• 한국수산과학회지
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• 제34권5호
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• pp.502-508
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• 2001

병원성 비브리오 속인 Vibrio furnissii의 용혈소를 분리정제하고 그 물리화학적 특성, 생물학적인 그리고 면역학적인 상호관련성을 검토하여, 다음과 같은 결과를 얻었다. 용혈소의 정제는 ammonium sulfate precipitation, DEAE-Sephadex A-50 (1st와 2nd)와 Sephaedx G-100 gel permeation chromatography를 거쳐 738배로 정제되었으며 회수율은 각각 $11.6\%$ 와 $5.2\%$로 나타났다 전기영동법으로 용혈소의 정제를 확인한 결과 단일 밴드의 형성을 관찰할 수 있었다. 분자량은 61 kDa이였으며 또한 SDS-PAGE 결과 subuint는 존재하지 않았다. 정제된 용혈소는 열에 불안정한 이열성 용혈소이며 $Cu^{2+},\;Zn^{2+}$와 cholesterol에 의해 용혈활성이 모두 저해되었다. 용혈기작에 있어서는 온도의존성 용혈기작으로서 최적 용혈의 온도는 $37^{\circ}C\sim47^{\circ}C$로 나타났다. 정제용혈소의 pH 안정성은 pH 6.0, 6.5, 10.0에서 각각 불안정하였다. 용혈활성에 있어서 최적 혈구 농도는 $1.0\sim2.0\%$였다. 또한 각종 동물 혈구의 감수성 시험 결과 두 용혈소 모두 토끼에서 가장 감수성이 높았으며 사람의 혈구에서는 B형 혈액형에서 다소 높은 감수성을 나타내었다.

• Journal of Microbiology and Biotechnology
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• 제17권1호
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• pp.44-51
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• 2007

Marine bacterial strains were isolated trom coastal regions of Goa and screened for the strains that produce the highest amount of mucous expolysaccharide (EPS). Our screening resulted in the identification of the strain Vibrio furnissii VB0S3 (hereafter called VB0S3), as it produced the highest EPS in batch cultures during the late logarithmic growth phase. The isolate was identified as VB0S3 based on morphological and biochemical properties. Growth and EPS production were studied in mineral salts medium supplemented with NaCl (1.5%) and glucose (0.2%). The exopolymer was recovered from the culture supernatant by using three volumes of cold ethanol precipitation and dialysis procedure. Chemical analyses of EPS revealed that it is primarily composed of neutral sugars, uronic acids, and proteins. Fourier-transform infrared (FT-IR) spectroscopy revealed the presence of carboxyl, hydroxyl, and amide groups, which correspond to a typical heteropolymeric polysaccharide, and the EPS also possessed good emulsification activity. The gas chromatographic analysis of an alditol-acetate derivatized sample of EPS revealed that it was mainly composed of galactose and glucose. Minor components found were mannose, rhamnose, fucose, ribose, arabinose, and xylose. EPS was readily isolated from culture supernatants, which suggests that the EPS was a slime-like exopolysaccharide. This is the first report of exopolysaccharide characterization that describes the isolation and characterization of an EPS expressed by Vibrio surnissii strain VB0S3. The results of the study contribute significantly and go a long way towards an understanding of the correlation between growth and EPS production, chemical composition, and industrial applications of the exopolysaccharide in environmental biotechnology and bioremediation.

