Incidence and properties of Bacillus cereus strains naturally present in cereals were evaluated by phenotypic characterization, antibiotic susceptibility testing, and pulsed-field gel electrophoresis. Of 293 cereal samples tested, 73 (25%) contained B. cereus strains. Incidence of B. cereus isolates varied with respect to sample; they were found in 15 (37%) of 83 brown rice samples, 23 (37%) of 63 glutinous rice samples, 16 (21%) of 76 barley samples, and 19 (27%) of 71 Job's tears samples. All B. cereus isolates from cereals were positive for diarrheal toxin genes. The isolates were susceptible to most of the antibiotics tested, but they were highly resistant to ampicillin, cefepime, oxacillin, and penicillin. Of the genes assayed by the PCR technique, a high frequency of nheA (99%) and hblDC (84%) was found in the genomic DNA of cereal-associated isolates, whereas cytK was less common (55%). From the strains carrying the hblDC genes, 93% produced enterotoxin HBL. B. cereus isolates did not have significant genetic homology. The genetic diversity and toxic potential differ among the strains isolated from cereals. These results provide important information on toxin gene profiles of cereal-associated B. cereus for population studies.
Bacillus cereus was divided into emetic toxin (cereulide)- and enterotoxin-producing strains, but emetic toxin-producing B. cereus is difficult to detect immunochemically. Screening methods for emetic toxin-producing B. cereus are needed. The objectives of this study were to identify and detect emetic toxin-producing B. cereus among 160 B. cereus strains, and to compare enterotoxin production and phenotypic characteristics between the emetic toxin-producing and enterotoxin-producing strains. Forty emetic toxin-producing B. cereus strains were determined with high-pressure liquid chromatography-mass spectrometry analysis. Among the emetic toxin-producing strains (n = 40), 31 (77.5%) and 3 (7.5%) strains produced nonhemolytic enterotoxin (NHE) and hemolysin BL (HBL) enterotoxins, respectively. In addition, 107 (89.2%) and 100 (83.3%) strains produced NHE and HBL enterotoxins among the enterotoxin-producing strains (n = 120). The number of strains positive for starch hydrolysis, salicin fermentation, and hemolysis among the emetic toxin-producing strains were 3 (7.5%), 3 (7.5%), and 26 (65.0%), respectively, and among enterotoxin-producing strains, these numbers were 101 (84.2%), 100 (83.3%), and 111 (92.5%), respectively. In particular, the three emetic toxin-producing B. cereus strains (JNHE 6, JNHE 36, and KNIH 28) produced the HBL and NHE enterotoxins and were capable of starch hydrolysis and salicin fermentation. The absence of HBL enterotoxin and certain phenotypic properties, such as starch hydrolysis and salicin fermentation, indicates that these properties were not critical characteristics of the emetic toxin-producing B. cereus tested in this study.
Bacillus cereus contamination is a major food safety problem for Korean fermented soybean products, but few studies have assessed its potential to cause foodborne illness. The objectives of this study were to investigate the prevalence and characteristics of B. cereus isolated from Korean fermented soybean products. B. cereus was detected in 110 of 162 (67.9%) samples. The highest B. cereus frequency was observed in deonjang (68 of 93 samples, 73.1%) and cheonggukjang (18 of 25, 72.0%); however, nonhemolytic enterotoxin was detected only in 22 of 162 samples (13.6%). Although the tested B. cereus isolates showed diverse pulsotypes according to repetitive sequence-PCR banding patterns, they displayed similar antibiotic sensitivity spectra. The low frequency of enterotoxin detection suggests that the potential risk of B. cereus foodborne illness associated with Korean fermented soybean products is lower than generally presumed. However, considering the prevalence of B. cereus and the high content of fermented soybean products in the Korean diet, it is necessary to constantly monitor the level of contamination with B. cereus and its toxins in such Korean food products.
Bacillus cereus can cause diarrheal and emetic types of food poisoning but little study has been done on emetic type of food poisoning in Korea. The objective of this study was to report on the emetic type of food poisoning associated with B. cereus in Korea. The toxin gene profile, toxin production, and antibiotic resistance of B. cereus isolates were investigated in this study. B. cereus was detected in three out of four samples, while the other food poisoning bacteria were not detected. All isolates (KUGH 10, 11, and 12) presented nhe A, B, and C diarrheal toxin genes (755, 743, and 683 bp), detected using NHA, NHB, and NHC primers, and ces emetic toxin gene (1271 bp), detected using CES primer, and produced nonhemolytic enterotoxin and emetic toxin (cereulide), detected using immunochemical assay and high performance liquid chromotography/mass spectrometry (HPLC/MS) analysis. All emetic-associated isolates were resistant to beta-lactam antibiotics. Most important finding in this study was that the risk of emetic-type B. cereus food poisoning has existed in Korea. This suggested that the food poisoning caused by B. cereus producing emetic and diarrheal toxins should be constantly evaluated to prevent misdiagnosis between emetic and diarrheal types of food poisoning.
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