• Clinical and Experimental Pediatrics
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• 제46권1호
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• pp.24-32
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• 2003

목 적 : 황색포도구균(S. aureus)은 건강인의 비강이나 피부에 정상 세균총으로 존재하지만, 병원 내 감염과 항생제 내성이 문제가 되고 있다. 본 연구는 신생아의 비강에서 S. aureus을 분리하여 항생제 감수성의 양상과 내성 관련 유전자를 검출하여 내성의 정도와 내성 관련 유전자의 상관관계를 규명해 보고자 하였으며, 병원성 인자의 검출을 실시하고, arbitrarily-primed polymerase chain reaction(AP-PCR)법에 의한 유전자형별을 통하여 역학적 해석을 시도하였다. 방 법 : 2001년 2월에서 5월 사이에 부산대학교병원 신생아 중환자실 및 신생아실에 입원한 28명을 대상으로 이들의 비강에서 총 120회의 nasal swab을 실시하여 51주의 S. aureus를 분리하였다. 분리균에 대한 항생제 감수성 검사, coagulase 검사과형별, plasmid DNA의 양상, mec 관련 유전자의 분포, 장독소의 유전자 및 TSST-1 독소의 유전자 보유 상태, AP-PCR법으로 유전자 형별을 조사하였다. 결 과 : 1) Methicillin 내성이 23주, vancomycin 내성이 6주였고, 3가지 이상의 약제에 대한 내성이 절반 이상을 차지하였다. 2) 42주가 coagulase 양성이었고, 이중 mecA 및 femA 유전 자를 보유한 12주를 대상으로 한 형별 검사에서 8종(coagulase I-VIII형)으로 분류되었다. 3) mecA 유전자가 22주에서, mecR1 유전자가 18주, mecI 유전자가 1주, femA 유전자가 17주에서 발견되었으며, 이들을 4가지의 유전자형으로 분류하였을 때 mecA형이 14주, mecR1형 10주, mecA-mecR1형 7주, mecA-mecR1-mecI형이 1주였다. mecA 및 mecA 관련 유전자와 항생제 다제내성과는 상관관계가 없었다. 4) 16주가 mecA와 femA를 모두 보유하였으며, 이들의 plasmid 양상은 I형이 7주, II형이 6주 그리고 III형이 2주였다. Oxacillin 및 vancomycin에 내성을 보이는 II형과 III형에서는 35.5kb의 plasmid DNA를 보유하였다. 5) 장독소 A, B, D, 및 E의 유전자는 51주 전부에서 검출되지 않았으나 장독소 C의 유전자는 64.7%, TSST-1 유전자는 80.4%가 양성이었다. 6) Coagulase 양성이며 mecA 및 femA 유전자를 보유한 MRSA 12균주에 대한 유전자 형별 검사에서 I형에서 X형으로 분류할 수 있었으며, II형 2주, X형 2주가 동일 유전자형이었고, 나머지는 다른 유전자형이었다. 결 론 : 입원 중인 신생아의 비강 내 S. aureus의 상재률이 아주 높았으며, 이들의 항생제 내성률, 특히 vancomycin의 내성률이 높아 이들에 의한 원내 감염에 대한 특별한 관리가 필요하며, 이들의 병원성 인자 및 관련 유전자에 대한 분석은 원내 감염에 대한 적절한 대책과 효과적인 치료에 많은 도움을 줄 수 있으리라 사료된다.

• 한국동물위생학회지
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• 제23권3호
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• pp.227-233
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• 2000

Salmonella enteritidis is the most prevalent etiologic agents of foodborne acute gastroenteritis. Direct isolation and identification of S enteritidis are time consuming work and not so highly sensitive. This study was conducted to develop for the specific detection of S enteritidis using polymerase chain reaction(PCR). PCR primers were selected to amplify a 351-base pair(bp) DNA fragment from the salmonella plasmid virulence A(spv A) gene of S enteritidis. With the primers, 351 bp DNA products were amplified from S enteritidis but not from other B, D, Cl serogroup Salmonella spp. It was sensitive to detect up to 40 pg of template DNA by agarose gel electrophoresis. This PCR assay is very rapid and specific method and less time consuming than the standard bacteriological methods.

