• Journal of Microbiology and Biotechnology
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• v.31 no.5
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• pp.710-716
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• 2021

A risk analysis of Shiga toxin (Stx)-encoding bacteriophage was carried out by confirming the transduction phage to non-Stx-producing Escherichia coli (STEC) and subsequent expression of the Shiga toxin genes. The virulence factor stx1 was identified in five phages, and both stx1 and stx2 were found in four phages from a total of 19 phage isolates with seven non-O157 STEC strains. The four phages, designated as ϕNOEC41, ϕNOEC46, ϕNOEC47, and ϕNOEC49, belonged morphologically to the Myoviridae family. The stabilities of these phages to temperature, pH, ethanol, and NaClO were high with some variabilities among the phages. The infection of five non-STEC strains by nine Stx-encoding phages occurred at a rate of approximately 40%. Non-STEC strains were transduced by Stx-encoding phage to become lysogenic strains, and seven convertant strains had stx1 and/or stx2 genes. Only the stx1 gene was transferred to the receptor strains without any deletion. Gene expression of a convertant having both stx1 and stx2 genes was confirmed to be up to 32 times higher for Stx1 in 6% NaCl osmotic media and twice for Stx2 in 4% NaCl media, compared with expression in low-salt environments. Therefore, a new risk might arise from the transfer of pathogenic genes from Stx-encoding phages to otherwise harmless hosts. Without adequate sterilization of food exposed to various environments, there is a possibility that the toxicity of the phages might increase.

• Journal of Environmental Health Sciences
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• v.38 no.6
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• pp.510-519
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• 2012

Objectives: This study was conducted in order to evaluate the microbiological contamination of the indoor air of the lunchrooms at child care centers and investigate the toxin genes and toxin production ability of food-borne pathogens. Methods: A total of 64 child care centers were sampled to test total aerobic bacteria, coliform bacteria, fungi, Staphylococcus aureus, Bacillus cereus and Salmonella spp. according to the Korea Food Code. All toxin genes of pathogens were detected using the Polymerase Chain Reaction method. The Sthaph. aureus enterotoxin was detected by a Staphylococcus aureus enterotoxin-reversed passive latex agglutination kit. The heamolysin BL (HBL) and non-heamolytic enterotoxin (NHE) produced by B. cereus were detected using a B. cereus enterotoxin-reversed passive latex agglutination kit and Bacillus diarrheal enterotoxin visual immunoassay kit, respectively. Results: The means of total aerobic bacteria and coliform bacteria were $1.91{\pm}1.84$ log CFU/plate and $0.47{\pm}0.62$ log CFU/plate, respectively. The mean of fungi also showed $0.59{\pm}0.71$ log CFU/plate. Among the pathogenic bacteria tested in this study, Staphy. aureus and B. cereus were detected in four (6.3%) and 21 (32.8%) out of 64 indoor air samples from lunchrooms in child care centers, respectively. All Staphy. aureus tested in this study possessed no toxin genes and did not produce enterotoxin. The detection rate of nheABC, hblCDA, entFM and ces toxin gene in B. cereus was 100, 57.1, 76.2 and 0%, respectively. B. cereus isolates were classified into four groups according to the presence or absence of toxin genes. The nheABC gene was the major toxin gene among B. cereus tested in this study. The HBL was detected in 11 out of 21 B. cereus isolates (52.4%) and three B. cereus isolates produced NHE (14.3%). Conclusion: The results indicated that the contamination by microorganisms in the indoor air of lunchrooms was unqualified to supply safe catering in child care centers. The ongoing control of indoor air quality is required.

• 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.

