• Title, Summary, Keyword: enterotoxin A

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Simultaneous Purification of Enterotoxin A and C by Fast Protein Liquid Chromatography (FPLC에 의한 Staphylococcal Enterotoxin A와 C의 동시분리)

  • Lee, Jung-Hee;Kim, Jong-Bae;Shin, Heuyn-Kil
    • Korean Journal of Food Science and Technology
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    • v.20 no.6
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    • pp.856-861
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    • 1988
  • A new method developed for simultaneous purification of enterotoxin A and C from Staphylococcus aureus strain L 350/1 consisted of chromatography on carboxymethyl (CM)-cellulose using a buffer of variable pH, gel filtration on Ultro gel, and fast protein liquid chromatography(FPLC) using a buffer of variable pH. The enterotoxin A and C were purified by three steps: batchwise adsorption from culture supernatant on Amberlite CG-50; chromatography on CM-cellulose using a buffer of constant pH and molarity; and gel filtration on Sephadex G-75. The purified enterotoxin appeared homogeneous by gel diffusion and polyacrylamide gel electrophoresis. Upon treatment with CM-cellulose using a elution of variable pH, enterotoxin A and C were so close that they were not separated completely. After elution from gels, the enterotoxins appeared as a single peak at the same position. Gel filtration gave a reaction of complete identity to enterotoxin A and C in Ouchterlony immunodiffusion. In FPLC using a CM-cellulose, enterotoxin A and C were simultaneously separated at pH 8.6 and 6.8. When each fraction was performed to gel immunodiffusion, at peak of enterotoxin A and C were not detected each other. In a method of elution by pH-gradient was to be more efficient as a simultaneous separation method in terms of speed, yields and simplicity. The purified toxin A and C were identical to type A and C reference enterotoxin on both disc electrophoresis and Ouchterlony gel diffusion.

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Comparison of Sensitivity for Detection of Heat-Labile Enterotoxin of Enterotoxienic Escherichia coli(EC 81) and Enterotoxin of Enterotoxigenic Clostridium perforngens type A (NCPC8238) by Means of a Polymerase Chain Reaction Assay (독소원성 대장균(EC81)이 생산하는 이열성장독소와 Clostridium perfringens A형 (NCTC8238)이 생산하는 장독소의 검색을 위한 중합효소 연쇄반응기법의 감도 비교)

  • 정희곤
    • The Korean Journal of Food And Nutrition
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    • v.13 no.1
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    • pp.1-5
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    • 2000
  • Detection for heat-labile enterotoxin(LT) of enterotoxigenic Escherichia coli(ETEC, EC81, O148:H28) and enterotoxin of enterotoxigentic Clostridium perfringents type A(CP, NCTC8238, Hobbs serotype 2) by use of a polymerase chain reaction (PCR) assay were positive reaction, which using LT gene-specific primers of ETEC with a detection limit equivalent from 100ng/${\mu}\ell$ to 1 pg of a DNA fragment of 417-bp in EC81 and enterotoxin gene-specific primers of CP with a detection limit equivalent from 100ng/${\mu}\ell$ to 10pg of a DNA fragment of 364-bp in NCTC8238. Detection for a LT gene of ETEC highly appeared 10-fold sensitivity than an enterotoxin gene of CP.

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A Multiplex PCR Assay for the Detection and Differentiation of Enterotoxin-producing and Emetic Toxin-producing Bacillus cereus Strains

  • Lee, Dae-Sung;Kim, Keun-Sung;Kwon, Ki-Sung;Hong, Kwang-Won
    • 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.

The Effect of Glucono delta Lactone, Starter Clulture and NaCl on the Production of Staphylococcal Enterotoxign A in the Processing of Fermented Sausage (발효 소세지의 숙성 중 Starter Culture, Glucono delta Lactone 및 소금첨가량이 Staphylococcal Enterotoxin의 생성에 미치는 영향)

  • Shin, Heuyn-Kil;Jin, Young-Ku;Lee, Young-Jin;Park, Woo-Moon;Kim, Jong-Bae
    • Korean Journal of Food Science and Technology
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    • v.23 no.2
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    • pp.150-156
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    • 1991
  • This research was conducted to investigate the effect of starter culture(Lactobacillus plantarum), glucono-delta-lactone(GdL), and NaCl on the production of staphylococcal enterotoxin A in the processing of fermented sausages. With the increasing amount of GdL(0, 0.23, 0.50 and 0.75%) added the production of enterotoxin was significantly decreased(p>0.01). Lactobacillus plantarum as starter culture were inoculated at the level of $10^6\;cells/g$. When GdL was not added, the amount of production of enterotoxin in the group with and without the starter culture were 40 and 80 ng/10g, respectively. With the addition of 0.5%, GdL, the maximum amount of enterotoxin produced in the group with and without starter culture were 30 and 50 ng/10g. These results showed the inhibiting effect of starter culture in the production of enterotoxin. When the amount of enterotoxin production was compared with the addition of 2.7 and 1.7% NaCl, the production of enterotoxin was higher at 2.7% NaCl level.

