Detection of lumpy skin disease virus reads in the human upper respiratory tract microbiome requires further investigation.

Lumpy skin disease virus (LSDV), a double-stranded DNA virus from the Capripoxvirus genus, primarily affects Bos indicus, Bos taurus breeds, and water buffalo. Arthropod vectors, including mosquitoes and biting flies, are the main LSDV transmitters. Although LSDV is not zoonotic, this study unexpectedly detected LSDV reads in the upper respiratory tract microbiome of humans from rural and urban areas in Maharashtra, India. Nasopharyngeal and oropharyngeal swab samples collected for SARS-CoV-2 surveillance underwent whole-genome metagenomics sequencing, revealing LSDV reads in 25% of samples. Split kmer analysis provided insights into sample relatedness despite the low coverage of LSDV reads with the reference genome. Our findings, which include the detection of LSDV contigs aligning to specific locations on the reference genome, suggest a common source for LSDV reads, potentially shared water sources, or milk/milk products. Further investigation is needed to ascertain the mode of transmission and reason for the detection of LSDV reads in human upper respiratory tract.

[Bacterial zoonoses of public health importance in Germany-incidence, distribution, and modes of transmission]. / Bakterielle Zoonosen mit Bedeutung für den öffentlichen Gesundheitsschutz in Deutschland ­ Vorkommen, Verbreitung und Übertragungswege.

Bacterial zoonotic pathogens are often the cause of diseases, sometimes with severe outcomes. They are mutually transferable between animals (both wild and domestic) and humans. The transmission paths are very variable and include oral intake via food, respiratory infection via droplets and aerosols, or infections via vectors such as tick bites or rodent contact. Furthermore, the emergence and spread of antibiotic-resistant bacterial pathogens is of paramount public health concern.The likelihood of further spread is influenced by various factors. These include the increase in international trade, the endangerment of animal habitats, and the increasingly closer contact between humans and wild animals. Additionally, changes in livestock and climate change may also contribute. Therefore, research into zoonoses serves to protect human and animal health and is of particular social, political, and economic importance.The aim of this review article is to present the range of infectious diseases caused by bacterial zoonotic pathogens in order to provide a better understanding of the important work in public health services, animal health services, and food safety control. The different transmission routes, epidemic potentials, and epidemiological measures of the exemplary selected diseases show the challenges for the public health system to monitor and control the spread of these bacterial pathogens in order to protect the population from disease.

Tracing the Source of Influenza A Virus Zoonoses in Interconnected Circuits of Swine Exhibitions.

BACKGROUND: Since 2011, influenza A viruses circulating in US swine exhibited at county fairs are associated with >460 zoonotic infections, presenting an ongoing pandemic risk. Swine "jackpot shows" that occur before county fairs each summer intermix large numbers of exhibition swine from diverse geographic locations. We investigated the role of jackpot shows in influenza zoonoses. METHODS: We collected snout wipe or nasal swab samples from 17 009 pigs attending 350 national, state, and local swine exhibitions across 8 states during 2016-2018. RESULTS: Influenza was detected in 13.9% of swine sampled at jackpot shows, and 76.3% of jackpot shows had at least 1 pig test positive. Jackpot shows had 4.3-fold higher odds of detecting at least 1 influenza-positive pig compared to county fairs. When influenza was detected at a county fair, almost half of pigs tested positive, clarifying why zoonotic infections occur primarily at county fairs. CONCLUSIONS: The earlier timing of jackpot shows and long-distance travel for repeated showing of individual pigs provide a pathway for the introduction of influenza into county fairs. Mitigation strategies aimed at curtailing influenza at jackpot shows are likely to have downstream effects on disease transmission at county fairs and zoonoses.

Increased human-animal interface & emerging zoonotic diseases: An enigma requiring multi-sectoral efforts to address.

