• Title/Summary/Keyword: staphylococcal

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Complete DNA Sequence and Analysis of a Cryptic Plasmid Isolated from Lactobacillus bifermentans in Kimchi

  • Jeon, Deok-Young;Lee, Sun-Ho
    • Journal of Microbiology and Biotechnology
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    • v.13 no.6
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    • pp.1018-1020
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    • 2003
  • The complete 1,486 nucleotide sequence of a cryptic plasmid separated from Lactobacillus bifermentans strain A02 isolated from Kimchi has been determined. The plasmid, designated as pA021, encodes a 33,488 Da putative Rep protein. Based on the sequence similarity, the protein shows homology with coding protein of pRS1, a previously reported plasmid of Oenococcus oeni and the replication initiation protein (Rep) of the Staphylococcal pT181 plasmid family.

Export of Human Proinsulin in E. coli : High Export of Proinsulin Fusion Protein but not of Proinsulin Itself (대장균에서 인체 프로인슐린의 분비 발현 : 프로인슐린 융합체의 고분비 발현과 프로인슐린의 저분비 발현)

  • Yup Kang
    • KSBB Journal
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    • v.11 no.2
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    • pp.165-172
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    • 1996
  • To obtain a correctly folded human proinsulin, export of proinsulin using Staphylococcal protein A signal sequence-mediated secretion pathway has been attempted in E.coli. A secretion operon for proinsulin was constructed by consecutively connecting T7 promoter, SPA ribosome binding site, SPA signal sequence gene, and human proinsulin gene. Little immunoreactive proinsulin was detected in the periplasmic space and. culture medium, and not even in cytoplasmic space. The qualitative analysis of transcribed proinsulin mRNA and the in vitro transcription/translation experiment suggests that the negligible level of proinsulin export appears to be due to intracellular degradation of proinsulin, rather than due to the blockage during translocation. However, expression of proinsulin fusion protein such as MBP-proinsulin could dramatically increase export of proinsulin in E.coli.

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Replication origin (ori) of R-plasmid pSBK203 Isolated from Staphylococcus aureus DHI (Staphylococcus aureus DH1에서 분리한 R-plasmid pSBK203상의 복제개시 부위 ori에 관한 연구)

  • Min, Kyung-Il;Byeon, Woo-Hyeon
    • Korean Journal of Microbiology
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    • v.32 no.3
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    • pp.186-191
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    • 1994
  • The origin of the leading strand replication (ori) and of lagging strand replication (M-O) of R-plasmid pSBK203 was identified and its base sequence was determined. About 50 bp of ori sequence residues overlapped with the structural gene of rep. Sequence comparison reveals that pSBK-ori shares obvious identities with those of pT181 family and consists of two regions, one is conserved and the other is variable region. Of two palindrome sequence located one after another in upstream region of rep gene, palA' instead of palA which shares sequence homology with diverse family of plasmids such as pOX6, pC194, and pE194 seems to act as a signal for conversion of primarily replicated ssDNA to dsDNA (minus origin (M-O)).

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Characteristics and Lytic Activity of Phage-Derived Peptidoglycan Hydrolase, LysSAP8, as a Potent Alternative Biocontrol Agent for Staphylococcus aureus

