• 대한수의학회지
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• 제44권4호
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• pp.581-586
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• 2004

The growth characteristics and the properties of the Enterococcus faecium (E. faecium) isolated from intestine of slaughting pig were examined in order to confirm whether it will be used practically as probiotics or not. E. faecium was identified by morphological and biochemical tests including of final confirming by API strep kit. Among 264 samples, 44 isolates (16.7%) of E. faecium were observed as an exellent growth as a range of $1{\times}10^7CFU/ml{\sim}3.3{\times}10^9CFU/ml$ in viable bacterial count on MRI medium. All isolates were shown non-haemolysis on the blood agar exception of 28 isolates shown ${\alpha}$-haemolysis and were identified as E. faecium by biochemical test using API strep enzyme kits. Eight strains (18.2%) were finally selected from which they were excellent sensitivity in showing of the acid-tolerance and the bile-tolerance in compare with reference strain.

• 한국어병학회지
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• 제20권1호
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• pp.49-60
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• 2007

우리 나라 남해안의 병든 넙치로부터 분리한 non-hemolytic Streptococcus parauberis의 지역적 특성을 구명하고자 균의 생리적, 생화학적 및 분자생물학적 분석을 실시하였다. 먼저, catalase-negative Gram-positive cocci를 대상으로 multiplex PCR을 실시하여 S. parauberis 12 균주를 선정하였으며, 표현형적 동정을 위해 시판되고 있는 API 20 Strep 및 API ZYM kit를 사용하였다. 분리 균주의 API porfile을 분석한 결과, Lactococcus lactis, S. constellatus 및 S. uberis 등으로 동정되었다. 또한, 터봇에서 분리된 S. parauberis와 비교해 볼 때 Voges-Proskauer, arginine, hippurate, alkiline phosphatase 및 pyrroidonyl arylamidase의 반응 결과는 동일하였으나, β-glucuronidase 활성은 다양한 것으로 나타났다. 약제 감수성 시험 결과에서는 florfenicol, ampicillin, ofloxacin 및 vancomycin에 대해 감수성이 있는 것으로 나타났으며, oxolinic acid, flumequine, nalidixic acid 및 sulfisoxazol에 대해 내성이 있는 것으로 나타났다. 그러나, 분리 균주의 16S rDNA 염기서열을 분석한 결과, 참조균주인 S. parauberis KCTC 3651 (AY584477)과 99%의 상동성을 나타내었으며, 넙치 분리 균주들이 유전적으로 동일한 그룹에 속하는 것으로 나타났다.

• 한국식품위생안전성학회지
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• 제22권4호
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• pp.294-299
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• 2007

서울시 가락동 수산물 도매시장에서 시판되는 건포류 164건에 대해 API strep kit를 사용하여 87주의 균주들을 분리하였고 이 가운데 28주에서 E. faecalis, and E. faecium가, 11주에서 Lactococcus lactis ssp. lactis가 분리되었다. 28주의 E. faecalis와 E. faecium에서는 내성을 나타내는 항생제들이 streptomycin(95.6%), kanamycin(84.5%), gentamycin(66.7%), cephaloxin(97.8%), ampicillin/sulbactam(88.9%), ticarcillin(66.7%), amikacin(97.8%), sulfonamides(97.8%), ceftriaxone(75.6%), nalidixic acid(100.0%), 그리고 cefoxitin(100.0%) 등이었고, 감수성을 나타내는 항생제들은 amoxicillin/clavulanic acid(97.8%), chloramphenicol(95.6%), solfamethoxazole/trimethoprim(97.8%), 그리고 tetracycline(71.1%)등으로 나타났다. 또한, 이 가운데 가장 많은 다제내성 형태로는 10 균주의 E. faecalis에서 나타난 $S-K-GM-CF-SAM-TIC-An-S_3-CRO-NA-FOX$에 대한 것이었다. 결국, 건포류에서 분리한 E. faecalis 균주에서 항생제 다제내성이 분포함으로 나타나 probiotics로 사용되고 있는 이러한 균에 대한 항생제 내성의 결과가 의의가 있다고 사료된다.

