in an 11 km2 area which, atypically, was initiated by local farmers, delivered by scientists and conservationists, and co‐funded by all three. Participating landholders cited controversies around culling and a desire to support neighbours as their primary reasons for adopting vaccination.The number of badgers vaccinated per km2 (5.6 km−2 in 2019) exceeded the number culled on nearby land (2.9 km−2 in 2019), and the estimated proportion vaccinated (74%, 95% confidence interval [CI] 40%–137%) exceeded the 30% threshold predicted by models to be necessary to control M. bovis. Farmers were content with how vaccination was delivered, and felt that it built trust with wildlife professionals.The percentage of badgers testing positive for M. bovis declined from 16.0% (95% CI 4.5%–36.1%) at the start of vaccination to 0% (95% CI 0%–9.7%) in the final year. With neither replication nor unvaccinated controls, this small‐scale case study does not demonstrate a causal link between badger vaccination and bTB epidemiology, but it does suggest that larger‐scale evaluation of badger vaccination would be warranted.Farmers reported that their enthusiasm for badger vaccination had increased after participating for 4 years. They considered vaccination to have been effective, and good value for money, and wished to continue with it.Synthesis and applications: Although small‐scale, this case study suggests that badger vaccination can be a technically effective and socially acceptable component of bTB control. A wider rollout of badger vaccination is more likely if it is led by the farming community, rather than by conservationists or government, and is combined with scientific monitoring. Read the free Plain Language Summary for this article on the Journal blog. Read the free Plain Language Summary for this article on the Journal blog [ABSTRACT FROM AUTHOR]

Bovine tuberculosis (bTB), caused by Mycobacterium bovis, poses significant zoonotic and economic challenges globally. The current prevention and treatment options are limited and increasingly complicated by the emergence of multidrug‐resistant strains. This study employs reverse vaccinology and immunoinformatics to design a multi‐epitope subunit vaccine targeting the MPB83, ArfA, DnaK, GrpE, and LpqH proteins of M. bovis. The T‐cell and B‐cell epitopes of the candidate vaccine were predicted and evaluated for antigenicity, allergenicity, and toxicity. The promising epitopes were then assembled into three vaccine constructs (bTBV1, bTBV2, and bTBV3) using appropriate adjuvants, pan HLA DR‐binding epitope (PADRE), and linkers. The constructs were analyzed for physicochemical properties, 3D structure, cytokines induction and stability, followed by molecular docking with bovine CD molecules and toll‐like receptor, TLR‐9. Among the candidates, bTBV3 was chosen as one of the most promising vaccine candidates due to its high aliphatic index (67.60), lowest instability score (27.26), and a strong binding affinity. Molecular dynamics simulations and the results of interactions between the vaccine–receptor complexes (eigenvalue 2.318704e‐06) show that the vaccine construct bTBV3 is stable. In silico immune simulation findings, such as elevated IgM levels and increased Th cell populations, suggest that the designed multi‐epitope vaccine candidate bTBV3 elicits robust humoral and cellular immune responses, confirming the vaccine's potential efficacy. Additionally, codon optimization (CAI: 0.997 and GC: 54.687%) and in silico cloning facilitated efficient expression in E. coli. This study highlights the potential of bioinformatics‐driven approaches in developing effective subunit vaccines against bTB, providing a foundation for experimental validation and future applications in combating this pervasive zoonotic disease, bovine tuberculosis. [ABSTRACT FROM AUTHOR]

This article from Veterinary Pathology provides images and descriptions of various pulmonary diseases in animals. It includes images and information on diseases such as bovine tuberculosis, bronchial cartilage hypoplasia, subpleural macrophage foci, pulmonary endometriosis, and enzootic calcinosis. The article aims to educate readers about these diseases and their characteristics. [Extracted from the article]

Simple Summary: Paratuberculosis (PTB) is a chronic disease that affects domestic and wild ruminants worldwide. This study was conducted in 12 diary caprine farms on the Canary Islands. The region counts with the fourth largest goat population in Spain and has "officially free" bovine tuberculosis status. Two sampling sessions were conducted, and 2774 serum samples were tested by an enzyme-linked immunosorbent assay. In the first session, a prevalence of 18.4% was obtained, varying from 2.5% up to 61.1%. In the second session, the effect of PTB vaccination was evaluated and both non-vaccinated (nV) and vaccinated (V) were included. Variable tendencies in antibody development were registered in farms with different initial seroprevalences. In farms in which up to 10% of the animals were positive, more adult goats had antibodies against PTB after vaccination. In farms with more than 10% of ELISA-positive animals, a heterogeneous response to vaccination was reported. We observed that in farms with higher initial prevalence, fewer goats that were V developed antibodies. Our work characterizes the caprine PTB situation on the Canary Islands and gives new insights on the effect of farm prevalence on the immune response to PTB vaccination, although further studies on a greater scale are needed. Paratuberculosis (PTB), caused by Mycobacterium avium subspecies paratuberculosis (MAP), is a chronic disease with economic impact on ruminant farming worldwide. The Canary Islands count with the fourth largest goat population in Spain and are "officially free" of bovine tuberculosis. Twelve farms were included with 2774 serum samples tested by an enzyme-linked immunosorbent assay (ELISA) for detection of anti-MAP antibodies in two sessions. In the first session, an overall apparent prevalence of 18.4% (2.5% up to 61.1%) was obtained. Farms with prevalences (0–10%], (10–20%] and >20% were identified, with differences in seroconversion in the same prevalence group between farms and age ranges. Non-vaccinated (nV) and vaccinated (V) animals were included in the second sampling session. Higher levels of antibodies were detected in V animals older than 12 months, with considerable variations between age ranges and farms. Our results describe the current PTB status of the Canary Islands' goat farming. Furthermore, new insights on the effect of the farm prevalence on seroconversion in V animals are provided, although further studies are needed to evaluate the multiple factors affecting the immune response to anti-MAP vaccination. [ABSTRACT FROM AUTHOR]