  • Yu, Jun-Hyeok;Lim, Jeong-A;Chang, Hyun-Joo;Park, Jong-Hyun
    • Journal of Microbiology and Biotechnology
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    • v.29 no.12
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    • pp.1916-1924
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    • 2019
  • Outbreaks of staphylococcal food poisoning (SFP) causing serious human diseases and economic losses have been reported globally. Furthermore, the spread of Staphylococcus aureus with increased resistance to multiple antimicrobial agents has become a major concern in the food industries and medicine. Here, we isolated an endolysin LysSAP8, as one of the peptidoglycan hydrolases, derived from the bacteriophage SAP8 infecting S. aureus. This endolysin was tagged with a 6×His at the C-terminal of the target protein and purified using affinity chromatography. LysSAP8 demonstrated lytic activity against a broad spectrum of bacteria, which included a majority of the staphylococcal strains tested in this study as well as the methicillin-resistant S. aureus (MRSA); however, no such activity was observed against other gram-positive or gram-negative bacteria. Additionally, LysSAP8 could maintain bactericidal activity until 0.1 nM working concentration and after heat treatment at 37℃ for 30 min. The ability of LysSAP8 to lyse cells under varying conditions of temperature (4-43℃), pH (3-9), and NaCl concentrations (0-1,000 mM), and divalent metal ions (Ca2+, Co2+, Cu2+, Mg2+, Mn2+, Hg2+, and Zn2+) was examined. At the optimized condition, LysSAP8 could disrupt approximately 3.46 log CFU/ml of the planktonic cells in their exponential phase of growth within 30 min. In this study, we have suggested that LysSAP8 could be a potent alternative as a biocontrol agent that can be used to combat MRSA.

Studies on Chronic Mastitis of Dairy Cattle in Kyungnam Province (경남지방(慶南地方)에서 발생(發生)한 젖소의 만성(慢性) 유방염(乳房炎)에 대한 연구(硏究))

  • Kim, Bong-Hwan;Kim, Jin-Koo;Choe, Sang-Yong
    • Korean Journal of Veterinary Research
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    • v.23 no.2
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    • pp.205-209
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    • 1983
  • Some investigations on chronic mastitis in dairy cattle in Kyungnam Province during the year 1982 were conducted with the special reference to the causative agents and their drug resistance. Milk samples from 46 isolated cases of chronic mastitis cattle were investigated bacteriologically and the organisms recovered were examined for their drug susceptibility against the major antibiotics used in this country by the use of disk diffusion susceptibility test. Four major causative agents involved in chronic mastitis were in order of prevalence Staphylococcus aureus (32.6%), Escherichia coli (28.3%), Streptococcus agalactiae (8.7%) and Candida albicans (8.7%). Staph. epidermidis, Streptococcus uberis, Klebsiella pneumoniae and Candida subtropicalis were found to be one of the minor agents. The majority of staphylococcal isolates were highly resistant to the most of antibiotics employed while 8% of them were resistant to gentamicin and 32% to chloramphenicol. The percentages of staphylococcal cultures resistant to penicillin, lincomycin. streptomycin, methicillin, oleandomycin, tetracycline, cephalothin, ampicillin and erythromycin were 100%, 96%, 96%, 92%, 84%, 84%, 80%, 76%, and 64% respectively. Streptococcal isolates were also highly resistant to the majority of the drugs used although 85.7% of them were susceptible to gentamicin. All Escherichia coli isolates were found to be resistant to erythromycin, lincomycin and penicillin while the majority of them were resistant to ampicillin (92.9%), carbenicillin (85.7%), oleandomycin (85.7%), streptomycin(85.7%), kanamycin (78.6%), methicillin (78.6%) and tetracycline (71.4%). The percentages of E. coli cultures resistant to gentamicin, nitrofurantoin, cephalothin and chloramphenicol were 21.4%, 21.4%, 35.7% and 50.0% respectively.

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An Improved Method for Detection of Salmonella Typhi O Antigen with Staphylococcal Protein A Using Enzyme Immunoassay (포도구균의 A단백질을 이용한 효소면역법으로 살모넬라 O항원 검출)

