• 한국해양바이오학회지
• /
• 제2권4호
• /
• pp.263-266
• /
• 2007

Vibrio vulnificusis a causative agent of serious diseases in humans resulting from the contact of wound with seawater or consumption of raw seafood. Several studies aimed at detecting V. vulnificus have targeted vvh as a representative virulence toxin gene belonging to the bacterium. In this study, we targeted the rpoS gene, a general stress regulator, to detect V. vulnificus. PCR specificity was identified by amplification of 8 V. vulnificus templates and by the loss of a PCR product with 36 non-V. vulnificus strains. The PCR assay had the 273-bp fragment and the sensitivity of 10 pg DNA from V. vulnificus. SYBR Green I-based real-time PCR assay targeting the rpoS gene showed a melting temperature of approximately $84^{\circ}C$ for V. vulnificus strains. The minimum level of detection by real-time PCR was 2 pg of purified genomic DNA, or $10^3$ V. vulnificus cells from pure cultured broth and $10^3$ cells in 1g of oyster tissue homogenates. These data indicate that real-time PCR is a sensitive, species-specific, and rapid method for detecting this bacterium using the rpoS gene in pure cultures and in infected oyster tissues.

• The Korean Journal of Physiology and Pharmacology
• /
• 제3권2호
• /
• pp.199-205
• /
• 1999

Vibrio vulnificus cytolysin caused platelet cytolysis and increased intracellular calcium concentration $([Ca^{2+}]_i)$ of rat platelets in a concentration-dependent manner. In the presence of V. vulnificus cytolysin (3 HU/ml), lactate dehydrogenase (LDH) activity was increased from $1.3{\pm}0.4%$ of control to $64.3{\pm}3.4%$ in platelet suspension buffer. In $Ca^{2+}-free$ platelet suspension buffer, however, V. vulnificus cytolysin did not induce $[Ca^{2+}]_i$ increase and LDH release. Addition of EGTA (2 mM) to suspension buffer after the initial $Ca^{2+}$ influx reversed $[Ca^{2+}]_i$ to the control level. However, a $Ca^{2+}$ channel blocker verapamil $(20\;{\mu}M)$ or mefenamic acid $(20\;{\mu}M)$ did not inhibit V. vulnificus cytolysin-induced $[Ca^{2+}]_i$ increase and LDH release. Divalent cations such as $Co^{2+},\;Cd^{2+}\;or\;Mn^{2+}$ (2 mM each) also did not alter V. vulnificus cytolysin-induced $[Ca^{2+}]_i$ increase and LDH release. V. vulnificus cytolysin (3 HU/ml)-induced calcium influx was completely blocked by lanthanum (2 mM). Lanthanum (2 mM) also completely blocked V. vulnificus cytolysin (3 HU/ml)-induced LDH release. Osmotic protectants such as, raffinose, sucrose or PEG600 (50 mM each) did not inhibit the lytic activity of V. vulnificus cytolysin. In conclusion, lanthanum sensitive $Ca^{2+}$ influx plays a significant role in Vibrio vulnificus cytolysin-induced platelet cytolysis and thrombocytopenia in V. vulnificus infection.

• Journal of Microbiology
• /
• 제44권2호
• /
• pp.226-232
• /
• 2006

Swarming has proven to be a good in vitro model for bacterial surface adherence and colonization, and the swarming differentiation of a bacterium has been shown to be coupled with changes in the expression of virulence factors associated with its invasiveness, particularly in the early stages of infection. In this study, we attempted to determine whether the expression of vvhA, which encodes for hemolysin/cytolysin (VvhA), is either upregulated or downregulated during the swarming differentiation of V. vulnificus. The insertional inactivation of vvhA itself exerted no detectable effect on the expression of V. vulnificus swarming motility. However, in our lacZ-fused vvhA transcriptional reporter assay, vvhA expression decreased in swarming V. vulnificus as compared to non-swarming or planktonic V. vulnificus. The reduced expression of vvhA in swarming V. vulnificus increased as a result of the deletional inactivation of luxS, a gene associated with quorum sensing. These results show that vvhA expression in swarming V. vulnificus is downregulated via the activity of the LuxS quorum-sensing system, suggesting that VvhA performs no essential role in the invasiveness of V. vulnificus via the adherence to and colonization on the body surfaces required in the early stages of the infection. However, VvhA may playa significant role in the pathophysiological deterioration occurring after swarming V. vulnificus is differentiated into planktonic V. vulnificus.