• Journal of Microbiology
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• 제38권2호
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• pp.93-98
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• 2000

Nine hundred patients diagnosed with diarrhea or hemorrhagic uremic syndrome in the Kyungpook Province, Korea, were examined from November 1998 to February 2000. One patient in Kumi appeared to possess the Escherichia coli O157:H7 strain, which is very important in clinical decision making and public health action. The isolated strain, an E. coli O157:H7 KM, contained a 60 MDa plasmid and typical virulence genes including the verotoxin 2 gene, ehxA gene (encoding enterohemorrhagic hemolysin), and eae (encoding attaching and effacing protein-intimin) gene. This strain produced only verotoxin 2. Pulsed field gel electrophoretic analysis showed that the genomic organization of the E. coli O157:H7 KM strain may differ greatly from those of representative strains previously reported in the United States and Japan.

• 방사선산업학회지
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• 제2권3호
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• pp.111-119
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• 2008

Microarrays can measure the expression of thousands of genes to identify the changes in expression between different biological states. To survey the change of whole Salmonella genes after a relatively high dose of gamma radiation (1 kGy), transcriptome dynamics were examined in the cells by using DNA microarrays. At least 75 genes were induced and 89 genes were reduced two-fold or more after irradiation. Several genes located in pSLT plasmid, cyo operon, and Gifsy prophage were induced along with many genes encoding uncharacterized proteins.While, the expression of genes involved in the virulence of Salmonella as well as metabolic functions were decreased. Although the radiation response as a whole could not be illustrated by using DNA microarrays, the data suggest that the response to high dose of irradiation might be more complex than the SOS response.

• 미생물학회지
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• 제35권3호
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• pp.205-210
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• 1999

Vibrio cholerae 는 사람에게 설사를 일으키는 병원성 세규닝며 본 연구에 이용된 V.cholerae KNIH002 는 국내의 설사질환 환자로부터 분리하였다. 콜레라 독소 검출용 프라이머를 이용하여 PCR 로 증폭한 산물을 탐침자로 이용하여 Southern hybridization을 실시한 결과 PstI 및 BglII로 이중절단된 4.5-kb 절편내에서 ctx 유전자가 존재함을 확인하였다. 따라서 염색체 DNA를 PstI 및 BglII로 절단 후 V. cholerae KNIH002 의 유전자 mini-libraries를 제조하였다. 그리고 동일 탐침자를 이용하여 colony hybridization을 실시한 결과 제조된 유전자 mini-libraries 로부터 신호를 나타내는 한 개의 클론을 선발하였다. 선발된 클로닝 지니는 플라스미드를 pCTX75 라 명명하였으며, 이 클론은 CHO 세포에 대한 세포 독력이 나타남을 확인하였다. 염기서열을 결정한 결과 클로닝된 플라스미드에는 ace 와 zot 유전자들은 각각 ATG 개시코돈과 TGA 종결코돈을 포함하여 291 bp와 1,200 bp 로 구성되어져 있었다. ace 유전자의 염기서열은 V.cholerae E7946 EI Tor Ogawa strain 이 것과 100% 일치하였다. 그러나 zot 유전자의 염기서열 및 아미노산 서열은 V. cholerae 395 Classical Ogawa strain 의 것과 각각 99% 및 98.8% 의 상동성을 보였다. 특히, V.cholerae 395 Classicale Ogawa strain 의 Zot 폴리펩타이드에서 100번, 272번, 281번째 alanine 은 V.cholerae KNIH002에서 모두 valine 으로 치환되어져 있었다.