• 한국수산과학회지
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• 제30권3호
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• pp.361-366
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• 1997

금강 하구의 물에서 분리된 sucrose 분해성 Vibrio 속 중에 용혈독소를 생산하는 균주의 독성을 시험한 결과와 기존에 알려진 12종의 병원성 Vibrio 녹과 비교한 결과는 다음과 같다. 1. 분리된 장소의 환경조건은 염도가 $4.7\%_{\circ}$, pH가 7.6, 수온이 $24^{\circ}C$ 및 conductivity가 $7800{\mu}MHOS$이었다. 2. 생리, 생화학적 특성과 염분 요구도를 비교한 결과 sucrose를 분해하는 병원성 Vibrio인 V. alginolyticus, V. cholerae, V. cincinnatiensis, V. fluvialis, V. furnissii 및 V. metschnikouii와는 확실히 구별되었다. 3. 생육가능한 염도는 $0.5\~7.5\%_{\circ}$이었으며 생육가능한 pH는 $4.5\~9.5$이었다. 4. TCBS 평판한천배지에서 균의 집락은 전형적인 황색집락이었으며, 전자현미경에 나타난 균의 형태은 콤마상 간균이었다. 5. 새앙쥐에 대한 치사독성이 확실하였으며, 사람과 면양 적혈구에 대한 용혈성이 확인되었고 rat 피부혈관 투과성 항진작용이 양성이었다. 이상의 결과로 이 균은 병원성이 확인되었고 기존에 알려지지 않은 Vibrio 속으로 확인되어 이 균을 Vibrio sp. D5로 명명하였다.

• 한국식품영양과학회지
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• 제30권2호
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• pp.222-227
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• 2001

The members of the genus Vibrio include harmless aquatic strain as well as strains capable of causing infections in human and fish. Pathogenic mechanisms are only understood for Vibrio cholerae O1 and O139 and not for the majority of Vibrio species. Twelve clinical and nonclinical strains were examined by in vitro and in vivo experiments for the importance of extracellular enzymes as a virulence determinant of Vibrio species. In vivo cytotoxicity assay was performed by injecting approximately $10^{8}$ cells/mL into mice (BALB/c). V. harvyi and V. vulnificus showed 100% lethality within 3hr after bacterial injection. V. fluvialis and four strains of V. parahaemolyticus showed 50% lethality within 4hr. V. mimicus, V. alginolyticus and V. furnissii revealed 30% lethality within 9hr. Nonclinical strains, V. campbellii and V. ordalii, did not show any lethality. In vitro protease and hemolytic activities were also good indicators for clinical and nonclinical strains of Vibrio species. The clinical strains showed much higher activities than nonclinical strains. The activity of some clinical strains of re-isolates was evidently increased. Most clinical strains had $\beta$ hemolytic activity. The results demonstrate that the prevalent distribution of extracellular proteases in pathogenic Vibrio sp. implies their importance as a virulence determinant.

• 대한미생물학회지
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• 제34권5호
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• pp.423-433
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• 1999

A novel bacteriophage, designated as VPP97, that infects the strains of Vibiro parahaemolyticus (hallophilic, Gram-negative bacterium) isolated most commonly from marine environments, has been discovered, and several of its properties have been determined. The plaques were clear and sized $0.6{\sim}1.0\;mm$ in diameter. The virion forms a single band on 70% sucrose gradient and ${\rho}1.50$ CsCl gradient by sucrose gradient centrifugation and CsCl gradient centrifugation respectively. It has a hexagonal head and a relatively long tail, as shown by electron microscopy. Vibrio alginolyticus, Vibrio fluvialis and Vibrio furnissii were also sensitive to this phage. It was almost totally inactivated at $70^{\circ}C$ and at pH below 5 or over 10. The nucleic acid of VPP97 is composed of DNA. The VPP97 had 9 specific structural proteins sized between 21.5 kDa and 97.4 kDa on SDS-PAGE. When V. parahaemolyticus cultures were treated with either phage VPP97 or one of the several antibiotics for 2 hours, the viable number of V. parahaemolyticus treated with the phage VPP97 is lower than that treated with chloramphenicol, erythromycin or penicillin, but not lower than that treated with tetracycline. Mice that have responded to the phage treatment revealed the lower numbers of V. parahaemolyticus in small intestine and less damage on small intestine compared to the untreated mice. Therefore, we suggest that the phage treatment appears effective to the infection by V. parahaemolyticus.

