AUTHOR=Bhutia Meera Ongmu , Thapa Namrata , Tamang Jyoti Prakash 

TITLE=Molecular Characterization of Bacteria, Detection of Enterotoxin Genes, and Screening of Antibiotic Susceptibility Patterns in Traditionally Processed Meat Products of Sikkim, India

JOURNAL=Frontiers in Microbiology

VOLUME=11

YEAR=2021

URL=https://www.frontiersin.org/journals/microbiology/articles/10.3389/fmicb.2020.599606

DOI=10.3389/fmicb.2020.599606

ISSN=1664-302X

ABSTRACT=<p>The lesser-known traditionally processed meat products such as beef <italic>kargyong</italic>, pork <italic>kargyong</italic>, <italic>satchu</italic>, and <italic>khyopeh</italic> are popular food items in the Himalayan state of Sikkim in India. The present study aimed to assess the microbiological safety of traditional meat products by identifying the potential spoilage or pathogenic bacteria, detecting the enterotoxins, and screening the antibiotic susceptibility patterns. The pH and moisture contents of the meat products varied from 5.3 to 5.9 and from 1.5 to 18%, respectively. The microbial loads of aerobic bacteria were 10<sup>5</sup> to 10<sup>7</sup> cfu/g, <italic>Staphylococcus</italic> 10<sup>3</sup> to 10<sup>6</sup> cfu/g, <italic>Bacillus</italic> 10<sup>4</sup> to 10<sup>6</sup> cfu/g, and total coliform 10<sup>2</sup> to 10<sup>7</sup> cfu/g, respectively. Based on 16S rRNA gene sequencing, the bacterial species isolated from traditionally processed meat products were <italic>Staphylococcus piscifermentans</italic>, <italic>Citrobacter freundii</italic>, <italic>Enterococcus faecalis</italic>, <italic>Salmonella enterica</italic>, <italic>Staphylococcus aureus</italic>, <italic>Citrobacter werkmanii</italic>, <italic>Klebsiella pneumoniae</italic>, <italic>Macrococcus caseolyticus</italic>, <italic>Klebsiella aerogenes</italic>, <italic>Staphylococcus saprophyticus</italic>, <italic>Pseudocitrobacter anthropi</italic>, <italic>Citrobacter europaeus</italic>, <italic>Shigella sonnei</italic>, <italic>Escherichia fergusonii</italic>, <italic>Klebsiella grimontii</italic>, <italic>Burkholderia cepacia</italic>, and <italic>Bacillus cereus</italic>. The enzyme-linked immunosorbent assay (ELISA) tests detected <italic>Salmonella</italic> spp. and enterotoxins produced by <italic>B. cereus</italic> well as <italic>Staphylococcus</italic> in a few tested samples. However, the PCR method did not detect the virulence genes of <italic>B. cereus</italic> and <italic>Salmonella</italic> in the isolates. Virulence gene (<italic>sea</italic>) was detected in <italic>S. piscifermentans</italic> BSLST44 and <italic>S. piscifermentans</italic> BULST54 isolated from beef <italic>kargyong</italic> and in <italic>S. aureus</italic> PSST53 isolated from pork <italic>kargyong</italic>. No enterotoxins were detected in <italic>khyopeh</italic> samples. The antibiotic sensitivity test showed that all bacterial strains were susceptible toward gentamicin, cotrimoxazole, norfloxacin, and trimethoprim. Gram-positive bacteria showed 100% sensitivity against clindamycin and erythromycin; however, 50% of the resistance pattern was observed against oxacillin followed by penicillin (33%) and ampicillin (27%).</p>