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Bacteremia by non-O1/non-O139 Vibrio cholerae is a rare entity associated with high mortality rates. We report a case of non-O1/non-O139 V. cholerae bacteremia confirmed by polymerase chain reaction and agglutination tests. The clinicoepidemiological characteristics and therapeutic options for this infection are also described.
La bacteriemia por Vibrio cholerae no-O1/no-O139 es una entidad poco frecuente que se asocia con altas tasas de mortalidad. Se reporta un caso de bacteriemia por V. cholerae no-O1/no-O139 confirmado por reacción en cadena de la polimerasa y test de aglutinación. Se describen las características clinicoepidemiológicas y las opciones terapéuticas para esta infección.
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Bacteriemia , Vibrio cholerae não O1 , Fatores de VirulênciaRESUMO
El cólera es una toxoinfección alimentaria ocasionada por la ingesta de agua y alimentos contaminados por el Vibrio cholerae. Es una de las enfermedades más antiguas de la humanidad y las primeras descripciones corresponden a Hipócrates. La primera epidemia documentada, sucedió en la India en 1817 y se extendió a Turquía y a los países árabes. En nuestro país, el primer brote ocurrió en 1856 en la ciudad de Bahía Blanca, asociada a la llegada de navíos con enfermos y a las deficientes condiciones sanitarias de la ciudad. Los sucesivos brotes se acompañaron de una alta mortalidad, a tal punto que el Dr. José María Penna señaló que costó más vidas a la nación que la guerra con Paraguay. En el presente artículo se analizan los sucesivos brotes de cólera en nuestro país
Cholera is a food poisoning caused by the ingestion of food and water contaminated by Vibrio cholerae. It is one of the oldest diseases of humanity and the first descriptions correspond to Hippocrates. The first documented epidemic occurred in India in 1817 and spread to Turkey and the Arab countries. In our country, the first outbreak occurred in 1856 in the city of Bahía Blanca, associated with the arrival of ships with patients and poor sanitary conditions in the city. The successive outbreaks were accompanied by high mortality, to the point that Dr. José María Penna pointed out that it cost the nation more lives than the war with Paraguay. This article analyzes the successive outbreaks of cholera in our country
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Humanos , Masculino , Feminino , Cólera/história , Cólera/epidemiologia , Epidemias/históriaRESUMO
Pullulanase is a starch debranching enzyme, which is difficult in secretory expression due to its large molecular weight. Vibrio natriegens is a novel expression host with excellent efficiency in protein synthesis. In this study, we achieved secretory expression of the full-length pullulanase PulA and its truncated mutant PulN2 using V. natriegens VnDX strain. Subsequently, we investigated the effects of signal peptide, fermentation temperature, inducer concentration, glycine concentration and fermentation time on the secretory expression. Moreover, the extracellular enzyme activities of the two pullulanases produced in V. natriegens VnDX and E. coli BL21(DE3) were compared. The highest extracellular enzyme activity of PulA and PulN2 in V. natriegens VnDX were 61.6 U/mL and 64.3 U/mL, which were 110% and 62% that of those in E. coli BL21(DE3), respectively. The results indicated that V. natriegens VnDX can be used for secretory expression of the full-length PulA with large molecular weight, which may provide a reference for the secretory expression of other large molecular weight proteins in V. natriegens VnDX.
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Escherichia coli/genética , Fermentação , Vibrio/genéticaRESUMO
OBJECTIVE@#This study aimed to investigate whether the VCA0560 gene acts as an active diguanylate cyclase (DGC) in Vibrio cholerae and how its transcription is regulated by Fur and HapR.@*METHODS@#The roles of VCA0560 was investigated by utilizing various phenotypic assays, including colony morphological characterization, crystal violet staining, Cyclic di-GMP (c-di-GMP) quantification, and swimming motility assay. The regulation of the VCA0560 gene by Fur and HapR was analyzed by luminescence assay, electrophoretic mobility shift assay, and DNase I footprinting.@*RESULTS@#VCA0560 gene mutation did not affect biofilm formation, motility, and c-di-GMP synthesis in V. cholerae, and its overexpression remarkably enhanced biofilm formation and intracellular c-di-GMP level but reduced motility capacity. The transcription of the VCA0560 gene was directly repressed by Fur and the master quorum sensing regulator HapR.@*CONCLUSION@#Overexpressed VCA0560 functions as an active DGC in V. cholerae, and its transcription is repressed by Fur and HapR.