Background: Bovine tuberculosis (bTB) is a chronic disease that results from infection with any member of the Mycobacterium tuberculosis complex. Infected animals are typically diagnosed with tuberculin-based intradermal skin tests according to World Organization of Animal Health which are presently in use. However, tuberculin is not suitable for use in BCG-vaccinated animals due to a high rate of false-positive reactions. Peptide-based defined skin test (DST) antigens have been identified using antigens (ESAT-6, CFP-10 and Rv3615c) which are absent from BCG, but their performance in buffaloes remains unknown. To assess the comparative performance of DST with the tuberculin-based single intradermal test (SIT) and the single intradermal comparative cervical test (SICCT), we screened 543 female buffaloes from 49 organized dairy farms in two districts of Haryana state in India. Results: We found that 37 (7%), 4 (1%) and 18 (3%) buffaloes were reactors with the SIT, SICCT and DST tests, respectively. Of the 37 SIT reactors, four were positive with SICCT and 12 were positive with the DST. The results show that none of the animals tested positive with all three tests, and 6 DST positive animals were SIT negative. Together, a total of 43 animals were reactors with SIT, DST, or both, and the two assays showed moderate agreement (Cohen's Kappa 0.41; 95% Confidence Interval (CI): 0.23, 0.59). In contrast, only slight agreement (Cohen's Kappa 0.18; 95% CI: 0.02, 0.34) was observed between SIT and SICCT. Using a Bayesian latent class model, we estimated test specificities of 96.5% (95% CI, 92–99%), 99.7% (95% CI: 98–100%) and 99.0% (95% CI: 97–100%) for SIT, SICCT and DST, respectively, but considerably lower sensitivities of 58% (95% CI: 35–87%), 9% (95% CI: 3–21%), and 34% (95% CI: 18–55%) albeit with broad and overlapping credible intervals. Conclusion: Taken together, our investigation suggests that DST has a test specificity comparable with SICCT, and sensitivity intermediate between SIT and SICCT for the identification of buffaloes suspected of tuberculosis. Our study highlights an urgent need for future well-powered trials with detailed necropsy, with immunological and microbiological profiling of reactor and non-reactor animals to better define the underlying factors for the large observed discrepancies in assay performance, particularly between SIT and SICCT. [ABSTRACT FROM AUTHOR]

The water buffalo (Bubalus bubalis) is susceptible to bovine tuberculosis (TB), which receives increased attention in areas where buffalo breeding is prevalent, such as in Southern Italy, especially in the Campania region, where 70% of the buffalo stock is bred. Since 2012, TB testing in buffalo herds has been conducted using the Single Intradermal Test (SIT), with the Comparative Intradermal test (CIT) used in cases of inconclusive results. From 2012 to 2016, the interferongamma (IFN-γ) test was occasionally employed experimentally in herds with TB outbreaks to expedite eradication efforts. A local TB eradication program was implemented in officially TB-free buffalo herds between 2017 and 2019. This program involves initial screening with SIT, followed by confirmatory tests, including CIT and IFN-γ, for positive reactions. Since June 2019, the IFN-γ test has replaced the CIT in officially TB-free herds upon positive SIT reactions. Additionally, in suspected and confirmed TB-outbreak herds, the IFN-γ test was used at the discretion of the competent authority. Between 2017 and 2019, approximately 295,000 buffaloes in Campania were screened annually with in vivo tests provided by TB eradication programs. During this period, 32,040 animals from 855 herds were tested using the IFN-γ test and 4,895 tested positive. Since 2020, the use of IFN-γ testing has increased, and has become a prerequisite for the acquisition of TB-free status and is being systematically applied for TB outbreak-extinction procedures. The test was performed in all breeding buffaloes in cases of doubtful SIT results in TB-free herds and when TB lesions are detected at slaughter in animals from TB-free herds. This combined approach helped detect more TB outbreaks, and thereby led to a reduction in the TB prevalence and incidence rates. By 2022, the prevalence had decreased to 1.56%, and the incidence had decreased to 0.73%, after the increased use of the IFN-γ test. This study highlights the effectiveness of implemented strategies in reducing TB in this region. Overall, the data demonstrate the successful impact of TB eradication measures and surveillance activities in reducing bubaline TB prevalence and incidence in the Campania region. [ABSTRACT FROM AUTHOR]

Bovine tuberculosis (bTB) is a chronic inflammatory disease primarily caused by Mycobacterium bovis. The infection affects domestic animals and wildlife, posing a zoonotic risk to humans. To understand the dynamics of transmission and genetic diversity in Italy's M. bovis population, we conducted whole-genome sequencing (WGS) analysis on two prevalent genotypes, belonging to Spoligotype SB0120, identified in different geographical and temporal contexts. By comparing these genomes with international M. bovis isolates, we identified a distinct clade within the lineage La1.2, encompassing the Italian SB0120 isolates, indicating a genomic segregation of Italian M. bovis from other European isolates. Within Italy, a significant level of genetic variability emerged across regions, while isolates within epidemiologically linked outbreaks exhibited minimal genetic diversity. Additionally, isolates derived from cattle and wild boars within a tuberculosis hotspot in Central Italy and from cattle and black pigs in Sicily formed unified clonal clusters. This indicates the presence of persistent strains circulating in the examined regions. The genetic diversity within herds was limited, as specific clones endured over time within certain herds. This research enhances our comprehension of the epidemiology and transmission patterns of bTB in Italy, thereby aiding the development of precise control strategies and disease management. Using WGS and implementing standardized protocols and databases will be pivotal in combating bTB and promoting One-Health approaches to address this noteworthy public health concern. [ABSTRACT FROM AUTHOR]