  • Rhyu, Mun-Gan;Kim, Gum-Ryong;Lee, Choong-Ki
    • The Journal of the Korean Society for Microbiology
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    • v.22 no.4
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    • pp.445-451
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    • 1987
  • Coagglutination method is widely used for the diagnosis of Salmonella infection. This test, however, has a disadvantage of false positive reaction due to the coagglutination of staphylococci with non-specific immune complexes or anti-staphylococci antibody in serum. Salmonell O antigen was detected by enzyme immunoassay with protein A-bearing Staphylococcus aureus as in the solid phase. Horse radish peroxidase was labeled to IgG specific against Salmonella O antigen. This enzyme immunoassay was much more sensitive than conventional coagglutination method without false poitive agglutination. To improve the sensitivity for detection of Salmonella O antigen in samples, we tried to determine the optimal concentration of normal IgG that inhibits non-specific binding of horse radish peroxidase labeled IgG to staphylococci, and to establish the optimal condition of reaction between antigen-antibody complex and staphylococci. Non-specific binding of horse radish peroxidase labeled specific IgG to staphylococci was almost blocked when the enzyme labeled IgG was 500-fold diluted with phosphate buffered saline containing 2mg/ml of normal IgG. When staphylococci coated with antibody to Salmonella O antigen were mixed with antigen-antibody complex and then incubated for 1 hour at room temperature, the minimal detectable concentration of Salmonella O antigen was 1ng/ml. The sensitivity of enzyme immunoassay was 100-fold greater than a conventional coagglutination method. This enzyme immunoassay could be expected as an improved method for detection of other infectious agents.

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Assessment of Bacterial Contaminations of the Surrounding Environment which Influences to Health (건강에 영향을 주는 주변환경의 미생물 오염 실태)

  • Kim, Mi-Jin;Lee, Do-Kyung;Jang, Seok;Kim, Jung-Rae;An, Hyang-Mi;Baek, Eun-Hye;Lee, Kang-Oh;Ha, Nam-Joo
    • YAKHAK HOEJI
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    • v.53 no.4
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    • pp.194-200
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    • 2009
  • Community-acquired antimicrobial resistant bacteria are an emerging problem whole world-wide. Generally, Hands are main mediator of pathogen transmission as compared with other body parts. So, the purpose of this research was to investigate the microbiological quality and antimicrobial susceptibilities of samples which were collected from the door knob and surface in public- and home-facilities, and also hand. Of total 191 samples, 71 samples (37%) were shown to be of high-level total bacterial count (>$10^8\;CFU/cm^2$). And presence of Staphylococcus and Enteric bacilli was observed in 61 samples (32%) and 76 samples (40%), respectively. Antimicrobial susceptibilities of staphylococcal isolates from the samples were tested for six different antimicrobial agents, which are in wide spread clinical use in Korea, as well as four new antimicrobials, daptomycin, linezolid, quinupristin/dalfopristin and tigecycline. Among staphylococcal isolates, antimicrobial resistance were observed in oxacillin (n=6), mupirocin (n=7), vancomycin (n=1), quinupristin/dalfopristin (n=2) and gentamicin (n=5). Fortunately, all the isolates were susceptible to new antimicrobial such as daptomycin, linezolid and tigecycline. Furthermore 4 Enterococcus faecalis were isolated from four hand samples, and all these isolates exhibited multidrug resistance to four different antimicrobial (oxacillin, quinupristin/dalfopristin, mupirocin, gentamicin). Also, 5 Escherichia coli were isolated from surface in home (n=3), door knob in public facilities (n=1) and hand (n=1). Escherichia coli isolated from hand was high-level resistant to tigecycline ($128{\mu}g$/ml) and gentamicin ($64{\mu}g$/ml).

Assessment of Bacterial Contamination and Antibiotic Resistance Patterns in Ready-To-Eat Side Dishes in Seoul Area, 2007 (2007년 서울 지역에서 판매되고 있는 반찬류의 세균 오염과 항생제 내성실태)