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Vibrio cholerae/genética , Biofilmes , Percepção de Quorum , Mutação , Regulação Bacteriana da Expressão Gênica , Proteínas de Bactérias/genéticaRESUMO
@#Vibrio cholerae is a gram-negative bacterium synonymous with its namesake disease, cholera. Thus, gastrointestinal symptoms are the norm and V. cholerae is very rarely associated with skin and soft tissue infections. We describe a case of a 63-year-old Chinese woman with multiple medical comorbidities on corticosteroid therapy who developed fever and a painful swelling on her left leg after being pricked by a branch while gardening. There was no abdominal pain, vomiting or diarrhea. A diagnosis of bullous cellulitis was made clinically, and blood was sent for bacteriological culture. A beta-hemolytic commashaped gram-negative bacillus was isolated from the blood. It was also oxidase-positive and produced an acid/alkaline (A/K) reaction on triple sugar iron agar. It was identified biochemically as Vibrio cholerae. After additional testing, it was found to be of the O1 serogroup and Ogawa serotype. The infection resolved following a 10-day course of high-dose co-trimoxazole therapy.
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ObjectiveIn this paper， the cause of an outbreak of foodborne disease in Huzhou City was analyzed， which may help avoid the recurrence of such incidents. MethodsThrough the field epidemiological investigation， the case definition was formulated and the questionnaire survey was carried out in the case group and the control group. In addition， the chi-square test and logistic regression method were used to identify the factors affecting the outbreak. The patient stool samples， food samples， environmental samples and water samples were collected and used for the laboratory test. The PFGE molecular typing was conducted on the isolated positive strains. ResultsThe number of people exposed during the same period was 410， and the number of possible cases was 18， with an incidence of 4.39%. Generally， the main symptoms were abdominal pain and diarrhea， accompanied by nausea， fatigue， fever and others. For case-control analysis， 17 of the 18 patients were included in the case group， and 19 non-patients were into control group. The results suggested that the risk factors were blanched deep-water shrimp（OR=19.42， 95%CI=1.06‒357.02， P=0.046）and steamed Ao Long （Australian lobster） with garlic and vermicelli （OR=22.01， 95%CI=1.24‒390.70， P=0.035）. According to the laboratory test results Vibrio parahaemolyticus （VP） was positive in 5 cases， and the serum type was is O10∶K4. In the reserved food， VP was positive in the samples of steamed Australian lobster with garlic vermicelli and lamb chops. The serum type was O5∶Kut. ConclusionThis incident was an outbreak of foodborne disease caused by the consumption of wedding food contaminated by VP. The dinner was served by Hotel B on September 17. Moreover， the suspicious foods include the blanched deep-water shrimp and steamed lobster with garlic vermicelli.
