• Title/Summary/Keyword: S aureus

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Amplified fragment length polymorphism fingerprinting analysis of Staphylococcus aureus isolated from bovine mastitis milk (소 유방염 유래 Staphylococcus aureus의 AFLP 지문분석)

  • Kim, Yeon-soo;Kim, Sang-kyun;Hwang, Eui-kyung
    • Korean Journal of Veterinary Research
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    • v.41 no.2
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    • pp.157-165
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    • 2001
  • Amplified fragment length polymorphism(AFLP) technique is based on the polymorphism detection through selective PCR amplification of restriction fragments from digested genomic DNA and thus includes the procedures of the total DNA digestion by endonucleases, ligation of adapters to the ends of the fragments, and following the selective amplification of the restricted DNA fragments. This study were aimed to : (1) determine the genetic variability of S aureus strains, (2) estimate genetic diversity within and among these strains, (3) compare phylogenetic relationships among these strains as genetic markers using AFLP techniques. Genomic DNA was digested with a particular combination of three restriction enzymes with specific recognition sites and the DNA fragments were ligated to restriction specific adapters and amplified using the selective primer combinations. In the S aureus strain, the number of scorable AFLP bands detected per each primer combination varied from 29 to 102, with an average of 61.59 using 27 primer combinations. A total of 1,663 markers were generated, 904 bands of which were polymorphic, showing a 33.48% level of polymorphism with these primer combinations. Among the primer combinations, E02/T02, E02/T03, E04/H02, E02/T01 and E04/H03 primer combinations showed a high level of polymorphism with 0.78, 0.76, 0.74, 0.71 and 0.70, respectively. But T03/H01, E01/T02 and E01/T03 primer combinations showed a low level of polymorphism with 0.38, 0.37 and 0.15, respectively, Therefore, the former primer combinations will be the most effective for AFLP analysis of S aureus. In SA1 sub-types the level of polymorphism of S aureus KCTC 1927 was similar to that of S aureus CU 01(0.825) and higher than those of other strains such as S aureus CU 02 (0.715), S aureus KCTC 2199(0.625), S aureus KCTC 1916(0.607) and S aureus KCTC 1621 (0.553). In SA2 sub-types the level of polymorphism of S aureus CU 07 was similar to that of S aureus CU 08(0.935) and higher than those of both S aureus CU 04(0.883) and S aureus CU 05(0.883) and lower than those of S aureus CU 03(0.583). In SA3 subtypes the level of polymorphism of S aureus CU 11 was similar to that of S aureus CU 12(0.913) and lower than that of S aureus CU 15(0.623). The results proved that AFLP marker analysis of S aureus strain could be used to study the epidemiology of mastitis and in addition, common genotype in geographic region could be useful for the development of an effective vaccine or DNA marker for easy diagnosis of mastitis caused by S aureus infection.

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Inhibitory Effect of Lactobacillus plantarum Extracts on HT-29 Colon Cancer Cell Apoptosis Induced by Staphylococcus aureus and Its Alpha-Toxin

  • Kim, Hangeun;Kim, Hye Sun;Park, Woo Jung;Chung, Dae Kyun
    • Journal of Microbiology and Biotechnology
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    • v.25 no.11
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    • pp.1849-1855
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    • 2015
  • Staphylococcus aureus plays an important role in sepsis, septic shock, pneumonia, and wound infections. Here, we demonstrate that Lactobacillus plantarum extracts inhibited S. aureus-induced cell death of a human epithelial cell line, HT-29. In particular, we have shown that S. aureus-induced cell death was abolished by neutralization of α-toxin, indicating that α-toxin is the major mediator of S. aureus-induced cell death. DNA fragmentation experiment and caspase assay revealed that the S. aureus-induced cell death was apoptosis. L. plantarum extracts inhibited the generation of effector caspase-3 and the initiator caspase-9 in S. aureus- or α-toxin-induced cell death. Moreover, expression of Bcl-2, an anti-apoptotic protein, was activated in L. plantarum extract-treated cells as compared with the S. aureus- or α-toxin-treated only cells. Furthermore, S. aureus-induced apoptosis was efficiently inhibited by lipoteichoic acid and peptidoglycan of L. plantarum. Together, our results suggest that L. plantarum extracts can inhibit the S. aureus-mediated apoptosis, which is associated with S. aureus spreading, in intestinal epithelial cells, and may provide a new therapeutic reagent to treat bacterial infections.

