Vibrio vulnificus was isolated from a blood culture of a 41-year-old male patient with liver cirrhosis. He had eaten raw fish one day prior to the onset of clinical symptoms which included fever, chills, diarrhea and hypotension. He also developed cellulitis of the right leg which developed into a necrotic ulcer. The isolate was a slightly curved gram-negative bacillus and the colony morphology on a TCBS plate was similar to that of V. parabaemolyticus. Acid production from lactose was detected after 2 days of incubation. Other biochemical tests showed typical reactions of V vulnificus. The isolate was susceptible to all of the tested antibiotics except to clindamycin, colistin and penicillin G.
Adult ; Human ; Liver Cirrhosis/complications* ; Male ; Microscopy, Electron ; Septicemia/microbiology* ; Vibrio/isolation & purification ; Vibrio/ultrastructure ; Vibrio Infections/microbiology*

OBJECTIVETo investigate the occurrence of important foodborne pathogens in shellstock Pacific oysters in the food markets in South China.

METHODSFrom July 2007 to June 2008, retail oysters were collected in different seasons from South China and analyzed for the prevalence and levels of Listeria monocytogenes, Vibrio vulnificus and Vibrio parahaemolyticus.

RESULTSNone of L. monocytogenes could be detected in any of the 202 oyster samples tested, while E vulnificus and V. parahaemolyticus could be detected in 67 (54.9%) and 109 (89.3%) of the 122 oyster samples analyzed, respectively, with an MPN (most probable number) value greater than or equal to 3. V. vulnificus and V. parahaemolyticus with a more than 102 MPN/g were found in 36 (29.5%) and 59 (48.4%) of the 122 oyster samples, respectively. The tdh and trh genes were detected in 4 (0.3%) and 8 (0.6%) of the 1 349 V parahaemolyticus isolates, respectively. Of the 122 samples, 4 (3.3%) was positive for either tdh or trh. The levels of V. vulnificus and total V. parahaemolyticus in oysters in South China varied in different seasons.

CONCLUSIONV. vulnificus and pathogenic V. parahaemolyticus are frequently found in oysters in south China, which may pose a potential threat to public health. Data presented here will be useful for the microbiological risk assessment in oysters in China.

Animals ; China ; Commerce ; Food Microbiology ; Listeria monocytogenes ; isolation & purification ; Ostreidae ; microbiology ; Vibrio parahaemolyticus ; isolation & purification ; Vibrio vulnificus ; isolation & purification

OBJECTIVETo investigate the serologic type, phage-biotype and toxic factor of Vibrio cholerae isolated from different sea products, analyze the relation between the Vibrio cholerae in sea products and cholera epidemiology, and provide references for forecasting cholera epidemic situation and drawing out a preventing plan.

METHODThe biotype of strains isolated was analyzed by using type and phage-biotype serological methods. The toxic gene was detected by PCR.

RESULTSThe constituent ratio of V. cholerae O139, Ogawa and Inaba were, respectively, 48.44%, 20.31% and 31.25% in 64 strains of V. cholerae. The result of phage-biotype showed that the 26 strains of V. cholerae O1 were all non-epidemic strains. The result of toxic gene detecting showed that positive rate of V. cholerae O139 was higher than those of Ogawa and Inaba.

CONCLUSIONThe positive rate of toxic gene in V. cholerae O139 was high and the V. cholerae O139 was mainly in turtle, breed aquatics water and crustacean, so these sea products were the important sectors in cholera prevention and control.

Animals ; Bacteriophage Typing ; DNA, Bacterial ; genetics ; Seafood ; microbiology ; Serotyping ; Vibrio cholerae ; classification ; genetics ; isolation & purification ; Vibrio cholerae O1 ; isolation & purification ; Vibrio cholerae O139 ; isolation & purification

Vibrio vulnificus causes very severe infections. The organism is isolated, for the most part, from the blood, and skin lesions. Isolation from other sources, including the urine, is very rate. Four cases of V. vulnificus septicemia were bacteriologically diagnosed in 1984 and 1985 at Severance Hospital. All of the patients were men, 42 years and older, with preexisting liver disease. All of them showed hypotension and secondary skin lesions, and all expired. The organism was isolated from the blood in all patients, from the peritoneal fluid in one, and from skin lesions in two. From one patient, isolation from a urine speAmen was also accomplished. All of the isolates were typical in their characteristics such as in their forming green colonies on Thiosulfate citrate bile sucrose (TCBS) agar, delayed acid production from lactose, and growth in broth with 6% NaCl.
Adult ; Human ; Middle Age ; Septicemia/diagnosis* ; Vibrio/isolation & purification ; Vibrio Infections/diagnosis*