• 한국식품영양학회지
• /
• 제10권3호
• /
• pp.320-324
• /
• 1997

유기산 처리법을 이용하여 15$^{\circ}C$ 저장 동안 생굴의 V. vulnificus strain 29307의 세균수 변화에 대한 영향을 조사하였다. 각각 3분 동안 0.5% 초산, 0.5% 유산, 또는 0.5% 구연산을 침지한 생굴은 저장 4일 후부터 V. vulnificus가 검출되지 않았다. 2% 초산 함유한 3% 알지닉산 처리구는 저장 2일 후부터 V. vulnificus가 검출되지 않았으며, 수돗물로 처리한 대조구는 저장 4일 동안 V. vulnificus가 분리되었다. 본 연구의 결과를 토대로 알지닉산과 초산의 조합은 각 유기산 처리구보다 V. vulnificus에 대한 항균력을 증진하였다.

• 한국환경보건학회지
• /
• 제50권1호
• /
• pp.66-72
• /
• 2024

Background: Vibrio vulnificus is a serious opportunistic human pathogen that has a worldwide distribution in a variety of marine and estuarine environments. Objectives: For this reason, we investigated the distribution of Vibrio vulnificus in coastal areas of Gyeonggido Province from 2018 to 2022. Also, we analyzed the correlation between V. vulnificus leading to infection and two marine environmental factors (water temperature and salinity). Methods: We collected a total of 266 samples from six coastal area points (i.e., seawater, mudflats). Specimens were isolated using selective plating media and isolated strains were identified by a VITEK 2 system. To find the relevance of the isolation rates of V. vulnificus and number of cases of V. vulnificus infection, we summarized the data on 48 cases of V. vulnificus infection from the open data of the Korea Disease Control and Prevention Agency. Results: Among the 266 samples taken during the investigation period, 47 strains were isolated, and the separation rates of V. vulnificus were 17.7%. The monthly isolation rates of V. vulnificus were ranked in the order of August (53.8%), September (33.3%), June (28.6%), and July (21.1%). There was a positive correlation with the temperature of seawater, but salinity was not significant. The number of cases of V. vulnificus infection reported in Gyeonggi-do Province were 18 (37.5%) in September, 14 (29.2%) in August, and eight (16.7%) in October. The proportion was 83.3%. It was relevant to the isolation rates of V. vulnificus in the marine environmental sources. Conclusions: Our data showed that the number of V. vulnificus infection cases could be affected by changes in the distribution of V. vulnificus due to rise the temperature of seawater in the marine environment.

• 환경생물
• /
• 제19권4호
• /
• pp.302-312
• /
• 2001

Immunoglobulin G was purified by 40% $(NH_4)_2SO_4$ precipitation, DEAE-Sephadex, Sephadex G-150 column chromatographies from rabbit antiserum against V. vulnificus ATCC 27562 O antigen and used for immunological test for V. vulnificus isolates. The profiles of cell lysate total protein and outer membrane protein from the isolates were analyzed by SDS-PAGE and densitometry. The overall profiles in all isolates were similar. Distict protein band was observed in comparison with V. parahaemolyticus. Western Blotting with rabbit Immunoglobulin G against cell lysates and OMP of V. vulnificus isolates showed a strong antigenic response to antigen 66, 60, 54, 48, 33 and 26 kDa which were common to all strains examined. The 26 kDa antigen showed V. vulnificus specific antigen in comparison with Vibrio parahaemolyticus. A sandwich enzyme-linked immunosorbent assay was developed by using rat anti-V. vulnificus ATCC 27562 polyclonal antibodies as capture antibody, a purified rabbit IgG antibody as detector antibody, and goat anti-rabbit IgG-alkaline phosphatase conjugate as developer antibody. When four V. vulnificus isolates were tested, the reactivity showed from 50 to 70% by sandwich ELISA.

• 한국식품영양과학회지
• /
• 제18권2호
• /
• pp.175-180
• /
• 1989

Vibrio 패혈증의 원인균인 V. vulnificus 에 대한 치사독소의 소재, 특성 및 독력을 밝히고저 해수에서 분리된 균(V. vulnificus S-1)과 환자에게 분리된 균(V. vulnificus M-1)을 새앙쥐 (institude cancer research mouse)에 복강 주사하여 실험하였다. 환자에서 분리된 균과 해수에서 분리된 균의 치사 독성은 차이가 없었으며 새앙쥐에 복강 주사하였을 때 $LD_{50}$은 $7.80{\times}10^6$ 이었다. V. vulnificus 의 조용혈소는 새앙쥐에 대한 치사독성이 없었으며 치사 독소는 균체 내에 존재하였으며 이 독소는 $80^{\circ}C$, 20분에 불활성화 되었다.

