• Title/Summary/Keyword: enterotoxin B

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Evaluation of the Probiotic Potential of Bacillus polyfermenticus CJ6 Isolated from Meju, a Korean Soybean Fermentation Starter

  • Jung, Ji Hye;Lee, Myung Yul;Chang, Hae Choon
    • Journal of Microbiology and Biotechnology
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    • v.22 no.11
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    • pp.1510-1517
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    • 2012
  • To evaluate the probiotic potential of Bacillus polyfermenticus CJ6 isolated from meju, a Korean traditional soybean fermentation starter, its functionality and safety were investigated. B. polyfermenticus CJ6 was sensitive to all antibiotics listed by the European Food Safety Authority. The strain was also non-hemolytic, carried no emetic toxin or enterotoxin genes, and produced no enterotoxins. The resistance of B. polyfermenticus CJ6 vegetative cells and spores to simulated gastrointestinal conditions was high (60-100% survival rate). B. polyfermenticus CJ6 produced high amounts (0.36 g as a purified lyophilized form) of ${\gamma}$-polyglutamic acid (PGA). We speculate that the improved cell viability and the production of ${\gamma}$-PGA have a significant correlation. Adhesion of the strain to Caco-2 and HT-29 cells was weaker than that of the reference strain (Lb. rhamnosus GG), but it was comparable to or stronger than those of reported Bacillus spp. When B. polyfermenticus CJ6 spores were given orally to mice, the number of cells excreted in the feces was 4-fold higher than the original inocula. This suggests the inoculated spores propagated within the intestinal tract of the mice. This idea was confirmed by field emission scanning electron microscopy, which revealed directly that B. polyfermenticus CJ6 cells germinated and adhered within the gastrointestinal tract of mice. Taken together, these findings suggest that B. polyfermenticus CJ6 has probiotic potential for both human consumption and use in animal feeds.

QUANTITATIVE ANALYSIS OF TRANSFORMING GROWTH $FACTOR-{\beta}_1$ IN HUMAN FIBROBLASTS INDUCED WITH STAPHYLOCOCCUS ENTEROTOXIN B AND LIPOPOLYSACCHARIDE (Staphylococcus enterotoxin B와 lipopolysaccharide를 작용시킨 사람 섬유아 세포에서 생성된 Transforming Growth $Factor-{\beta}_1$의 정량적 분석)

  • Lee, Seong-Geun;Kim, Kwang-Hyuk;Kim, Uk-Kyu;Kim, Jong-Ryoul;Chung, In-Kyo;Yang, Dong-Kyu
    • Maxillofacial Plastic and Reconstructive Surgery
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    • v.22 no.2
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    • pp.123-132
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    • 2000
  • $TGF-{\beta}_1$ is a potent chemotactic factor for inflammatory cells and fibroblasts. It also stimulates the celluar source and components of extracellular matrix and the production of proteinase inhibitors. Collectively, these biologic activities lead to the accumulation and stabilization of the nascent matrix, which is vital to infection control. The objective of this study is to investigate production of $TGF-{\beta}$ in vitro fibroblast culture in the presence of Staphylococcus enterotoxin B(SEB) and/or lipopolysaccharide(LPS) and to elucidate the role of $TGF-{\beta}_1$ which may be responsible for infection control. The fibroblasts were originated from gingiva and facial dermis in 26 year-old male patient. In the presence of LPS($0.01{\mu}g$, $0.1{\mu}g$, $1.0{\mu}g$), SEB($0.01{\mu}g$, $0.l{\mu}g$, $1.0{\mu}g$) respectively, $cells(5{\times}10^3ml)$ were cultivated in vitro. At 1, 3, and 5 days after incubation, cells were counted. Also, $cells(2.5{\times}10^5ml)$ were cultivated in EMEM with LPS(0.01, 0.1 and $1.0{\mu}g$), SEB(0.01, 0.1 and $1.0{\mu}g$) respectively and $LPS(0.1{\mu}g)$ and $SEB(0.1{\mu}g)$ in combination for 24, 48, and 72 hours respectively. Culture supernatants were harvested at 1, 2, and 3 days after incubation period and triplicate culture supernatants were pooled and $TGF-{\beta}_1$ was assayed in duplicate. The results were as follows. 1. In gingival fibroblast induced with SEB and LPS respectively or in combination, the suppression of cell Proliferation occurred very significantly since 3 days after incubation, compared with the control and the production of $TGF-{\beta}_1$ occurred very significantly at 1 day after incubation, compared with the control. 2. In facial dermal fibroblast induced with SEB and LPS respectively or in combination, the suppression of cell proliferation occurred very significantly at 1 day after incubation, compared with the control. In SEB exposure, the production of $TGF-{\beta}_1$ was decreased very significantly at 1 day after incubation, compared with the control. However, in LPS, SEB and LPS exposure, the production of $TGF-{\beta}_1$ was increased very significantly at 1 day after incubation, compared with the control. In conclusion, the concentration of bacterial toxins and the incubation period correlated with cell proliferation and production of $TGF-{\beta}_1$ very significantly. The gingival and facial dermal fibroblasts have different phenotype each other The orchestrated understanding of fibroblast proliferation and $TGF-{\beta}_1$ production play an important part in host defense against the bacterial Infection and may prevent tissue necrosis such as necrotizing fasciitis and life-threatening syndrome such as multiple organ failure.