• 한국어병학회지
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• 제17권2호
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• pp.91-98
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• 2004

본 연구의 목적은 국내 어류 연쇄구균증 병원체 동정에 있어서 Streptococcus iniae에 대한 구체적인 국내 연구 보고가 없어, 기존에 수행되고 있는 동정방법을 근간으로 16S-23S intergenic spacer region (ISR)의 염기서열을 분석하여 보다 명확한 동정을 실시하고자 하였다. API 20 strep system에 의한 생화학적 성상을 분석해본 결과 정확한 종 동정은 이루어지지 않았지만 기존의 연구에서 보고된 API 20 strep system에 의한 S. iniae의 생화학적 성상과 높은 유사성을 보이는 10균주를 분리할 수 있었다. S. iniae 16S rRNA gene 서열 유래 종 특이적 primer Sin-1 5'-CTAGAGTACACATGTACT(AGCT)AAG-3'와 Sin-2 5'-GGATTTTC CACTCCCATTAC-3'에 의한 PCR-assay 결과 시험균주 10 균주 모두 동일하게 약 300bp의 증폭산물이 관찰되었고, 비 특이적인 반응은 관찰되지 않았다. 시험균주 모두 3개의 16S-23S ISR operon 구조가 관찰되었으나 S. iniae 표준균주 (KCTC 3657)의 경우 단일 구조가 관찰되었다. 16S-23S intergenic spacer region (ISR)의 염기서열을 분석해본 결과 기존에 보고된 S. iniae (Genbank accession number AF 048773)와 96%의 상동성을 보였으며 전체서열에서 상동성을 보이는 근연종이나 근연속은 검색되지 않아 최종적으로 S. iniae로 동정하였다.

• 한국수산과학회지
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• 제36권6호
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• pp.654-660
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• 2003

To evaluate the biological diversity of fish pathogenic streptococci, 35 strains isolated from diseased olive flounder (Paralichtys olivaceus), were analyzed using a random amplified polymorphic DNA (RAPD) technique with the oligonucleotide commercial primer 6 (Amersham Biosciences). Api 20 Strep test, drug resistance and artificial infection were carried out for further characterization of the isolates. RAPD fingerprints showed similar pattern in 25 strains (about $71.4\%$ of 35 isolates) and these strains were designed as RA group 1. Similarities greater than $44\%$ were obtained when the Dice coefficient was applied among the isolates of RA 1. On the other hand, the reference Streptococcus iniae showed a similar RAPD profile to the isolates with similarity levels of $40-93.3\%.$ Rh I was suggested to be the dominant group isolated from olive flounder suffering from streptococcosis. However, the isolates of Rh 1 group were not classified into the same species by the Api 20 Strep identification system. There was no peculiarity in drug resistance patterns of Rh I group isolates against 7 antibacterial agents. However, only 3 of 25 isolates $(0.12\%)$ showed oxytetracycline (OTC) resistance and OTC might be a useful chemotherapeutic agent in controlling the streptococcosis by strains of RA I group in olive flounder. Fish injected intraperitoneally with $10^5$ CFU of an isolate of Rh I and RA III group showed $60\%\;and\;50\%$ accumulative mortality for 20 days, respectively ($20\%$ in control or Rh II). However luther comparative studies about differences in virulence between isolates are needed.

• 한국어병학회지
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• 제16권1호
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• pp.51-59
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• 2003

Streptococcal infection is one of the most serious disease of cultured olive flounder, Paralychthys olivaceus in Korea and caused by more than one species. However, there has been considerable confusions about the taxonomic position of the fish pathogenic streptococci. In this study, We performed the randomly amplified polymorphic DNA(RAPD) pattern analysis to evaluate the possible classification in 8 streptococci isolated from diseased olive flounder and reference strains based on their DNA structure. RAPD PCR with DNA solution prepared by simple boiling and 10-mer random primer was appeared to be a good tool for discrimination of different streptococcal strains. Phenotypical characters by simple biological test and API 20 Strep corresponded well to the specific profiles of RAPD in streptococcal isolates of this study. Therefore, the RAPD profile was considered as one of differential characters to discriminate the streptococcal isolates from diseased olive flounder.

• Restorative Dentistry and Endodontics
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• 제20권2호
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• pp.577-609
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• 1995

Bacteria have been regarded as one of the most important factors in pulpal and periapical diseases. Streptococci are frequently isolated facultative anaerobes in infected root canals. Recently molecular biological techniques have been rapidly progressed. This study was designed to apply the molecular biological tools to the identification and classification of streptococci in the endodontic microbiology. Streptococci isolated from infected root canals were identified with both Vitek Systems and API 20 STREP. Identification results were somewhat different in several strains of streptococci. Eighteen streptococci and enterococcal was difficult so to digest plasmid DNA using Hind III and EcoRI to differentiate strains by restriction enzyme analysis of plasmid DNA. 16S rDNA of chromosome was amplified by polymerase chain reaction(PCR) and then restricition fragment length polymorphism(RFLP) using several restriction enzymes was observed. The molecular mass of 16S rDNA of chromosomal DNA was approximately 1.4kb. There were three to five RFLP patterns using eight restriction enzymes. RFLP patterns digested with CfoI which recognizes four base sequences were identical in all stains. Hind III which recognizes six base sequences could not digest the 16S rDNA. Restriction enzymes which recognize five base sequences were suitable for RFLP pattern analysis. At least three different restriction enzymes were needed to compare each strains. 16S rDNA PCR-RFLP was simple and rapid to differentiate and classify strains and could be used in the epidemiological study of root canal infections.

