Genotype ; Humans ; Seafood ; microbiology ; Vibrio Infections ; epidemiology ; microbiology ; Vibrio parahaemolyticus ; drug effects ; genetics ; pathogenicity

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*

Shewanella algae infections are rare in humans. Previously reported cases of S. algae have mainly been associated with direct contact with seawater. We report a case of primary S. algae bacteremia occurring after the ingestion of raw seafood in a patient with liver cirrhosis that presented a fulminent course of necrotizing fasciitis.
Bacteremia/*blood ; Fasciitis, Necrotizing/*microbiology ; Fatal Outcome ; Humans ; Korea ; Liver Cirrhosis/physiopathology ; Male ; Middle Aged ; Seafood/microbiology ; Sepsis/*microbiology ; Shewanella/*pathogenicity ; Vibrio/*pathogenicity ; Vibrio Infections/*blood

Vibrios are universal conditioned-pathogenic bacteria in marine culture environment, and the outbreak of vibrio disease resulted in a serious damage to aquaculture. Considering that vibrio disease in aquatic species, especially fishes, usually originated from mixed infection of different species (serotypes or subspecies) of vibrios, it is important to select the potential cross-protective protein antigens as candidates of polyvalent or combined vaccines. In present research, several strains of vibrios were isolated from infected large yellow croaker (Pseudosciaena crocea) and subsequently identified as six strains of V. harveyi, one V. parahaemolyticus and one V. alginolyticus by physiological, biochemical and molecular biological methods. Their outer membrane proteins (OMPs) were extracted and the SDS-PAGE and Western blotting results show that three immuno-blots with common molecular weight presented at approximate 45 kDa, 35 kDa and 22 kDa on their OMP electrophoretogram, indicating the existence of antigens with cross-protection in their OMPs. With the aids of combination of two-dimensional electrophoresis (2-D) and Western blotting and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS), a deduced porin (GenBank Accession No. ZP_01260407) from V. alginolyticus and a maltoporin precursor (GenBank Accession No. NP_801154) from V. parahaemolyticus were able to react with polyclonal antibody to whole V. harveyi, suggesting these two proteins could act as the cross-protective antigens and the vaccines prepared with these porins would be probable to bring cross protection to three different vibrios.
Animals ; Antigens, Bacterial ; immunology ; Bacterial Outer Membrane Proteins ; immunology ; Cross Reactions ; Fish Diseases ; microbiology ; Perciformes ; microbiology ; Vibrio ; classification ; immunology ; isolation & purification ; pathogenicity ; Vibrio Infections ; microbiology

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*

OBJECTIVESTo investigate the characteristics of virulence gene in Vibrio parahaemolyticus strains isolated from clinical patients and environment in Hangzhou, China.

METHODSThermostable direct hemolysin gene (tdh) and thermostable direct hemolysin-related hemolysin gene (trh) were determined in a total of 174 strains of V. parahaemolyticus isolated from patients and environment (seafood) in Hangzhou area by PCR.

RESULTSThe tdh was found in 92 out of 94 V. parahaemolyticus strains from food poisoning patients and in 33 out of 34 strains from sporadic diarrhea patients, and trh was not detected in all above clinical strains. Meanwhile the tdh was negative in all V. parahaemolyticus strains from environment, and the trh was also negative except one strain with urease activity. All strains with trh negative had no the activity of urease.

CONCLUSIONSThe V. parahaemolyticus strains from food poisoning patients and sporadic diarrhea patients are tdh positive and trh negative. The V. parahaemolyticus strains with tdh negative and almost trh positive in environment might be a potential pathogen in Hangzhou.

Bacterial Proteins ; genetics ; Bacterial Toxins ; genetics ; China ; Environmental Microbiology ; Foodborne Diseases ; microbiology ; Hemolysin Proteins ; genetics ; Humans ; Shellfish ; microbiology ; Urease ; genetics ; Vibrio Infections ; microbiology ; Vibrio parahaemolyticus ; classification ; genetics ; isolation & purification

BACKGROUNDVibrio vulnificus (Vv) is an estuarine bacterium that can cause primary septicemia as well as serious wound infections. However, little is known about the mechanisms by which Vv infects dendritic cells (DCs) and its effects on cytoskeleton. In this study, we aimed to investigate the invasion, internalization, and the organelles damage of the cultured dendritic cells (a DC 2.4 strain) during Vv infection.

