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Molecular Speciation of Mycobacterial Isolates from Raw Cow Milk Reveals Predominance of Mycobacterium chelonae

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Proceedings of the National Academy of Sciences, India Section B: Biological Sciences Aims and scope Submit manuscript

Abstract

Owing to the fact that raw milk intake is still practiced in selected communities globally, milk borne diseases need to be addressed as some mycobacterial species use milk as a ploy for disseminating the disease. An approach was made in the present study for screening the mycobacterial species in raw cow milk. For mycobacterial isolation, a total of 200 milk samples were collected from three districts of Northeast India—Dimapur district of Nagaland, East Khasi Hills and Ri-Bhoi districts of Meghalaya. Samples were collected by trained personnel and transported in cold chain. Following decontamination, samples were inoculated in both Lowenstein Jensen (LJ)-glycerol and LJ-pyruvate and incubated up to 12 weeks with regular interval check. Colonies were examined for acid-fastness and confirmed by genus-specific hsp65 PCR. Amplicons were sequenced, analyzed in Basic Local Alignment Search Tool (BLAST) and phylogenetically explored. A total of 22 (11%) isolates were recovered. Sequencing of genus-specific hsp65 gene identified 17 (80.95%) isolates to be Mycobacterium chelonae, 03 (14.28%) as M. lentiflavum, 01 (4.76%) M. peregrinum and 01 (0.5%) as a member of Mycobacterium tuberculosis complex. The dendrogram presented vivid species distinction as the isolates clustered into their respective phyletic line. Significant isolation of M. chelonae from raw milk in this study is noteworthy as it is an opportunistic human pathogen and one of the most dominant species isolated from human nontuberculous mycobacteria infections.

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Acknowledgements

The authors thank the Director, Indian Council of Agricultural Research Research Complex for North Eastern Hill Region, Meghalaya, for extending necessary facilities and funds to undertake the research (PIMS-IXX05508); Vice Chancellor, Lovely Professional University, Punjab, for consenting the collaborative work; and The University Grants Commission- statutory body of the government of India for awarding fellowship to embark on the Ph.D. research.

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Authors and Affiliations

  1. Division of Animal Health, Indian Council of Agricultural Research (ICAR) Research Complex for North Eastern Hill Region, Umiam, Meghalaya, 793103, India

    Esther Vise, Sandeep Ghatak, Amarjit Karam, Uttaran Bhattacharjee, Arnab Sen, Ingudam Shakuntala, Kekungu Puro, Rajkumari Sanjukta & Samir Das

  2. School of Bioengineering and Biosciences, Lovely Professional University, Phagwara, Punjab, 144401, India

    Esther Vise, Akshay Garg & Neeta Raj Sharma

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  1. Esther Vise
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  2. Akshay Garg
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Correspondence to Samir Das.

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Significance statement

Presence of significant number of viable nontuberculous mycobacteria in raw milk requires scrutinization, as they are known to cause infections in immunocompromised individuals. Findings of this study is important with regard to food safety.

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Vise, E., Garg, A., Ghatak, S. et al. Molecular Speciation of Mycobacterial Isolates from Raw Cow Milk Reveals Predominance of Mycobacterium chelonae . Proc. Natl. Acad. Sci., India, Sect. B Biol. Sci. 88, 1623–1628 (2018). https://doi.org/10.1007/s40011-017-0911-7

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  • DOI: https://doi.org/10.1007/s40011-017-0911-7

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