• 한국어병학회지
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• 제26권3호
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• pp.289-294
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• 2013

제주도에서 좌초된 뱀머리돌고래, Steno bredanensis에서 Enterococcus faecium이 분리 되었다. E. faecium은 간, 비장, 신장, 심장, 폐에서 $1.54{\times}10^6$ cfu/g까지 분리 되었다. E. faecium KCCM 12118과 분리균주 사이의 효소 활성은 차이가 없었고 생화학적 특성은 같거나 유사하였다. 모든 분리균주는 6 종류 항균제에 대해 다재내성을 갖고 있었으나 chloramphenicol과 vancomycin에 대한 내성은 없었다.

• 한국어병학회지
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• 제36권2호
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• pp.277-286
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• 2023

Korean rockfish, Sebastes schlegelii, is a representative bony fish that belongs to the family Scorpaenidae and the order Scorpaeniformes. It has high ecological and economic value and is widely cultivated in many East Asian countries, including South Korea, Japan and China. One of streptococci, Streptococcus iniae, is Gram-positive cocci with a negative reaction for catalase and oxidase. The Korean rockfish shows clinical signs when infected with S. iniae, such as body darkening, bleeding, enlarged kidneys, blurred eyes, abdominal distension, etc., ultimately leading to death. The Korean rockfish causes significant economic losses every year in South Korea due to streptococcosis. In this study, we identified bacteria from the fish using polymerase chain reaction and conducted analyses of hemolytic activity and biochemical tests using API 20 STREP and API ZYM systems. Results of confirming the hemolytic activity (n=4) observed in alpha-type hemolysis (25%), beta-type hemol- ysis (50%), and gamma-type hemolysis (25%) of isolates. The biochemical test results exhibited sig- nificant variation among S. iniae. Additionally, we performed intraperitoneal injection with S. iniae in the fish and analyzed the phylogenetic tree using housekeeping genes of S. iniae, including cpsD, arcC, glnA, groEL, gyrB, mutS, pheT, prkC, rpoB, and tkt, via multilocus sequence typing (MLST). The lethal dose (LD₅₀) showed strong pathogenicity, such as 3.34 × 10 colony-forming unit (CFU)/ml for 23FBStr0601 strain and 7.16 × 10 CFU/ml for 23FBStr0602 strain. 23FBStr0603 strain showed relatively low pathogenicity at 1.73 × 10⁵ CFU/ml. The strains 23FBStr0601 and 23FBStr0602, which showed strong pathogenicity, clustered into one monophyletic group. The 23FBStr0603 strain showed weak pathogenicity and formed a monophyletic group with KCTC 3657.

• 한국미생물·생명공학회지
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• 제26권6호
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• pp.523-528
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• 1998

본 연구에서는 토마토에서 bacteriocin 생산균주를 분리하여 균주동정 및 bacteriocin 정제를 행하였다. 분리균주는 Gram 염색, catalase test, 포자생성유무, API 20 STREP. kit 등을 통하여 Enterococus faecium으로 잠정 동정하였다. 분리균주가 생산하는 항균성물질은 Gram 양성 세균인 Listeria manocytogmes, Leuonostoc mesenteroides, Lactobacillus plantarum, Streptococus agalartiae, Streptococus pyrogenes, Gram 음성 세균인 Pseudomonas aeruginosa등에 대해서 항균력을 나타내었다. 단백질 분해 효소인 proteinase K에 의해서 그 활성이 소실되고, catalase에 의해서는 항균력이 유지되므로 단백질성 bacteriocin임을 확인할 수 있었다. Bacteriocin 정제를 위하여 ethanol침전, CM Sepharose CL-6B 양이온교환 chromatography, Sephacryl S-100 HR gel filtration의 순으로 정제하여 초기 배양액의 bacteriocin활성 대비 35.8배의 정제도를 얻었다. 13% SDS-polyacrylamide gel전기 영동을 통하여 단일 band를 확인할 수 있었으며 분자량은 약 51 kDa 정도_로 추정되었다. Crude bacteriocin의 열 안정성은 10$0^{\circ}C$에서 1시간 처리하였을 때의 잔존활성이 3.3%로 비교적 낮았으며, pH에 대해서는 pH2~7에서 비교적 안정하였다. Bacteriocin의 항균작용을 알아보기 위하여 피검균인 Leu. mesenteroides를 초기농도 1.0$\times$$10^{9}$ CFU/$m\ell$로 접종한 배지에 bacteriocin을 첨가하여 배양하였을 때 4시간 이후에 완전히 사멸되므로 bacteriocidal 작용을 하는 것으로 추정되었다.