• Proceedings of the PSK Conference
• /
• 2003.04a
• /
• pp.226.2-226.2
• /
• 2003

The FimH subunit of type 1-fimbriated Escherichia coli has been determined as a major cause of urinary tract infection. To produce a possible vaccine antigen against urinary tract infection, the fimH gene was genetically linked to the Itxa2b gene, which was then cloned into the pMAL -p2E expression vector. The chimaeric construction of pMALfimH/Itxa2b was transformed into Escherichia coli TB1 and its N-terminal amino acid sequence was analyzed. (omitted)

• Korean Journal of Veterinary Service
• /
• v.29 no.2
• /
• pp.97-102
• /
• 2006

A total of 1,848 samples was taken from bovine and porcine from January 2003 to November 2005. They were examined for the presence of Clostridiuml perfringens. The properties of the isolates were characterized for Gram staining, biochemical features and enterotoxin production. Forty-one strains (2.2%) of C perfringens were isolated from the 1,848 bovine and porcine carcass using selective media. 30 (3.2%) C perfringens were isolated from the 925 of bovine cacasses, and 11(1.2%) were isolated from the 923 of porcine cacasses. In TSC agar, all isolates showed lecithinase activity. The isolation rate was higher in spring and summer than in autumn and winter. Among 41 isolates, only 1 isolate detected the cpe gene in PCR. The PCR amplified band was observed 233 bp.

• The Korean Journal of Food And Nutrition
• /
• v.33 no.2
• /
• pp.142-148
• /
• 2020

The purpose of this study was to investigate the prevalence, toxin gene profiles, and enterotoxin producing ability of Bacillus cereus isolated from environment-friendly vegetables and good agricultural practices (GAP) vegetables. A total of 49 vegetables including 40 environment-friendly vegetables and 9 GAP vegetables were tested. The Vitek 2 system was used to identify B. cereus and the PCR was used to detect 6 toxin genes, respectively. B. cereus was detected in 34 (69.3%) of 49 vegetables and the prevalence of B. cereus in GAP vegetables (44.4%) was lower than in the environment-friendly vegetables (75.0%). The detection rates of entFM, nheA, hblC, and cytK enterotoxin genes, respectively, among all isolates were 100%, 97.0%, 88.2%, and 73.5%, respectively. All of the isolates had at least one or more enterotoxin gene and 20 isolates (58.8%) had hemolysin BL enterotoxin producing ability. The risk of food poisoning from the environment-friendly vegetables and the GAP vegetables has been shown as constant. Thus, it is necessary to expand the supply of GAP vegetables showing lower B. cereus contamination than the environment-friendly vegetables. The characteristics of the environment-friendly vegetables and the GAP vegetables that must be consumed after cleaning should be disseminated to consumers regarding food poisoning prevention.

• Journal of Microbiology and Biotechnology
• /
• v.25 no.6
• /
• pp.872-879
• /
• 2015

Many strains of Bacillus cereus cause gastrointestinal diseases, and the closely related insect pathogen Bacillus thuringiensis has also been involved in outbreaks of diarrhea. The diarrheal diseases are attributed to enterotoxins. Sixteen reference strains of B. cereus and nine commercial and 12 reference strains of B. thuringiensis were screened by PCR for the presence of 10 enterotoxigenic genes (hblA, hblC, hblD, nheA, nheB, nheC, cytK, bceT, entFM, and entS), one emetogenic gene (ces), seven hemolytic genes (hlyA, hlyII, hlyIII, plcA, cerA, cerB, and cerO), and a pleiotropic transcriptional activator gene (plcR). These genes encode various enterotoxins and other virulence factors thought to play a role in infections of mammals. Amplicons were successfully generated from the strains of B. cereus and B. thuringiensis for each of these sequences, except the ces gene. Intriguingly, the majority of these B. cereus enterotoxin genes and other virulence factor genes appeared to be widespread among B. thuringiensis strains as well as B. cereus strains.

• Journal of Microbiology
• /
• v.43 no.4
• /
• pp.354-360
• /
• 2005

Heat-labile enterotoxin B subunit (LTB) of enterotoxigenic Escherichia coli (ETEC) is both a strong mucosal adjuvant and immunogen. It is a subunit vaccine candidate to be used against ETEC-induced diarrhea. It has already been expressed in several bacterial and plant systems. In order to construct yeast expressing vector for the LTB protein, the eltB gene encoding LTB was amplified from a human origin enterotoxigenic E. coli DNA by PCR. The expression plasmid pLTB83 was constructed by inserting the eltB gene into the pYES2 shuttle vector immediately downstream of the GAL1 promoter. The recombinant vector was transformed into S. cerevisiae and was then induced by galactose. The LTB protein was detected in the total soluble protein of the yeast by SDS-PAGE analysis. Quantitative ELISA showed that the maximum amount of LTB protein expressed in the yeast was approximately $1.9\%$ of the total soluble protein. Immunoblotting analysis showed the yeast-derived LTB protein was antigenically indistinguishable from bacterial LTB protein. Since the whole-recombinant yeast has been introduced as a new vaccine formulation the expression of LTB in S. cerevisiae can offer an inexpensive yet effective strategy to protect against ETEC, especially in developing countries where it is needed most.