Screening of Inducible Resistance Genes to Macrolide-Lincosamide-Streptogramin B(MLS) Antibiotics (마크로라이드-린코사마이드-스트렙토그라민 B(MLS)계 항생물질에 대한 유도 내성)

  • Kwon, Ae-Ran;Choi, Sung-Sook;Kim, Sook-Kyung;Chung, Young-Ja;Choi, Eung-Chil;Kim, Byoung-Kak
    • YAKHAK HOEJI
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    • v.38 no.3
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    • pp.293-299
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    • 1994
  • Forty nine clinical isolates of S. aureus showing resistance to erythromycin(EM) were selected from 83 strains isolated recently in Korea. Fourteen strains of S. aureus showing inducible resistance to MLS antibiotics were selected by disc agar diffusion method. Colony hydridization was executed using two MLS inducible resistance genes, ermA and ermC, identified previously from S. aureus as probes. S. aureus 375 and S. aureus 507 whose genes were not homologous to those probes were finally selected. It was confirmed that the resistance genes of S. aureus 375 and S. aureus 507 had no homology with those probes in southern hybridization test using ermA, ermC and ermAM as probes. It was determined that S. aureus 375 had a plasmid whose size was about 35 kb. To know if the plasmid may have the genes related to inducible resistance to MLS antibiotics, it was attempted to transform Bacillus subtillis BR151 and S. aureus RN4220 with the plasmid isolated from S. aureus 375. It was shown that the gene related to inducible resistance to MLS antibiotics did not exist in this plasmid. These results indicate that two clinical isolates of S. aureus showing inducible resistance to MLS antibiotics have novel genes that have no homology with MLS resistance genes identified so far. It is assumed that these genes may exist in chromosomal DNA.

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Panton-Valentine Leukocidin Positive Methicillin-Susceptible Staphylococcus aureus: A Case Report of Two Pediatric Patients with Thrombotic Complications

  • Ng, David Chun Ern;Anand, L. Alexis;Khiu, Fu Lung;Tan, Kah Kee
    • Pediatric Infection and Vaccine
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    • v.25 no.1
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    • pp.50-53
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    • 2018
  • Staphylococcus aureus is a well-recognized human pathogen that causes a wide range of infections as a result of its extensive virulence factors. One of these factors is Panton-Valentine leukocidin (PVL), a potent pore-forming cytotoxin that has been linked to invasive S. aureus infections. PVL is one of the important virulence factors for S. aureus and has been largely recognized as one of the markers for community-acquired methicillin-resistant S. aureus. However, the presence of PVL in methicillin-susceptible S. aureus infections is not widely reported in the literature. Thrombotic sequelae of S. aureus infections associated with PVL expression are uncommon in children. We hereby report two children with thrombotic complications associated with PVL-producing methicillin-susceptible S. aureus. Both patients responded well to antibiotic and anticoagulant therapies, and survived without any long-term sequelae.

