• Title/Summary/Keyword: staphylococcal toxin

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Production and partial purification of Staphylococcus aureus alpha toxin

  • Park, Hee-myung;Oh, Tae-ho;Han, Hong-ryul
    • Korean Journal of Veterinary Research
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    • v.39 no.5
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    • pp.1028-1032
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    • 1999
  • Alpha toxin of S aureus has cytolytic activity respectively. This antigen has been received the most attention since it is a major virulence factor in pathogenesis of staphylococcal mastitis. Thus, alpha toxin has been focused as potential candidate of vaccine to minimize mastitis in cows. The purpose of this study was to develop a simple, efficient production and purification methods of sufficient amount of alpha toxin antigen from S aureus. Alpha toxin production measured by hemolytic activity was the highest at 18 hrs postinoculation in yeast extract culture medium supplemented with thiamine, nicotinic acid and casamino acid. Alpha toxin was purified by ammonium sulfate precipitation (65%) and ultrafiltration. Molecular weight of the toxin was 33 kDa in the analysis with SDS-PAGE. Conclusionally, when alpha toxin was included in the vaccine, the optimal harvest time of alpha toxin was at 18 hrs after inoculation in yeast extract medium supplemented with thiamine and nicotinic acid.

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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.

Gene typing of staphylococcal superantigens produced by Staphylococcus aureus isolates from meat (식육유래의 Staphylococcus aureus으로부터 산생되는 staphylococcal superantigens의 유전자형 분석)

  • Yoon, Jang-won;Jung, Suk-chan;Yang, Soo-jin;Jung, Byong-youel;Seo, Keon-suk;Kim, So-hyun;Park, Yong-ho
    • Korean Journal of Veterinary Research
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    • v.38 no.3
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    • pp.553-558
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    • 1998
  • Using the previous established multiplex PCR (mPCR), the presence of six kinds of staphylococcal superantigen (SAg) genes was investigated for nineteen Staphylococcus aureus isolates from the commercialized meat sources. As a result, only one isolate from pork among 19 S aureus isolates (5.3%) was confirmed as a potential SAg producer and harbored sec gene. The results in this study suggest that meat may not be major contagion of staphylococcal food poisoning (SFP) in Korea and that staphylococcal enterotoxin type C may be associated with the disease. Also, the mPCR method in this study can be a useful genotypic method which can overcome the typical disadvantages of conventional antibody-based methods due to antigenic homology, and furthur survey on food-borne S aureus isolates can provide the important epidemiological data for SFP in Korea.

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A Comparison of Adult and Pediatric Methicillin-Resistant Staphylococcus aureus Isolates Collected from Patients at a University Hospital in Korea

  • Park, Jin-Yeol;Jin, Jong-Sook;Kang, Hee-Young;Jeong, Eun-Hee;Lee, Je-Chul;Lee, Yoo-Chul;Seol, Sung-Yong;Cho, Dong-Taek;Kim, Jung-Min
    • Journal of Microbiology
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    • v.45 no.5
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    • pp.447-452
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    • 2007
  • In this study, we compared the phenotypic and genotypic characteristics of 138 MRSA isolates obtained from adult and pediatric patients (adult, 50; children, 88). The resistance rates against gentamicin, clindamycin, and ciprofloxacin were much higher in the adult MRSA isolates than in the pediatric MRSA isolates. The ermC gene, which is responsible for inducible clindamycin resistance, was detected in 52(59.1%) of the 88 pediatric MRSA isolates but in only 5(10.0%) of the 50 adult MRSA isolates. MRSA isolates of clonal type ST5 with an integration of SCCmec type II/II variants was the most predominant clone among the adult isolates, while clonal type ST72 with an integration of SCCmec IV/IVA was the most predominant clone among the pediatric MRSA isolates. Staphylococcal enterotoxin A and toxic shock syndrome toxin-1 were prevalent among the adult MRSA isolates but not among the pediatric MRSA isolates. The results of this study demonstrated remarkable differences between adult and pediatric MRSA isolates in terms of their antimicrobial susceptibility profiles, SCCmec type, multilocus sequence type, staphylococcal toxin genes, and erythromycin resistance genes.