• Microbiology and Biotechnology Letters
• /
• v.33 no.3
• /
• pp.207-214
• /
• 2005

To characterize the genotypic traits of clinical methicillin-resistant Staphyiococus aureus (MRSA) isolates (n=49), major virulence-associated genes were detected by using PCR-based methods. All the MRSA isolates possessed coagulase gene and showed four polymorphism types [500bp ($6{\%}$),580bp ($27{\%}$), 660bp ($65{\%}$) and 740bp ($2{\%}$)] due to variable numbers of tandem repeats present within the gene. The four or five different loci of hemolysin gene family were dominant in the MRSA isolates,25 of which($51{\%}$) possessed a combination of hla / hlb / hld/ hlg / hlg-2 genes as the most prevalent type. The prevalence of enterotoxin genes was varied among the MRSA isolates. sea and seb genes were detected from all the MRSA isolates. But sei, tsst-1, seg, sec, and seh genes were detected from 31 ($63{\%}$), 16 ($33{\%}$), 14 ($29{\%}$), 8 ($16{\%}$), and 5 ($10{\%}$) isolates, respectively. sed and sej genes were detected from 1 ($2{\%}$) isolate, respectively. see, eta, and etb genes were not detected at all. sea / seb genes were co-detected from 11 ($23{\%}$) isolates, sea / seb / sei genes from 9 ($19{\%}$) isolates, and sea / seb / seg / sei / tsst-1 genes from 5 ($10{\%}$) isolates. Other genes were co-detected with below $10{\%}$ frequencies.

• Journal of Food Hygiene and Safety
• /
• v.6 no.3
• /
• pp.147-155
• /
• 1991

Enterotoxigenic E. coli is one of the major causative agents of the infantile diarrhea and traveler's diarrhea. The heat-stable enterotoxin(ST) is thought to be a virulence factor in the pathogenesis of the diarrhea and to be a maker for identification of the enterotoxingeic E. coli from non pathogenic E. coli. The isolate of enterotoxigenlc E. coli was isolated from swine during 1989 year(from 5 to 10 month) in the Kyong-gi and Chung-Cheong provinces, and three strains(KM-4, KM-7 and KM-12) was selected from 189 isolates of ST producing E. coli. The detection of a ST produced of the isolated E. coli was performed by the infant mouse assay(IMA). This study was designed to know optimal conditions for the production of the ST and the molecular properties of plasmids of the enterotoxigenic E. coli. Amount of ST produced were the most at initial pH 8.5~9.0 of succinate salts medium culture. The cultural time of the same medium was accumulated the highest level of ST was at the 14 to 16 hours, and then stationary phase was at the 20 hours. From this experiment the KM-7 strain was selected among ST producing strains by IMA. Partial plasmid-curing experiment was done to select plasmid encoding for ST among other plasmids and then comparing the plasmid pattern of ST producing strain(KM-7) with those of other ST non-producing strains, it is found that ST gene exists on the about 80 Kbp plasmid. Each fragment of this plasmid digested with EcoRl was ligated to vector pBR 322 and transformed into E. coli K-12. A clone producing ST(eKT 53) was selected by IMA. The EcoRl digestion pattern of the isolated plasmid(pKD 37) from the ST producing clone it is indicated that the size of the inserted fragment in eKT 53 strain is 16 Kbp. The cultured supernatant of eKT 53 strain was positive result of ST production in IMA.

• Korean Journal of Food Science and Technology
• /
• v.38 no.2
• /
• pp.295-303
• /
• 2006

To provide microbial information for the safety of agricultural production, the presence of enterotoxin genes and antibiotic susceptibility of 14 isolated Staphylococcus aureus (11.7%) strains were investigated using PCR-based methods and disk diffusion method, respectively. Among enterotoxin-encoding genes, sea was detected from two isolates (14.3%), sea and sed genes were co-detected from three isolates (21.4%), and sea, sed, and see genes in seven isolates (50.0%), whereas seb, sec, and tsst were not detected in any isolate. Nine (64.3%), eight (57.1%), six (42.9%), two (14.3%), and one (7.2%) isolates were resistant to penicillin, novobiocin, amphicillin, erythromycin and oxacillin, and doxycycline and kanamycin, respectively. Methicilline-resistant S. aureus was found in roller of B farm and in hydroponic solution of D farm.

• Journal of Microbiology and Biotechnology
• /
• v.25 no.12
• /
• pp.2043-2048
• /
• 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 Microbiology and Biotechnology
• /
• v.27 no.4
• /
• pp.709-717
• /
• 2017

Mucosal tissues are the initial site through which most pathogens invade. As such, vaccines and adjuvants that modulate mucosal immune functions have emerged as important agents for disease prevention. Herein, we investigated the immunomodulatory mechanisms of the B subunit of Escherichia coli heat-labile enterotoxin type IIa ($LT-IIa-B_5$), a potent non-toxic mucosal adjuvant. Alternations in gene expression in response to $LT-IIa-B_5$ were identified using a genome-wide transcriptional microarray that focused on dendritic cells (DC), a type of cell that broadly orchestrates adaptive and innate immune responses. We found that $LT-IIa-B_5$ enhanced the homing capacity of DC into the lymph nodes and selectively regulated transcription of pro-inflammatory cytokines, chemokines, and cytokine receptors. These data are consistent with a model in which directional activation and differentiation of immune cells by $LT-IIa-B_5$ serve as a critical mechanism whereby this potent adjuvant amplifies mucosal immunity to co-administered antigens.