Development of a Sampling and Real-time PCR Method for the Quantitative Detection of Campylobacter spp. in Retail Chicken Meat Without DNA Extraction.

Campylobacter food poisoning is caused by consumption of the contaminated foods, especially poultry meat. Continuous quantitative measurement of Campylobacter spp. in contaminated foods is crucial to develop preventive measures. We developed a direct-qPCR method for determining the viable cell counts of Campylobacter spp. using qPCR without DNA extraction from enriched food samples and a sampling method (the wrap procedure) in which the sample is wrapped in a sheet, different from the conventional homogenization procedure. The viable cell counts of Campylobacter spp. before and after enrichment of the samples sampled using the wrap and homogenization procedures from chicken samples inoculated with Campylobacter jejuni were determined using the culture method, and the cycle threshold (CT) values after enrichment were determined using the direct-qPCR. An enrichment regression equation was generated from the viable cell counts obtained before and after enrichment, and a direct-qPCR regression equation was generated from the CT values and viable cell counts obtained after enrichment, enabling the viable cell counts before enrichment to be estimated from the CT values. Estimated viable cell counts were similar for the culture method when sampled by the homogenization procedure, but lower for the wrap procedure. However, the detection rate of direct-qPCR was 37.5% for liver and 89.7% for breast fillet using the homogenization procedure, whereas using the wrap procedure, it was 100% for both samples. The detection rate of direct-qPCR for retail chicken was 30.4-35.7% for the homogenization procedure, and 85.7-100% for the wrap procedure. Colonies were observed using the culture method, but their quantification was difficult due to swarming or their low number. However, estimating viable cell counts using the combination of wrap procedure and direct-qPCR methods is possible. The developed method can provide baseline data for the risk assessment Campylobacter food poisoning.

Distribution of Antimicrobial Resistance in Campylobacter Strains Isolated from Poultry at a Slaughterhouse and Supermarkets in Japan.

Fifty strains of Campylobacter jejuni/coli were detected in 108 specimens of chicken meat and organs sampled at six supermarkets and one poultry slaughterhouse (large scale) between April and October 2013 (isolation rates: 84.8% from the slaughterhouse, 29.3% from the supermarkets). 46/50 strains were successfully recovered and subjected to the E-test to examine their susceptibility to three fluoroquinolone antibacterial agents authorized for use in poultry in Japan: enrofloxacin (ERFX), ofloxacin (OFLX), and norfloxacin (NLFX). 29 isolates (63%) were resistant to all three agents and 2 isolates (4.3%) were resistant to two agents (ERFX and OFLX). The resistance rates of strains isolated fom the supermarkets and slaughterhouse were 61.9% and 72.0%, respectively. Because the chickens processed at the slaughterhouse were raised without the use of fluoroquinolone, the results did not suggest a positive relationship between the use of these agents and the distribution of antimicrobial-resistant bacteria. Susceptibility to macrolide antibiotics (erythromycin [EM]) was also tested in 42 strains, and one strain (2.4%), C. coli from a retailer sample, showed resistance. Previous studies have detected high rates of fluoroquinolone-resistant strains, suggesting an expanding distribution of resistant bacteria. The detection of EM-resistant bacteria downstream in the food distribution chain (i.e., closer to consumers) is a concern for human health.

Ribotyping and a study of transmission of Staphylococcus aureus collected from food preparation facilities.

Food poisoning from Staphylococcus aureus is sometimes caused by improper handling of food items in food preparation facilities. Prevention of contamination by employees is particularly important in facilities where a significant amount of food preparation is performed by hand. Some experiments have been performed to describe bacterial cross-contamination in the food preparation process, but there have been few studies of cross-contamination in actual food preparation facilities. Aiming to shed light on the transmission of S. aureus in food preparation facilities, this study collected samples of 66 strains of this bacterium from the fingers of food preparation staff, foodstuffs, prepared foods, cooking utensils, and cooking equipment and typed them with the ribotyping method. S. aureus from the same ribogroup was detected on the hands of a study participant, a faucet, knife, frying pan, and a salad, indicating that bacteria found on the hands of the study participant was transmitted to cooking utensils and prepared foods. Transmission (from a faucet to a frying pan handle) of bacteria by another person, a third party, was also detected.