Genotypic characterization, toxigenic and spoilage features and multi drug resistance profile of Bacillus cereus from Milk Co-operative societies of Mizoram, India
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Authors
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Gowtham Gowtham M
M
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DEVAJANI DEKA
Keywords:
Bacillus cereus, Genotypic characterization, Toxigenic and Spoilage features, MDR, Milk Co operative societies, Mizoram, Phylogenetic analysisAbstract
Bacillus cereus is a food borne pathogenic and food spoiling bacterium commonly found in nature. Emphasizing the recent rise of Mizoram in the small scale dairy farming and alongside probable compromising quality of milk produced in land locked and potable water scarce hilly state, the research investigated the level of contamination, genotypic characteristic, toxigenic gene distribution, spoilage features and multi drug resistance pattern of B. cereus isolated from different of the state. A total of 108 samples of raw, bulk, and retail milk, six samples each, were collected from six Milk Co-operative societies, namely Durtlang, Sihphir, Durtlang-leitan, Thuampui, Phunchawng, and Mualpui. The B. cereus group count exceeded the regulatory limit of 5.7x104cfu/ml in farm raw, bulk, and retail milk, by a significant margin. In PCR assay, using the house keeping panC gene, seven (6.48%) B. cereus strains were confirmed from farm raw milk but the organism could not be detected from bulk and retail milk. All seven B. cereus strains had at least one enterotoxigenic gene with the exception of the haemolytic hblA gene. However, the isolated strains lacked the emetic (ces) gene. The study strains belonged to Clade III with 99.72 percent homology, which are mesogenic and cytotoxic in nature. A proportion of 85.71-100 percent of B. cereus strains exhibited lipolytic and proteolytic spoilage features and none of the strains was psychro-tolerant. Two B. cereus strains with NCBI accession no. OR786090 and OR786089 had highest three enterotoxigenic genes. Upon phylogenetic analysis, the study strains were homologous to the reference strains sourced from milk product, soil and patient’s vomit. Antibiotic sensitivity test upon the organism revealed highest resistance (97.05%) to β-lactam antibiotics and sensitivity (97.05%) to ciprofloxacin and erythromycin. Five B. cereus strains were multi drug resistant, with a distribution to 3-4 classes of antibiotics of different combination.
The prevalence of mesophilic enterotoxigenic B. cereus with the count above legal limit, toxigenic and spoilage features and MDR in raw milk from different Milk Co-operative societies of Mizoram, indicated the significance of the organism from food safety and shelf-life point of view.
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