Increased human-animal interfaces impose threats on human life by creating scope for the emergence and resurgence of many infectious diseases. Over the last two decades, emergence of novel viral diseases such as SARS, influenza A/H1N1(09) pdm; MERS; Nipah virus disease; Ebola haemorrhagic fever and the current COVID-19 has resulted in massive outbreaks, epidemics and pandemics thereby causing profound losses of human life, health and economy. The current COVID-19 pandemic has affected more than 200 countries, reporting a global case load of 167,878,000 with 2 per cent mortality as on May 26, 2021. This has highlighted the importance of reducing human- animal interfaces to prevent such zoonoses. Rapid deforestation, shrinking of boundaries between human and animal, crisis for natural habitation, increasing demands for wildlife products and threat of extinction compounded by biodiversity narrowing compel to increased human-animal conflict and contact. Large quantities of animal waste generated due to animal agriculture may also allow rapid selection, amplification, dissemination of zoonotic pathogens and facilitate zoonotic pathogen adaptation and hinder host evolution for resistance. Public health system faces challenges to contain such epidemics due to inadequate understanding, poor preparedness, lack of interdisciplinary approach in surveillance and control strategy and deficient political commitments. Because the management measures are beyond the purview of health system alone, policy-level adaptation in the transdisciplinary issues are required, emphasizing the engagement of multiple stakeholders towards wildlife protection, alternative land use, community empowerment for natural resource management and regulation on business of wildlife products to ensure comprehensive one health practice.

Evolutionary Dynamics of Influenza A Viruses in US Exhibition Swine.

The role of exhibition swine in influenza A virus transmission was recently demonstrated by >300 infections with influenza A(H3N2) variant viruses among individuals who attended agricultural fairs. Through active influenza A virus surveillance in US exhibition swine and whole-genome sequencing of 380 isolates, we demonstrate that exhibition swine are actively involved in the evolution of influenza A viruses, including zoonotic strains. First, frequent introduction of influenza A viruses from commercial swine populations provides new genetic diversity in exhibition pigs each year locally. Second, genomic reassortment between viruses cocirculating in exhibition swine increases viral diversity. Third, viral migration between exhibition swine in neighboring states demonstrates that movements of exhibition pigs contributes to the spread of genetic diversity. The unexpected frequency of viral exchange between commercial and exhibition swine raises questions about the understudied interface between these populations. Overall, the complexity of viral evolution in exhibition swine indicates that novel viruses are likely to continually reemerge, presenting threats to humans.

H9N2 influenza A viruses found to be enzootic in Punjab Pakistan's bird markets with evidence of human H9N2 nasal colonization.

OBJECTIVES: This study sought to detect and characterize influenza A (IAV) and influenza D (IDV) viruses circulating among commercial birds and shop owners in Pakistan's live bird markets. METHODS: Oropharyngeal swabs (n = 600; n = 300 pools) collected from poultry and nasopharyngeal swabs (n = 240) collected from poultry workers were studied for molecular evidence of IAV and IDV using real-time and conventional real-time reverse transcription polymerase chain reaction protocols. RESULTS: Nineteen (6.3%) poultry pools were positive for IAV and 73.9% of these were positive for H9N2 subtypes. Two (0.83%) poultry workers had evidence of IAV, and both were also H9N2 subtypes. The poultry and human IAV-positive specimens all clustered phylogenetically by Sanger and next-generation sequencing with previously detected H9N2 poultry isolates. No field specimens were positive for IDV. CONCLUSION: H9N2 IAV is likely enzootic in Punjab Province Pakistan's live bird markets and may be colonizing the noses of workers and market visitors. Regular monitoring for avian influenza-associated human illness in Punjab seems to be a needed public measure.

Human Seasonal Influenza Viruses in Swine Workers in Lagos, Nigeria: Consequences for Animal and Public Health.