  • Kim, Mi-Jin;Lee, Do-Kyung;Jang, Seok;Yang, Hwan-Jin;An, Hyang-Mi;Baek, Eun-Hye;Ha, Nam-Joo
    • YAKHAK HOEJI
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    • v.52 no.6
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    • pp.434-440
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    • 2008
  • The purpose of this research was to investigate the microbiological quality and antimicrobial susceptibilities of the ready-to-eat side dishes, which are were collected from the grocery stores, big markets, and department stores in Seoul throughout November, 2007. Of total 124 samples, presence of staphylococci and enteric bacilli was observed in 38 samples (31%) and 53 samples (43%), respectively. And 30 samples (24%) were shown to be of unsatisfactory quality for total bacterial count (>$10^5$). Antimicrobial susceptibilities of the staphylococci isolated from the side dishes were tested for six different antimicrobial agents, which are in widespread clinical use in Korea, as well as four new antimicrobials, daptomycin, linezolid, quinupristin/dalfopristin and tigecycline. As a result, the staphylococcal isolates were found to be resistant to oxacillin (${MIC}_{90}$, >128 ${\mu}g$/ml), teicoplanin (${MIC}_{90}$, >128 ${\mu}g$/ml), mupirocin (${MIC}_{90}$, >128 ${\mu}g$/ml), linezolid (${MIC}_{90}$, 128 ${\mu}g$/ ml) and quinupristin/dalfopristin (${MIC}_{90}$, 32 ${\mu}g$/ml). Especially, some of the staphylococcal isolates exhibited high level and multi-drug resistance. Moreover, these bacteria were also resistant to new antimicrobials, except tigecycline.

Isolation and identification of culturable bacteria from human skin (배양가능한 피부세균의 분리 및 동정)

  • Bae, Young-Min
    • Journal of the Korean Applied Science and Technology
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    • v.37 no.6
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    • pp.1698-1705
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    • 2020
  • Bacteria were collected from the thumb surface of the twenty young adults that are 20 to 25 years old and cultured on the Luria-Bertani agar. The 16S rDNA of the cultured bacteria was amplified by polymerase chain reaction(PCR) and DNA sequence of the PCR products analyzed. Total 14 different bacterial species were identified by comparing their 16S rDNA sequence with the data in genbank. It appears that each individual has 2.5 different bacterial species in average. Staphylococcal species were the most abundant among the identified bacteria and Micrococcus luteus was the second. Staphylococcal species were isolated at similar frequency between male and female donors but Micrococcus luteus was isolated more frequently from female than male donors. The result obtained in this study might be useful in research of dermatic diseases, searching for new drugs for those diseases and development of new cosmetics.

Detection and characterization of potential virulence determinants in Staphylococcus pseudintermedius and S. schleiferi strains isolated from canine otitis externa in Korea

  • Gi Yong Lee;Soo In Lee;Ji Heon Park;Sun Do Kim;Geun-Bae Kim;Soo-Jin Yang
    • Journal of Veterinary Science
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    • v.24 no.6
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    • pp.85.1-85.13
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    • 2023
  • Background: A recent increase in the occurrence of canine skin and soft tissue infections, including otitis externa and pyoderma, caused by antimicrobial-resistant Staphylococcus pseudintermedius and S. schleiferi has become a significant public and veterinary health issues. Objective: We investigated the virulence potentials associated with the occurrence of canine otitis externa in S. pseudintermedius and S. schleiferi. Methods: In this study, the prevalence of genes encoding leukocidins, exfoliative toxins, and staphylococcal enterotoxins (SEs) was investigated using previously characterized S. pseudintermedius (n = 26) and S. schleiferi (n = 19) isolates derived from canine otitis externa. Susceptibility to cathelicidins (K9CATH and PMAP-36) and hydrogen peroxide (H2O2) was also examined in both staphylococcal species. Results: A high prevalence of genes encoding leukocidins (lukS/F-I, lukS1/F1-S, and lukS2/F2-S), exfoliative toxins (siet, expB, and sset), and SEs was identified in both S. pseudintermedius and S. schleiferi isolates. Notably, S. pseudintermedius isolates possessed higher number of SE genes, especially newer SE genes, than S. schleiferi isolates harboring egc clusters. Although no significant differences in susceptibility to K9CATH and H2O2 were observed between the two isolate groups, S. pseudintermedius isolates exhibited enhanced resistance to PMAP-36 compared to S. schleiferi isolates. Conclusions: These findings suggest that high a prevalence of various toxin genes together with enhanced resistance to cathelicidins may contribute to the pathogenicity of S. pseudintermedius and S. schleiferi in canine cutaneous infections.