METHODSThe study model was the cultured DCs infected by a Vv 1.758 strain. Electron microscopy was used to observe the localization of bacteria at the different time points of infection, cell morphology, and the process of organelles changes. The cytoskeleton structure including the microfilaments and the microtubules rearrangement was examined under a fluorescence microscope.

RESULTSThe Vv were pinocytosised into the DC cells through double-sides, and localized at 1 - 2 mm of the inner side membrane. It took 1.3, 1.9, and 3.4 hours to reach the infection ratio of 25%, 50%, and 75%, respectively. Using electron microscopy, the DCs had been observed to have developed chromatin aggregation within 4.0 hours, and significant cytoskeleton structure disruption was noted within 6.0 hours.

CONCLUSIONThe high lethality of Vv infection may be associated with the direct disruption of the DCs cytoskeleton structure.

Animals ; Apoptosis ; physiology ; Cells, Cultured ; Cytoskeleton ; metabolism ; ultrastructure ; DNA Fragmentation ; Dendritic Cells ; metabolism ; microbiology ; ultrastructure ; Mice ; Microscopy, Electron ; Microscopy, Electron, Transmission ; Vibrio Infections ; metabolism ; Vibrio vulnificus ; pathogenicity

OBJECTIVETo explore the phenetic and genetic features of clinical Vibrio parahaemolyticus strains from 2007-2009 in China.

METHODSA total of 135 clinical Vibrio parahaemolyticus strains, isolated from Zhejiang, Jiangsu, Sichuan, Guangxi, Liaoning Provinces during 2007 to 2009, were selected for the research. The occurrence of virulence genes thermostable direct hemolysin (tdh) and TDH-related hemolysin (trh), species-specific genes thermolabile hemolysin (tlh), toxR, VPM and gyrB, the pandemic clone gene markers(GS-PCR, PGS-PCR, orf8 and HU-α) in 135 Vibrio parahaemolyticus strains was detected by PCR. The antimicrobial susceptibilities to eight antimicrobial agents of the experimental strains were determined by the broth microdilution method. All strains were serotyped and underwent the cluster analysis with pulsed-field gel electrophoreses.

RESULTSThe results of PCR methods claim that all experiment strains carry species-specific genes such as tlh, toxR, gyrB, VPM. Among clinical strains, 85.9% (116/135) carry tdh and/or trh. 85.2% (115/135) were positive for tdh, and 3.0% (4/135) were positive for trh; while 3 strains carried both.66.7% (90/135) , 80.7% (109/135) , 65.2% (88/135) , 66.7% (90/135) clinical strains carried the genes of GS-PCR, PGS-PCR, orf8, HU-α, respectively. The results of antibiotics susceptibility test showed that 8.1% (11/135) strains were resistant to at least one agent, including 9 strains were resistant to ampicillin, 2 strains were resistant to trimethoprim and sulphamethoxazole, and 1 strain were resistant to tetracycline. All clinical strains were sensitive to cefotaxime, ceftazidime, gentamicin, ciprofloxacin and chloromycetin.Serological analysis of the O and K antigens claimed that a total of 29 serotypes were identified for clinical strains, predominantly O3, O4 and O1 groups, accounting for 89.6% (121/135). O3: K6 was dominant serotype, accounting for 56.3% (76/135). The pandemic flora in China included O3: K6, O4: K68, O1: K36, O1: K25, O1: K5 and O3: K29 serotypes.Genomic DNAs of 135 clinical strains were digested with SfiI and NotI, the molecular size of PFGE restriction fragments used for analysis mainly ranged from 30-700 kb.When subjected to UPGMA clustering, 6 and 9 clusters were grouped by SfiI and NotI, and the minimal similarity was 52.6% and 58.7%, and pandemic flora were located in C groups and D group, respectively.

CONCLUSIONMost of Vibrio parahaemolyticus strains isolated from clinical sources in China were pathogenic. The pandemic clone, especially O3: K6 was prevalent. The GS-PCR and HU-α genes were reliable markers to identify the pandemic flora. The serotype by PFGE was reliable to distinguish the pandemic flora and the sporadic strains.

China ; epidemiology ; Drug Resistance, Bacterial ; Genes, Bacterial ; Humans ; Molecular Epidemiology ; Vibrio Infections ; epidemiology ; microbiology ; Vibrio parahaemolyticus ; genetics ; isolation & purification ; pathogenicity ; Virulence ; genetics

OBJECTIVETo investigate the influence of genetic polymorphism in NF-E2-related factor-2 (nrf2) gene promoter locus at 336 in alcoholic liver disease (ALD) with Vibrio vulnificus (VV) sepsis.