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ObjectiveTo ascertain the causes of a food poisoning incident and provide references for the prevention of similar incidents in the future. MethodsCase investigation was conducted through field epidemiological investigation methods， and suspicious meals and foods were searched by the analytical epidemiological method. A food hygiene investigation was conducted in the establishment involved and samples of suspicious food， processing steps， and cases were collected for laboratory testing. ResultsA total of 91 individuals meeting the case definition were identified， resulting in an attack rate of 14.97% （91/608）. The main clinical manifestations included abdominal pain （97.80%）， diarrhea （84.62%）， nausea （62.64%）， vomiting （72.53%）， fever （12.09%）， and increased white blood cells （90.11%）. The peak incidence occurred from 16：00 to 18：00 on June 15. The epidemic curve showed a point-source exposure pattern， with an incubation period of 1 h minimum and 10 h maximum， and an average of 5 h. Analytical epidemiological studies indicated that lunch on June 15 was the suspicious meal （χ2=38.78， P<0.001）， and those who consumed cold-dressed tofu with preserved eggs had a significantly higher risk of falling ill compared to non-consumers （χ2=105.21， P<0.001）. Laboratory testing results revealed Vibrio parahaemolyticus detected in 1 employee’s anal swab and 18 cases’ anal swabs. Staphylococcus aureus was detected in 1 food ingredient and 1 case’s anal swab. The remaining samples tested negative. ConclusionThe cause of this food poisoning incident is Vibrio parahaemolyticus. The cause is the canteen’s supply of cold-dressed tofu with preserved eggs beyond its permissible business scope， potentially leading to cross-contamination during food processing. Regulatory authorities should strengthen routine law enforcement inspections and monitoring. Food service establishments should strengthen food safety awareness， standardize operational procedures in strict accordance with relevant national laws and regulations and food safety standards， and strive to reduce the occurrence of foodborne disease incidents at their source.
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OBJECTIVE@#This study investigated how the natural phytophenol and potent SIRT1 activator resveratrol (RSV) regulate necroptosis during Vibrio vulnificus (V. vulnificus)-induced sepsis and the potential mechanism.@*METHODS@#The effect of RSV on V. vulnificus cytolysin (VVC)-induced necroptosis was analyzed in vitro using CCK-8 and Western blot assays. Enzyme-linked immunosorbent assays and quantitative real-time polymerase chain reaction, western blot, and immunohistochemistry and survival analyses were performed to elucidate the effect and mechanism of RSV on necroptosis in a V. vulnificus-induced sepsis mouse model.@*RESULTS@#RSV relieved necroptosis induced by VVC in RAW264.7 and MLE12 cells. RSV also inhibited the inflammatory response, had a protective effect on histopathological changes, and reduced the expression level of the necroptosis indicator pMLKL in peritoneal macrophages, lung, spleen, and liver tissues of V. vulnificus-induced septic mice in vivo. Pretreatment with RSV downregulated the mRNA of the necroptosis indicator and protein expression in peritoneal macrophages and tissues of V. vulnificus-induced septic mice. RSV also improved the survival of V. vulnificus-induced septic mice.@*CONCLUSION@#Our findings collectively demonstrate that RSV prevented V. vulnificus-induced sepsis by attenuating necroptosis, highlighting its potency in the clinical management of V. vulnificus-induced sepsis.
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Animais , Camundongos , Necroptose , Resveratrol/uso terapêutico , Vibrio vulnificus , Sepse/tratamento farmacológico , Western BlottingRESUMO
Objective To prepare a biomimetic nano carrier macrophage membrane hybrid liposome by heterozygous macrophage membrane and liposome, which could be used for the clearance and toxicity inhibition of Vibrio vulnificus hemolysin A (VvhA). Methods Macrophage membrane was extracted and hybridized with liposome by thin-film evaporation combined extrusion method. The hybridized liposome of macrophage membrane was constructed and characterized. The in vitro detoxification ability of the hybridized vector was evaluated by hemolysis test and cytotoxicity test. The detoxification ability of the vector was evaluated by mouse skin infection model. Results Anti toxoid studies in vivo and in vitro showed that the anti-hemolysis rate of macrophage membrane heterozygous liposomes in vitro reached 97.03%, which could effectively inhibit the skin ulceration in subcutaneous infected mice and make the survival rate of abdominal infected mice reach 80%. Conclusion The constructed macrophage membrane hybrid liposome had high detoxification ability, which could provide a potential solution and research basis for the prevention and treatment of Vibrio vulnificus infection.