• 한국어병학회지
• /
• 제35권1호
• /
• pp.57-63
• /
• 2022

Vibrio parahaemolyticus와 V. vulnificus 같은 인체 위해성 세균들은 수산물을 통하여 인체에 감염되는 것으로 알려져 있음으로 생선회로 선호도가 높은 넙치에서 이들 세균이 어느 정도 생존 가능한지 알아보고자 하였다. 먼저 온도와 염분 농도가 다른 배지에서 이들 세균의 생장도를 조사하였다. 37℃ 에서 보다 25℃에서 V. parahaemolyticus와 V. vulnificus는 약 50~60% 감소한 생장률을 보였다. 그러나 1%, 2% 및 3% NaCl을 함유한 배지로 25℃에서 배양하였을 때의 생장은 3% NaCl을 함유한 배지에서 V. vulnificus가 증가된 생장을 보인 것을 제외하고 차이는 없었다. V. parahaemolyticus와 V. vulnificus에 대한 넙치의 감수성을 알아보기 위하여 1×10⁶CFU/fish로 복강 주사하여 인위 감염을 하였으나 1주일간 사망 개체는 없었다. V. parahaemolyticus와 V. vulnificus에 대한 넙치의 방어능력을 알아보기 위하여 넙치의 혈청 및 체표 점액에서의 세균 생존력과 신장에서 분리한 macrophage의 식균 활성을 알아보았다. 혈청에서는 3시간 이내에 각각 85%, 99% 이상의 균이 제거되었고 체표 점액에서는 12시간까지 세균 수가 감소하였으며 신장백혈구는 약 70% 이상에서 식균 작용을 나타냈다. 따라서 넙치는 환경 수에 있는 V. parahaemolyticus와 V. vulnificus에 대한 단순 이동매개체로 작용한다 할지라도 발달된 자체 항균력으로 이들 세균의 수를 감소시켜 인체 감염 가능성을 저하하는 효과가 있음을 시사하였다.

• 대한미생물학회지
• /
• 제22권4호
• /
• pp.393-402
• /
• 1987

The halophilic bacterium Vibrio vulnificus, previously called lactose-positive(L+) Vibrio and Beneckea vulnifica, causes acute, fulminating wound infections and septicemia in humans. Septicemia is very serious infection with a fatality rate of about 50%. Most patients with primary septicemia due to V. vulnificus have preexisting liver disease. V. vulnificus also cause severe wound infections usually after trauma and exposure to marine animals or the marine environment. The mortality rate is not nearly as high as in primary septicemia caused by this organism. In most cases human disease results from ingestion of contaminated seafood or from infection of a wound, frequently of seawater or crab origin. The author made an attempt to isolation of the V vulnificus from seawater, seamud, fish, shellfish, and algae on the southern sea of Korea from January to September in 1987, using for the purpose of vaccine preparation. The author investigated for bacteriological identification, hemolysis and determination of serotypes of isolated V. vulnificus strains. Eighty-five strains(5.9%) out of 1450 specimens collected of V. vulnificus were isolated. The distribution of the 85 isolates were as follows: 21 strains from seawater, 11 strains from seamud, 28 strains from fish, 19 strains from shellfish, and 6 strains from algae, respectively. All 85 isolates were positive reaction on human blood agar. The distribution of serotypes of V. vulnificus isolates were O1 to O8: 13 strains of O1, 6 strains of O2, 11 strains of O3, 9 strains of O4, 10 strains of O5, 7 strains of O6, 15 strains of O7, and 10 strains of O8, respectively. Eighty-one strains showed agglutination with O antisera, but 4 strains failed to show agglutination. In this study, the author suspected that serotypes of V. vulnificus isolates distributed also in the seaside of Korea as well as in most seaside of the world, and new serotypes were in existence in the seaside of Korea except reported up to now.

• 약학회지
• /
• 제43권6호
• /
• pp.802-808
• /
• 1999

Vibrio vulnificus cytolysin has been incriminated as one of the important virulence determinants in V. vulnificus infection. In the present study, the effects of Vibrio vulnificus cytolysin on platelets were examined. Vibrio vulnificus cytolysin induced platelet aggregation and increased intracellular calcium concentration ($[Ca^{2+}]_i$) of rat platelets. These effects were abolished in $Ca^{2+}-free$ buffer (2 mM EGTA). Cytolysin also potentiated ADP-and collagen-induced platelet aggregation. Lanthanum (2 mM) inhibited cytolysin-diduced platelet aggregation. However, another $Ca^{2+}$ channel blockers, verapamil ($20{\;}{\mu}M$) or mefenamic acid ($20{\;}{\mu}M$) did not block cytolysin-induced platelet aggregation. Osmotic protectants, sucrose (50 mM) and raffinose (50 nM) suppressed platelet aggregation by 35.9% and 63.4%, respectively. V. vulnificus cytolysin increased membrane conductances of platelet membranes. These results suggest that cytolysin-induced platelet aggregation is mediated via lanthanum sensitive-calcium influx which resulted from the pore formation by V. vulnificus cytolysin.