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      Prevalence of bovine tuberculosis and characterization of the members of the Mycobacterium tuberculosis complex from slaughtered cattle in Rwanda

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          Abstract

          Background

          Bovine tuberculosis (bTB) is an endemic disease in Rwanda, but little is known about its prevalence and causative mycobacterial species. The disease causes tremendous losses in livestock and wildlife and remains a significant threat to public health.

          Materials and methods

          A cross-sectional study employing a systematic random sampling of cattle (n = 300) with the collection of retropharyngeal lymph nodes and tonsils (n = 300) irrespective of granulomatous lesions was carried out in six abattoirs to investigate the prevalence and identify mycobacterial species using culture, acid-fast bacteria staining, polymerase chain reaction, and GeneXpert assay. Individual risk factors and the origin of samples were analysed for association with the prevalence.

          Findings

          Of the 300 sample pools, six were collected with visible TB-like lesions. Our findings demonstrated the presence of Mycobacterium tuberculosis complex (MTBC) in 1.7% (5/300) of sampled slaughtered cattle. Mycobacterium bovis was isolated from 1.3% (4/300) animals while one case was caused by a rifampicin-resistant (RR) M. tuberculosis. Non-tuberculous mycobacteria were identified in 12.0% (36/300) of the sampled cattle. There were no significant associations between the prevalence and abattoir category, age, sex, and breeds of slaughtered cattle.

          Conclusions

          This study is the first in Rwanda to isolate both M. bovis and RR M. tuberculosis in slaughtered cattle indicating that bTB is present in Rwanda with a low prevalence. The isolation of RR M. tuberculosis from cattle indicates possible zooanthroponotic transmission of M. tuberculosis and close human-cattle contact. To protect humans against occupational zoonotic diseases, it is essential to control bTB in cattle and raise the awareness among all occupational groups as well as reinforce biosafety at the farm level and in the abattoirs.

          Author summary

          Tuberculosis in cattle (bTB) causes financial losses to livestock owners and is a disease transmissible to humans especially those with an occupational risk through exposure to infected animals and animal products. This study aimed to identify the prevalence of bTB and characterize the mycobacterial species from cattle slaughtered in the six abattoirs in Rwanda. Four M. bovis, as well as one rifampicin-resistant (RR) M. tuberculosis, were identified from slaughtered cattle and, thus, the apparent bTB prevalence was 1.7% (5/300). Likely, the RR M. tuberculosis isolate was mostly likely of human origin and transmitted to cattle during close human-cattle contact. It is therefore essential to control bTB in cattle and reinforce the protection of farmworkers and abattoir workers who are always exposed to infected animals.

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          Most cited references50

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          A new evolutionary scenario for the Mycobacterium tuberculosis complex.

          The distribution of 20 variable regions resulting from insertion-deletion events in the genomes of the tubercle bacilli has been evaluated in a total of 100 strains of Mycobacterium tuberculosis, Mycobacterium africanum, Mycobacterium canettii, Mycobacterium microti, and Mycobacterium bovis. This approach showed that the majority of these polymorphisms did not occur independently in the different strains of the M. tuberculosis complex but, rather, resulted from ancient, irreversible genetic events in common progenitor strains. Based on the presence or absence of an M. tuberculosis specific deletion (TbD1), M. tuberculosis strains can be divided into ancestral and "modern" strains, the latter comprising representatives of major epidemics like the Beijing, Haarlem, and African M. tuberculosis clusters. Furthermore, successive loss of DNA, reflected by region of difference 9 and other subsequent deletions, was identified for an evolutionary lineage represented by M. africanum, M. microti, and M. bovis that diverged from the progenitor of the present M. tuberculosis strains before TbD1 occurred. These findings contradict the often-presented hypothesis that M. tuberculosis, the etiological agent of human tuberculosis evolved from M. bovis, the agent of bovine disease. M. canettii and ancestral M. tuberculosis strains lack none of these deleted regions, and, therefore, seem to be direct descendants of tubercle bacilli that existed before the M. africanum-->M. bovis lineage separated from the M. tuberculosis lineage. This observation suggests that the common ancestor of the tubercle bacilli resembled M. tuberculosis or M. canettii and could well have been a human pathogen already.
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            Differentiation of Mycobacterium tuberculosis complex by PCR amplification of genomic regions of difference.