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@#Abstract: Objective To investigate the molecular characteristics and drug resistance of non-O1/non-O139 Vibrio cholerae in Zhongshan City, and to provide laboratory basis for cholera prevention and control. Methods The strains of non-O1/non-O139 Vibrio cholerae isolated from sporadic patients and aquatic products from 2015 to 2021 in Zhongshan city were collected. The identification and cluster analysis of the strains were analyzed by matrix assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF-MS), the ctxA virulence gene of strains were detected by real-time fluorescence quantitative PCR, the cluster analysis of the strains was analyzed by pulsed-field gel electrophoresis (PFGE), and the drug resistance of the strains were analyzed by microbroth dilution method. Results From 2015 to 2021, 33 strains of non-O1/non-O139 Vibrio cholerae were isolated from Zhongshan City, including 28 strains from sporadic patients and 5 strains from aquatic products. Through MALDI-TOF-MS identification, 33 strains of non-O1/non-O139 Vibrio cholera can be identified to the level of species, and the identification results were all Vibrio cholerae. Among 33 non-O1/non-O139 Vibrio cholerae strains, 1 strain carried the ctxA virulence gene. The drug-resistant strains accounted for 69.7% (23/33), and the multidrug resistant strains accounted for 18.2% (6/33). A total of 7 kinds of drug resistance spectrum were produced, including 3 kinds of multidrug resistant spectrum, and showed drug resistance to 8 antibiotics, among which the resistance rates to streptomycin, cefazolin and compound sulfamethoxazole were above 30%. The 33 strains of non-O1/non-O139 Vibrio cholerae were divided into 32 PFGE fingerprints with a similarity ranging from 61.7% to 100%. MALDI-TOF-MS cluster analysis divided 33 non-O1/non-O139 Vibrio cholerae strains into two clusters. Conclusions The results of molecular typing of non-O1/non-O139 Vibrio cholerae in Zhongshan City presented diversity, and no significant correlation was found between PFGE and MALDI-TOF-MS cluster analysis. The strains demonstrated various degrees of resistance to certain antibiotics, and there were multidrug-resistant and toxigenic strains. Therefore, it is necessary to alert to the harmfulness of non-O1/non-O139 Vibrio cholerae and enhance monitoring.
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Objective To evaluate the risk of disease of Vibrio parahaemolyticus in aquatic products of raw food animals for population in guangzhou,and determine risk management points. Methods VP quantitative detection was carried out in aquatic products of raw food animals sold in Guangzhou from 2009 to 2022.sQMRA was applied to assess Vibrio parahaemolyticus risk of aquatic products of raw food animals. According to stratified analysis based on the pollution of Vibrio parahaemolyticus and evaluation results,carry out risk management and analysis. Results Among the 98 samples were detected positive of VP from 1 343 samples from 2009 to 2022 , with an overall positive rate of 7.30%.The number of Vibrio parahaemolyticus infection cases caused by eating aquatic products of raw food animals in Guangzhou was 3012. If the proportion of raw food is reduced , the number of Vibrio parahaemolyticus infection cases will be significantly reduced. The number of cases caused by eating raw fash will be reduced from 2128 to 217.The detection rate of Vibrio parahaemolyticus in raw fresh water products was much higher than that in marine products. The probability of infection in the population was higher. The number of cases caused by eating raw fash was the highest.The detection rate of Vibrio parahaemolyticus was higher in raw crustaceans and molluscs. The incidence of Vibrio parahaemolyticus infection cases caused by eating raw fash in the four quarters varied from high to low as such sequence ,4.93×10-5 in the three quarters , 2.53×10-5 in the second quarter , 2.40×10-5 in the first quarter ,1.77×10-5 in the fourth quarter . Conclusion The risk of disease of Vibrio parahaemolyticus in aquatic products of raw food animals was higher. The public health education should be done well. Aquatic products should be cooked thoroughly before eating . Reduce the intake of raw aquatic products and avoid cross contamination. Focus on the risks of summer and autumn seasons and seafood such as crustaceans and molluscs. Concentrate on scientific research on Vibrio parahaemolyticus pollution of fresh water products.
