• Korean Journal of Food Science and Technology
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• v.52 no.6
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• pp.678-684
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• 2020

The objective of this study was to identify the fermentation characteristics of Bacillus thuringiensis and emetic, non-emetic Bacillus cereus using analytical profile index (API) test. Ten strains of B. thuringiensis and 10 strains of B. cereus including 3 strains of emetic type were used at the same concentrations. The differences of fermentation characteristics between the B. thuringiensis and B. cereus was not obvious, but the differences between the non-emetic and emetic B. cereus were distinctive. Seven among 50 substrates were negative for all non-emetic B. cereus strains and positive for all emetic strains, and three substrates among additional 12 substrates had the same tendency. From these differences, 3 emetic B. cereus strains were not indicated as B. cereus by API test. These results indicate that API test is not a suitable method to identify some strains of emetic B. cereus, and the distinctive differences in substrate utilization can be used to improve selective media.

• Korean Journal of Food Science and Technology
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• v.53 no.6
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• pp.815-818
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• 2021

The objective of this study was to develop a differential medium with improved selectivity for the isolation of Bacillus cereus. Mannitol egg yolk polymyxin medium supplemented with D-galactose allowed the differentiation of diarrheal- and emetic-type B. cereus through pH monitoring. The pH of the medium decreased significantly when incubating the emetic-type B. cereus, whereas the pH change was not significant when incubating the diarrheal-type. The addition of pH indicators, such as methyl red and phenol red, to the medium allowed visual differentiation between diarrheal- and emetic-type B. cereus. A solid agar medium was also developed by optimizing the concentrations of medium components such as monosaccharides, agar, egg yolk enrichment, pH indicators, and antibiotics. This study indicates the possibility of applying selective media for the differentiation of diarrheal- and emetic-type B. cereus.

• Korean Journal of Food Science and Technology
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• v.42 no.3
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• pp.361-367
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• 2010

Bacterial contamination of cooked rice was analyzed to evaluate the microbial safety. Thirty raw rice samples were collected in Korea and cooked in an electric rice cooker. Mesophilic aerobe, food-poisoning Bacillus cereus group, and their toxin genes were determined on cooked rice. The percentage of total mesophilic aerobe based on 1-3 log CFU/g was 27% among the samples. Bacillus spp. in MYP selective medium was similar to the number of mesophilic aerobe, whileas Bacillus spp. was detected in most samples after enrichment. Thirty-seven isolates from 30 cooked rices were identified as B. thuringiensis, B. cereus, B. valismortis, B. pumilus, B. coagulans, B. licheniformis, Geobacillus stearothermophilus, and Brevibacillus laterosporus. Twenty isolates (54%), more than half of the isolates, were B. thuringiensis while nine (27%) were identified as B. cereus. All B. thuringiensis isolates possessed non-hemolytic toxin genes and interestingly, seven B. cereus among nine isolates possessed emetic toxin genes. More B. thuringiensis was present on the cooked rice than B. cereus and most B. cereus possessed emetic toxin genes rather than diarrheal toxin genes. Therefore, food-borne outbreak due to B.cereus on the cooked rice kept at room temperature might be examples of emetic food-poisoning.

• Food Science and Biotechnology
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• v.17 no.4
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• pp.761-765
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• 2008

Bacillus cereus causes two different types of food poisoning syndromes: diarrhea and emesis. The diarrheal syndrome is attributed to various enterotoxins, including nonhemolytic enterotoxin, hemolytic enterotoxin, and enterotoxin-T, whereas the emetic syndrome is caused by the dodecadepsipeptide toxin cereulide. A multiplex polymerase chain reaction (PCR) assay was developed to rapidly detect and identify B. cereus strains. Three primer pairs specific to regions within genes encoding nonhemolytic enterotoxin (nheA), molecular chaperonin (groEL), and cereulide synthetase (ces) were used to identify and differentiate between the enterotoxin-producing and emetic toxin-producing B. cereus strains. The cereulide-producing emetic B. cereus showed 3 PCR products of 325, 405, and 685 bp for the groEL, ces, and nheA genes, respectively, whereas the enterotoxin-producing B. cereus showed 2 PCR products without a ces gene specific DNA fragment. Specific amplifications and differentiations by multiplex PCR assay were obtained using 62 B. cereus strains and 13 strains' of other bacterial species. The detection limit of this assay for enterotoxin-producing strain and emetic toxin-producing strain from pure cultures were $2.4{\times}10^1$ and $6.0{\times}10^2\;CFU/tube$, respectively. These results suggest that our multiplex PCR method may be useful for the rapid detection and differentiation of B. cereus strains in foods.

• Applied Biological Chemistry
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• v.51 no.4
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• pp.263-268
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• 2008

Seventy-one Bacillus cereus strains (12 references and 59 food isolates) were analyzed for the occurrence of five different enterotoxin genes (nheABC, hblCDA, entFM, cytK, and bceT) and one emetic toxin cereulide synthetase gene (ces) by PCR (polymerase chain reaction). PCR analysis revealed eight toxigenic patterns in all B. cereus strains tested; they all carried both entFM and nheABC. The presence of hblCDA, cytK, and bceT varied according to the enterotoxin-producing strains, among which hblCDA was the least frequently detected in the food-isolated strains. Only five B. cereus strains harbored ces, associated with the emetic type of food poisoning; however, these strains were devoid of hblCDA, cytK, and bceT.