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Analysis of the Diversity of Methicillin Resistant Staphylococcus aureus (MRSA) Strains Isolated from Diarrhea Patients in Busan Area (부산지역 설사환자에서 분리한 MRSA 균주의 다양성 분석)

  • Choi, Seong-Hwa;Park, Eun-Hee;Park, Yon-Koung;Kim, Joung-A;Kim, Nam-Ho;Lee, Young-Suk;Bin, Jae-Hun;Park, Ho-Kuk
    • Journal of Life Science
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    • v.18 no.8
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    • pp.1083-1089
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    • 2008
  • We investigated the molecular epidemiological characteristic of methicillin resistant Staphylococcus aureus (MRSA) strains isolated from stool samples in Busan from 2004 to 2006. Among 142 isolates of S. aureus, 49 isolates (34.5%) were confirmed as MRSA. With the antimicrobial susceptibility tests, 37 isolates (75.5%) showed multiple resistance to more than 10 antibiotics, but all isolates were sensitive to vancomycin. All of MRSA had enterotoxin A in 30.6%, B 4.1%, C 8.3%, D, C/G, A 2.0% and None 51%. PFGE of SmaI-digested chromosomal DNA was performed on 49 sporadic MRSA isolates. Restriction fragment patterns consisted of 8 to 14 fragments ranged in size from 48.5 to 630.5 kbp. We could divided the isolates into 7 groups ($I{\sim}VII$) by analyzing PFGE patterns. Group I subdivided as 2 subgroups and 17 (34.7%) strains belong to the group I. Dendrogram of PFGE patterns showed that MRSA strains in Busan were heterogeneous but we could find out minor homogeneity in hospital.

Prevalence of Staphylococcus aureus in Kimbap (김밥 중 황색포도상구균의 분포조사)

  • 김창민;강윤숙;윤선경;좌승협;이동하;우건조;박영식
    • Journal of Food Hygiene and Safety
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    • v.17 no.1
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    • pp.31-35
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    • 2002
  • According to food poisoning statistic data of 2000, the food poisoning outbreaks have occurred mainly by meat (27.9%), shellfish and its processed products (26.0%), and ready-to eat meals (24.0%) such as Kimbap and packed lunch boxes. The major causative flood poisoning bacteria were Salmonella spp. (35.6%), Staphylococcus aureus(11.3%) and Vibrio parahaemolyticus (3.2%). In this study, we conducted the isolation and enumeration for S. aureus in Kimbap. This monitoring data will be applied to the following study, risk assessment. The Kimbap samples were collected from department stores, convenient stores and snack bars located in Seoul, Busan, Daejeon, and Gwangju. The overall isolation rate of S. aureus from 214 Kimbap samples was 34.1% and the average count was 623 cells. Enterotoxin typing test for isolates showed 42.5%, 4.1% and 2.7% for type, A, B and C, respectively. There was no significant seasonal difference in S. aureus isolation, but the average count in summer(793 cfu/g) was 1.8 times higher than that of winter(446 cfu/g).

Mucosal Immune Response and Adjuvant Activity of Genetically Fused Escherichia coli Heat-Labile Toxin B Subunit

  • Lee, Yung-Gi;Kang, Hyung-Sik;Lee, Cheong-Ho;Paik, Sang-Gi
    • Journal of Microbiology and Biotechnology
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    • v.14 no.3
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    • pp.490-497
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    • 2004
  • Although the E. coli heat-labile enterotoxin B subunit (LTB) is known to be a potent mucosal adjuvant towards co-administrated unrelated antigens and immunoregulator in T-helper 1-type-mediated autoimmune diseases, a more efficient and useful LTB is still required for prospective vaccine adjuvants. To determine whether a novel chimeric LTB subunit would produce an enhanced mucosal adjuvant activity and immune response, a number of LTB subunits were genetically fused with chimeric proteins using the epitope genes of the envelope glycoprotein E2 (gp51-54) from the classical swine fever virus (CSFV). It was found that the total serum immunoglobulin (Ig) levels of BALB/c mice orally immunized with chimeric proteins containing an N-terminal linked LTB subunit (LE1, LE2, and LE3) were higher than those of mice immunized with LTB, E2 epitope, and chimeric proteins that contained a C-terminal linked LTB subunit. In particular, immunization with LE1 markedly increased both the total serum Ig and fecal IgA level compared to immunization with LTB or the E2 epitope. Accordingly, the current results demonstrated that the LTB subunit in a chimeric protein exhibited a strong mucosal adjuvant effect as a carrier molecule, while the chimeric protein containing the LTB subunit stimulated the mucosal immune system by mediating the induction of antigen-specific serum Ig and mucosal IgA. Consequently, an LE1-mediated mucosal response may contribute to the development of effective antidiarrhea vaccine adjuvants.