The influenza A virus has been scarcely investigated in pigs in Africa, with rare detection prior to 2009. The spread of A(H1N1)pdm09 changed the epidemiology due to frequent human-to-swine transmission and the emergence of various new reassortants. This study therefore aimed at estimating the level of circulation and characterizing influenza A viruses at the interface between swine workers, who are crucial players in the inter-species transmission of influenza A viruses, and their animals in several farms in Nigeria, a hub for pig production in Africa. This cross-sectional study showed that 24.6% (58/236) of the pig serum samples collected in 2013-2014 had anti-influenza A antibodies in the absence of vaccination programs, but none of the pig swabs (n = 1193) were positive according to RT-qPCR. Viral RNA was detected in 0.9% (2/229) of swine workers sampled at their place of work, and the strains were characterized as A(H1N1)pdm09 and seasonal A(H3N2). Our results highlight that more awareness of swine workers regarding the consequences of reverse zoonosis for animal and public health is warranted. Annual vaccination and the wearing of masks when experiencing influenza-like symptoms would help decrease influenza inter-species transmission, while surveillance should be adequately supported for early detection.

Unbiased Virus Detection in a Danish Zoo Using a Portable Metagenomic Sequencing System.

Metagenomic next-generation sequencing (mNGS) is receiving increased attention for the detection of new viruses and infections occurring at the human-animal interface. The ability to actively transport and relocate this technology enables in situ virus identification, which could reduce response time and enhance disease management. In a previous study, we developed a straightforward mNGS procedure that greatly enhances the detection of RNA and DNA viruses in human clinical samples. In this study, we improved the mNGS protocol with transportable battery-driven equipment for the portable, non-targeted detection of RNA and DNA viruses in animals from a large zoological facility, to simulate a field setting for point-of-incidence virus detection. From the resulting metagenomic data, we detected 13 vertebrate viruses from four major virus groups: (+)ssRNA, (+)ssRNA-RT, dsDNA and (+)ssDNA, including avian leukosis virus in domestic chickens (Gallus gallus), enzootic nasal tumour virus in goats (Capra hircus) and several small, circular, Rep-encoding, ssDNA (CRESS DNA) viruses in several mammal species. More significantly, we demonstrate that the mNGS method is able to detect potentially lethal animal viruses, such as elephant endotheliotropic herpesvirus in Asian elephants (Elephas maximus) and the newly described human-associated gemykibivirus 2, a human-to-animal cross-species virus, in a Linnaeus two-toed sloth (Choloepus didactylus) and its enclosure, for the first time.

Molecular survey of Zika virus in the animal-human interface in traditional farming.

Backyard animal husbandry is common in rural communities in developing countries and, given the conditions in which it occurs, it can increase the risk of disease transmission, such as arboviruses. To determine the presence of the Zika virus (ZIKV) and abundance of its arthropod vectors we evaluated the socioeconomic implications involved in its transmission in two highly vulnerable Mayan communities in the state of Yucatan that practice backyard farming. An analytical cross-sectional study was carried out throughout 2016 to understand socioeconomic variables and seasonal patterns in mosquito populations. We selected 20 households from each community. Social exclusion indicators were analyzed, human and domestic animals were sampled, and mosquitoes were collected and identified. Four out of eight indicators of social exclusion were higher than the reported national averages. We captured 5,825 mosquitoes from 16 species being Culex quinquefasciatus and Aedes aegypti the most abundant. The presence of chickens and human overcrowding in dwellings were the most significant factors (P = 0.026) associated with the presence of Ae. aegypti. Septic tanks (odds ratio = 6.64) and chickens (odds ratio = 27.41) in backyards were the main risk factors associated with the presence of immature states of Ae. aegypti in both communities. Molecular analysis to detect ZIKV was performed in blood samples from 416 humans, 1,068 backyard animals and 381 mosquito pools. Eighteen humans and 10 pig pools tested positive for ZIKV. Forty-three mosquito pools tested positive for flavivirus. Ten of the 43 pools of positive mosquitoes were sequenced, corresponding 3/10 to ZIKV and 1/10 to Dengue virus type 2. The findings obtained indicate the continuous circulation of Flavivirus (including ZIKV) in backyard environments in vulnerable communities, highlighting the importance of studying their transmission and maintenance in these systems, due that backyard animal husbandry is a common practice in these vulnerable communities with limited access to health services.