A clinical review of community acquired methicillin resistant staphylococcal scalded skin syndrome (지역사회 획득 메치실린 내성 포도알균에 의한 포도알균 열상 피부 증후군의 임상적 고찰)

  • Heo, Soon Young;Song, Yoon Jeong;Kim, Sung Jun;Park, Sun Young;Kang, Du Cheul;Ma, Sang Hyuk
    • Pediatric Infection and Vaccine
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    • v.14 no.1
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    • pp.83-90
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    • 2007
  • Purpose : Staphylococcal scalded skin syndrome (4S) is a well known disease defined by clinical, microbiological and histological criteria caused by Staphylococcus aureus. This disease is uncommon but has been increasingly recognized. We investigated the clinical features of staphylococcal scalded skin syndrome. Methods : We reviewed retrospectively medical records of 53 patients diagnosis of staphylococcal scalded skin syndrome who were admitted to Changwon Fatima hospital from February 2002 to December 2005. These patients were divided into 3 clinical types; generalized type, intermediate type, abortive type. Age, sex ratio, clinical manifestations, laboratory findings, response to therapy and prognosis were investigated. Result : 1)The mean age of patients was 2.8 years, ranging from 20 days to 7 years. Male-to-female ratio was 1.9:1. 2) By clinical types, 6 patients were in the generalized type (11%), 29 patients in the intermediate type (55%), 18 patients in the abortive type (34%). The coexisting diseases were variable, including conjunctivitis (25 cases), atopic dermatitis (11 cases), otitis media (1 case). On laboratory findings, most of patients didn't have leukocytosis or increased C-reactive protein. 4) A total of fifteen Methicillin Resistant Staphylococcal Aureus (MRSA) strains were isolated from September 2003 through December 2005. Fourteen strains were positive for exfoliative toxin B gene by PCR and negative for enterotoxin, toxic shock syndrome toxin and Panton-Valentine leukocidin genes. 5) The mean duration of admission was 7.3 days. Patients were treated with vancomycin or amoxacillin/clavulanate or ampicillin/sulbactam or cefuroxime without significant sequelaes. Conclusion : Recently, Staphylococcal scalded skin syndrome caused by exfoliative toxin B produced by MRSA in the Changwon area has been increasing.

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Development of cellulose nano beads based a rapid detection kit to detect staphylococcal enterotoxin B

  • Kim, Giyoung;Yoo, Jinyoung;Park, Saetbyeol
    • Korean Journal of Agricultural Science
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    • v.46 no.3
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    • pp.549-557
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    • 2019
  • Staphylococcal enterotoxin is a very common cause of food poisoning. Conventional detection methods for the toxin including enzyme-linked immunosorbent assays (ELISAs), chemiluminescence (ECL), and polymerase chain reaction (PCR) assays require a lot of time, efforts, and expert technicians. Lateral flow strip kits have shown great potential for the rapid detection of foodborne pathogens. The lateral flow strip kit is widely used in clinical settings because it is easy to use, fast, and cost effective. A typical lateral flow strip kit uses colloidal gold to generate a visual signal. However, the lateral flow strip kit based on colloidal gold has limited sensitivity to fulfill food safety regulation requirements. This study was performed to develop a rapid test kit for pathogenic staphylococcal enterotoxin B (SEB) in food samples. The rapid detection kit was fabricated based on a nitrocellulose lateral-flow strip. Cellulose nano beads and SEB antibodies were used as the tag and receptor, respectively, to improve the detection performance. Manually spotted SEB antibody and anti-rabbit antibody on the surface of the nitrocellulose membrane were used as test and control spots, respectively. The feasibility of the rapid test kit to detect SEB in samples was evaluated. The sensitivity of the kit was 10 ng/mL SEB spiked in PBS. Additionally, the rapid test kit could detect 1 ng/mL of SEB in chicken meat extract.

Antimicrobial Activity of Licochalcone E Against Staphylococcus aureus and Its Impact on the Production of Staphylococcal Alpha-Toxin

  • Zhou, Tiezhong;Deng, Xuming;Qiu, Jiazhang
    • Journal of Microbiology and Biotechnology
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    • v.22 no.6
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    • pp.800-805
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    • 2012
  • Licochalcone E was firstly isolated from licorice root in 2005, which belongs to the retrochalcone family. Studies on the biological activities of licochalcone E were in the initial stage. In the study, we demonstrated that licochalcone E has potent antimicrobial property against Staphylococcus aureus. Furthermore, via hemolysis, Western blot, and real-time RT-PCR assays, we have shown that subinhibitory concentrations of licochalcone E dose-dependently reduces the production of ${\alpha}$-toxin in both methicillin-sensitive S. aureus (MSSA) and methicillin-resistant S. aureus (MRSA). The data suggest that licochalcone E may deserve further investigation as a potential therapeutic against S. aureus infections, or the structure of licochalcone E may be used as a basis for chemical synthesis of novel anti-S. aureus compounds.

