• Journal of Food Hygiene and Safety
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• v.14 no.4
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• pp.327-332
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• 1999

The prevalence and serotype of food-borne pathogens was investigated from 888 samples of chilled meat, 222 samples of packed frozen meat and 117 samples of imported frozen meat during the period from March 1996 to October 1998. Isolation rates of pathogens associated with food poisoning were revealed in order of Staphyloccus aureus, Campylobacter jejuni /coli, Listeria monocytogenes and Salmonella spp, but Escherichia coli O157:H7 was not isolated in all of the meat samples. Amusingly, Campylobacter jejuni /coli were isolated highly in refrigerated meat, but was not isolated in packed frozen meat. L. monocytogenes was encounted higher isolation frequency in packed frozen chicken meat than in refrigerated chicken meat. In the distribution of serotypes of isolates, most isolates of Sta. aureus classified as enterotoxin type C and D. All of the Salmonella spp. isolated from pork were diagnosed group A and most of isolates from chicken meat were grouped B and D. Most of L. monocytogenes isolated from chicken meat were grouped type 1 and a few number of isolates classified as type 4.

• Journal of Microbiology and Biotechnology
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• v.20 no.3
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• pp.550-557
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• 2010

Escherichia coli (E. coli) heat-labile enterotoxin B subunit (LTB) was regarded as one of the most powerful mucosal immunoadjuvants eliciting strong immunoresponse to coadministered antigens. In the research, the high-level secretory expression of functional LTB was achieved in P. pastoris through high-density fermentation in a 5-1 fermentor. Meanwhile, the protein was expressed in E. coli by the way of inclusion body, although the gene was cloned from E. coli. Some positive yeast and E. coli transformants were obtained respectively by a series of screenings and identifications. Fusion proteins LTB-6$\times$His could be secreted into the supernatant of the medium after the recombinant P. pastoris was induced by 0.5% (v/v) methanol at $30^{\circ}C$, whereas E. coli transformants expressed target protein in inclusion body after being induced by 1 mM IPTG at $37^{\circ}C$. The expression level increased dramatically to 250-300 mg/l supernatant of fermentation in the former and 80-100 mg/l in the latter. The LTB-6$\times$His were purified to 95% purity by affinity chromatography and characterized by SDS-PAGE and Western blot. Adjuvant activity of target protein was analyzed by binding ability with GMI gangliosides. The MW of LTB-6$\times$His expressed in P. pastoris was greater than that in E. coli, which was equal to the expected 11 kDa, possibly resulted from glycosylation by P. pastoris that would enhance the immunogenicity of co-administered antigens. These data demonstrated that P. pastoris producing heterologous LTB has significant advantages in higher expression level and in adjuvant activity compared with the homologous E. coli system.

• Korean Journal of Veterinary Research
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• v.41 no.2
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• pp.177-188
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• 2001

Mastitis is one of the most significant cause of economic loss to the dairy industry. Especially, Staphylococcus aureus is a major contagious mastitis-causing pathogen in dairy cattle. Because of its high transmission rate and resistance to antibiotic therapy, staphylococcal mastitis presents a constant threat to the dairy industry. Staphylococcal enterotoxin C(SEC) produced by S aureus has been known as one of superantigens which are able to stimulate a large proportion of T lymphocytes independently of their antigenic specificity. In this experiment, we have conducted preliminary studies with mice and lactating cows to evaluate the immunogenicity and safety of the experimental vaccine consists of SEC mutant antigen on controlling the bovine mastitis associated with S aureus infections. The average value of somatic cell counts in quarter milk, isolation rate of S aureus were consistently decreased in SEC-SER vaccinated groups, whereas antibody titers were highly increased in SEC-SER vaccinated groups. Peripheral blood were also collected from the lactating cows to determine the proportion of leukocyte subpopulation associated with humoral immunity(HI) and cell mediated immunity(CMI). Proportion of leukocyte subpopulation expressing $BoCD2^+$(total T lymphocyte), $BoCD4^+$(T helper cell), $BoCD8^+$(T cytotoxic/suppressor cell) and NonT/NonB lymphocyte which are involved in CMI in SEC-SER vaccinated groups were decreased for the initial stage after first vaccination and then increased from ten weeks after first vaccination maintaining elevated level till 14 weeks after vaccination. In contrast, proportion of monocyte, MHC class II and B lymphocyte which are associated with the production of primary immune response in SEC-SER vaccinated groups were increased for the initial period and then decreased from ten weeks after first vaccination. We present evidence that vaccination of SEC-SER mutant antigen in lactating cows induced a significant proliferation of bovine T lymphocytes. These results suggest that SEC-SER mutant antigen used in this experiment might be one of potential immunogen in developing innovative vaccine against bovine IMI associated with S aureus. Additional challenge trials should be carried out to evaluate substantial protection against S aureus under the commercial farm conditions.

