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Genotypic characterization by spoligotyping and VNTR typing of Mycobacterium bovis and Mycobacterium caprae isolates from cattle of Tunisia

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Abstract

This work is an approach to the molecular epidemiology of Mycobacterium tuberculosis complex (MTBC) bovine infections in Tunisia. A total of 35 MTBC isolates from both lateral retropharyngeal lymph node samples of cattle slaughtered in different Tunisian regions were genotyped by spoligotyping and variable number tandem repeat typing (VNTR)-typing. Spoligotyping allowed to identify two profiles not previously registered, namely SB2024, a Mycobacterium caprae isolate from Nabeul Region (North East Tunisia), the first description of this species in the country, and SB2025 (Mycobacterium bovis) from Sfax Region (Southern Tunisia). A second M. caprae isolate with a spoligotyping profile previously described in Europe mainland, SB0418, was also isolated from a bovine of Sfax region. Both isolates suggest the possibility of a widespread distribution of this species in the country. The predominant spoligotype was SB0120, present in all Tunisian regions selected for the study but Nabeul. Molecular typing also allowed to describe a mixed infection caused by two different M. bovis isolates (SB0120 and SB0848) in the same animal. VNTR typing was highly discriminant by testing a panel of six loci. Loci QUB3232 and QUB11b were the most discriminant, whereas ETR-D and QUB11a had the lower diversity index. The value of allelic diversity can significantly vary among countries; thus, it is important to standardize a panel of loci for future inter-laboratory comparisons. Although VNTR typing proved to be useful for an efficient discrimination among MTBC isolates, especially in combination with spoligotyping, further studies are needed in order to assess the genetic diversity of the MTBC in Tunisia.

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Acknowledgments

The study was supported by the Ministry of Higher Education and Scientific Research of Tunisia, and funded by the Universidad de Extremadura (Cáceres, Spain). Special thanks to Dr. Mansouri Riadh and Dr. Soussi Houda at the Institute of Veterinary Research of Tunis, Mr. Znati Abdel Razzark for his assistance in providing lymph node samples, Dr. Jemli Mohammed Habib at the University of Veterinary Medicine, and Dr. Ben Sassi Hatem at the Regional Agricultural Development Offices of Béja (Tunisia). Mr. Benítez-Medina and Mr. García-Jiménez acknowledge the Gobierno de Extremadura and FSE for his research fellowship (PRE08042 and PRE07024, respectively).

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Authors and Affiliations

  1. Laboratory of Animal Health, Institute of Veterinary Research of Tunisia, Tunis, Tunisia

    Hela Lamine-Khemiri, Mohammed Salah Abassi & Imed Khazri

  2. Red de Grupos de Investigación Recursos Faunísticos, Facultad de Veterinaria, Universidad de Extremadura, Avenida de la Universidad s/n, 10003, Cáceres, Spain

    Remigio Martínez, Waldo Luis García-Jiménez, Jose Manuel Benítez-Medina, Maria Cortés, Inés Hurtado & Javier Hermoso-de-Mendoza

  3. Laboratories of Pathology and Infectious Diseases, National School of Veterinary Medicine of Sidi Thabet, Sidi Thabet, Tunisia

    Mohammed Benzarti

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  1. Hela Lamine-Khemiri
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Correspondence to Javier Hermoso-de-Mendoza.

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Lamine-Khemiri, H., Martínez, R., García-Jiménez, W.L. et al. Genotypic characterization by spoligotyping and VNTR typing of Mycobacterium bovis and Mycobacterium caprae isolates from cattle of Tunisia. Trop Anim Health Prod 46, 305–311 (2014). https://doi.org/10.1007/s11250-013-0488-y

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