Characterization of Tetracycline Resistance Plasmids of Staphylococcus aureus (황색포도상구균의 테트라사이클린 내성 플라스미드 동정)

  • Park, Jung-Hee;Lee, Jae-Yoon;Moon, Kyung-Ho
    • YAKHAK HOEJI
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    • v.52 no.4
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    • pp.279-282
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    • 2008
  • Plasmids were isolated from 15 tetracycline (Tc) resistant S. aureus. Two small tetracycline resistance plasmids, pKH16 and pKH17, have been isolated from Staphylococcus aureus JY10 and Staphylococcus aureus JY22, respectively and the complete nucleotide sequences of those plasmids have been determined. pKH16 consisted of 4,442 bp and showed high identity to pKH6 (99% matching percentage) isolated in 1989 from S. aureus SA2. pKH17 consisted of 4,441 bp and showed less identity to pKH6 (95% matching percentage) than pKH16. PCR analysis showed that tetK and tetM did not exist in ten large plasmids isolated from ten Tc resistant S. aureus. Twelve Tc resistant S. aureus showed reistance both to Tc and Mn and we might analogize that twelve Tc resistant S. aureus had tetM in their chromosome.

Antibacterial Activity of Glycyrrhizae Radix against Methicillin-resistant Staphylococcus (감초의 메티실린 내성 황색포도구균에 대한 생육억제 효과)

  • 김미랑;정병무;신정인;윤철호;정지천;서운교
    • The Journal of Korean Medicine
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    • v.23 no.3
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    • pp.223-232
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    • 2002
  • Objectives : Methicillin-resistant Staphylococcus aureus (MRSA) CCARM 3251 and S. aureusKCTC 1928 have been known to be resistant to many kinds of antibiotics. The extract of Glycyrrhizae Radix showed antibacterial activity against MRSA and antibiotics-resistant S. aureus. Methods : We examined the effects of the water-soluble extract and the methanol-soluble extract of Glycyrrhizae Radix on MRSA and antibiotic-resistant S. aureus. The methanolic extract was further fractionated with organic solvents such as hexane, chloroform, and ethyl acetate in that order. Results and Conclusions : The methanol-soluble extract of Glycyrrhizae Radix showed relatively high antibacterial activity against MRSA and antibiotic-resistant S. aureus. However, the water-soluble extract of Glycyrrhizae Radix showed no antibacterial activity against MRSA and antibiotic-resistant S. aureus. Among the fractions tested, the chloroform fraction showed the highest antibacterial activity against MRSA and antibiotic-resistant S. aureus. The methanol-soluble extract of Glycyrrhizae Radix minimal inhibitory concentrations (MICs) against MRSA and antibiotics-resistant S. aureus were $5{\;}mg/m{\ell}$ in both. The methanol-soluble extract of Glycyrrhizae Radix was separated using thin-layer chromatography and detected with UV -detector. Further study should be carried out to identify which effects cell growth inhibition of MRSA and antibiotics-resistant S. aureus.

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Epidemiological Investigation of Staphylococcus aureus Isolated from Bovine Mastitis Based on the Polymorphism of Coagulase Gene (젖소 유방염 유래 Staphylococcus aureus의 Coagulase Gene 유전형 분석에 의한 감염경로 규명)

  • Moon, Jin-San;Lee, Ae-Ri;Lym, Suk-Kyung;Joo, Yi-Seok;Kang, Hyun-Mi;Kim, Jong-Man;Kim, Mal-Nam
    • Korean Journal of Veterinary Research
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    • v.43 no.1
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    • pp.95-102
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    • 2003
  • Because Staphylococcus aureus (S. aureus) has variable number of short sequence repeat region in coagulase gene, it has been used to investigate the relatedness of S. aureus isolates. In this study, we isolated S. aureus strains from 20 dairy farms with bovine mastitis from September 2000 to August 2001. PCR-RFLP analysis of coagulase gene revealed 10 different patterns. Most of the S. aureus isolates showed only one coagulase gene RFLP pattern per farm. However, there were several S. aureus clones spreading between dairy farms. All the farms showed poor management conditions of milking machine and milker, indicating that managements for mastitis control program include use of proper milking matching, premilking sanitation, and segregation in the S. aureus infection herd. Our data suggest that PCR-RFLP analysis of coagulase gene might be applicable for the epidemiological investigations of S. aureus isolated from bovine mastitis cows.