Recently, two successive epidemics of cholera were observed in Korea. The first one started in Suhchun-Goon of Choong-Chung-Namdo in August 1969, and the 2nd in Changyoung-Goon of Kyung Sang-Namdo in August 1970. With stool specimens collected from patients in Suhchun, Ko-Chang, Seoul, Inchun, Youngkwang, Chang-hang and Wooljin were epidemic areas in l969, and from patients in Chang-Young, Pusan, Taegu, and Seoul which were epidemic areas in l970, studies were carried out in 1) the isolation and identification of cholera vibrio strains 2) the differentiation of E1 Tor vibrio from classic cholera strains 3) the liberation test of Kappa-type phages and 4) El Tor phage typing. Five strains, which were isolated in the epidemic area of the Philippines in l969 were included for a comparative study. The results are summarized as follows. 1) The epidemic strains of 1969 were identified as Vibrio cholerae, Celebes type El Tor and those of 1970 epidemic as Vibrio cholerae, biotype El Tor, El Tor phage type IV. 2) Korean strains and Philippine strains of 1969 epidemic appeared to be identical in biochemical and serological tests and phage susceptibility tests.
Cholera/epidemiology ; Cholera/microbiology* ; Human ; Korea ; Philippines ; Vibrio/isolation & purification*

We describe a case of septic shock due to Vibrio alginolyticus presenting with fever and bilateral leg pain. Despite intensive management with antibiotics and inotropic agents, the patient died from septic shock 1 day after hospitalization. V. alginolyticus was isolated from both leg wounds and a blood culture. To the best of our knowledge, this is the first reported case of V. alginolyticus bacteremia in Korea.
Bacteremia/etiology/pathology ; Humans ; Korea ; Male ; Middle Aged ; Shock, Septic/*etiology/pathology ; Vibrio Infections/*complications/pathology ; Vibrio alginolyticus/*isolation & purification

Vibrio fetus was isolated from blood specimens of a subacute bacterial endocarditis patient. The 38 year old male patient was admitted to Severance Hospital in January 1970 for 11 days and again in July 1970 for 13 days. Subacute bacterial endocarditis was the major condtion. Aortic insufficiency and cholestatic hepatitis were the accessory diagnosis. The organism was isolated during the second admission. V. fetus human infection is known to be very rare, and the present case appears to be the first case in Korea. V. fetus grows very slowly with increased carbon dioxide tension which favours the growth. It is a slightly curved, S-shaped and spiral gram-negative organism. Many antibiotics, effective to gram negative organisms, inhibit the growth of the organism. V. fetus is an animal pathogen causing disease in ruminants. The patient enjoyed raw beef dishes. He could be infected with the organism by eating raw beef.
Adult ; Campylobacter fetus/isolation & purification ; Endocarditis, Subacute Bacterial/etiology ; Endocarditis, Subacute Bacterial/microbiology* ; Human ; Male ; Vibrio/isolation & purification* ; Vibrio Infections/microbiology*

OBJECTIVETo develop a real-time SYBR Green polymerase chain reaction (PCR) for detection of Vibrio cholerae serogroups O1 and O139, and to evaluate its reliability through detection of estuary water samples.

METHODSO antigen rfb genes specific for O1 and O139 were used for the design of PCR primers. The real-time SYBR Green PCR system in detecting O1 and O139 specific rfb genes in one tube was developed, and its sensitivity, specificity and reproducibility were evaluated. The ability of the real-time PCR in detection of estuary water samples was compared with the routine PCR and bacteria isolation.

RESULTSThe amplification of O1 or O139 specific target gene could be detected according to the melt curve temperature of amplicons. No amplification was observed in the templates of other 10 non-cholerae vibrios. When comparing to the real-time PCR to bacteria isolation in detection of 524 estuary water samples, it showed high sensitivity, plus also positive in real-time PCR detection among all the samples in which bacteria of O1 or O139 were isolated.

CONCLUSIONThe real-time SYBR Green PCR could be used as the first step of rapid environment screen of V. cholerae in water samples thus might enhance the efficiency of isolation in screening of large amount of water samples.

