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Isolation and molecular characterization of Mycobacterium bovis from Kafue lechwe (Kobus leche kafuensis) from Zambia

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Abstract

Bovine tuberculosis (BTB) is a chronic bacterial disease caused by Mycobacterium bovis. Infections due to M. bovis, which serves as a stable reservoir, can pose serious challenge to control and eradicate in both wildlife and livestock at the interface. This study aimed at isolating and characterizing M. bovis from Kafue lechwe (Kobus leche kafuensis) and black lechwe (Kobus leche smithemani) at the animal/human interface in Zambia. The samples with lesions compatible with BTB collected during the hunting seasons of 2009 and 2010 were cultured for isolation of mycobacteria using Stonebrink with pyruvate (BD Diagnostics, MD, USA) and Middlebrook 7H10 (BD Diagnostics) slants. Isolated mycobacteria were identified using IS6110 polymerase chain reaction and deletion analysis. Molecular characterization of the isolates was performed using spoligotyping and mycobacteria interspersed repetitive unit–variable number tandem repeat (MIRU–VNTR) with nine loci. Data was analyzed using BioNumerics software 6.1. Out of the 39 samples, acid fast bacilli were detected in 27 (69.2 %) based on smear microscopy. Seven isolates were found to belong to Mycobacterium tuberculosis complex, and all were identified as M. bovis based on deletion analysis. All seven isolates were identical on spoligotyping as belonging to the SB0120 (SIT 482). MIRU–VNTR differentiated the isolates into five different patterns. This study has confirmed that M. bovis circulates in the Kafue lechwe, and non-tuberculous mycobacteria were detected in the black lechwe in Zambia which represents a wildlife reservoir, with a potential to spillover to cattle and humans. Isolates of M. bovis from lechwe antelopes are much conserved as only one spoligotype was detected. The study has shown that three loci differentiated fairly well. This option is cheap and less laborious, and hence a better option in resource-strained country like Zambia. The study further showed that some of the loci recommended by the European Reference Laboratory are not suitable for typing M. bovis in Zambia.

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Acknowledgments

This work was carried out with financial support from the Norwegian Education and loan fund as quota scholarship to Sydney Malama, as well as some core funding from the Norwegian School of Veterinary Science and the Norwegian Veterinary Institute. The authors acknowledge the assistance from Dr. Musso Munyeme in sample collection, funded by the Norwegian Education and Research Fund (NUFU). The authors are further grateful to the field and laboratory technical support in Zambia and to the Norwegian Veterinary Institute in Norway.

Conflict of interest

The authors declared no potential conflicts of interest with respect to the research, authorship, and/or publication of this article.

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Authors and Affiliations

  1. Institute of Economic and Social Research, University of Zambia, P.O. Box 30900, Lusaka, Zambia

    Sydney Malama

  2. Norwegian Veterinary Institute, P.O. Box 750, 0106, Oslo, Norway

    Tone Bjordal Johansen & Berit Djønne

  3. Department of Disease Control, School of Veterinary Medicine, University of Zambia, P.O. Box 32379, Lusaka, Zambia

    John Bwalya Muma

  4. Department of Biomedical Science, Tropical Diseases Research Centre, P.O. Box 71769, Ndola, Zambia

    Sydney Mwanza

  5. Department of Food Safety and Infection Biology, Section of Epidemiology and Biostatistics, Norwegian School of Veterinary Science, P.O. Box 8146, Oslo, Norway

    Jacques Godfroid

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  1. Sydney Malama
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Correspondence to Sydney Malama.

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Malama, S., Johansen, T.B., Muma, J.B. et al. Isolation and molecular characterization of Mycobacterium bovis from Kafue lechwe (Kobus leche kafuensis) from Zambia. Trop Anim Health Prod 46, 153–157 (2014). https://doi.org/10.1007/s11250-013-0466-4

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