High throughput qPCR analyses suggest that Enterobacterales of French sheep and cow cheese rarely carry genes conferring resistances to critically important antibiotics for human medicine

Int J Food Microbiol. 2023 Oct 16:403:110303. doi: 10.1016/j.ijfoodmicro.2023.110303. Epub 2023 Jun 24.

Abstract

Bacteria present in raw milk can carry acquired or intrinsic antimicrobial resistance genes (ARGs) and mobile genetic elements (MGEs). However, only a few studies have evaluated raw milk cheese as a potential reservoir of ARGs. This study thus aimed at providing new data regarding resistance markers present in raw milk cheese. Sheep (n = 360) and cow (n = 360) cheese samples produced in France were incubated in buffered peptone water supplemented with acriflavin or novobiocin; as corroborated by 16S metabarcoding, samples were enriched in Gram-negative bacteria since Escherichia coli and Hafnia alvei respectively accounted for 40 % and 20 % of the samples' microbiota. Screening of the samples for the presence of 30 ARGs and 16 MGEs by high throughput qPCR array showed that nine ARGs conferring resistances to 1st-generation beta-lactams, aminoglycosides, trimethoprim/sulfonamides and tetracyclines occurred in >75 % of both sheep and cow samples. This is neither surprising nor alarming since these resistance genes are widely spread across the One Health human, animal and environmental sectors. Conversely, genes conferring resistances to last-generations cephalosporins were rarely identified, while those conferring resistances to carbapenems or amikacin, which are restricted to human use, were never detected. Multiple MGEs were detected, the most frequent ones being IncF plasmids, confirming the potential transmission of ARGs. Our results are in line with the few studies of the resistome of milk or milk cheese showing that genes conferring resistances to 1st-generation beta-lactams, aminoglycosides and tetracyclines families are widespread, while those conferring resistances to critically important antibiotics are rare or absent.

Keywords: Antimicrobial resistance genes; Dairy products; Enterobacterales; High-throughput qPCR; Metabarcoding; Raw milk cheese; Resistome.

MeSH terms

  • Aminoglycosides
  • Animals
  • Anti-Bacterial Agents / pharmacology
  • Cattle
  • Cheese* / microbiology
  • Escherichia coli
  • Genes, Bacterial
  • Humans
  • Sheep
  • beta-Lactams

Substances

  • Anti-Bacterial Agents
  • Aminoglycosides
  • beta-Lactams