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Objective To investigate the etiological characteristics of food poisoning isolates of Vibrio parahaemolyticus (VP) from 2019 to 2021 in Zhongshan City. Methods A total of 37 strains of Vibrio parahaemolyticus isolated from 8 food poisoning incidents in Zhongshan City from 2019 to 2021 were collected, including 1 residual food isolate and 36 human isolates. The genetic correlation of Vibrio parahaemolyticus food poisoning isolates in this region was analyzed by serological typing, virulence gene detection (TLH, TDH, and TRH), drug sensitivity test, pulsed field gel electrophoresis (PFGE) and multipoint sequence typing (MLST). Results The 37 strains of Vibrio parahaemolyticus were divided into 4 serotypes: O3:K6, O10:K4, O4:K8, and O4:KUT. The tdh+ and trh- were the main virulence genotypes, accounting for 97.30% (36/37). The drug resistance rate of cefazolin was 40.54% (15 strains R, 22 strains I), and no multidrug-resistant strains were found. The 37 VP strains were divided into 23 PFGE types and 6 cluster groups, with correlation coefficients ranging from 60.4%-100%. The multipoint sequencing typing showed that the 37 VP strains were divided into 9 ST types and 3 complex groups, of which ST3 type was the main type (23 strains, 62.1%). Conclusion This study has found that the dominant virulence types of Vibrio parahaemolyticus food poisoning isolates in Zhongshan City from 2019 to 2021 are tdh+ and trh-, and 37 representative strains can be divided into 6 PFGE clusters and 9 ST types with MLST type being mainly ST3. This study has identified the rare serotype O10:K4 which has caused an increase in the proportion of food poisoning events, suggesting that we should strengthen detection and be alert to the risk of continued local epidemics of new rare serotype strains.
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Wound infection is becoming a considerable healthcare crisis due to the abuse of antibiotics and the substantial production of multidrug-resistant bacteria. Seawater immersion wounds usually become a mortal trouble because of the infection of Vibrio vulnificus. Bdellovibrio bacteriovorus, one kind of natural predatory bacteria, is recognized as a promising biological therapy against intractable bacteria. Here, we prepared a B. bacteriovorus-loaded polyvinyl alcohol/alginate hydrogel for the topical treatment of the seawater immersion wounds infected by V. vulnificus. The B. bacteriovorus-loaded hydrogel (BG) owned highly microporous structures with the mean pore size of 90 μm, improving the rapid release of B. bacteriovorus from BG when contacting the aqueous surroundings. BG showed high biosafety with no L929 cell toxicity or hemolysis. More importantly, BG exhibited excellent in vitro anti-V. vulnificus effect. The highly effective infected wound treatment effect of BG was evaluated on mouse models, revealing significant reduction of local V. vulnificus, accelerated wound contraction, and alleviated inflammation. Besides the high bacterial inhibition of BG, BG remarkably reduced inflammatory response, promoted collagen deposition, neovascularization and re-epithelization, contributing to wound healing. BG is a promising topical biological formulation against infected wounds.
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@#Abstract: Objective To understand the contamination status, drug resistance, virulence gene carrying status, and molecular typing characteristics of Vibrio parahaemolyticus (VP) in aquatic products sold in Nanchang City. Methods A total of 170 commercial crayfishes, freshwater fish frogs and related smears samples were collected from various farmers' markets in Nanchang from March to September 2021. The strains of V. parahaemolyticus were detected and isolated from the samples. Antibiotic resistance test, virulence gene test, and pulsed-field gel electrophoresis (PFGE) molecular typing analysis were carried out. Results Among the collected samples, V.parahaemolyticus was only isolated from crayfish and crayfish smear samples, with a total of 35 strains of VP isolated. No V.parahaemolyticus strain was isolated from other freshwater fish, frogs, and their smear samples. Among the 17 common antibiotics tested, only two trains showed resistance to ampicillin, and one strain to streptomycin, , and all were sensitive to other antibiotics; all 35 strains of V. parahaemolyticus carried the gene, but only one strain carried the heat-resistant related hemolysin gene trh, and no direct heat-resistant hemolysin gene tdh positive strain was found; PFGE pattern clustering showed that there was no strain with the same PFGE pattern, and there was no obvious dominant cluster among these strains, and their genetic relationship was relatively distant. Conclusions The contamination of V. parahaemolyticus in small and medium-sized crayfish sold in the market in Nanchang City is relatively serious. The V. parahaemolyticus isolates in these polluted crayfish generally do not carry key virulence genes such as tdh, are sensitive to common antibiotics, and only have low-level resistance to ampicillin and streptomycin. PFGE pattern clustering showed that V. parahaemolyticus does not have no obvious dominant cluster, and these strains have rich genetic diversity, indicating that they may have different sources.