Bovine tuberculosis (bTB), caused by Mycobacterium bovis (M. bovis), represents a significant problem for the agriculture industry as well as posing a risk for human health. Current diagnostic tests for bTB target the cell-mediated immune (CMI) response to infection with M. bovis, primarily through screening of animals with the tuberculin skin test. Epigenetic modifications have been shown to alter the course of the immune response and differentially methylated regions (DMRs) might also influence the outcome of the skin test in cattle. Whole Genome Bisulphite Sequencing (WGBS) was used to profile DNA methylation levels from peripheral blood of a group of cattle identified as test positive for M. bovis (positive for the single intradermal comparative tuberculin test (SICTT) and/or the interferon-γ release assay compared to a test negative control group [n = 8/group, total of 16 WGBS libraries]. Although global methylation profiles were similar for both groups across the genome, 223 DMRs and 159 Differentially Promoter Methylated Genes (DPMGs) were identified between groups with an excess of hypermethylated sites in SICTT positive cattle (threshold > 15% differential methylation). Genes located within these DMRs included the Interleukin 1 receptor (IL1R1) and MHC related genes (BOLA and BOLA-DQB). KEGG pathway analysis identified enrichment of genes involved in Calcium and MAPK signalling, as well as metabolism pathways. Analysis of DMRs in a subset of SICTT negative cattle that were IFN-γ positive showed differential methylation of genes including Interleukin 10 Receptor, alpha (IL10RA), Interleukin 17 F (IL17F) and host defence peptides (DEFB and BDEF109). This study has identified a number of immune gene loci at which differential methylation is associated with SICTT test results and the degree of methylation could influence effective host immune responses. [ABSTRACT FROM AUTHOR]

Interferon gamma (IFN‐γ) released by T lymphocytes, plays an important role in immune regulation and is also an important biomarker for the early diagnosis of bovine tuberculosis. In this work, an electrochemical immunosensor based on gold nanoparticles (AuNPs)/chitosan composite was developed for highly sensitive determination of bovine IFN‐γ. AuNPs/chitosan composite shows high surface area and good hydrophilicity. The bovine IFN‐γ immunosensor was prepared by immobilizing IFN‐γ antibody on AuNPs/chitosan composite‐modified glassy carbon electrode. The immunosensors were characterized by scanning electron microscopy, electrochemical impedance spectroscopy, and cyclic voltammetry (CV). Using horseradish peroxidase (HRP)‐labeled IFN‐γ antibody as amplification probe, the concentration of bovine IFN‐γ can be monitored via the increasing current signal, which can be attributed to the specific immune binding between bovine IFN‐γ antigen and HRP‐labeled IFN‐γ antibody, as well as the efficient catalysis reaction between thionine and HRP with the presence of H2O2. The developed electrochemical immunosensor exhibited a wide linear range of 0.01 to 30 ng/mL, and low detection limit of 0.003 ng/mL with good specificity and stability. This study provides a simple and promising method for diagnosis of bovine tuberculosis, which can be expanded to determine other biomarkers. [ABSTRACT FROM AUTHOR]

Simple Summary: Bovine tuberculosis is caused by Mycobacterium bovis, which causes tuberculosis in humans and animals. Diagnosis of bovine tuberculosis has relied on examination of immune responses to M. bovis; however, using these methods, disease detection during the earliest phases of infection has been difficult, allowing a window for cattle-to-cattle transmission to occur within a herd. Alternative means of diagnosis could include methods to detect M. bovis or M. bovis DNA in bodily fluids such as nasal secretions, saliva, or blood rather than the animal's response to infection. DNA-from M. bovis was detected in nasal swabs and saliva from a small number of experimentally infected calves during the first 8 weeks after experimental infection. Although DNA from M. bovis could be detected, no culturable M. bovis was recovered from nasal swabs or saliva. Moreover, M. bovis DNA was not found in blood samples collected weekly. Successful infection of all calves was demonstrated using an interferon gamma release assay. Identification of infected animals through the detection of M. bovis will require the use of alternative samples or alternative assays. Bovine tuberculosis is caused by Mycobacterium bovis, a member of the M. tuberculosis complex of mycobacterial species that cause tuberculosis in humans and animals. Diagnosis of bovine tuberculosis has relied on examinations of cell-mediated immune responses to M. bovis proteins using tuberculin skin testing and/or interferon gamma release assays. Even when using these methods, disease detection during the earliest phases of infection has been difficult, allowing a window for cattle-to-cattle transmission to occur within a herd. Alternative means of diagnosis could include methods to detect M. bovis or M. bovis DNA in bodily fluids such as nasal secretions, saliva, or blood. During the first 8 weeks after experimental aerosol infection of 18 calves, M. bovis DNA was detected in nasal swabs from a small number of calves 5, 6, and 8 weeks after infection and in samples of saliva at 1, 7, and 8 weeks after infection. However, at no time could culturable M. bovis be recovered from nasal swabs or saliva. M. bovis DNA was not found in blood samples collected weekly and examined by real-time PCR. Interferon gamma release assays demonstrated successful infection of all calves, while examination of humoral responses using a commercial ELISA identified a low number of infected animals at weeks 4–8 after infection. Examination of disease severity through gross lesion scoring did not correlate with shedding in nasal secretions or saliva, and calves with positive antibody ELISA results did not have more severe disease than other calves. [ABSTRACT FROM AUTHOR]