            Differentiation of members of the Mycobacterium tuberculosis complex by conventional mycobacteriological methods is time consuming, making surveillance of species-specific disease difficult. A two-step, multiplex polymerase chain reaction (PCR) method based on genomic regions of difference (RD1, RD1(mic), RD2(seal), RD4, RD9 and RD12) was developed for the differentiation of M. canettii, M. tuberculosis, M. africanum, M. microti, M. pinnipedii, M. caprae, M. bovis and M. bovis BCG. The size of the respective multiplex PCR amplification products corresponded to the presence of the different M. tuberculosis complex members. This method allows for rapid differentiation, making it suitable for routine laboratories and surveillance purposes.
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              High prevalence and increased severity of pathology of bovine tuberculosis in Holsteins compared to zebu breeds under field cattle husbandry in central Ethiopia.

              A comparative study on the prevalence and pathology of bovine tuberculosis (TB) was conducted on 5,424 cattle (2,578 zebus, 1,921 crosses, and 925 Holsteins), which were kept on pasture in the central highlands of Ethiopia, using a comparative intradermal tuberculin test, postmortem examination, and bacteriology. The overall prevalence of bovine TB was 13.5%; prevalence was higher in Holsteins than either zebus (22.2% versus 11.6%, chi(2) = 61.8; P < 0.001) or crosses (22.2% versus 11.9%, chi(2) = 50.7; P < 0.001). Moreover, the severity of pathology in Holsteins (mean +/- standard error of the mean [SEM], 6.84 +/- 0.79) was significantly higher (P = 0.018) than the severity of pathology in zebus (5.21 +/- 0.30). In addition, the risk of TB in Holsteins was more than twice (odds ratio [OR] = 2.32; 95% confidence interval [CI] = 1.89, 2.85) that in zebus. Animals between 5 and 9 years of age were at higher (OR = 2.37; 95% CI = 1.80, 3.12) risk of bovine TB than those 2 years of age or below. A significant difference (chi(2) = 351; P < 0.001) in the occurrence of TB lesions in lymph nodes was recorded; the mesenteric lymph node (mean pathology score +/- SEM, 1.95 +/- 0.08) was most severely affected, followed by the retropharyngeal (0.80 +/- 0.05) and caudal mediastinal (0.8 +/- 0.06) lymph nodes. Fifty-six percent (n = 145) of the animals with gross TB lesions were culture positive; the lowest culture positivity was recorded in the skin lesions (27.3%) and the lesions of the mesenteric lymph node (31.5%). Both the skin test response and the postmortem findings suggested a higher susceptibility to bovine TB in Holsteins than zebus under identical field husbandry conditions (on pasture). In the light of increased numbers of Holstein cattle introduced into this area to raise milk production to satisfy the needs of Addis Ababa's growing population, these findings highlight the need for a control program in these herds.
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                Author and article information