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Vibrio parahaemolyticus is a major pathogen frequently found in seafood. Rapid and accurate detection of this pathogen is important for the control of bacterial foodborne diseases and to ensure food safety. In this study, we established a one-pot system that combines uracil-DNA glycosylase (UDG), loop-mediated isothermal amplification (LAMP), and clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated protein 12b (Cas12b) for detecting V. parahaemolyticus in seafood. This detection system can effectively perform identification using a single tube and avoid the risk of carry-over contamination.
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Vibrio parahaemolyticus/genética , Uracila-DNA Glicosidase/genética , Temperatura Alta , Sistemas CRISPR-Cas , Inocuidade dos AlimentosRESUMO
Resumen Vibrio vulnificus es una de las especies de Vibrio más virulentas que se conocen. Es una bacteria de distribución universal. El primer caso registrado en Uruguay se produjo en 2001, y desde entonces ocurren varias infecciones por año. Recientemente, en ese país, V. vulnificus fue responsable de una infección de partes blandas de curso letal. Aunque no han sido comunicados casos de infección humana por esta especie en Argentina, se ha identificado recientemente Vibrio vulnificus en muestras asociadas con microplancton en el estuario del Río Negro. Presentamos el caso de una infección grave de piel y partes blandas por V. vulnificus a partir de una herida abierta en un paciente en contacto con medio acuático marino en la costa de Uruguay del Río de la Plata. El aislamiento de vibrios en muestras de heridas puede causar un daño en los tejidos con rápida progresión, en particular V. vulnificus, que tiene una alta mortalidad sin la precoz y apropiada intervención. En nuestro caso, la rápida identificación del microorganismo permitió avalar el tratamiento empírico utilizado, con una buena evolución clínica.
Abstract Vibrio vulnificus is one of the most virulent Vibrio species known. It is a bacterium with universal distribution. The first case registered in Uruguay occurred in 2001 and, since then, several infections have occurred per year. Recently, in this country, V. vulnificus was responsible for a fatal soft tissue infection. Although no cases of human infection with this species have been reported in Argentina, researchers have recently identified V. vulnificus in samples associated with microplankton in the Rio Negro estuary. We present the case of a severe skin and soft tissue infection by V. vulnificus from an open wound in a patient in contact with a marine aquatic environment on the coast of the River Plate, in Uruguay. Isolation of vibrios from wound specimens can cause rapidly progressing tissue damage, particularly V. vulnificus which has a high mortality rate without early and appropriate intervention. In our case, the rapid identification of the microorganism allowed us to support the empirical treatment used, which a good clinical evolution.
