Characterization of Bacillus cereus isolates from local dairy farms in China

FEMS Microbiol Lett. 2016 Jun;363(12):fnw096. doi: 10.1093/femsle/fnw096. Epub 2016 Apr 27.

Abstract

Bacillus cereus is an important opportunistic foodborne pathogen. In the present work, a total of 306 milk and environmental samples were collected from 10 local dairy farms in Beijing, China. Of the 92 B. cereus-like isolates, 88 and 4 belonged to B. cereus and B. thuringiensis, respectively. The prevalence of B. cereus isolates in bedding, feces, feed, liquid manure and raw milk was 93.3%, 78.9%, 41.2%, 100.0% and 9.8%, respectively. Three main toxin genes nhe, hbl and ces were detected with rates of 100.0%, 78.3% and 1.1%, but no strain harbored cytK1 The production of Nhe, Hbl and cereulide could be confirmed by specific monoclonal antibodies-based enzyme immunoassays in 94.6%, 70.7% and 1.1% of all isolates, respectively. Cytotoxicity tests were used to further corroborate the results of genetic and protein-based assays; 91.3% of the isolates showed cytotoxicity to Vero cells. All isolates were tested for antimicrobial resistance against 17 antibiotics. All isolates were resistant to lincomycin, retapamulin, tiamulin and valnemulin, while two strains were susceptible to ampicillin and ceftiofur. A total of 16 isolated strains were resistant to tetracycline. Since spores of B. cereus are not inactivated during manufacturing of most milk products, contamination of milk with B. cereus on the farm level may represent a potential hazard, particularly with respect to emetic toxin-producing strains.

Keywords: Bacillus cereus; antimicrobial susceptibility; dairy; toxin.

MeSH terms

  • Animal Feed / microbiology
  • Animals
  • Anti-Bacterial Agents / pharmacology
  • Bacillus cereus / drug effects
  • Bacillus cereus / genetics
  • Bacillus cereus / isolation & purification*
  • Bacillus cereus / physiology*
  • Bacillus thuringiensis / drug effects
  • Bacillus thuringiensis / genetics
  • Bacillus thuringiensis / isolation & purification
  • Bacterial Proteins / genetics
  • Bacterial Proteins / toxicity
  • Bacterial Toxins / genetics
  • Bacterial Toxins / toxicity
  • Cattle
  • Chlorocebus aethiops
  • Colony Count, Microbial
  • DNA, Bacterial / genetics
  • Dairying*
  • Farms*
  • Feces / microbiology
  • Food Microbiology*
  • Hemolysin Proteins / genetics
  • Hemolysin Proteins / toxicity
  • Manure / microbiology
  • Milk / microbiology*
  • Raw Foods / microbiology
  • Spores, Bacterial
  • Vero Cells

Substances

  • Anti-Bacterial Agents
  • Bacterial Proteins
  • Bacterial Toxins
  • DNA, Bacterial
  • Hemolysin Proteins
  • Manure
  • hemolysin BL protein, Bacillus