Quantifying human-animal contact rates in Malaysian Borneo: Influence of agricultural landscapes on contact with potential zoonotic disease reservoirs.

Changing landscapes across the globe, but particularly in Southeast Asia, are pushing humans and animals closer together and may increase the likelihood of zoonotic spillover events. Malaysian Borneo is hypothesized to be at high risk of spillover events due to proximity between reservoir species and humans caused by recent deforestation in the region. However, the relationship between landscape and human-animal contact rates has yet to be quantified. An environmentally stratified cross-sectional survey was conducted in Sabah, Malaysia in 2015, collecting geolocated questionnaire data on potential risk factors for contact with animals for 10,100 individuals. 51% of individuals reported contact with poultry, 46% with NHPs, 30% with bats, and 2% with swine. Generalised linear mixed models identified occupational and demographic factors associated with increased contact with these species, which varied when comparing wildlife to domesticated animals. Reported contact rates with each animal group were integrated with remote sensing-derived environmental data within a Bayesian framework to identify regions with high probabilities of contact with animal reservoirs. We have identified high spatial heterogeneity of contact with animals and clear associations between agricultural practices and high animal rates. This approach will help inform public health campaigns in at-risk populations and can improve pathogen surveillance efforts on Malaysian Borneo. This method can additionally serve as a framework for researchers looking to identify targets for future pathogen detection in a chosen region of study.

Prioritizing zoonotic diseases utilizing the One Health approach: Jordan's experience.

BACKGROUND: Zoonotic diseases constitute a threat to humans and animals. The Middle East Region is a hotspot for such a threat; given its geographic location under migratory birds' flight paths, mass gatherings, political conflicts, and refugee crises. Thus, prioritizing zoonotic diseases of national significance is critical for preventing and controlling such threats and optimizing limited resources. Using a multi-sectoral One Health (OH) approach, this study aimed at prioritizing zoonotic diseases of national significance to Jordan and identifying future recommendations and action plans. METHODS: Zoonotic diseases of national significance to Jordan were initially identified (n = 27 diseases). In December 2019, national staff from governmental and non-state sectors were invited to develop ranking criteria, including questions and answers choices, and to weigh each criterion. Then, the national staff were asked to assess zoonotic diseases' priority using the developed criteria and provide recommendations and action plans to strengthen multi-sectoral collaboration. RESULTS: Seven zoonotic diseases were identified as being of great significance. Rabies was ranked as the number one priority disease, followed by middle east respiratory syndrome, avian influenza, brucellosis, leishmaniasis, rickettsiosis, and salmonellosis. The highest weighted criteria used to rank diseases were disease severity, outbreaks profile, and potential human-to-human transmission. Establishing a one-health platform, surveillance, laboratory, preparedness planning, outbreak response, and workforce were suggested as recommendations for approaching the priority diseases. Respondents identified data sharing, coordination, event-based surveillance, and effective communication channels as vital areas to enhance prevention and control strategies, conduct joint outbreak investigations, and improve multi-sectoral collaboration. CONCLUSIONS: This study represents the first attempt to prioritize zoonotic diseases of national significance in Jordan using the OH approach and a semi-qualitative, transparent, and comparative method. Study results can be used as a decision-making guide for policymakers and stakeholders and a cornerstone for combating zoonotic disease threats.

SARS-CoV-2 at the human-animal interphase: A review.