Prevalence of Enterotoxigenic Staphylococcus aureus in Retail Ready-to-eat Korean Kimbab Rolls

  • Yoon, Sun-Kyoung;Kang, Yun-Sook;Sohn, Mun-Gi;Kim, Chang-Min;Park, Ji-Yong
    • Food Science and Biotechnology
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    • v.16 no.4
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    • pp.621-625
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    • 2007
  • Staphylococcus aureus in Korean kimbab rolls was monitored seasonally in 4 major cities of Korea to investigate the risk of S. aureus in a pre-prepared meal. Thirty-five (28.6%) of 105 kimbab rolls purchased in winter were contaminated with S. aureus with an average level of 2.6 log CFU/g. Thirty-six (33.0%) of 109 kimbab rolls purchased in summer and autumn were contained S. aureus with an average level of 2.9 log CFU/g. Kimbab purchased in snack bars showed higher S. aureus contamination rates with the maximum level of 4.7 log CFU/g than that purchased in convenience stores. Of the raw materials in kimbab, uncooked perilla leaf had the highest contamination rate of S. aureus. Less than 50% of S. aureus isolated from kimbab produced enterotoxin and most of the staphylococcal enterotoxin produced by S. aureus in kimbab was type A.

Autolysis of Pseudomonas aeruginosa Quorum-Sensing Mutant Is Suppressed by Staphylococcus aureus through Iron-Dependent Metabolism

  • Shin-Yae Choi;In-Young Chung;Hee-Won Bae;You-Hee Cho
    • Journal of Microbiology and Biotechnology
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    • v.34 no.4
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    • pp.795-803
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    • 2024
  • Microorganisms usually coexist as a multifaceted polymicrobial community in the natural habitats and at mucosal sites of the human body. Two opportunistic human pathogens, Pseudomonas aeruginosa and Staphylococcus aureus commonly coexist in the bacterial infections for hospitalized and/or immunocompromised patients. Here, we observed that autolysis of the P. aeruginosa quorum-sensing (QS) mutant (lasRmvfR) was suppressed by the presence of the S. aureus cells in vitro. The QS mutant still displayed killing against S. aureus cells, suggesting the link between the S. aureus-killing activity and the autolysis suppression. Independent screens of the P. aeruginosa transposon mutants defective in the S. aureus-killing and the S. aureus transposon mutants devoid of the autolysis suppression revealed the genetic link between both phenotypes, suggesting that the iron-dependent metabolism involving S. aureus exoproteins might be central to both phenotypes. The autolysis was suppressed by iron treatment as well. These results suggest that the interaction between P. aeruginosa and S. aureus might be governed by mechanisms that necessitate the QS circuitry as well as the metabolism involving the extracellular iron resources during the polymicrobial infections in the human airway.

Anti-Biofilm Activity of Cell-Free Supernatant of Saccharomyces cerevisiae against Staphylococcus aureus

  • Kim, Yeon Jin;Yu, Hwan Hee;Park, Yeong Jin;Lee, Na-Kyoung;Paik, Hyun-Dong
    • Journal of Microbiology and Biotechnology
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    • v.30 no.12
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    • pp.1854-1861
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    • 2020
  • Staphylococcus aureus is one of the most common microorganisms and causes foodborne diseases. In particular, biofilm-forming S. aureus is more resistant to antimicrobial agents and sanitizing treatments than planktonic cells. Therefore, this study aimed to investigate the anti-biofilm effects of cell-free supernatant (CFS) of Saccharomyces cerevisiae isolated from cucumber jangajji compared to grapefruit seed extract (GSE). CFS and GSE inhibited and degraded S. aureus biofilms. The adhesion ability, auto-aggregation, and exopolysaccharide production of CFS-treated S. aureus, compared to those of the control, were significantly decreased. Moreover, biofilm-related gene expression was altered upon CFS treatment. Scanning electron microscopy images confirmed that CFS exerted anti-biofilm effects against S. aureus. Therefore, these results suggest that S. cerevisiae CFS has anti-biofilm potential against S. aureus strains.