Understanding how bovine tuberculosis (TB) is maintained in wildlife reservoirs is critical for the management of this disease impacting cattle in many countries. For the primary wildlife reservoir of the disease in New Zealand, the brushtail possum (Trichosurus vulpecula), transmission of this contagious bacterial disease among possums is often assumed to be linked to home range overlap. Identifying drivers of possum home range, and home range overlap between individuals, is thus important for informing wildlife reservoir TB management in New Zealand. We monitored four sub-populations of free-living possums in New Zealand native forests during 10 consecutive months using live trapping, to give the first direct insight into how the frequency and area of overlaps alters with density, sex and age. A total of 832 individuals were captured (average 9.3, range from 1 to 40 captures per animal with a median value of 7) and 35,820 home range overlaps were recorded. Both the number and area of overlaps were significantly associated with age class, with 66.6% of overlaps occurring between adults, 30% between adults and juveniles, and only 3.4% between juveniles. Overall, adult males showed significantly higher numbers of overlaps than expected, while adult and juvenile females showed significantly lower numbers of overlaps than expected and no differences were observed in juvenile males. In addition, males exhibited more and larger overlaps than females. The number and size of overlaps per individual decreased with increasing local population density. Understanding shared areas of activity among individuals can provide insights into the interactions occurring and potential pathways for diseases transmitted by contact such as TB. These results can inform to develop effective strategies for the control of diseases carried and dispersed by possums. [ABSTRACT FROM AUTHOR]

Long noncoding RNAs (lncRNAs) are RNA molecules with a length greater than 200 nucleotides that do not code for functional proteins. Although, genes play a vital role in immune response against a disease, it is less known that lncRNAs also contribute through gene regulation. Bovine tuberculosis is a significant zoonotic disease caused by Mycobacterium bovis (M. bovis) in cattle. Here, we report the in-silico analysis of the publicly available transcriptomic data of calves infected with M. bovis. A total of 51,812 lncRNAs were extracted across all the samples. A total of 216 genes and 260 lncRNAs were found to be differentially expressed across all the 4 conditions—infected vs uninfected at 8- and 20-week post-infection (WPI), 8 vs 20-WPI of both infected and uninfected. Gene Ontology and Functional annotation showed that 8 DEGs were annotated with immune system GOs and 2 DEGs with REACTOME immune system pathways. Co-expression analysis of DElncRNAs with DEGs revealed the involvement of lncRNAs with the genes annotated with Immune related GOs and pathways. Overall, our study sheds light on the dynamic transcriptomic changes in response to M. bovis infection, particularly highlighting the involvement of lncRNAs with immune-related genes. The identified immune pathways and gene–lncRNA interactions offer valuable insights for further research in understanding host–pathogen interactions and potential avenues for genetic improvement strategies in cattle. [ABSTRACT FROM AUTHOR]

Inhalable microparticle-based anti TB drug delivery systems are being investigated extensively for Tuberculosis [TB] treatment as they offer efficient and deep lung deposition with several advantages over conventional routes. It can reduce the drug dose, treatment duration and toxic effects and optimize the drug bioavailability. Yeast derived β-glucan is a β-[1–3/1–6] linked biocompatible polymer and used as carrier for various biomolecules. Due to presence of glucan chains, particulate glucans act as PAMP and thereby gets internalized via receptor mediated phagocytosis by the macrophages. In this study, β-glucan microparticles were prepared by adding l-leucine as excipient, and exhibited 70% drug [Rifabutin] loading efficiency. Further, the sizing and SEM data of particles revealed a size of 2–4 µm with spherical dimensions. The FTIR and HPLC data confirmed the β-glucan composition and drug encapsulations efficiency of the particles. The mass median aerodynamic diameter [MMAD] and geometric standard deviation [GSD] data indicated that these particles are inhalable in nature and have better thermal stability as per DSC thermogram. These particles were found to be non-toxic upto a concentration of 80 µg/ml and were found to be readily phagocytosed by human macrophage cells in-vitro as well as in-vivo by lung alveolar macrophage. This study provides a framework for future design of inhalable β-glucan particle based host-directed drug delivery system against pulmonary TB. [ABSTRACT FROM AUTHOR]

Tuberculosis (TB) remains a global public health threat. Understanding the dynamics of host-pathogen interactions within TB granulomas will assist in identifying what leads to the successful elimination of infection. In vitro TB models provide a controllable environment to study these granuloma dynamics. Previously we developed a biomimetic 3D spheroid granuloma model that controls bacteria better than a traditional monolayer culture counterpart. We used agent-based simulations to predict the mechanistic reason for this difference. Our calibrated simulations were able to predict heterogeneous bacterial dynamics that are consistent with experimental data. In one group of simulations, spheroids are found to have higher macrophage activation than their traditional counterparts, leading to better bacterial control. This higher macrophage activation in the spheroids was not due to higher counts of activated T cells, instead fewer activated T cells were able to activate more macrophages due to the proximity of these cells to each other within the spheroid. In a second group of simulations, spheroids again have more macrophage activation but also more T cell activation, specifically CD8+ T cells. This higher level of CD8+ T cell activation is predicted to be due to the proximity of these cells to the cells that activate them. Multiple mechanisms of control were predicted. Simulations removing individual mechanisms show that one group of simulations has a CD4+ T cell dominant response, while the other has a mixed/CD8+ T cell dominant response. Lastly, we demonstrated that in spheroids the initial structure and movement rules work synergistically to reduce bacterial load. These findings provide valuable insights into how the structural complexity of in vitro models impacts immune responses. Moreover, our study has implications for engineering more physiologically relevant in vitro models and advancing our understanding of TB pathogenesis and potential therapeutic interventions. Author summary: Tuberculosis (TB) is a respiratory infection that is associated with granulomas, organized clusters of immune cells that form around the bacteria. Recent work has focused on making in vitro models that better approximate these granulomas, such as our spheroid granuloma model. We found that this spheroid was better able to control bacteria than its traditional culture counterpart, so, in this work, we use agent-based modeling to explore the causes of this differential control. The simulated spheroids have increased macrophage activation. Surprisingly, this is not due to larger numbers of activated T cells, which activate these macrophages. Rather, the increase in activated macrophages is due to their proximity to fewer activated T cells. We also see a subset of simulations where the spheroids have increased cytotoxic CD8+ T cell activation. The simulation is further used to simulate changes in the structure of the in vitro models to explore its impact on immune response and bacterial control. These findings reveal the importance of structural complexity in in vitro models and offer insights into TB pathogenesis. Ultimately, our work paves the way for more realistic in vitro models and a deeper understanding of how our bodies combat TB. [ABSTRACT FROM AUTHOR]