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In the coastal areas of the world, most Vibrio species have been incriminated as notorious agents causing foodborne, wound and other infections. These pathogens are known to be associated with the consumption of raw or undercooked seafoods or the exposure of wounds to warm seawater. Aim: Therefore, this research work was designed with the aim of assessing the microbiological quality of the water bodies as well as the seafoods consumed in Cross River State (CRS). Study Design: The Study was designed using the completely randomized block design and the data was analyzed using of two-way analysis of variance, Generalized Linear Model Univariate analysis. Significant means were separated using the Least significant difference (LSD). Place and Duration of Study: This study was done in the Department of Microbiology, University of CRS, Calabar, CRS, Nigeria, between 2016-2019. Methodology: we evaluated a variety of seafoods viz; crayfish, blue crabs, Periwinkles, apple nails, red lobsters etc. collected from major Beaches, markets and other sale points and water sources (rivers streams sea and gutters) in Calabar, CRS of Nigeria, using standard bacteriological techniques, for the prevalence of Vibrio species. Results: The mean percentage mean viable cell counts obtained ranged from 1.79�45 (seawater)-9.15�79CFU/mL (gutter water) and 7.68�58 (Blue Crab)- 11.37�82 CFU/g (fish) in the Rainy season. The counts for the Dry season Ranged from 1.79 �42 (Seawater)-8.94� 4.51(gutter water), and 5.83 7.21 CFU/g (apple snail) -12.64 5.95 CFU/g (Fish). The total percentage mean counts obtained were 8.09�91 CFU/mL in the Rainy Season to 7.61�58 CFU/mL in the dry Season. From both seasons, the overall total mean count was 11.09�94 CFU/ml. From the nine locations evaluated in this study, it was observed that the Mean percentage counts for the Northern Senatorial District (NSD) ranged from 2.81� 3.49 (Ogoja)- 3.14 �07CFU/mL (Obudu). For the Central (CSD) the range was from 3.34 �20 (Boki)- 9.89 �15 (Ikom), while for the Southern (SSD) it was from12.01� 6.52 (Akamkpa)- 14.47 �44 (Calabar). The overall Total percentage mean counts from all the three Senatorial Districts was 14.03�86 CFU/mL. From the Northern Senatorial District, the total Percentage mean was 3.01�77 CFU/mL, 7.05�79 CFU/mL from the Central and 13.49� 5.72 CFU/mL from the Southern Senatorial District. The Vibrio pathotypes isolated include Vibrio cholerae (V. cholerae) (both O1 and non-O1 serotypes) 1155 (31.61%), Vibrio parahaemolyticus (V. parahaemolyticus), 752 (20.58%), Vibrio fluvialis (V. fluvialis) 480 (13.14%), V. vulnificus 473 (12.94%) Vibrio mimicus (V. mimicus) 400 (10.95%) and Other Vibrios 394 (10.78%). Out of the 3654 Vibrio isolates, the greatest number 663�31 (18.14%) were from Seawater, while the least 133�.84 (3.64%) were from the Gutter Water. Also, the highest number 1245�61 (34.07%) came from Calabar, and the least 102�.65 (2.79%) from Obanlikwu. The NSD had the least number 327 (8.95%), followed by the CSD with 570 (15.59%) and then the SSD with 2757 (75.45%) as the highest number of isolates. Conclusion: The presence of these pathogenic bacterial species in common seafoods in this area is of great public health concern. It is therefore important that serious emphasis be laid on proper cooking of these seafoods as well as the establishment of regular hygiene surveillance strategies in the state.
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@#Some of Vibrio species is well known as pathogenic bacteria in aquaculture and the marine industry. Its infection is able to generate a massive outbreak and affect the fish population, especially for net caged fish such as seabass. This study was conducted to investigate the prevalence of Vibrio spp. isolated from seabass (Lates calcarifer) in Sri Tujuh Lagoon, Tumpat, Kelantan. Then, to determine the antibiotic resistance in Vibrio isolates. Polymerase chain reaction (PCR) was used to detect Vibrio species using specific primer VR169 and VR744 with estimation base pair size band, 597 bp and further identified by sequencing. On the other hand, antibiotic susceptibility tests were continued by using 13 types of antibiotics; kanamycin (K30), chloramphenicol (C30), neomycin (N10), ampicillin (AMP10), nitrofurantoin (F300), tetracycline (TE30), streptomycin (S10), norfloxacin (NOR10), ciprofloxacin (CIP5), nalidixic acid (NA30), gentamicin (CN10), doxycycline (DO30) and sulfamethoxazole (SXT100). As a result, 14 Vibrio isolates were identified, including Vibrio fluvialis (n=6), Vibrio parahaemolyticus (n=3), Vibrio harveyi (n=2) and each isolate for Vibrio vulnificus, Vibrio alginolyticus and Vibrio spp. The results showed that all isolates were sensitive to most antibiotics except ampicillin, neomycin and streptomycin. The MAR index value was ranging from 0 to 0.31. This study demonstrates the prevalence of Vibrio spp. in seabass and the report on multidrug resistance strains that could be of concern to the fish farmers. In addition, data from this study can be further used in fish disease management plans.