• Korean Journal of Veterinary Service
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• v.46 no.1
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• pp.35-45
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• 2023

Pathogenic Escherichia coli is the cause of a wide range of diseases in pigs, including diarrhea, edema disease, and septicemia. Diarrhea caused E. coli may result in significant economic losses, making pathogenic E. coli an important pathogen for the swine industry. This study investigated the prevalence of virulence factor genes, antimicrobial resistance phenotypes, and resistance genes in E. coli isolated from feces of piglets in Korea between 2017 and 2020. As a result, 119 pathogenic E. coli isolates were obtained from 601 fecal samples. The F4 adhesin gene and the STb enterotoxin gene were commonly present in E. coli isolated from diarrhea samples. The dominant virulotypes of isolates from diarrhea samples were STb, Stx2e, and F4:LT:STb. More than 80% of the screened isolates were resistant to ampicillin, sulfisoxazole, chloramphenicol, or tetracycline. To confirm the resistance mechanisms for β-lactam or quinolone, we investigated the genotypic factors of resistance. Each of the ceftiofur-resistant E. coli produced an extended-spectrum β-lactamase encoded by bla_CTX-M-14, bla_CTX-M-27, and bla_CTX-M-55. And all ciprofloxacin-resistant E. coli harbored mutations in quinoloneresistance-determining-regions. In addition, some of the ciprofloxacin-resistant E. coli contained the plasmid-mediated-quinolone-resistance genes such as qepA, qnrB1, or qnrD. This study has confirmed that the F4 fimbria and the STb enterotoxin are the most predominant in pathogenic E. coli isolated from piglets with diarrhea in Korea and there is a great need for responsible and prudent use of antimicrobials to treat colibacillosis.

• Journal of Microbiology
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• v.44 no.3
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• pp.293-300
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• 2006

The Escherichia coli heat-labile enterotoxin B subunit (HLT-B) is one of the most powerful mucosal immunogens and known mucosal adjuvants. However, the induction of high levels of HLT-B expression in E. coli has proven a difficult proposition. Therefore, in this study, the HLT-B gene was cloned from pathogenic E. coli and expressed as a fusion protein with GST (glutathion S-transferase) in E. coli BL2l (DE3), in an attempt to harvest a large quantity of soluble HLT-B. The culture conditions, including the culture media used, temperature, pH and the presence of lactose as an inducer, were all optimized in order to obtain an increase in the expression of soluble GST-rHLT-B. The biological activity of the purified rHLT-B was assayed in a series of GMI-ELISA experiments. The findings of these trials indicated that the yield of soluble recombinant GST-rHLT-B could be increased by up to 3-fold, as compared with that seen prior to the optimization, and that lactose was a more efficient alternative inducer than IPTG. The production of rHLT-B, at 92 % purity, reached an optimal level of 96 mg/l in a 3.7 L fermentor. The specific GM1 binding ability of the purified rHLT-B was determined to be almost identical to that of standard CTB.

• Journal of Dairy Science and Biotechnology
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• v.32 no.2
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• pp.101-104
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• 2014

Food safety is a global health goal, and food-borne disease are a significant public health threat throughout the world. Dairy products are susceptible to contamination through a wide variety of physical, microbial, and chemical hazards. Risks of microbiological hazards are of immediate and serious concern to human health. Milk was inoculated with Bacillus cereus and stored at 10, 15, 20, and $30^{\circ}C$ for 7 days. We monitored the effect of the temperature on growth rate and variance of toxin production. The growth rate of B. cereus was suppressed in low temperature. We confirmed that the growth rate and the toxin production were accelerated when the storage temperature was increased. B. cereus began to produce toxins when the number of bacteria was higher than $10^7CFU/mL$. Therefore, managing the storage temperature of milk is important to inhibit the growth and the toxin production of B. cereus.

• Food Science of Animal Resources
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• v.31 no.1
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• pp.40-46
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• 2011

The growth profile of Bacillus cereus in ready-to-eat (RTE) food products of animal origin was examined under different temperature and incubation conditions. In sandwiches and Kimbab, B. cereus did not grow or exhibited only minimal growth at 4 and $10^{\circ}C$, but it grew rapidly at ambient temperature. In sandwiches, B. cereus did not grow efficiently at $25^{\circ}C$, however, in ham, the main ingredient of sandwiches, B. cereus growth was observed at the same temperature, with bacterial levels reaching 7.94 Log CFU/g after incubation for 24 h at $25^{\circ}C$. Toxigenicity of B. cereus was observed only at temperatures above $25^{\circ}C$. In Kimbab, B. cereus produced toxin after 9 h at $30^{\circ}C$ and after 12 h at $25^{\circ}C$. Ingredients of sandwiches and Kimbab were collected from 3 different Korean food-processing companies to investigate the source of contamination by B. cereus. Among the 13 tested food items, 6 items including ham were found to be contaminated with B. cereus. Of these ingredients, B. cereus isolates from 3 items produced enterotoxins. None of these isolates harbored the emetic toxin-producing gene. The findings of the present study can be used for risk assessments of food products, including ham and cheese, contaminated with B. cereus.