The current study was designed to evaluate the reliability of the interferon-gamma release assay (IGRA) as a screening test compared to intensive post-mortem inspection using agreement analyses. This study also aimed to determine the prevalence and risk factors of bovine tuberculosis (bTB) among bovines presented for slaughter in the largest abattoir in Lahore, Pakistan. After antimortem inspection, a total of 102 animals were randomly selected for sample and data collection. Selected animals were slaughtered and a thorough post-mortem examination was done for all carcasses to find TB-like lesions. Blood samples were processed by IGRA. Prevalence estimates were generated and Cohen's Kappa test was done for agreement analyses to compare the reliability of the two tests for bTB diagnosis. A substantial agreement (κ = 0.79) was estimated between the IGRA and intensive post-mortem inspection. The apparent prevalence was computed as 5.88% (95% CI; 2.59-11.97) and the true prevalence was estimated as 3.92% (95% CI; 1.35-9.47). A parallel testing strategy with IGRA and intensive post-mortem inspection is a useful approach for screening bTB. [ABSTRACT FROM AUTHOR]

The traditional way of thinking about human diseases across clinical and narrow phenomics silos often masks the underlying shared molecular substrates across human diseases. One Health and planetary health fields particularly address such complexities and invite us to think across the conventional disease nosologies. For example, tuberculosis (TB) and lung cancer (LC) are major pulmonary diseases with significant planetary health implications. Despite distinct etiologies, they can coexist in a given community or patient. This is both a challenge and an opportunity for preventive medicine, diagnostics, and therapeutics innovation. This study reports a bioinformatics analysis of publicly available gene expression data, identifying overlapping dysregulated genes, downstream regulators, and pathways in TB and LC. Analysis of NCBI-GEO datasets (GSE83456 and GSE103888) unveiled differential expression of CEACAM6, MUC1, ADM, DYSF, PLOD2, and GAS6 genes in both diseases, with pathway analysis indicating association with lysine degradation pathway. Random forest, a machine-learning-based classification, achieved accuracies of 84% for distinguishing TB from controls and 83% for discriminating LC from controls using these specific genes. Additionally, potential drug targets were identified, with molecular docking confirming the binding affinity of warfarin to GAS6. Taken together, the present study speaks of the pressing need to rethink clinical diagnostic categories of human diseases and that TB and LC might potentially share molecular substrates. Going forward, planetary health and One Health scholarship are poised to cultivate new ways of thinking about diseases not only across medicine and ecology but also across traditional diagnostic conventions. [ABSTRACT FROM AUTHOR]

Understanding how disease moves through wildlife communities is essential to managing outbreaks of zoonotic diseases across the globe. Bovine tuberculosis is a disease caused by the bacterium Mycobacterium bovis that can threaten domestic and wildlife species. The mechanism by which Mycobacterium bovis is spread between species is still poorly understood. Previous reviews are limited in the breadth of species considered and are primarily concerned with transmission from wildlife to domestic species.We conducted a review and analysis of Mycobacterium bovis prevalence rates in European wildlife species to identify species of concern for the transmission of bovine tuberculosis in a wildlife community. We subsequently conducted a narrative review of these species assessing the risk of Mycobacterium bovis transmission in a wildlife community based on available literature.We calculated weighted mean disease prevalence rates to be highest in fallow deer (Dama dama, 20%), Eurasian badgers (Meles meles, 11%), wild boar (Sus scrofa, 9%) and red foxes (Vulpes vulpes, 4%). We considered these species to be of particular concern for the transmission of Mycobacterium bovis and selected them as the focus of our narrative review and risk assessment. Our risk assessment considered disease pathology, spatiotemporal activity patterns and animal behaviour as factors affecting the likelihood of Mycobacterium bovis transmission between wildlife species.We found that prior research has principally focused on a few individual species, but that Mycobacterium bovis transmission through a wildlife community is likely more complex. We determined that disease transmission between multiple species may compound the severity of an outbreak of bovine tuberculosis. Broad, multi‐species sampling campaigns and standardised Mycobacterium bovis testing protocols should be implemented in future studies. We also determined that an in‐depth analysis of spatiotemporal overlap between species was needed to better assess the risk of transmission between wildlife species. [ABSTRACT FROM AUTHOR]

This study aims to conduct a comparative analysis of the uterotubal junction in two distinct mammalian species, the bovine (Bos taurus) and the dromedary camel (Camelus dromedarius), focusing on histological and histomorphometric parameters. Uterotubal junction (UTJ) was dissected from 8 cows and 12 camels with dominant follicles, and processed for H&E staining for histology, and histomorphometry examination. The results showed that the camel uterotubal junction papillae (UTJP) existed only in camels and was completely absent in cattle. Histologically, the cow UTJ appears with a star‐shaped lumen, and the mucosa is lined by a simple columnar epithelium with ciliated and non‐ciliated cells. Superficial (SG) and deep glands (DG) were abundant in the submucosa. Camel UTJP is a conical structure, it has a pale yellowish colour, 0.5 ± 0.2 cm height, and 0.3 ± 0.1 cm width, and it has a distinct sphincter at the tip towards the uterine lumen. In the camel UTJP, the lumen is wider, and the mucosa showed large multiple folds lined with ciliated and non‐ciliated cells in a pseudostratified columnar epithelium. The submucosa showed no mucosal glands. Lumen area, lumen epithelial height, luminal epithelial density, the thickness of the muscular layer, number of folds, fold height, epithelial height, fold area and epithelial perimeter were higher in camel UTJP (p <.001) than cow UTJ, while the total endosalpinx area and glandular epithelial density were lower in camel UTJP (p <.001) than in cow UTJ. In conclusion, this study elucidates potential variations in the uterotubal junction between bovines and dromedary camels, providing valuable insights into their reproductive adaptations. The epithelial lining, absence of glands and the thick layer of tunica muscularis might indicate that camel UTJP could play a mechanical role in selecting spermatozoa and assisting the hatching of blastocysts during their passage into the uterus. [ABSTRACT FROM AUTHOR]