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Objective:To study the virulence-related functions and the pathogenic molecular mechanism of the transcription factor OmpR in Vibrio vulnificus. Methods:The Vibrio vulnificus ompR mutant (Δ ompR) strain was constructed using a markerless gene deletion system. Changes in the tolerance to osmotic pressure, swarming mobility and biofilm synthesis were detected by growth curve, swimming assay and crystal violet assay, respectively. Colony counting methods were used to analyze the cytoadherence of the Δ ompR strain. The cytotoxicity of the Δ ompR strain was measured by lactate dehydrogenase (LDH) releasing test. Real-time fluorescent quantitative RT-PCR (qRT-PCR) was used to detect the changes in the expression of target genes including ompN, ompU, ompA and hcp2 at mRNA level in the Δ ompR. Results:The Δ ompR strain was constructed successfully. Compared with the wild-type strain, the mutant strain showed decreased tolerance to high osmotic pressure, suppressed capability of biofilm formation and reduced cytoadherence and cytotoxicity, whereas no significant difference in motility was detected. The expression of ompN gene at mRNA level in the Δ ompR strain was down-regulated ( P<0.05), while the expression of other target genes showed no significant changes. Conclusions:The transcription factor OmpR regulated the tolerance to high osmotic pressure, biofilm formation, cytoadherence and cytotoxicity in Vibrio vulnificus.
RESUMO
Vibrio parahaemolyticus, the main pathogen causing seafood related food poisoning worldwide, has strong biofilm formation ability. ToxR is a membrane binding regulatory protein, which has regulatory effect on biofilm formation of V. parahaemolyticus, but the specific mechanism has not been reported. c-di-GMP is an important second messenger in bacteria and is involved in regulating a variety of bacterial behaviors including biofilm formation. In this study, we investigated the regulation of ToxR on c-di-GMP metabolism in V. parahaemolyticus. Intracellular c-di-GMP in the wild type (WT) and toxR mutant (ΔtoxR) strains were extracted by ultrasonication, and the concentrations of c-di-GMP were then determined by enzyme linked immunosorbent assay (ELISA). Three c-di-GMP metabolism-related genes scrA, scrG and vpa0198 were selected as the target genes. Quantitative real-time PCR (q-PCR) was employed to calculate the transcriptional variation of each target gene between WT and ΔtoxR strains. The regulatory DNA region of each target gene was cloned into the pHR309 plasmid harboring a promoterless lacZ gene. The recombinant plasmid was subsequently transferred into WT and ΔtoxR strains to detect the β-galactosidase activity in the cellular extracts. The recombinant lacZ plasmid containing each of the target gene was also transferred into E. coli 100λpir strain harboring the pBAD33 plasmid or the recombinant pBAD33-toxR to test whether ToxR could regulate the expression of the target gene in a heterologous host. The regulatory DNA region of each target gene was amplified by PCR, and the over-expressed His-ToxR was purified. The electrophoretic mobility shift assay (EMSA) was applied to verify whether His-ToxR directly bound to the target promoter region. ELISA results showed that the intracellular c-di-GMP level significantly enhanced in ΔtoxR strain relative to that in WT strain, suggesting that ToxR inhibited the production of c-di-GMP in V. parahaemolyticus. qPCR results showed that the mRNA levels of scrA, scrG and vpa0198 significantly increased in ΔtoxR strain relative to those in WT strain, suggesting that ToxR repressed the transcription of scrA, scrG and vpa0198. lacZ fusion assay showed that ToxR was able to repress the promoter activities of scrA, scrG and vpa0198 in both V. parahaemolyticus and E. coli 100λpir. EMSA results showed that His-ToxR was able to bind to the regulatory DNA regions of scrA and scrG, but not to the regulatory DNA region of vpa0198. In conclusion, ToxR inhibited the production of c-di-GMP in V. parahaemolyticus via directly regulating the transcription of enzyme genes associated with c-di-GMP metabolism, which would be beneficial for V. parahaemolyticus to precisely control bacterial behaviors including biofilm formation.