• Korean Journal of Food Science and Technology
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• v.36 no.1
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• pp.44-49
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• 2004

Changes in quality properties of Kochujang prepared with Paecilomyces japonica powder and extract using different solvents were investigated during 90 days of fermentation at $20^{\circ}C$. Although moisture contents were not significantly different, pH of P. japonica-added Kochujang was lower than that of control group without P. japonica, and decreased with increasing fermentation time. Amino nitrogen content increased up to 60 days of fermentation and decreased slightly after 90 days, with that of P. japonica-added Kochujang showing highest on 30 and 60 days at 179.2 and 282.2 mg%, respectively, higher than control gruup. L, a, and b values decreased in proportion to fermentation period, with P. japonica-added Kochujang, particularly P. japonica powder-added Kochujang, lower than those of control g개up. Sensory evaluation test showed color of control group was 'clear red', whereas that of P. japonica powder-added Kochujang was 'dark reddish brown' and P. japonica extract-added Kochujang was darker than control group; consumer preference for dark color was low, Textures of all samples were 'glossy and smooth', showing high consumer preference. Salt content of P. japonica-added Kochujang was higher than that of control group, with P. japonica extract-added Kochujang higher than that made with powder Hot taste or P. japonica-added Kochujang was weaker, whereas its flavor higher, than control group, with P. japonica powder-added Kochujang showing highest flavor score. Overall preference was higher for P. japonica-added Kochujang than control group, with P. japonica water extract-added Kochujang showing the highest score.

• Journal of Food Hygiene and Safety
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• v.28 no.2
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• pp.138-145
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• 2013

This study was conducted to evaluate the microbiological contamination on commonly used hand towels in the child care centers and to investigate the toxin gene and toxin production ability of food-borne pathogens. A total of 22 commonly used hand towels including 7 for before use and 15 for during use were tested. The average number of total aerobic bacteria and fungi were 6.2 log CFU/100 $cm^2$ and 4.1 log CFU/100 $cm^2$. Coliform bacteria were detected in 4 out of 7 before used towels (57.1%) and all of during used towels (100%). These results showed that the sanitary conditions of hand towels in the child care centers should be improved promptly. Among the pathogenic bacteria, Staph. aureus and B. cereus without Salmonella spp. were detected in 5 (22.7%) and 11 (50.0%) out of 22 hand towels. All of Staphy. aureus isolated in this study did not possess any toxin genes and did not produce enterotoxin. The detection rate of hblC, hblD, and hblA toxin genes in B. cereus was 72.7, 72.7, and 54.5% respectively. The possession rate of nheA, nheB, and nheC toxin genes showed 81.8, 72.7, and 54.5% respectively. The cytK and entFM toxin genes were presented at 45.5 and 90.0% in B. cereus. The HBL was detected in 8 out of 11 B. cereus isolates (72.7%) and 5 B. cereus isolates produced NHE (45.5%). Ten out of eleven B. cereus isolates (90.9%) produced one or more enterotoxin such as HBL and NHE. From the results, using a private hand towel or paper towel is required to prevent the cross-contamination between commonly used hand towel and children's hands in the child care center.

• Korean Journal of Food Science and Technology
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• v.53 no.6
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• pp.799-808
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• 2021

This study was performed to investigate the extent of microbial contamination, the presence of enterotoxin genes, and the antibiotic susceptibility of Bacillus cereus in 58 red pepper plants and 43 environmental samples (soil, irrigation water, and gloves) associated with the plant cultivation. The detected counts of total aerobic bacteria, coliform bacteria, Escherichia coli, Bacillus cereus, and Staphylococcus aureus were lower in these samples, as compared to the regulations of standards for foods; moreover, pathogens, such as E. coli, E. coli O157:H7, Listeria monocytogenes, and Salmonella spp., were not detected. Genes encoding hemolysin BL enterotoxins (hblA, hblC, and hblD) as well as non-hemolytic enterotoxins (nheA, nheB, and nheC) were detected in 23 B. cereus specimens that were isolated from the test samples and had β-hemolytic activity. Interestingly, B. cereus is resistant to β-lactam and susceptible to non-β-lactam antibiotics. However, in this case, the isolated B. cereus specimens exhibited a shift from resistant to intermediate in response to cefotaxime and from susceptible to intermediate in case of rifampin, trimethoprim-sulfamethoxazole, vancomycin, clindamycin, and erythromycin. Therefore, the levels of B. cereus should be monitored to detect changes in antibiotic susceptibility and guarantee their safety.