Since its emergence in China in December 2019, COVID-19 remains the recent leading disease of concern drawing the public health attention globally. The disease is known of viral origin and zoonotic nature originating from animals. However, to date neither the source of the spillover nor the intermediate hosts are identified. Moreover, the public health situation is intermittently aggravated by identification of new animals susceptible to the SARS-CoV-2 infection, potentially replicating the virus and maintaining intra and interspecies spread of the disease. Although the role of a given animal and/or its produce is important to map the disease pattern, continuous efforts should be undertaken to further understand the epidemiology of SARS-CoV-2, a vital step to establish effective disease prevention and control strategy. This manuscript attempted to review updates regarding SARS-CoV-2 infection at the human-animal interface with consideration to postulations on the genetic relatedness and origin of the different SARS-CoV-2 variants isolated from different animal species. Also, the review addresses the possible role of different animal species and their produce in transmission of the disease. Also, the manuscript discussed the contamination potentiality of the virus and its environmental stability. Finally, we reviewed the currently instituted measures to prevent and manage the spread of SARS-CoV-2 infection. The manuscript suggested the One Health based control measures that could prove of value for the near future.

The importance of long-term studies on wildlife diseases and their interfaces with humans and domestic animals: A review.

Long-term wildlife disease research (LTWDR) and its interfaces with humans and domestic animals provide perspective to understand the diseases' main drivers and how they operate. In a systematic review, we analysed the temporal trend of the studies on LTWDR, their aims, and the hosts, pathogens and geographic areas studied. We also evaluated the added value that such studies provide. For analysis, we selected a total of 538 articles from 1993 to 2017 with a study period greater than or equal to 4 consecutive years. A marked increase in the number of studies published during the last 20 years was observed that reflects a growing awareness of the outstanding role of wildlife as a reservoir of diseases. The most studied pathogen agents were viruses (39.2%), bacteria (38.5%) and protozoans (15.8%). Concerning the hosts, mammals (84.9%), particularly ungulates (40%) and carnivores (30.9%), and birds (12.5%) were the most represented in these long-term studies. Most articles reached conclusions concerning the effect of the disease on the infection/host dynamics (98.7%) and over 40% considered the economic consequences or proposed management and control measures. The research was mainly located in the Northern Hemisphere. While the definition of LTWDR is not only determined by the duration of the monitoring, the study must be long enough to: (a) address ecological and epidemiological questions that cannot be resolved with short-term observations or experiments, and (b) clarify the effects of different drivers. This review demonstrates that LTWDR has provided information about the causes and consequences of disease change that otherwise could not have been obtained. It may be used to inform decisions related to the emergence of disease and might help to design early warning systems of disease based on retrospective investigations.

Reconsideration of the plague transmission in perspective of multi-host zoonotic disease model with interspecific interaction.

The human-animal interface plays a vital role in the spread of zoonotic diseases, such as plague, which led to the "Black Death", the most serious human disaster in medieval Europe. It is reported that more than 200 mammalian species including human beings are naturally infected with plague. Different species acting as different roles construct the transmission net for Yersinia pestis (plague pathogen), in which rodents are the main natural reservoirs. In previous studies, it focused on individual infection of human or animal, rather than cross-species infection. It is worth noting that rodent competition and human-rodent commensalism are rarely considered in the spread of plague. In order to describe it in more detail, we establish a new multi-host mathematical model to reflect the transmission dynamics of plague with wild rodents, commensal rodents and human beings, in which the roles of different species will no longer be at the same level. Mathematical models in epidemiology can clarify the interaction mechanism between plague hosts and provide a method to reflect the dynamic process of plague transmission more quickly and easily. According to our plague model, we redefine the environmental capacity K with interspecific interaction and obtain the reproduction number of zoonotic diseases RZ0, which is an important threshold value to determine the zoonotic disease to break out or not. At the same time, we analyze the biological implications of zoonotic model, and then study some biological hypotheses that had never been proposed or verified before.

Mycobacterium bovis and you: A comprehensive look at the bacteria, its similarities to Mycobacterium tuberculosis, and its relationship with human disease.