• Journal of the FoodService Safety
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• v.1 no.1
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• pp.9-18
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• 2020

B. cereus-produced cereulide as an emetic toxin is commonly isolated in starch-based cooked foods. This study examined the prevalence of B. cereus from ready-to-eat foods in markets by polymerase chain reaction analysis and determined the relationship between the level of B. cereus and the quantity of cereulide in the sample after different storage times and temperatures. The prevalence of general B. cereus in 43 starch foods was 32.6%, and the level of B. cereus ranged from 0.5 to 1.95 log cfu/g, meeting the Korea Food Code Specifications of 3 log CFU/g of B. cereus. No samples revealed emetic B. cereus. Fried rice samples were inoculated with a cereulide-producing reference strain, B. cereus NCCP 14796, to determine the level of B. cereus and the quantity of cereulide in the samples after storage for 0, 4, 6, 8, 20, 24, 30, 48, 72, and 96 h at 7, 25, 35, and 57℃. The average levels of B. cereus at 7, 25, 35, and 57℃ were 4.38, 7.31, 7.88, and 3.82 log cfu/g, and the levels of cereulide were 150.41, 1680.70, 2652.65, and 77.83 ㎍/mL, respectively, showing a significant difference according to the incubation time (P<0.05) and temperature (P<0.001).

• Journal of Microbiology and Biotechnology
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• v.20 no.7
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• pp.1107-1113
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• 2010

Because conventional methods for detecting emetic-toxin-producing B. cereus are laborious and costly, various PCR assays, which are easy and cheap, have recently been reported. Therefore, this study estimated and compared the ability of various PCR assays to detect emetic-toxin-producing B. cereus strains isolated in Korea. The PCR assays were performed on 160 B. cereus strains, including 40 emetic-toxin-producing strains. Although the species-specific PCR assays were all shown to be highly specific, the sensitivities varied greatly. The accuracies of the primers were 97.5% (CER), 95.6% (EM1), 96.3% (RE234), 89.4% (CES), and 83.1% (Ces3R/CESR2). Moreover, the CER primer had a higher sensitivity (100%) than all the other primers tested, and a specificity of 96.7%. Thus, the CER primer was shown to be the most effective for screening the emetic-toxin-producing B. cereus strains tested in this study. However, the ability of these PCR assays to identify emetic-toxin-producing B. cereus should also be confirmed using other methods.

• Journal of Microbiology and Biotechnology
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• v.22 no.5
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• pp.649-653
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• 2012

A temperate phage was isolated from emetic Bacillus cereus NCTC 11143 by mitomycin C and characterized by transmission electron microscopy and DNA and protein analyses. Whole genome sequencing of Bacillus phage 11143 was performed by GS-FLX. The phage has a dsDNA genome of 39,077 bp and a 35% G+C content. Bioinformatic analysis of the phage genome revealed 49 putative ORFs involved in replication, morphogenesis, DNA packaging, lysogeny, and host lysis. Bacillus phage 11143 could be classified as a member of the Siphoviridae family by morphology and genome structure. Genomic comparisons at the DNA and protein levels revealed homologous genetic modules with patterns and morphogenesis proteins similar to those of other Bacillus phages. Thus, Bacillus phages might have a mosaic genetic relationship.

• Food Science and Biotechnology
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• v.18 no.3
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• pp.594-603
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• 2009

The genus Bacillus includes a variety of diverse bacterial species, which are widespread throughout the environment due to their ubiquitous nature. A well-known member of the genus, Bacillus cereus, is a food poisoning bacterium causing both emetic and diarrhoeal disease. Other Bacillus species, particularly B. subtilis, B. licheniformis, B. pumilus, and B. thuringiensis, have also recently been recognized as causative agents of food poisoning. However, reviews and research pertaining to bacilli have focused on B. cereus. Here, we review the literature regarding the potentially toxigenic Bacillus species and the toxins produced that are associated with food poisoning.

• Journal of Science Criminal Investigation
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• v.11 no.3
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• pp.210-217
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• 2017

In the forensic microbiology laboratories, microorganism analyses from food are requested. There have been several cases of Bacillus cereus isolated from the samples requested to the National Forensic Service. B. cereus is an important pathogenic bacterium which can cause food-borne outbreaks. Therefore, we isolated B. cereus from anchovy aekjeot recently requested for microbial examination and identified using MSId based on the 16S rDNA sequence and real-time PCR method. We also conducted PCR for detection of diarrheal toxin genes and an emetic toxin gene and found the presence of nheABC, bceT and entFM diarrheal toxin genes in the B. cereus isolate. There are several clinically important food-poisoning bacteria that should be noted during inspection. In particular, B. cereus can cause food poisoning even when cooked foods are ingested, because B. cereus forms endo-spore which confers strong environmental resistance and heat resistance to the bacteria, and the bacterial emetic toxin also has heat resistance. Here we highlight the importance to distinguish clinically important bacteria such as B. cereus from food specimens, and we expect this study will provide procedures for identification of B. cereus and detection of the bacterial toxin genes for future cases in the forensic microbiology laboratories.