• Journal of Microbiology and Biotechnology
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• v.26 no.1
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• pp.44-55
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• 2016

Bacillus cereus is a gram-positive, rod-shaped, spore-forming bacterium that has been isolated from contaminated fermented soybean food products and from the environment. B. cereus produces diarrheal and emetic toxins and has caused many outbreaks of foodborne diseases. In this study, we investigated whether B. amyloliquefaciens RD7-7, isolated from rice doenjang (Korean fermented soybean paste), a traditional Korean fermented soybean food, shows antimicrobial activity against B. cereus and regulates its toxin gene expression. B. amyloliquefaciens RD7-7 exhibited strong antibacterial activity against B. cereus and inhibited the expression of B. cereus toxin-related genes (groEL, nheA, nheC, and entFM). We also found that addition of water extracts of soybean and buckwheat soksungjang (Korean fermented soybean paste made in a short time) fermented with B. amyloliquefaciens RD7-7 significantly reduced the growth and toxin expression of B. cereus. These results indicate that B. amyloliquefaciens RD7-7 could be used to control B. cereus growth and toxin production in the fermented soybean food industry. Our findings also provide a basis for the development of candidate biological control agents against B. cereus to improve the safety of fermented soybean food products.

• Journal of Food Hygiene and Safety
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• v.35 no.1
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• pp.6-12
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• 2020

Prevalence of toxin genes and profiles of antibiotic resistance in Vibrio vulnificus were investigated for prevention of Vibrio sepsis and selection of effective antibiotics. A total of 23 V. vulnificus strains were isolated from Vibrio sepsis patients, fish, and water samples collected from fish tanks in restaurants in Jeonnam province during 2015-2017 period. Prevalence of toxin genes including, RtxA, viuB and vvhA were assessed and susceptibilities to 15 different antibiotics were determined. As a result of the toxin gene profile, the RtxA toxin gene was detected in 19 (82.6%) out of 23 strains, and vvhA and viuB toxin genes were positive in all strains. These results showed that V. vulnificus tested in this study possessed at least one more toxin gene, and the toxin gene detection rate was higher than in previous reports. Therefore, there is always a risk of Vibrio sepsis through eating fish or having contact with aquarium water at seafood restaurants. Especially, it was deemed necessary to provide preventive education about Vibrio sepsis for workers in such restaurants. The results of antibiotic susceptibility tests presented 94.4% resistance to cepoxitin antibiotics but all strains showed susceptibility to 14 kinds of antibiotics including chloramphenicol and tetracycline. The currents antibiotic therapy using chloramphenicol and teteracycline against Vibrio sepsis was judged to be useful.

• Journal of Microbiology and Biotechnology
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• v.25 no.9
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• pp.1460-1466
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• 2015

In this study, the prevalence of Shiga toxin-producing Escherichia coli (STEC) was investigated among raw meat or meat products from slaughterhouses and retail markets in South Korea, and their potential for antibiotic resistance and virulence was further analyzed. A total of 912 raw meats, including beef, pork, and chicken, were collected from 2008 to 2009. E. coli strains were frequently isolated in chicken meats (176/233, 75.9%), beef (102/217, 42.3%), and pork (109/235, 39.2%). Putative STEC isolates were further categorized, based on the presence or absence of the Shiga toxin (stx) genes, followed by standard O-serotyping. Polymerase chain reaction assays were used to detect the previously defined virulence genes in STEC, including Shiga toxins 1 and Shiga toxin 2 (stx1 and 2), enterohemolysin (ehxA), intimin (eaeA), STEC autoagglutination adhesion (saa), and subtilase cytotoxin (subAB). All carried both stx1 and eae genes, but none of them had the stx2, saa, or subAB genes. Six (50.0%) STEC isolates possessed the ehxA gene, which is known to be encoded by the 60-megadalton virulence plasmid. Our antibiogram profiling demonstrated that some STEC strains, particularly pork and chicken isolates, displayed a multiple drug-resistance phenotype. RPLA analysis revealed that all the stx1-positive STEC isolates produced Stx1 only at the undetectable level. Altogether, these results imply that the locus of enterocyte and effacement (LEE)-positive strains STEC are predominant among raw meats or meat products from slaughterhouses or retail markets in Korea.