• 한국동물위생학회지
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• 제21권4호
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• pp.413-429
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• 1998

Escherichia coli is recovered from a wide variety of infections in many animals species. It may be a primary or secondary agent. Nursing and young animals are particularly susceptible, and urinary tract infections are frequent. The various serotypes of E coli are intestinal inhabitants of animals including humans and probably infect most mammals and birds : therefore, they have a cosmopolitan distribution. Colibacillosis refers to any totalized or systemic infection caused entirely or partly by E coli. Collibacillosis in mammals is most often a primary enteric disease, whereas collibacillosis in poultry is typically a secondary located or systemic disease occurring when host defenses have been impaired or overwhelmed. Other opportunistic bacteria, which can be identified by culture, may play a similar role to that of I coli in secondary infections. Collectively, infections caused by E coli are responsible for significant economic losses to the animal performance. From the standpoint of pathogenic mechanisms and diseases, four major categories of E coli are recognized : enterotoxigenic(ETEC), enteropathogenic (EPEC), enteroinvasive(EIEC), and enterohemorrhagic(EHEC). In addition, two less-well-defined E coli categories are recognized in animals and humans : enteroaggregative and cytotoxin necrotizing factor-positive. The aforementioned categories are represented by different serotypes. Certain serotypes show a host preference and are encountered more frequently in some disease syndromes. Of the four major categories, ETEC is the most common cause of diarrhea in calves, lambs, and pigs. Strains in the other categories cause the less-common diarrhea and other disease syndromes. Enterotoxins and pilus antigens are the two most prominent virulence factors thus far identified for ETEC. Two enterotoxins, one heat-stable(ST) and one heat-labile(LT), are produced by enterotoxigenic strains of E coli : not all culture produce both of these plasmid-based enterotoxins.

• 대한수의학회지
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• 제39권2호
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• pp.327-337
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• 1999

The transposon TnphoA was used to generate avirulent mutants from a type A Pasteurella multocida. A suicide vector plasmid pRT733 carrying TnphoA, having the kanamycin resistant gene and harbored in Escherichia coli K-12 strain SM10(${\lambda}pir$), was mated with streptomycin resistant P. multocida P-1059 strain as recipient. This resulted in the generation of two TnphoA insertion mutants (transconjugants, tc95-a and tc95-b) which were resistant both to kanamycin ($Km^{R}$) and streptomycin ($Sm^{R}$), secreted alkaline phosphatase, and were avirulent to turkeys. Southern blot hybridization using two probes derived from internal fragments of TnphoA, confirmed the insertion of TnphoA into 12.9kb or 13.7kb DNA fragment from the EcoRV digested genomic fragments of transconjugants. The two transconjugants, tc95-a and tc95-b, were distinguishable from their parent strains by differences in ribotypes, and outer membrane protein profiles. TnphoA insertion in both transconjugants also resulted in constitutive expression of a 33Kd iron regulated outer membrane protein (IROMP). The gene encoding $Sm^{R}$ was also located within the same 12.9kb EcoRV genomic fragment from both transconjugants. Furthermore, our finding that the recipient P. multocida P-1059 $Sm^{R}$ strain and both transconjugants were avirulent to turkeys suggest that the either 12.9kb or 13.7kb genomic DNA contains the virulence gene and speculate that the presence of $Sm^{R}$ gene or TnphoA insertion may be responsible for regulating and inactivating the gene(s) encoding virulence in P. multocida.

• 대한미생물학회지
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• 제22권2호
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• pp.155-162
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• 1987

