Korean fermented soybean products, such as doenjang, kochujang, ssamjang, and cho-kochujang, can harbor foodborne pathogens such as Bacillus cereus sensu lato (B. cereus sensu lato). The aim of this study was to characterize the toxin gene profiles, biochemical characteristics, and antibiotic resistance patterns of B. cereus sensu lato strains isolated from Korean fermented soybean products. Eighty-eight samples of Korean fermented soybean products purchased from retails in Seoul were tested. Thirteen of 26 doenjang samples, 13 of 23 kochujang samples, 16 of 30 ssamjang samples, and 5 of 9 cho-kochujang samples were positive for B. cereus sensu lato strains. The contamination level of all positive samples did not exceed 4 log CFU/g of food (maximum levels of Korea Food Code). Eighty-seven B. cereus sensu lato strains were isolated from 47 positive samples, and all isolates carried at least one enterotoxin gene. The detection rates of hblCDA, nheABC, cytK, and entFM enterotoxin genes among all isolates were 34.5%, 98.9%, 57.5%, and 100%, respectively. Fifteen strains (17.2%) harbored the emetic toxin gene. Most strains tested positive for salicin fermentation (62.1%), starch hydrolysis (66.7%), hemolysis (98.9%), motility test (100%), and lecithinase production (96.6%). The B. cereus sensu lato strains were highly resistant to β-lactam antibiotics such as ampicillin, penicillin, cefepime, imipenem, and oxacillin. Although B. cereus sensu lato levels in Korean fermented soybean products did not exceed the maximum levels permitted in South Korea (<10 4 CFU/g), these results indicate that the bacterial isolates have the potential to cause diarrheal or emetic gastrointestinal diseases.
The present study was undertaken to determine the prevalence of Salmonella in 100 chicken carcass samples from five integrated broiler operation brands in Korea. Serotypes, antibiotic resistance patterns, extended-spectrum β-lactamase (ESBL) genotype, and clonal divergence using multilocus sequence typing of the isolated strains were analyzed. A total of 42 chicken samples were contaminated with nontyphoidal Salmonella (NTS) isolates: 16 isolates (38%) were Salmonella Virchow, 9 (21%) were Salmonella Bareilly, and 8 (19%) were Salmonella Infantis. A multidrug resistance (MDR; resistant to more than three classes of antibiotics) phenotype was observed in 29% of the isolates, which were resistant to five or more classes of antibiotics. The dominant MDR type was resistance to classes of penicillin, cephalosporins, aminoglycosides, quinolones, and tetracyclines. All the MDR isolates were positive for ESBL producers, and all but one (with the CTX-M-1 genotype) had the CTX-M-15 genotype. Multilocus sequence typing of the isolates revealed ST16 as the dominant sequence type; Salmonella Virchow, Salmonella Infantis, and Salmonella Richmond were all ST16, indicating a close genetic relationship between these serovars. This is the first study in Korea showing the CTX-M-1 type of NTS and the prevalence of ESBL-producing strains among NTS isolated from retail chicken meat. Our findings suggest that MDR Salmonella contamination is widely prevalent in retail chicken meat, and consumption of inadequately cooked products could lead to dissemination of NTS, which is hazardous to human health.
Kefir is a unique fermented dairy product that is produced by a mixture of lactic acid bacteria, acetic acid bacteria and yeast. In this study, a culture-independent quantitative real-time polymerase chain reaction (PCR) technique was used to enumerate lactic acid bacteria, acetic acid bacteria and yeast in kefir grain and kefir milk using group-specific primers. The most abundant microbial groups were lactic acid bacteria and acetic acid bacteria, followed by yeast (P < 0.05) in both kefir grain and kefir milk. The most leading genera of lactic acid bacteria and yeast were Lactobacillus/Lactococcus (P < 0.05) and Candida, respectively. All microbial groups were present in lower numbers in kefir milk than in kefir grain (P < 0.05), except for Enterococcus and Saccharomyces groups. This is the first report to characterize the number of specific microbial populations in kefir grain and kefir milk using quantitative real-time PCR. In conclusion, we successfully established the methods for the quantification of microorganisms in kefir grain and kefir milk using group-specific quantitative real-time PCR and provided quantitative microbiological information for quality control of kefir. PRACTICAL APPLICATIONSThis report describes the enumerative methods of microorganisms in the kefir grain and kefir milk using group-specific quantitative real-time PCR assay. It could facilitate the microbiological quality control of kefir-fermented milk and grain and can be used in further studies to understand the fermentation process of kefir milk. In addition, the results provide basic information on the number of lactic acid bacteria, acetic acid bacteria and yeast in kefir grain and kefir milk.
In South Korea, few reports have indicated the occurrence and characteristics of extended-spectrum β-lactamase (ESBL)-producing Escherichia coli in food-producing animals, particularly in poultry slaughterhouses. In this study, we investigated the occurrence and antibiotic resistance of ESBL-producing E. coli from whole chicken carcasses (n=156) and fecal samples (n=39) of chickens obtained from 2 slaughterhouses. Each sample enriched in buffered peptone water was cultured on MacConkey agar with 2 mg/L cefotaxime and ESBL agar. ESBL production and antibiotic susceptibility were determined using the Trek Diagnostics system. The ESBL genotypes were determined by polymerase chain reaction (PCR) using the bla(SHV), bla(TEM), and bla(CTX-M) gene sequences. Subtyping using a repetitive sequence-based PCR system (DiversiLab™) and multilocus sequence typing (MLST) were used to assess the interspecific biodiversity of isolates. Sixty-two ESBL-producing E. coli isolates were obtained from 156 samples (39.7%). No bla(SHV) genes were detected in any of the isolates, whereas all contained the bla(TEM) gene. Twenty-five strains (40.3%) harbored the CTX-M group 1 gene. The most prevalent MLST sequence type (ST) was ST 93 (14.5%), followed by ST 117 (9.7%) and ST 2303 (8.1%). This study reveals a high occurrence and β-lactams resistance rate of E. coli in fecal samples and whole chickens collected from slaughterhouses in South Korea.
The current study was conducted to assess Salmonella spp. contamination in chicken carcasses produced at major poultry processing plants in South Korea. In total, 120 chicken carcasses were collected through 12 individual trials (10 chickens per trial) from six poultry processing plants in the summer of 2014 and the winter of 2015. Eighteen chicken samples (15%) were contaminated with Salmonella, with a higher rate of contamination observed during summer (14 isolates, 11.7%) than during winter (four isolates, 3.3%). Salmonella enterica serotype Typhimurium was the most prevalent, followed by Salmonella Hadar, Salmonella Rissen, Salmonella Bareilly, and Salmonella Virchow. Among five multidrug resistant isolates, a single strain was resistant to 10 antibiotics, including third-generation cephalosporins. This cephalosporin-resistant strain exhibited the extended-spectrum β-lactamase (ESBL) phenotype and harbored the gene encoding CTX-M-15, the most prevalent ESBL enzyme worldwide. Herein, repetitive-sequence-based polymerase chain reaction (rep-PCR) subtyping was conducted to discriminate the isolated Salmonella spp. and the ESBL-producing Salmonella isolate was distinguished by rep-PCR molecular subtyping, showing low genetic similarity in their rep-PCR-banding patterns. Given that poultry processing plants are the last stage in the chicken-production chain, the occurrence of Salmonella spp. including ESBL-producing strain in individually packaged chicken products highlights the necessity for regular monitoring for Salmonella in poultry processing plants.
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