While Mycobacterium tuberculosis is the primary cause of tuberculosis in people, multiple other mycobacteria are capable of doing so. With the World Health Organization's goal of a 90% reduction in tuberculosis by 2035, all tuberculous mycobacteria need to be addressed. Understanding not only the similarities, but importantly the differences between the different species is crucial if eradication is ever to be achieved. Mycobacterium bovis, while typically thought of as a disease of cattle, remains a possible source of human infection worldwide. Although this species' genome differs from Mycobacterium tuberculosis by only 0.05%, significant differences are present, creating unique challenges to address. This review focuses on features which distinguish this bacterium from Mycobacterium tuberculosis, including differences in origin, structure, environmental persistence, host preferences, infection and disease, host immune response, diagnostics and treatment.

Human-Animal Interaction and the Emergence of SARS-CoV-2.

The COVID-19 pandemic has affected all sectors of society, from health and economics to socialization and travel. The level and extent of this impact are unprecedented. Although the cause of COVID-19 was quickly identified to be a new coronavirus (SARS-CoV-2), the world was poorly prepared for preventing its spread. One important pillar of preparedness is surveillance of the sources of emerging pathogens and responding appropriately to prevent their spread in the human population. The ever-increasing interaction between humans and animals is one leading factor in facilitating the emergence of new pathogens. In this viewpoint, we discuss the possibility of the zoonotic origin of SARS-CoV-2, highlight the importance of understanding human-animal interaction to improve preparedness for future outbreaks, and outline recommendations for prevention.

One Health operational framework for action for the Eastern Mediterranean Region, focusing on zoonotic diseases.

Human health is intrinsically linked to the health of animals and to the environment, and efforts by just one sector alone cannot prevent or adequately address the complex problems at the human-animal-environment interface. Countries of the World Health Organization Eastern Mediterranean Region, as any other region, face the threat of emerging and remerging zoonoses. However, the challenges in this Region are high given the lack of resources, poor health systems, and political factors. Hence, adopting the One Health approach becomes urgent to assist those countries. Subsequently, based on analysis of One Health capacities in the Region and in close consultation with representatives and subject matter experts from countries in the Region, a framework for action towards effectively implementing the One Health approach was developed. The framework capitalizes on current opportunities in the region and provide countries with a list of practical key activities towards optimal use of their resources and strengthening their capabilities to tackle concurrent and future health challenges at the interface. Strong governance structures and building on existing mechanisms are crucial for achieving effective disease surveillance and response. Additionally, using intersectoral approaches for risk assessment and risk mitigation for health issues at the human-animal-environment interface can improve efficiency and result in more successful outcomes.

A multifaceted risk model of brucellosis at the human-animal interface in Egypt.

Brucellosis is a highly contagious zoonosis affecting humans and a wide range of domesticated and wild animal species. An important element for effective disease containment is to improve knowledge, attitudes and practices (KAP) of afflicted communities. This study aimed to assess the KAP related to brucellosis at the human-animal interface in an endemic area of Egypt and to identify the risk factors for human infection. A matched case-control study was conducted at the central fever hospitals located in six governorates in northern Egypt. Face-to-face interviews with cases and controls were conducted using a structured questionnaire. In total, 40.7% of the participants owned farm animals in their households. The overall mean practice score regarding animal husbandry, processing and consumption of milk and dairy products were significantly lower among cases compared with controls (-12.7 ± 18.1 vs. 0.68 ± 14.2, respectively; p < .001). Perceived barriers for notification of animal infection/abortion were predominate among cases and positively correlated with participants' education. The predictors of having brucellosis infection were consumption of unpasteurized milk or raw dairy products and practicing animal husbandry. Applying protective measures against infection significantly reduced its risk. A model predicting risk factors for brucellosis among those who own animal showed that frequent abortions per animal increased the chance for brucellosis infection among human cases by 50-fold (95% CI: 8.8-276.9), whereas the use of protective measures in animal care reduced the odds (OR = 0.11 [95% CI: 0.03-0.45]). In conclusion, consumption of unprocessed dairy products was equally important as contact with infected/aborted animals as major risk factors for Brucella spp. infection among humans in Egypt. There is poor knowledge, negative attitudes and risky behaviours among villagers which can perpetuate the risk of brucellosis transmission at the human-animal interface. This supports the need for integrating health education into the national brucellosis control programme.