Environmental Monitoring ; methods ; Genes, Bacterial ; Polymerase Chain Reaction ; methods ; Reproducibility of Results ; Rivers ; microbiology ; Sensitivity and Specificity ; Vibrio cholerae O1 ; isolation & purification ; Vibrio cholerae O139 ; isolation & purification

OBJECTIVEThrough systematic monitoring of the number and strain types of O1 and O139 Vibrio cholerae in the Pearl River estuary waters to analyze it's relevance with the temperature of environment, and the relevance between strains in water and isolates during outbreaks and epidemics as well as to estimate the methods used for environmental water detection and the potential role in cholera surveillance program.

METHODSTwenty-four stations along the Pearl River were selected and the water samples were collected monthly from March 2006 to February 2007. V. cholerae O1 and O139 strains were isolated from the samples. Real-time PCR established in our laboratory was used to detect V. cholerae O1 and O139. Air temperature and water temperature were collected during sampling. Pulsed field gel electrophoresis (PFGE) was applied in molecular typing of the isolates.

RESULTS862 water samples were collected during the study period. A total number of 77 O1 and O139 V. cholerae were isolated in 67 water samples and the positive rates were 7.77% for isolation and 26.33% for real-time PCR. Seasonal trend of positive rates by month were approximately coincident with the change of water temperature. The positive rates in the stations in urban area were higher than those in other areas. Toxigenic O139 strains were found in one station located in downstream of a marine market. Most of the O1 and O139 isolates were non-toxigenic. No trend of seasonal variation of the strains was noticed. Within these 75 isolates, 49 PFGE patterns were identified and the patterns differed widely with the similarity of 57.4% - 100%.

CONCLUSIONV. cholerae existed as the natural habitat in estuary water of the Pearl River and showed obvious genetic diversity. Data from monitoring waters might show the separation of strains with certain seasonal association. But the crowd did not show the relationship between the infections. Results from water surveillance program might provide indicators on the appearance of cholera pathogen which might be used in assessing the environmental risk of cholera epidemics as well as the alert of cholera.

Electrophoresis, Gel, Pulsed-Field ; Environmental Monitoring ; Phylogeny ; Polymerase Chain Reaction ; Seasons ; Temperature ; Vibrio cholerae O1 ; classification ; genetics ; isolation & purification ; Vibrio cholerae O139 ; classification ; genetics ; isolation & purification

OBJECTIVETo establish a duplex nested PCR assay system which is capable for detecting O1 and O139 groups of Vibrio cholerae simultaneously, and is applicable to environmental specimens from routine cholera surveillance.

METHODSBased on nucleic acid sequences available in GenBank, six sets of primers were designed by PrimerSelect program of DNAStar, targeting the rfb gene that encodes the O antigens of O1 and O139 V. cholerae, respectively. The specificity of several primer combinations was tested. A duplex nested PCR assay system for simultaneously detecting O1 and O139 V. cholerae was established, subsequently, its sensitivity, specificity, reproducibility and field evaluation were tested. The sensitivity of this assay was evaluated by comparing detection limits of nested PCR and conventional PCR. Its reproducibility was tested by 32 positive samples (11 samples positive for O1, 21 samples positive for O139) from environmental surveillance. In addition, the selected amplicons from positive samples were sequenced and analyzed with relevant sequences.

RESULTSThis newly-established duplex nested PCR assay might distinguish O1 V. cholerae from O139 V. cholerae, based on fragment lengths of amplicons, with reliable reproducibility, and no specific amplification was observed as compared with other vibrio species. The sensitivity of this nested PCR was (15 000) higher than conventional PCR, and there was no interference observed with multiple primers and complicated templates in the same vial. In its field evaluation, 32 positive DNA samples were detected and be further confirmed with double or triple tests, implying reliable reproducibility and consistency of this system. These results indicated that this assay had reliable reproducibility. No amplification was observed in all negative specimens and also suggested the acceptable specificity of this assay. Sequence analysis of the selected amplification products revealed 100% homogeneous with relevant genes from V.cholerae, indicating that these amplicons were originated from V. cholerae.

CONCLUSIONThis duplex nested PCR assay system should be rapid, sensitive and especially applicable to small laboratories, and be suitable for dynamic environmental surveillance.

DNA, Bacterial ; Environmental Monitoring ; methods ; Polymerase Chain Reaction ; methods ; Sequence Analysis, DNA ; Vibrio cholerae O1 ; genetics ; isolation & purification ; Vibrio cholerae O139 ; genetics ; isolation & purification