Distribution and characteristics of Staphylococcus aureus subtypes isolated from dairy herds (젖소 목장에서 분리된 황색포도상구균의 아형 분포와 특성)

  • Yoo, Jong-hyun;Park, Hee-myung;Oh, Tae-ho;Sohn, Dae-ho;Han, Hong-ryul
    • Korean Journal of Veterinary Research
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    • v.39 no.5
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    • pp.995-1005
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    • 1999
  • Staphylococcus aureus is one of most prevalent intramammary pathogens and have characteristics which are not easily eradicated. Recently, to understand the sources and transmission of S aureus, many studies have focused on the subtyping of field isolate. This study was preformed to investigate the distribution pattern and characteristics of the isolates using phenotyping and genotyping. Samples were collected from milk of each udder, cow bodies (perianal region, vagina, tail, udder skin, sole) and environment (floor, liner, milker's hands, water, towel, insect) from 6 herds located in Kyung-gi province. Forty five strains of S aureus were isolated from 3 dairy herds (A, B, C) and were typed by hemolytic pattern, antibiotic resistant pattern, enterotoxin typing and PCR-based DNA fingerprinting. Slime productivity was also compared by each subtype to examine potential infectiousness. Of 45 strains, 41 were isolated from milk samples and 4 were isolated from liners. No strains isolated in the bodies and environment. Forty five strains isolated were classified as 18 subtypes by phenotyping and genotyping. There was common subtype between A and B herd, but the subtype of C herd showed different pattern. Among predominant subtypes, 60% of S aureus strain isolated from A and B herd showed subtype I and 50% of S aureus strain isolated from C herd belonged to subtype VI and X II. Neither somatic cell count (SCC) nor slime production was significantly different between predominant and minor subtypes. In summary, the study revealed that liners play more important roles in the mode of transmission than environmental sources. Several subtypes can be found in a herd, only a few subtype, however, was largely associated with the majority of infection.

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Application of Engineered Zinc Finger Proteins Immobilized on Paramagnetic Beads for Multiplexed Detection of Pathogenic DNA

  • Shim, Jiyoung;Williams, Langley;Kim, Dohyun;Ko, Kisung;Kim, Moon-Soo
    • Journal of Microbiology and Biotechnology
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    • v.31 no.9
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    • pp.1323-1329
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    • 2021
  • Micro-scale magnetic beads are widely used for isolation of proteins, DNA, and cells, leading to the development of in vitro diagnostics. Efficient isolation of target biomolecules is one of the keys to developing a simple and rapid point-of-care diagnostic. A zinc finger protein (ZFP) is a double-stranded (ds) DNA-binding domain, providing a useful scaffold for direct reading of the sequence information. Here, we utilized two engineered ZFPs (Stx2-268 and SEB-435) to detect the Shiga toxin (stx2) gene and the staphylococcal enterotoxin B (seb) gene present in foodborne pathogens, Escherichia coli O157 and Staphylococcus aureus, respectively. Engineered ZFPs are immobilized on a paramagnetic bead as a detection platform to efficiently isolate the target dsDNA-ZFP bound complex. The small paramagnetic beads provide a high surface area to volume ratio, allowing more ZFPs to be immobilized on the beads, which leads to increased target DNA detection. The fluorescence signal was measured upon ZFP binding to fluorophore-labeled target dsDNA. In this study, our system provided a detection limit of ≤ 60 fmol and demonstrated high specificity with multiplexing capability, suggesting a potential for development into a simple and reliable diagnostic for detecting multiple pathogens without target amplification.

Isolation and Growth Pattern of Bacillus cereus from Ready-to-Eat Foods. (즉석조리식품에서의 Bacillus cereus오염실태조사 및 생육 pattern 분석)