                Contributors
                Role: ConceptualizationRole: Data curationRole: Formal analysisRole: InvestigationRole: MethodologyRole: SoftwareRole: Writing – original draftRole: Writing – review & editing
                Role: ConceptualizationRole: MethodologyRole: SupervisionRole: ValidationRole: Writing – review & editing
                Role: SupervisionRole: Writing – review & editing
                Role: ResourcesRole: Writing – review & editing
                Role: ResourcesRole: Writing – review & editing
                Role: ConceptualizationRole: Funding acquisitionRole: MethodologyRole: Project administrationRole: ResourcesRole: SupervisionRole: Writing – review & editing
                Role: Editor
                Journal
                PLoS Negl Trop Dis
                PLoS Negl Trop Dis
                plos
                PLoS Neglected Tropical Diseases
                Public Library of Science (San Francisco, CA USA )
                1935-2727
                1935-2735
                3 August 2022
                August 2022
                : 16
                : 8
                : e0009964
                Affiliations
                [1 ] Bovine Tuberculosis and Brucellosis Research Programme, Department of Veterinary Tropical Diseases, Faculty of Veterinary Science, University of Pretoria; Pretoria, South Africa
                [2 ] Department of Veterinary Medicine, College of Veterinary Medicine, University of Rwanda, Kigali, Rwanda
                [3 ] Department of Human Medicine and Device assessment and Registration, Rwanda Food and Drug Administration, Kigali, Rwanda
                [4 ] National Reference Laboratory Division, Department of Biomedical Services, Rwanda Biomedical Centre, Kigali, Rwanda
                [5 ] Department of Clinical Biology, School of Medicine and Pharmacy, College of Medicine and Health Sciences, University of Rwanda, Kigali, Rwanda
                NIH-National Institute for Research in Tuberculosis-ICER, INDIA
                Author notes

                The authors have declared that no competing interests exist.

                [¤]

                Current address: Department of Human Medicine and Device assessment and Registration, Division of Human Medicine, Assessment and registration, Rwanda Food and Drug Administration, Kigali, Rwanda

                ‡ AB and CD also contributed equally to this work.

                Author information
                https://orcid.org/0000-0002-9404-2214
                Article
                PNTD-D-21-01588
                10.1371/journal.pntd.0009964
                9377585
                35921351
                68bd8061-a680-42fe-822a-fce03c4325a6
                © 2022 Ntivuguruzwa et al

                This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

                History
                : 5 November 2021
                : 8 July 2022
                Page count
                Figures: 2, Tables: 2, Pages: 13
                Funding
                Funded by: Directorate-General for Development Cooperation
                Award ID: FA DGD-ITM 2017 – 2021
                Award Recipient :
                This research was funded by grant FA DGD-ITM 2017 – 2021 awarded to HvH by the Belgian Directorate-General for Development Cooperation, through its Framework Agreement with the Institute of Tropical Medicine ( https://www.itg.be). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.
                Categories
                Research Article
                Biology and Life Sciences
                Organisms
                Eukaryota
                Animals
                Vertebrates
                Amniotes
                Mammals
                Bovines
                Cattle
                Biology and Life Sciences
                Zoology
                Animals
                Vertebrates
                Amniotes
                Mammals
                Bovines
                Cattle
                Biology and Life Sciences
                Organisms
                Eukaryota
                Animals
                Vertebrates
                Amniotes
                Mammals
                Ruminants
                Cattle
                Biology and Life Sciences
                Zoology
                Animals
                Vertebrates
                Amniotes
                Mammals
                Ruminants
                Cattle
                Biology and Life Sciences
                Organisms
                Bacteria
                Actinobacteria
                Mycobacterium Tuberculosis
                Biology and Life Sciences
                Agriculture
                Animal Management
                Livestock
                Biology and Life Sciences
                Organisms
                Bacteria
                Actinobacteria
                Mycobacterium Bovis
                Biology and Life Sciences
                Molecular Biology
                Molecular Biology Techniques
                Artificial Gene Amplification and Extension
                Polymerase Chain Reaction
                Research and Analysis Methods
                Molecular Biology Techniques
                Artificial Gene Amplification and Extension
                Polymerase Chain Reaction
                People and Places
                Geographical Locations
                Africa
                Rwanda
                Biology and Life Sciences
                Organisms
                Bacteria
                Actinobacteria
                Nontuberculous Mycobacteria
                Biology and Life Sciences
                Veterinary Science
                Veterinary Diseases
                Custom metadata
                vor-update-to-uncorrected-proof
                2022-08-15
                All relevant data are within the manuscript and its Supporting Information files.

                Infectious disease & Microbiology
                Infectious disease & Microbiology

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