Genotypic characterization of enterotoxigenic Bacillus cereus isolates associated with spoilage and drug resistance from milk in Mizoram

A S  TAMCHUILA; PARIMAL  ROYCHOUDHURY; M  GOWTHAM; DEVAJANI  DEKA

doi:10.56093/ijans.v95i7.169544

Authors

A S TAMCHUILA Department of Veterinary Public Health and Epidemiology, College of Veterinary Sciences and Animal Husbandry, Central Agricultural University, Imphal
PARIMAL ROYCHOUDHURY Department of Veterinary Public Health and Epidemiology, College of Veterinary Sciences and Animal Husbandry, Central Agricultural University, Imphal
M GOWTHAM Department of Veterinary Public Health and Epidemiology, College of Veterinary Sciences and Animal Husbandry, Central Agricultural University, Imphal
DEVAJANI DEKA Department of Veterinary Public Health and Epidemiology, College of Veterinary Sciences and Animal Husbandry, Central Agricultural University, Imphal

https://doi.org/10.56093/ijans.v95i7.169544

Keywords:

Bacillus cereus, Genotypic characterization, MDR, Milk Co operative societies, Phylogenetic analysis

Abstract

Bacillus cereus, a ubiquitous food borne pathogen and spoilage bacterium, poses significant risk to dairy safety as it’s spores can withstand pasteurization and forms persistent biofilms on dairy equipment and utensils. With the recent rise of small-scale dairy farming in Mizoram, this study assessed contamination levels, genotypic traits, toxigenic gene distribution, spoilage potential, and multidrug resistance of B. cereus isolated from dairy milk across the state. A total of 108 samples of farm raw, bulk, and retail milk, six samples each, were collected from six different milk co-operative societies, namely Durtlang, Sihphir, Durtlang-leitan, Thuampui, Phunchawng, and Mualpui. The cereus group count exceeded the regulatory limit of 5.7x10⁴cfu/ml or 4.76 log cfu/ml in farm raw, bulk, and retail milk without significant variations among different milk co-operative societies. In PCR based detection of housekeeping panC gene, the prevalence of B. cereus was found to be 19.45 percent (7 isolates) in farm raw milk, however, the organism could not be confirmed from bulk and retail milk. In terms of pathogenic traits, all the seven cereus isolates carried at least one enterotoxigenic gene except hblA and the emetic ces gene was absent. The study strains belonged to Clade III with 99.72 percent homology, which are mesogenic and cytotoxic in nature. Two B. cereus strains bearing NCBI accession numbers, OR786090 (AS09LM) and OR786089 (AS09TM) from Durtlang - leitan and Thuampui, respectively carried highest numbers of enterotoxigenic genes, three each. Upon phylogenetic analysis, these two strains were homologous to the reference strains sourced from milk product, soil and patient’s vomit. A proportion of 85.71-100 percent of B. cereus strains exhibited lipolytic and proteolytic spoilage potential and none of the strains were psychrotolerant. The B. cereus isolates showed 97.05percent resistance to β-lactams and 97.05percent sensitivity to ciprofloxacin and erythromycin. Five B. cereus strains were multidrug-resistant, spanning 3-4 antibiotic classes. The prevalence of B. cereus in farm raw milk from Mizoram, with counts exceeding permissible limits and exhibiting enterotoxigenic, spoilage, and multi drug resistance potential, underscores the need for scientific interventions in dairy farming suitable to local environment of the land locked hilly state to ensure dairy safety and shelf-life stability.

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