• 대한미생물학회지
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• 제34권6호
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• pp.513-518
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• 1999

A total of 113 Vibrio sp. strains were examined for plasmid content which were subjected to digestion with restriction enzymes. About the 55% Vibrio spp. have the plasmid more than one. Most of these plasmid various derivatives ranged from $2.4\;kb{\sim}23\;kb$, especially two strains of V. mimicus and one strain of V. furnissii carried one high-molecular weight plasmid (molecular weight ranging between $70\;kb{\sim}100\;kb$). Results of restriction analysis for plasmid of this three strains were by no means the rule. For detection of tdh and ctx gene, the virulence factor involved in the pathogenesis, we carried out the TDH and CT assay, PCR amplification, and hybridization. A total 11 strains were produced TDH, involved in 9 strains of V. parahaemolyticus and 1 strain of V. alginolyticus from clinical isolates and 1 strains of V. mimicus from environmental isolates. In the experiments of tdh gene detection, in all, 3 strains of V. parahaemolyticus from clinical isolates and 2 strains from environmental isolates could be successfully amplified in 400 bp by PCR. The PCR results were consistent with DNA hybridization tests. In the experiments of CT assay, in all, 3 strains of V. cholerae from clinical isolate and 1 strain of V. cholerae from environmental isolates were observed CT-producing. These CT-producing strains amplified in 302 bp by PCR for the detection of ctx gene. All CT-producing strains hybridized with digoxigenin-labeled DNA probe, while CT-negative strains did not hybridize. Also hybridization tests results for detection of ctx gene consistent with PCR.

• 한국어병학회지
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• 제27권1호
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• pp.17-24
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• 2014

질병 치료를 위하여 사용되는 항균제는 효과적이고 사용이 편리한 장점이 있지만, 내성균 발생이나 항균물질 잔류와 같은 문제점들을 안고 있다. 따라서 어류 질병 치료 및 예방을 위해 어류와 인체에 안전한 생약제 개발과 함께 어류 질병 원인균의 과다 발생을 억제하며 어체의 건강을 유지시키기 위하여 유용 균주를 이용하고자 하는 연구가 다양하게 진행되고 있다. 본 연구는 이전 연구에서 분리되어 Listonella anguillarum에 대하여 생장 억제효과가 있는 것으로 밝혀진 Pseudomonas aeruginosa MB I-3 (MB I-3)를 이용하여 넙치의 비브리오병에 대한 생물학적 방제효과를 검토하였다. Double layered plate assay와 co-culture을 통하여 MB I-3의 L. anguillarum에 대한 생장 억제능력을 조사하였고, MB I-3 균주 배양액을 ethyl acetate로 추출하여 disk 확산법으로 추출물의 항균 효과를 확인하였다. 액체 및 고체 배양에서 생장이 억제된 L. anguillarum을 전자현미경으로 관찰하였으며, 넙치 치어 사육 수조에 L. anguillarum과 MB I-3 균주를 동시에 첨가하여 폐사율을 비교하였다. MB I-3는 8종의 병원성 비브리오균에 대하여 항균력을 나타내었으며, 96시간 동안 실시한 co-culture에서 L. anguillarum은 배양 후 9시간까지 생장 증가를 보였으나, 그 후 감소하는 경향을 보였다. 한편 MB I-3 배양액 추출물 또한 L. anguillarum에 항균활성을 보여 항균 물질이 ethyl acetate로 추출됨을 알 수 있었다. 전자현미경 관찰에서 L. anguillarum은 세포질의 밀도 감소 및 세포막의 swelling에 의한 세포 용해 현상을 보였다. 한편 MB I-3를 L. anguillarum과 함께 투여한 넙치 치어는 대조군에 비해 누적 폐사율이 약 20% 감소되는 결과를 보였으므로, MB I-3를 이용한 수산용 probiotics 개발 가능성을 시사하였다.