• Journal of Food Hygiene and Safety
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• v.35 no.3
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• pp.219-224
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• 2020

The purpose of this study was to provide basic data necessary for the prevention of food poisoning and safe food management. We examined 872 food samples for B. cereus in accordance with the MFDS Food Code and investigated characteristics of their harboring toxin genes. We detected and isolated 113 strains of B. cereus from 78 food samples (8.9%), and the average detection level was 48 CFU/g. B. cereus isolates carried at least 1 toxin gene among the emetic toxins and 5 enterotoxin genes. The toxin gene profiles of B. cereus were classified into 18 different types of isolates showing genetic diversity. Among the strains, 34 (30.1%) had all 5 enterotoxin genes (Cytk-nheA-entFM-bceT-hblC), accounting for the highest percentage. The entFM and nheA genes were major enterotoxin genes, while the emetic toxin gene, CER, was the least detected in B. cereus isolated from food samples.

• Journal of Life Science
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• v.21 no.2
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• pp.257-264
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• 2011

Seventy five methicillin- resistant Staphylococcus aureus (MRSA) strains and 24 methicillin- susceptible S. aureus (MSSA) were isolated from clinical specimens obtained from a hospital in Suncheon, Jeonnam province, Korea, from July to December, 2009. Antibiotic resistance was determined using the disc diffusion method. Genes encoding enterotoxin (SE), toxic shock syndrome toxin-1 (TSST-1), exfoliative toxin (ET) and Panton-Valentine leukocidin (PVL) were detected by multiplex PCR-mediated amplification using specific primers. Sixty (80%) MRSA isolates possessed either one or more toxin genes and the most common pattern that coexisted in MRSA was seb, sec, seg, sei and tst (22.7%) followed by coexistence of sec, seg, sei and tst genes (18.7%). Gene pvl encoding leukocidin was not found. Significant correlation between the production of sec, seg, sei and tst genes was found. MRSAs were resistant to erythromycin (89% of the isolates), gentamicin (70.7%), ciprofloxacin (69.3%), clindamycin (61.3%) and tetracycline (58.7%), while MSSAs were susceptible to the antibiotics with the exception of erythromycin. Toxin genes seb, sec and tst were related to the tetracycline resistance of MRSA.

• Journal of Microbiology and Biotechnology
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• v.25 no.12
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• pp.2043-2048
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• 2015

Bacillus thuringiensis microbial insecticide products have been applied worldwide. Although a few cases of B. thuringiensis foodborne illness have been reported, little is known about the toxigenic properties of B. thuringiensis isolates. The aims of this study were to estimate the pathogenic potential of B. thuringiensis selected from microbial insecticide products, based on its possession of toxin genes and production of enterotoxins. Fifty-two B. thuringiensis strains selected from four kinds of microbial insecticide products were analyzed. PCR assay for detection of toxin genes and immunoassay for detection of enterotoxins were performed. The hemolysin BL complex as a major enterotoxin was produced by 17 (32.7%), whereas the non-hemolytic enterotoxin complex was detected in 1 (1.9%) of 52 B. thuringiensis strains. However, cytK, entFM, and ces genes were not detected in any of the tested B. thuringiensis strains. The potential risk of food poisoning by B. thuringiensis along with concerns over B. thuringiensis microbial insecticide products has gained attention recently. Thus, microbial insecticide products based on B. thuringiensis should be carefully controlled.

• Journal of Life Science
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• v.27 no.1
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• pp.67-71
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• 2017

Foulbrood disease is infected by Paenibacillus larvae on larval stage of honeybee, and is lethal disease to result in population death. This disease was manifested in 2008 in Korea, is still suffered by the secondary damages. In this study, we are to examine diagnostic PCR approaches to manage the Foulbrood disease. PCR amplification of 16S rRNA is generally using for microbial infection, but the specificity is little poor for the correct diagnosis. Therefore, we are to identify specific genes expressed in Paenibacillus larvae, and perform PCR analysis. We selected five distinct genes from literature references. Those genes are commonly known as toxic genes for host infection, and include Toxin1, Toxin2A & 2B, SplA, CBP49, and SevA&SevB. PCR amplification for these genes is difficult to detect at the first time. So, we performed the second PCR using the first PCR product as a template. This approach using the nested PCR was very useful for detecting large marker genes. When Paenibacillus larvae was cultured in the medium containing plant extracts, PCR amplification of the identified genes is correlated with the microbial growth inhibition. Therefore, these results suggest that the identified genes might be useful to study diagnostic PCR markers for honeybee Foulbrood disease.