Prevalence and Characterization of Listeria monocytogenes Isolated from Beef and Sheep Carcasses in Turkey with Characterization of Locally Isolated Listeriophages as a Control Measure

Abstract

Swab samples from cattle and sheep carcasses (120 of each) were tested for Listeria monocytogenes, and 120 slaughterhouse wastewater samples were tested for listeriophages over 12 months (10 samples per month) to note the seasonal distribution. L. monocytogenes and bacteriophage isolates were characterized, and the biocontrol of L. monocytogenes was investigated in meatballs with a phage cocktail. L. monocytogenes was found in 3.4 and 2.5% of cattle and sheep carcasses, respectively. All the isolates were found to harbor hlyA, actA, inlA, inlB, inlC, inlJ, plcA, plcB, fbpA, and fri genes with varied mRNA expression levels by real-time reverse transcriptase PCR analysis. Five isolates did not harbor the vip gene. According to enterobacterial repetitive intergenic consensus PCR, L. monocytogenes isolates were classified into four different groups based on their DNA patterns. The L. monocytogenes isolates were characterized for antibiotic susceptibility; one strain was found to be resistant to five different antibiotic classes. Of 11 lytic listeriophages, two were selected for the cocktail based on their DNA restriction profiles, efficiency of plating, transmission electron microscopy, and in vitro and in vivo analyses. In the biocontrol study, we used a food model that consisted of a novel bacteriophage cocktail in raw meatballs. The highest reduction of L. monocytogenes was recorded as 2.2 log CFU/g at a multiplicity of cellular infection of 4.7 at the end of 1 h. In conclusion, the new bacteriophage cocktail in this study can be considered an efficient biocontrol agent of L. monocytogenes in meatballs.

Keywords: Bacteriophage; Cattle; Characterization; Sheep; Slaughterhouse wastewater.