The Mycobacterial growth inhibition assay (MGIA) is an ex-vivo assay used to measure the overall functional immune response elicited by infection or vaccination. In tuberculosis (TB) vaccine development, MGIA is a potentially important tool for preclinical evaluation of early-stage vaccine candidates to complement existing assays, and to potentially reduce the need for lengthy and costly pathogenic Mycobacterium tuberculosis (Mtb) animal challenge experiments. The conventional method of MGIA in mice entails directly infecting mixed cell cultures, most commonly splenocytes, from immunised mice with mycobacteria. However, this direct infection of mixed cell populations may yield unreliable results and lacks sufficient sensitivity to discriminate well between different vaccines due to the low number of mycobacteria-permissive cells. Here, we modified the assay by inclusion of mycobacteria-infected congenic murine macrophage cell lines as the target cells, and by measuring the total number of killed cells rather than the relative reduction between different groups. Thus, using splenocytes from Mycobacterium bovis BCG immunised mice, and J774 and MH-S (BALB/c background) or BL/6-M (C57Bl/6 background) macrophage cell lines, we demonstrated that the modified assay resulted in at least 26-fold greater mycobacterial killing per set quantity of splenocytes as compared to the conventional method. This increased sensitivity of measuring mycobacterial killing was confirmed using both the standard culture forming unit (CFU) assay and luminescence readings of luciferase-tagged virulent and avirulent mycobacteria. We propose that the modified MGIA can be used as a highly calibrated tool for quantitating the killing capacity of immune cells in preclinical evaluation of vaccine candidates for TB. [ABSTRACT FROM AUTHOR]

Six cattle heads which tested positive against bovine tuberculosis (bTB) in Morocco were investigated to confirm the disease and to determine the source(s) of infection. Polymerase Chain Reaction (PCR) was directly performed on tissue samples collected from slaughtered animals. All investigated animals tested positive to PCR for the Mycobacterium bovis sub-type. Bacteriological isolation was conducted according to the technique recommended by WOAH for the cultivation of the Mycobacterium tuberculosis Complex (MBTC). Whole genome sequencing (WGS) was carried out on six mycobacterial isolates and the phylogenic tree was constructed. The six Moroccan isolates fit with clades II, III, IV, V and VII and were confirmed to belong to the clonal complexes Eu2, Unknown 2 and 7 as well as to sublineages La1.7.1, La1.2 and La1.8.2. The significant Single Nucleotide Polymorphism (SNPs) ranged from 84 to 117 between the isolates and the reference M. bovis strain and from 17 to 212 between the six isolates. Considering the high resolution of WGS, these results suggests that the source of infection of the bTB could be linked to imported animals as five of the investigated reactor animals were imported a few months prior. WGS can be a useful component to the Moroccan strategy to control bTB. [ABSTRACT FROM AUTHOR]

Bovine tuberculosis (bTB) continues to have significant economic and veterinary health impacts on cattle herds where the disease remains endemic. The continual tailoring of policies to address such maintenance requires an in-depth analysis of national data, underpinning new control strategies. In Ireland, when outbreaks occur, ancillary testing of herd mates deemed to be at the highest risk of exposure to reactors is undertaken using the interferon gamma (GIF) test. This highest risk cohort was hypothesised to be of a higher future risk despite this ancillary testing. We used a dataset from Ireland to model bovine test failure to the comparative tuberculin skin test using a survival analysis (observations: 39,248). Our primary exposure of interest was whether an animal that tested negative had a GIF test after the disclosure of infection within a herd during a bTB breakdown. There was evidence that animals with a negative GIF test during a breakdown had an increased risk of failing a test relative to other animals from the same herds without this exposure. The time to failure was 48.8% (95%CI: 38.3–57.5%) shorter for the exposed group relative to the unexposed group during a two-year follow-up period (2019–2022; time ratio: 0.51; 95%CI: 0.43–0.62; p < 0.001). The results from this study suggest that animals who were GIF-tested, having been deemed to have a higher risk of exposure, subsequently had shorter time-to-test failure periods. The absolute numbers of failure are small (only 2.5% of animals go on to fail during 2-year follow-up). Importantly, however, a high proportion of these high-risk herds included in the dataset failed at least one test at the follow-up (21/54 herds), impacting breakdown duration or recurrence. Such risk-informed targeting of animals could be utilised in future control policies, though further research is warranted. [ABSTRACT FROM AUTHOR]

Background: Tuberculosis (TB) persists as a global health challenge, with its treatment hampered by the side effects of long-term combination drug therapies and the growing issue of drug resistance. Therefore, the development of novel therapeutic strategies is critical. This study focuses on the role of immune checkpoint molecules (ICs) and functions of CD8+ T cells in the search for new potential targets against TB. Methods: We conducted differential expression genes analysis and CD8+ T cell functional gene analysis on 92 TB samples and 61 healthy individual (HI) samples from TB database GSE83456, which contains data on 34,603 genes. The GSE54992 dataset was used to validated the findings. Additionally, a cluster analysis on single-cell data from primates infected with mycobacterium tuberculosis and those vaccinated with BCG was performed. Results: The overexpression of LAG-3 gene was found as a potentially important characteristic of both pulmonary TB (PTB) and extrapulmonary TB (EPTB). Further correlation analysis showed that LAG-3 gene was correlated with GZMB, perforin, IL-2 and IL-12. A significant temporal and spatial variation in LAG-3 expression was observed in T cells and macrophages during TB infection and after BCG vaccination. Conclusion: LAG-3 was overexpressed in TB samples. Targeting LAG-3 may represent a potential therapeutic target for tuberculosis. [ABSTRACT FROM AUTHOR]