  • 김순한;김미경;강민철;손영욱;이창희;김인복;이영자;최수영
    • Journal of Life Science
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    • v.14 no.4
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    • pp.664-669
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    • 2004
  • The contamination of Bacillus cereus was investigated in 240 RTE (ready-to-eat) food samples including 118 seafoods, 82 Korean packaged meals and 40 other RTE foods. Many B. cereus presumptive strains were isolated from the enrichment culture in Tryptic Soy Broth (TSB) added polymyxin, followed by selective culture in Mannitol Egg Yolk Polymyxin (MYP) agar and Gram staining. A total of 36 strains (16 in seafoods, 17 in Korean pack-aged meals and 3 in other RTE foods) were identified as B. cereus by the analysis of 61 biochemical tests of the API 50CHB/20E system test and supplementary tests of $\beta$-hemolysis, rhizoid growth, motility and oxidase activity. The 28 strains out of 36 B. cereus isolates produced diarrhoeal enter-otoxin in Brain Heart Infusion (BHI) broth. All isolates were resistant to ampicillin and penicillin antibiotics, and most of them were susceptible to gentamicin, vancomycin, bacitracin, chloram-phenicol, kanamycin and streptomycin. The growth of B. cereus was affected by environmental temperature and incubation time. Culture with temperature under 1$0^{\circ}C$ effectively restricted the growth of B. cereus.

Association of Specific Immunoglobulin E to Staphylococcal Enterotoxin with Airway Hyperresponsiveness in Asthma Patients

  • Kim, Seong Han;Yang, Seo Yeon;You, Jihong;Lee, Sang Bae;You, Jin;Chang, Yoon Soo;Kim, Hyung Jung;Ahn, Chul Min;Byun, Min Kwang;Park, Hye Jung;Park, Jung-Won
    • Tuberculosis and Respiratory Diseases
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    • v.79 no.4
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    • pp.295-301
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    • 2016
  • Background: Specific immunoglobulin E (IgE) sensitization to staphylococcal enterotoxin (SE) has been recently considered to be related to allergic disease, including asthma. Despite studies on specific IgE (sIgE) to SE and its relationship to asthma diagnosis and severity, the association of sIgE to SE with airway hyperresponsiveness (AHR) remains unclear. Methods: We enrolled 81 asthma patients admitted to the Severance Hospital in Korea from March 1, 2013, to February 28, 2015 and retrospectively reviewed the electronic medical records of the enrolled subjects. The serum levels of sIgE to SE (A/B) of all subjects was measured using the ImmunoCAP 250 (Phadia) system with SE-sIgE positive defined as >0.10 kU/mL. Results: The SE-sIgE level was not significantly correlated with asthma severity (forced expiratory volume in 1 second [$FEV_1$], $FEV_1$/forced vital capacity, sputum eosinophils, and serum eosinophils), whereas the SE-sIgE level in patients with positive AHR ($mean{\pm}standard$ error of the mean, $0.606{\pm}0.273kU/mL$) was significantly higher than that in patients with negative AHR ($0.062{\pm}0.015kU/mL$, p=0.034). In regression analysis, SE sensitization (sIgE to SE ${\geq}0.010kU/mL$) was a significant risk factor for AHR, after adjustment for age, sex, $FEV_1$, and sputum eosinophils (odds ratio, 7.090; 95% confidence interval, 1.180-42.600; p=0.032). Prevalence of SE sensitization was higher in patients with allergic rhinitis and non-atopic asthma patients, as compared to patients without allergic rhinitis and atopic asthma patients, respectively, but without statistical significance. Conclusion: SE sensitization is significantly associated with AHR.

Characteristics of Staphylococcus aureus Isolated from Patients with Diarrhea. (설사 환자에서 분리된 Staphylococcus aureus의 특성)

  • Park Eun Hee;Min Sang Gi;Lee Ju Hyeoun;Park Yon Koung;Jeong Gu Young;Bin Jae Hun
    • Journal of Life Science
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    • v.15 no.4 s.71
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    • pp.647-651
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    • 2005
  • The major causative bacteria of food poisoning were Salmonella spp. $(35.6\%)$, Staphylococcus aureus $(11.3\%)$ and Vibrio parahaemolyticus $(3.2\%)$ in our country. In this study we attempted isolation of S. aureus from stools of patients with diarrhea. Sixty-four strains $(9.1\%)$ were isolated from 704 the stools of patients with diarrhea. The enterotoxin was detected from 29 isolates $(45.3\%)$: 24 isolates $(37.5\%)$, 3 isolates $(4.7\%)$ and 2 isolates $(3.1\%)$ were A, B and C type, respectively. In the antibiotic susceptibility, 63 isolates $(98.4\%)$ were resistant to penicillin, 60 isolates $(93.8\%)$ to ampicillin, 35 isolates $(54.7\%)$ to erythromycin, 32 isolates $(50.0\%)$ to gentamycin, 22 isolates $(34.4\%)$ to tetracycline and 20 isolates $(31.3\%)$ to oxacillin. All of S. aureus isolates were susceptible to chloramphenicol and vancomycin, 20 isolates $(31.3\%)$ were methicillin-resistance S. aureus (MRSA). MRSA isolation rate was higher in male $(35.7\%)$ than female $(26.3\%)$. With the exception of two isolates which were resistant only to penicillin, sixty-one isolates were multiple antibiotic resistance.