METHODSThrough the simple random sampling method, C57B6 male mice were divided into normal feeding group (group A, 10 mice), alcoholic liver disease group (group B, 10 mice), normal feeding group infected with VV through intraperitoneal injection (group C, 8 mice), alcoholic liver disease group infected with VV (group D, 110 mice). Through gene sequencing method, nrf2 gene promoter 336 polymorphism in D group was analyzed and grouped into: non-mutation group (336T) (group D1, 7 mice) and mutation group (336C) (group D2, 10 mice). Through RT-PCR, Western-blotting and ELISA method, expressions of nrf2, tumor necrosis factor-α (TNF-α), interleukin-10 (IL-10), high mobility group protein 1 (HMGB(1)) gene and protein of liver were measured. The pathological changes in liver were recorded with light microscope.

RESULTSAfter infected with VV for 48 hours for A, B, C, D1, D2 group, the expression medians of nrf2 mRNA in liver were 0.115, 0.173, 0.211, 0.764, 0.352, respectively (χ(2) = 40.64, P < 0.05), the expression medians of IL-10 mRNA in liver were 0.338, 0.637, 1.002, 1.825, 1.403, respectively (χ(2) = 41.05, P < 0.05), the expression medians of TNF-α mRNA in liver were 0.140, 0.254, 0.372, 0.399, 0.699, respectively (χ(2) = 38.16, P < 0.05), the expression medians of HMGB(1) mRNA in liver were 0.230, 0.410, 0.668, 0.508, 1.021, respectively (χ(2) = 31.45, P < 0.05). After infected with VV 48 hours for mice in A, B, C, D1, D2 group, the expression medians of nrf2 protein in liver were 0.908, 1.461, 2.061, 3.982, 2.243, respectively (χ(2) = 33.72, P < 0.05), the expression medians of IL-10 protein in liver were 13.97, 22.54, 30.14, 57.98, 41.53, respectively (χ(2) = 37.31, P < 0.05), the expression medians of TNF-α protein in liver were 114.07, 142.94, 175.44, 174.60, 266.11, respectively (χ(2) = 32.29, P < 0.05), the expression medians of HMGB(1) protein in liver were 2.01, 6.05, 9.62, 6.24, 12.89, respectively (χ(2) = 36.94, P < 0.05). Compared with group A, there were large amount of fat drops, fatty changes in group B, inflammatory cell infiltration, disorder of hepatic cell in group C, and extension of hepatic duct and vein, edema of liver cells and disorder of hepatic cells in group D.

CONCLUSIONThe nrf2 gene promoter of T336C mutation in C57B6 mouse of ALD can significantly decrease the expression of nrf2, and intensify organ inflammation and damage when they were infected by VV.

Animals ; Liver Diseases, Alcoholic ; complications ; genetics ; metabolism ; microbiology ; Male ; Mice ; Mice, Inbred C57BL ; NF-E2-Related Factor 2 ; genetics ; metabolism ; Polymorphism, Single Nucleotide ; Promoter Regions, Genetic ; Sepsis ; complications ; genetics ; microbiology ; Vibrio Infections ; complications ; genetics ; Vibrio vulnificus

Vibrio cholerae non-O1 have caused several well-studied food-borne outbreaks of gastroenteritis and also have been responsible for sporadic cases of otitis media, wound infection, and bacteremia. Few cases of liver abscess caused by Vibrio cholerae non-O1 have been reported. A 73-year-old man with underlying diabetes mellitus was admitted with nausea, vomiting, dyspepsia and febrile sensation. We identified Vibrio cholerae non-O1 in his blood cultures and multiple hepatic microabscess on abdominal computed tomography. He was treated with systemic antibiotics and fluid therapy, but died due to septic shock on sixth day. We report here, a case of liver abscess with bacteremia due to Vibrio cholerae non-O1 in a patient with diabetes mellitus.
Aged ; Anti-Bacterial Agents/therapeutic use ; Bacteremia/drug therapy/*microbiology ; Ceftriaxone/therapeutic use ; Humans ; Liver Abscess/*diagnosis/drug therapy/microbiology ; Male ; Metronidazole/therapeutic use ; Shock, Septic/diagnosis ; Tomography, X-Ray Computed ; Vibrio Infections/drug therapy/*microbiology ; Vibrio cholerae non-O1/*isolation & purification