During tuberculosis (TB), migration of dendritic cells (DCs) from the site of infection to the draining lymph nodes is known to be impaired, hindering the rapid development of protective T-cell- mediated immunity. However, the mechanisms involved in the delayed migration of DCs during TB are still poorly defined. Here, we found that infection of DCs with Mycobacterium tuberculosis (Mtb) triggers HIF1A-mediated aerobic glycolysis in a TLR2-dependent manner, and that this metabolic profile is essential for DC migration. In particular, the lactate dehydrogenase inhibitor oxamate and the HIF1A inhibitor PX-478 abrogated Mtb-induced DC migration in vitro to the lymphoid tissue-specific chemokine CCL21, and in vivo to lymph nodes in mice. Strikingly, we found that although monocytes from TB patients are inherently biased toward glycolysis metabolism, they differentiate into poorly glycolytic and poorly migratory DCs compared with healthy subjects. Taken together, these data suggest that because of their preexisting glycolytic state, circulating monocytes from TB patients are refractory to differentiation into migratory DCs, which may explain the delayed migration of these cells during the disease and opens avenues for host-directed therapies for TB. [ABSTRACT FROM AUTHOR]

Mycobacterium tuberculosis, the causative agent of tuberculosis, is a pathogenic bacterium that has claimed millions of lives since the Middle Ages. According to the World Health Organization's report, tuberculosis ranks among the ten deadliest diseases worldwide. The presence of an extensive array of genes and diverse proteins within the cellular structure of this bacterium has provided us with a potent tool for diagnosis. While the culture method remains the gold standard for tuberculosis diagnosis, it is possible that molecular diagnostic methods, emphasis on the identification of mutation genes (e.g., rpoB and gyrA) and single nucleotide polymorphisms, could offer a safe and reliable alternative. Over the past few decades, as our understanding of molecular genetics has expanded, methods have been developed based on gene expansion and detection. These methods typically commence with DNA amplification through nucleic acid targeted techniques such as polymerase chain reaction. Various molecular compounds and diverse approaches have been employed in molecular assays. In this review, we endeavor to provide an overview of molecular assays for the diagnosis of tuberculosis with their properties (utilization, challenges, and functions). The ultimate goal is to explore the potential of replacing traditional bacterial methods with these advanced molecular diagnostic techniques. [ABSTRACT FROM AUTHOR]

A quarter of humanity is estimated to have been exposed to Mycobacterium tuberculosis (Mtb) with a 5–10% risk of developing tuberculosis (TB) disease. Variability in responses to Mtb infection could be due to host or pathogen heterogeneity. Here, we focused on host genetic variation in a Peruvian population and its associations with gene regulation in monocyte-derived macrophages and dendritic cells (DCs). We recruited former household contacts of TB patients who previously progressed to TB (cases, n = 63) or did not progress to TB (controls, n = 63). Transcriptomic profiling of monocyte-derived DCs and macrophages measured the impact of genetic variants on gene expression by identifying expression quantitative trait loci (eQTL). We identified 330 and 257 eQTL genes in DCs and macrophages (False Discovery Rate (FDR) < 0.05), respectively. Four genes in DCs showed interaction between eQTL variants and TB progression status. The top eQTL interaction for a protein-coding gene was with FAH, the gene encoding fumarylacetoacetate hydrolase, which mediates the last step in mammalian tyrosine catabolism. FAH expression was associated with genetic regulatory variation in cases but not controls. Using public transcriptomic and epigenomic data of Mtb-infected monocyte-derived dendritic cells, we found that Mtb infection results in FAH downregulation and DNA methylation changes in the locus. Overall, this study demonstrates effects of genetic variation on gene expression levels that are dependent on history of infectious disease and highlights a candidate pathogenic mechanism through pathogen-response genes. Furthermore, our results point to tyrosine metabolism and related candidate TB progression pathways for further investigation. Author summary: Tuberculosis (TB), caused by Mycobacterium tuberculosis (Mtb), is a leading cause of mortality globally. Mtb-exposed individuals have heterogeneous outcomes following infection with Mtb, where some progress to TB, while many remain asymptomatic. We hypothesized that genetic variation could partly explain risk of TB, by regulating the expression of genes involved in the control of Mtb infection. In this study in Peru, we recruited former TB patients and Mtb-exposed household contacts of TB patients who remained asymptomatic to define how genetic variation regulates expression of genes in the innate immune system. We identified 4 genetic variants that have different effects on gene expression based on the TB history of the participant. The lead variant regulated expression of a key enzyme (Fumarylacetoacetate Hydrolase; FAH) in tyrosine catabolism. Knocking out the gene in myeloid cells increased susceptibility to Mtb infection. The results implicate FAH as a candidate host factor involved in TB progression. [ABSTRACT FROM AUTHOR]