To determine the prevalence of antibiotic resistance in fecal E. coli and to investigate possible associations between antibiotic resistance and other plasmid-mediated virulence properties, antibiotic disk susceptibility tests for nine antibiotics were done on 141 strains of E. coli isolated from diarrheal children and well controls. Eighty two percent of the test strains were resistant to one or more antibiotics. Antibiotics to which the test strains were most resistant in descending order were ampicillin (85%), trimethoprim/sulfamethoxazol (60%), and cephalothin (55%). Seventy nine percent of these resistant strains were resistant to two or more antibiotics. All 141 test strains were sorted into enterotoxigenic E. coli (ETEC), enteropathogenic E. coli (EPEC), enteroadherent E. coli (EAEC) and non-pathogenic E. coli and the percentages of strains resistant to multiple antibiotics were compared. Among ETEC regardless of its source, multiple drug resistance was more frequent in strains producing heatstable enterotoxin (ST) only than in strains producing only heat-labile enterotoxin (LT) or both. In EAEC, multiple resistance was more frequently associated with strains isolated from diarrheal patients than with those from well controls. The major antibiotic resistance patterns possessed by multiple resistant enteropathogenic strains were $SXT^R$ $AM^R$, $CR^R$, and $SXT^R$ $AM^R$ $CR^R$. Of 28 ST- producing $SXT^R$ ETEC, 26(96%) were also resistant to ampicillin and 17 (61%) were resistant to cephalothin. The similar pattern was observed in EAEC and EPEC as well. This study has important implications for the treatment of E. coli diarrhea with antibiotics because it is possible that dissemination of virulence could occur under the force of selective antibiotic pressure. In addition, this study suggests that the in vivo efficacy of SXT in treating diarrheal illness be reevaluated.

• Journal of Microbiology and Biotechnology
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• 제31권10호
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• pp.1420-1429
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• 2021

The safety of the probiotic strain Q180, which exerts postprandial lipid-lowering effects, was bioinformatically and phenotypically evaluated. The genome of strain Q180 was completely sequenced, and single circular chromosome of 3,197,263 bp without any plasmid was generated. Phylogenetic and related analyses using16S rRNA gene and whole-genome sequences revealed that strain Q180 is a member of Lactiplantibacillus (Lp., formerly Lactobacillus) plantarum. Antimicrobial resistance (AMR) genes were bioinformatically analyzed using all Lp. plantarum genomes available in GenBank, which showed that AMR genes are present differently depending on Lp. plantarum strains. Bioinformatic analysis demonstrated that some mobile genetic elements such as prophages and insertion sequences were identified in the genome of strain Q180, but because they did not contain harmful genes such as AMR genes and virulence factor (VF)- and toxin-related genes, it was suggested that there is no transferability of harmful genes. The minimum inhibition concentrations of seven tested antibiotics suggested by the European Food Safety Authority guidelines were slightly lower than or equal to the microbiological cut-off values for Lp. plantarum. Strain Q180 did not show hemolytic and gelatinase activities and biogenic amine-producing ability. Taken together, this study demonstrated the safety of strain Q180 in terms of absence of AMR genes and VF- and toxin-related genes as a probiotic strain.

• 한국동물위생학회지
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• 제46권1호
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• pp.35-45
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• 2023

Pathogenic Escherichia coli is the cause of a wide range of diseases in pigs, including diarrhea, edema disease, and septicemia. Diarrhea caused E. coli may result in significant economic losses, making pathogenic E. coli an important pathogen for the swine industry. This study investigated the prevalence of virulence factor genes, antimicrobial resistance phenotypes, and resistance genes in E. coli isolated from feces of piglets in Korea between 2017 and 2020. As a result, 119 pathogenic E. coli isolates were obtained from 601 fecal samples. The F4 adhesin gene and the STb enterotoxin gene were commonly present in E. coli isolated from diarrhea samples. The dominant virulotypes of isolates from diarrhea samples were STb, Stx2e, and F4:LT:STb. More than 80% of the screened isolates were resistant to ampicillin, sulfisoxazole, chloramphenicol, or tetracycline. To confirm the resistance mechanisms for β-lactam or quinolone, we investigated the genotypic factors of resistance. Each of the ceftiofur-resistant E. coli produced an extended-spectrum β-lactamase encoded by bla_CTX-M-14, bla_CTX-M-27, and bla_CTX-M-55. And all ciprofloxacin-resistant E. coli harbored mutations in quinoloneresistance-determining-regions. In addition, some of the ciprofloxacin-resistant E. coli contained the plasmid-mediated-quinolone-resistance genes such as qepA, qnrB1, or qnrD. This study has confirmed that the F4 fimbria and the STb enterotoxin are the most predominant in pathogenic E. coli isolated from piglets with diarrhea in Korea and there is a great need for responsible and prudent use of antimicrobials to treat colibacillosis.