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Virulence Factors of Staphylococcus aureus Isolated from Korean Pork bulgogi: Enterotoxin Production and Antimicrobial Resistance

  • Jung, Byeong Su;Lee, Yong Ju;Lee, Na-Kyoung;Kim, Hyoun Wook;Oh, Mi-Hwa;Paik, Hyun-Dong
    • Food Science of Animal Resources
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    • v.35 no.4
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    • pp.502-506
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    • 2015
  • The aim of this study was to investigate the antimicrobial resistance profiles of and the enterotoxin gene distribution in 4 strains of Staphylococcus aureus (S10-2, S10-3, S12-2, and S13-2) isolated from 90 bulgogi samples. The S. aureus enterotoxin H gene (seh) was found in all the strains, while the S. aureus enterotoxin A gene (sea) was found only in 3 of the 4 strains. The S10-2 strain expressed a combination of enterotoxin genes - seg, seh, sei, sej, selm, and seln. The strains S10-2 and S13-2 were resistant to ampicillin and penicillin G, and all the isolated strains were resistant to tetracycline. The S10-2 strain was the only mecA-positive strain; it was also resistant to β-lactam antibiotics. Thus, genes encoding enterotoxin as well as those conferring antibiotic resistance were identified in the S. aureus strains isolated from pork bulgogi. These results represents the potential occurrence of MRSA in pork bulgogi, and the need for a monitoring system for pork bulgogi in order to prevent an outbreak of staphylococcal food poisoning.

Expression of Enterotoxin Genes in Staphylococcus aureus Isolates Based on mRNA Analysis

  • Lee, Young-Duck;Moon, Bo-Youn;Park, Jong-Hyun;Chang, Hyo-Ihl;Kim, Wang-June
    • Journal of Microbiology and Biotechnology
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    • v.17 no.3
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    • pp.461-467
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    • 2007
  • Staphylococcus aureus strains are important foodborne pathogens that produce various toxins. To evaluate the risk of the enterotoxins, four S. aureus strains from kimbap and two clinical samples were isolated and identified, and their expression of the enterotoxin genes were analyzed using a reverse transcription real-time PCR. Various enterotoxin genes were detected, including sea, seg, seh, sei, sen, seo, and sem, where each isolate contained one or two. When the mRNA detection of the enterotoxin genes was analyzed using a reverse transcriptase PCR, various levels of expression were found depending on the species and enterotoxin gene. Therefore, it is reasonable to suggest that the poisoning risk of S. aureus can be effectively evaluated based on the gene expression at the mRNA level.

Effect of Gamma-Irradiation on the Cell Proliferating and Interleukin-2 Producing Activity of Mouse Splenocytes of Staphylococcal Enterotoxin B (감마선 조사가 Staphylococcal Enterotoxin B의 비장세포 증식률 및 Interleukin-2 분비능에 미치는 영향)

  • Park, Jong-Heum;Sung, Nak-Yun;Byun, Eui-Baek;Song, Du-Sup;Kim, Jae-Kyung;Song, Beom-Seok;Kim, Jae-Hun;Lee, Ju-Woon;Yoo, Young-Choon
    • Journal of Radiation Industry
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    • v.7 no.2_3
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    • pp.161-166
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    • 2013
  • The purpose of this study was to investigate the cell proliferating and interleukin-2 producing activity of staphylococcal enterotoxin B by gamma-irradiation. Staphylococcal enterotoxin B was gamma-irradiated with the various doses of 0, 2, 20 and 50 kGy. SDS-PAGE analysis showed that gamma-irradiation caused the sharp decrease of the content of staphylococcal enterotoxin B and the effect was irradiating dose-dependent. Non-irradiated staphylococcal enterotoxin B increased the cell proliferation of splenocytes isolated from female Balb/c mouse, whereas 2 kGy-irradiated toxin significantly decreased the activity. 20 and 50 kGy-irradiated staphylococcal enterotoxin B was no effect. A similar effect on the interleukin-2 production of mouse splenocytes was observed with non-irradiated and irradiated staphylococcal enterotoxin B. It was considered to be due to the decrease of the antigenicity of staphylococcal enterotoxin B by gamma-irradiation. Therefore, these results suggest that gamma-irradiation can be effective for the decrease of the antigenicity of staphylococcal enterotoxin B as superantigen.