Mycobacterium tuberculosis infects two billion people across the globe, and results in 8–9 million new tuberculosis (TB) cases and 1–1.5 million deaths each year. Most patients have no known genetic basis that predisposes them to disease. Here, we investigate the complex genetic basis of pulmonary TB by modelling human genetic diversity with the Diversity Outbred mouse population. When infected with M. tuberculosis, one-third develop early onset, rapidly progressive, necrotizing granulomas and succumb within 60 days. The remaining develop non-necrotizing granulomas and survive longer than 60 days. Genetic mapping using immune and inflammatory mediators; and clinical, microbiological, and granuloma correlates of disease identified five new loci on mouse chromosomes 1, 2, 4, 16; and three known loci on chromosomes 3 and 17. Further, multiple positively correlated traits shared loci on chromosomes 1, 16, and 17 and had similar patterns of allele effects, suggesting these loci contain critical genetic regulators of inflammatory responses to M. tuberculosis. To narrow the list of candidate genes, we used a machine learning strategy that integrated gene expression signatures from lungs of M. tuberculosis-infected Diversity Outbred mice with gene interaction networks to generate scores representing functional relationships. The scores were used to rank candidates for each mapped trait, resulting in 11 candidate genes: Ncf2, Fam20b, S100a8, S100a9, Itgb5, Fstl1, Zbtb20, Ddr1, Ier3, Vegfa, and Zfp318. Although all candidates have roles in infection, inflammation, cell migration, extracellular matrix remodeling, or intracellular signaling, and all contain single nucleotide polymorphisms (SNPs), SNPs in only four genes (S100a8, Itgb5, Fstl1, Zfp318) are predicted to have deleterious effects on protein functions. We performed methodological and candidate validations to (i) assess biological relevance of predicted allele effects by showing that Diversity Outbred mice carrying PWH/PhJ alleles at the H-2 locus on chromosome 17 QTL have shorter survival; (ii) confirm accuracy of predicted allele effects by quantifying S100A8 protein in inbred founder strains; and (iii) infection of C57BL/6 mice deficient for the S100a8 gene. Overall, this body of work demonstrates that systems genetics using Diversity Outbred mice can identify new (and known) QTLs and functionally relevant gene candidates that may be major regulators of complex host-pathogens interactions contributing to granuloma necrosis and acute inflammation in pulmonary TB. Author summary: We investigated the genetic basis of susceptibility to Mycobacterium tuberculosis using Diversity Outbred mice, a mouse population suited for studying complex genotype-phenotype relationships. We found five new genetic loci and three known loci. Three loci associated with multiple correlated disease traits likely contain genes that are major regulators of host inflammatory responses which favor M. tuberculosis growth. Further, these three loci contain four gene candidates with single nucleotide polymorphisms that are predicted to have deleterious effects upon protein functions. [ABSTRACT FROM AUTHOR]

Background: Zoonoses are infectious diseases that are transmitted from animals to humans. Studying the knowledge, perceptions and practices of communities related to zoonoses and the associated risk factors is crucial for effective control and prevention. This study aimed to assess the knowledge, perceptions, and practices of respondents on zoonoses and the associated risk factors in and around Chiro town, Ethiopia. Zoonotic diseases, such as rabies, anthrax, bovine tuberculosis, and brucellosis, pose a direct threat to health and livelihoods in the communities where they occur. These diseases emerge due to a combination of human-animal interactions, migration, and contact with wildlife and their respective parasites and vectors. Hence, recognizing residents' perceptions, knowledge, and practices is crucial for effectively minimizing risks. Methods: A cross-sectional study was conducted from January 2020 to July 2021 in and around Chiro town using a pretested close-ended questionnaire. A total of 350 respondents were selected using simple random sampling methods. The questionnaire included information on the sociodemographic status of the respondents and questions concerning the knowledge, perceptions, and practices of the participants regarding the selected zoonotic diseases. The associations of knowledge, perceptions, and practices related to zoonoses with zoonotic risk factors were analysed using chi-square tests. Results: The study revealed that 82.9% of the respondents had knowledge of bovine tuberculosis, followed by knowledge of rabies (80%), knowledge of anthrax (45.1%), and knowledge of brucellosis (24.3%). Males had greater knowledge of bovine tuberculosis (84.8%), followed by rabies (79.8%) and anthrax (48.6%), while females had greater knowledge of brucellosis (23.6%). The most cited source of information was radio (68%). Most respondents mentioned the outbreaks of rabies (62.5%), bovine tuberculosis (53.2%), anthrax (35.6%), and brucellosis (15.7%). Respondents with higher educational levels and urban residents had more knowledge of zoonoses. More than 75% of respondents had a good perception of the transmission of zoonotic disease from animals, and the practice of consuming raw milk or raw/undercooked meat and sharing the same house with animals was high. Conclusion: The majority of respondents reported that they had knowledge of bovine tuberculosis and rabies, but lower knowledge and perceptions were reported for anthrax and brucellosis. These findings illustrate the need for collaboration among animal, human and environmental health offices in one health approach to prevent and control zoonotic disease. [ABSTRACT FROM AUTHOR]

This study aimed to determine the sensitivity (Se) and specificity (Sp) of a circulating pathogen-specific biomarker (polyketide synthetase 5, Pks5)-based enzyme-linked immunosorbent assay (ELISA) independently or in conjunction with a caudal fold tuberculin (CFT) test for bovine tuberculosis (bTB) screening in dairy cattle. We enrolled 987 dairy cows from 34 herds in Chiang Mai province, Thailand. A conditionally independent Bayesian model with a single population was inferred from the test results. The percentage of positive results for the Pks5-ELISA using 0.4 OD cutoff test and CFT test were 9.0% (89/987) and 10.5% (104/987), respectively. The median of posterior estimates of Se for the Pks5-ELISA test was 90.2% (95% posterior probability interval [PPI] = 76.6-97.4%), while the estimated Sp was slightly higher (median = 92.9, 95% PPI = 91.0-94.5%). The median estimated Se of the CFT test was 85.9% (95% PPI = 72.4-94.6%), while the estimated Sp was higher, with a median of 90.7% (95% PPI = 88.7-92.5%). The posterior estimate for true disease prevalence was 2.4% (95% PPI = 1.2-3.9%). The Pks5-ELISA test yielded characteristics at or above the acceptable standards for bTB detection. Therefore, the pathogen-specific biomarker, Pks5, is a potential detection system for bTB screening and may be applied as an ancillary test together with the currently applied standard method (CFT test) to reinforce the bTB control and eradication programs. [ABSTRACT FROM AUTHOR]