Inhibitory Effect of Carbamylated Staphylococcal Enterotoxins B on Inflammatory Response in HL-60 Cells

  • Chang, Jeong Hyun
    • Biomedical Science Letters
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    • v.20 no.2
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    • pp.96-102
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    • 2014
  • Staphylococcal enterotoxin B (SEB) is bacterial toxin that induces the activation of immune cells. Because the inhibition of pro-inflammatory effect of SEB can resolve the inflammation, I determined the influence of functional or structural change of SEB on immune cells. The post translational modification of protein occurs through carbamylation. Carbamylation can change the structure of proteins and can modify the biological activity of protein. In the present study, I investigated the effect of carbamylated SEB (CSEB) on the inflammatory response mediated by LPS in HL-60 cells. To determine the anti-inflammatory effect of CSEB, I produced carbamylated SEB using potassium cyanate (KCN) and then examined whether CSEB involved in cytokine releases and apoptosis of LPS-stimulated HL-60 cells. Although CSEB had not any effect on the LPS-stimulated HL-60 cells, the protein levels of IL-8, TNF-${\alpha}$ and IL-$1{\beta}$ were significantly decreased by CSEB without cytotoxicity. CSEB also blocked Akt and NF-${\kappa}B$ activation. These results indicate that the suppressive effect of CSEB in LPS-stimulated cytokine releases is occurred by inhibition of Akt and NF-${\kappa}B$ activity. Through further studies, CSEB may be used as anti-inflammatory molecule that makes the immune system more efficient.

Efficacy of Bovine Staphylococcal Mastitis Vaccine Composed of Alpha toxin, Capsular Polysaccharide and Fibronectin Binding Protein in Lactating Cows and Heifers (비유우와 처녀우에서 황색포도구균의 alpha toxin, capsular polysaccharide와 fibronectin binding protein으로 구성된 유방염 백신의 효능)

  • 한홍율;박희명
    • Journal of Veterinary Clinics
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    • v.17 no.1
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    • pp.152-158
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    • 2000
  • capsular polysaccharide(CPS), alpha toxin과 재조합 fibronectin binding protein (FnBP)으로 구성된 유방염 백신을 개발하여 야외효능실험을 수행하였다. 이 subunit 백신은 비유우 23두와 처녀우 20두를 대상으로 10개월동안 각각 수행되었다. 비유우는 매 3주간격으로 2회 백신접종을 하였으며 대조군은 PBS를 상유방림조절 주위에 피하주사하였다. 처녀우를 대상으로 한 실험에서는 분만 8주전부터 시작하여 매 3주간격으로 2회 비유우 백신접종부위와 동일하게 접종하였다. 접종후 비유우에서는 황색포도상구균에 의한 총유선내 감염율이 대조균에 비해 유의성있게 감소하였으며 (p<0.001) 처녀우에서도 총유선내 감염이 백신 접종균(1.6%)이 대조군(11.7%)에 비해 다음 비유기동안 유의성있게 낮았다. (p<0.001). 비유기동안 백신접종한 젖소의 체세포수는 변화가 없었으며 처녀우에서는 추가접종후 백신접종군에서 대조군의 체세포수에 비해 낮았지만 통계학적으로 유의성은 없었다(p>0.05). 본 실험결과 황색포도구균에 대한 유방염 아단위 백신은 비유우와 처녀우에서 체세포수를 증가시키지 않고 총 유선내 감염율을 낮추어주었다. 하지만 본 실험은 1군데의 목장을 대상으로 하였기 때문에 향후 대규모 목장을 대상으로 하는 유방염 백신의 야외효능실험이 심도있게 이루어져야 한다고 생각된다.

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Detection and Identification of $\beta$-lactamase, Enterotoxin and Other Exotoxins Genes of Staphylococcus aureus by PCR

  • Yoon, Y.H.;Kim, K.I.
    • Asian-Australasian Journal of Animal Sciences
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    • v.16 no.3
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    • pp.425-429
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    • 2003
  • Staphylococcus aureus is a major pathogen for cattle, causing various forms of subclinical and clinical mastitis and could be a causative agent of food poisoning, it produces various superantigenic exotoxins which have a great public health significance. A total of 72 S. aureus clinical isolates from dairy farms located in Kyunggi Province Korea were examined for the species identification by biochemical method, and for the detection of $\beta$-lactamase, enterotoxin and other exotoxins genes by PCR. The results of species identification by biochemical method agreed with those of PCR done with species specific primer STA-AU. $\beta$-lactamase is an enzyme closely associated with the resistance to antibiotic penicillin, which is an important means of treatment of mastitis, all the isolates were positive for the presence of genes encoding $\beta$-lactamase, which were reproduced in penicillin susceptibility disc assay. Six types of toxin genes, Staphylococcal enterotoxin (SE)A, SEB, SEC, SEE, toxic shock syndrome toxin (TSST-1) and exfoliative toxin A (ET A) were detected in 72 isolates by PCR associated genotypic method in this study, none of the isolates carried the genes for enterotoxin D (SED) and exfoliative toxin B (ETB). The occurrence rate of exotoxin genes rated as 12.5%, and the precision of the PCR identification results has been confirmed using the reference strains.