Mechanism of Heat-Libile E. coli Enterotoxin Production (대장균의 이열성장독소 생산기전)

  • Choi, Myoung-Sik;Rhee, Kwang-Ho;Chang, Woo-Hyun;Lee, Seung-Hoon
    • The Journal of the Korean Society for Microbiology
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    • v.17 no.1
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    • pp.35-41
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    • 1982
  • Enterotoxigenk E. coli is one of the major causative agents of the infantile diarrhea and traveler's diarrhea. The heat-labile enterotoxin is thought to be a virulence factor in the pathogenesis of the diarrhea and to be a marker for identification of the enterotoxigenic E. coli from non pathogenic E. coli. Therefore knowledge about the heat-labile enterotoxin is essential not only for understanding the pathogenesis but also for the diagnosis of the diarrhea. However the in-vitro heat-labile enterotoxin production is reported to be greatly affected by the cultural condition. In this regards, this study was designed to know the optimal conditions for the production of the heat-labile enterotoxin by assaying the permeability factor in the 18 hours culture supernatant of E. coli 08K25(B2) H9 and of E. coli 015 H11. Results obtained were summerized as follows: 1. Amounts of heat-labile enterotoxin produced were greater at initial pH 8.5 than at 7.0 of CYES-2 broth culture. However, the bacterial growth itself was more abundant at 7.0 than at 8.5. 2. Heat-labile enterotoxin per unit volume of culture supernatant was greater at shaking culture than at standing culture condition, but ratio of the enterotoxin produced over the unit mass of E. coli calculated was greater at standing culture than shaking culture condition, indicating that the greater yields of the toxin produced at shaking culture was due to increase in E. coli cell mass compared to the standing culture condition: 3. The enterotoxin produced in the lincomycin(128 microgram/ml) supplemented media was 5 or 11 times greater on the basis of enterotoxin per unit mass of E. coli, compared to the lincomycin-non-supplemented media, indicating that lincomycin itself increases the enterotoxin production. 4. Treatment of 18 hours culture of E. coli with polymyxin B(0.2 mg/ml) for 1 hour increased the yields of enterotoxin amounting to 2 or 5 times of the non-treated control cultures.

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Characterization and Enterotoxigenicity of Staphylococcus aureus Isolated form Patient and Healthy Human (환자 및 건강인 유래 Staphylococcus aureus의 특성과 Enterotoxin 산생성)

  • 최홍근;손원근;강호조
    • Journal of Food Hygiene and Safety
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    • v.6 no.2
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    • pp.89-93
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    • 1991
  • The present study was conducted to investigate the prevalence, the biochemical properties and the enterotoxin types of Staphylococcus aureus in healthy human and patient. A total of 61 S. aureus strains were isolated from 142 samples. The prevalence of S. aureus isolated from healthy human and patient were 17.7% and 14.0%, respectively. All the isolates showed the production of coagulase, lecithinase and hemolysis (${\alpha}-hemolysis;\;32.8%,\;{\beta}-hemolysis;\;67.2%$) on sheep blood agar. Coagulase type VII (38.4%) and type 111 (26.0%) were dominant among coagulase types I through VIII. Twenty-four (52.2%) of 46 strains tested produced one or more enterotoxin; enterotoxin A, Band C were produced by 3, 9 and 12 strains, respectively. Enterotoxins were produced by 100% of type 11 strains, 75% of type 111 and 39.1 % of type VII. Commonly, coagulase type II produced enterotoxin B or C, and type VII produce enterotoxin C.

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Molecular typing and detection of enterotoxin by multiplex PCR of Staphylococcus aureus isolated from bovine mastitis (유방염 유즙에서 분리한 포도구균의 분자생물학적 typing과 multiplex PCR을 이용한 장독소의 검출)

  • Kim, Sin;Hong, Hyon-Pyo;Kim, Sang-Yun;Kwon, Heon-Il;Lee, Hee-Moo
    • Korean Journal of Veterinary Service
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    • v.25 no.3
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    • pp.275-283
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    • 2002
  • Forty strains of Staphylococcus aureus were isolated from mastitic milk. As a result of antimicrobial susceptibility test, the strains of S aureus revealed 47.5% were resistant to ampicillin and penicillin, and 7.5% to gentamicin. But 45% of isolates were sensitive to antimicrobial agents tested. In case of enterotoxin production, 56.3% of 16 strains produced enterotoxin D. Two strain of enterotoxin D producers produced both enterotoxin B and D. According to isolation date, 15 representative strains were selected. As a results of pulsed field gel eletrophoresis analysis of the 15 representative strains, 14 strains were identical. Therefore we consider the identical strains of S aureus have caused continuously bovine mastitis in this dairy farm. If autogenous vaccine can be made by the strains, it will work well for the prevention of bovine mastitis caused by S aureus.