Complete genome sequencing of H1N1pdm09 swine influenza isolates from Nigeria reveals likely reverse zoonotic transmission at the human-animal interface in intensive piggery.

Prevailing agro-ecological conditions and intermingling of human and animals in intensive farms in urban and peri-urban areas in Africa favour cross species transmission of pathogens at the human-animal interface. However, molecular epidemiology studies of zoonotic swine influenza viruses in this region are limited. In this study, isolates of pandemic influenza virus (H1N1pdm09) obtained from pigs in Nigeria were fully sequenced. BLAST of swine influenza virus genes from Nigeria was carried out in GenBank and gene alignment was done using MEGA version 7. Maximum likelihood method (PhyML program) was used to determine gene evolutionary relationships with other viruses and phylogenetic trees were constructed to infer genomic clusters and relationship. Swine influenza viruses isolated and sequenced in this study were monophyletic and 99% congenetic with human isolates from Nigeria, Cameroon, Ghana and USA suggesting reverse zoonotic transmission from humans to pigs in intensive husbandry. A Q240R and S31N substitution among others were detected in the haemagglutinin and matrix genes, respectively, indicating potentials for mutations during interspecies co-mingling and transmission. The A/H1N1pdm09 viruses circulating in pigs that are also exposed to avian influenza in the same epidemiological zones could engender emergence of novel viruses with zoonotic or pandemic potential requiring enhanced surveillance and monitoring.

Isolation and Potential for Transmission of Mycobacterium bovis at Human-livestock-wildlife Interface of the Serengeti Ecosystem, Northern Tanzania.

Mycobacterium bovis, the causative agent of bovine tuberculosis (bTB), is a multihost pathogen of public health and veterinary importance. We characterized the M. bovis isolated at the human-livestock-wildlife interface of the Serengeti ecosystem to determine the epidemiology and risk of cross-species transmission between interacting hosts species. DNA was extracted from mycobacterial cultures obtained from sputum samples of 472 tuberculosis (TB) suspected patients and tissue samples from 606 livestock and wild animal species. M. bovis isolates were characterized using spoligotyping and Mycobacterial Interspersed Repetitive Units-Variable Tandem Repeats (MIRU-VNTR) on 24 loci. Only 5 M. bovis were isolated from the cultured samples. Spoligotyping results revealed that three M. bovis isolates from two buffaloes (Syncerus caffer) and 1 African civet (Civettictis civetta) belonged to SB0133 spoligotype. The two novel strains (AR1 and AR2) assigned as spoligotype SB2290 and SB2289, respectively, were identified from indigenous cattle (Bos indicus). No M. bovis was detected from patients with clinical signs consistent with TB. Of the 606 animal tissue specimens and sputa of 472 TB-suspected patients 43 (7.09%) and 12 (2.9%), respectively, yielded non-tuberculous mycobacteria (NTM), of which 20 isolates were M. intracellulare. No M. avium was identified. M. bovis isolates from wildlife had 45.2% and 96.8% spoligotype pattern agreement with AR1 and AR2 strains, respectively. This finding indicates that bTB infections in wild animals and cattle were epidemiologically related. Of the 24 MIRU-VNTR loci, QUB 11b showed the highest discrimination among the M. bovis strains. The novel strains obtained in this study have not been previously reported in the area, but no clear evidence for recent cross-species transmission of M. bovis was found between human, livestock and wild animals.