Search results for: Vibrio parahaemolyticus

Authors: Ming-Lun Chiang, Hsi-Chia Chen, Chieh Wu, Yu-Ting Tseng, Ming-Ju Chen

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In this study, three strains of Vibrio parahaemolyticus (690, BCRC 13023 and BCRC 13025) were subjected to acid adaptation at pH 5.5 for 90 min. The survival of acid-adapted and non-adapted V. parahaemolyticus strains under simulated gastric condition and their protein expression profiles were investigated. Results showed that acid adaptation increased the survival of the test V. parahaemolyticus strains after exposure to simulated gastric juice (pH 3). Additionally, acid adaptation also affected the protein expression in these V. parahaemolyticus strains. Nine proteins, identified as atpA, atpB, DnaK, GroEL, OmpU, enolase, fructose-bisphosphate aldolase, phosphoglycerate kinase and triosephosphate isomerase, were induced by acid adaptation in two or three of the test strains. These acid-adaptive proteins may play important regulatory roles in the acid tolerance response (ATR) of V. parahaemolyticus.

Keywords: Acid adaptation, protein expression, simulated gastric juice, Vibrio parahaemolyticus

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Authors: Ming-Lun Chiang, Chieh Wu, Ming-Ju Chen

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Three strains of Vibrio parahaemolyticus (690, BCRC 13023 and BCRC 13025) implicated in food poisoning outbreaks in Taiwan were subjected to acid adaptation at pH 5.5 for 90 min. The growth behaviors of acid-adapted and non-adapted V. parahaemolyticus in the media supplemented with various nitrogen and carbon sources were investigated. The effects of acid adaptation on the thermostable direct hemolysin (TDH) secretion and fatty acid profiles of V. parahaemolyticus were also examined. Results showed that acid-adapted and non-adapted V. parahaemolyticus 690, BCRC 13023 and BCRC 13025 grew similarly in TSB-3% NaCl and basal media supplemented with various carbon and nitrogen sources during incubation period. Higher TDH secretion was noted with V. parahaemolyticus 690 among the three strains. However, acid-adapted strains produced less amounts of TDH than non-adapted strains when they were grown in TSB-3% NaCl. Additionally, acid adaptation increased the ratio of SFA/USFA in cells of V. parahaemolyticus strains.

Keywords: Vibrio parahaemolyticus, acid adaptation, thermostable direct hemolysin, fatty acid profile.

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Authors: E. Binaeian, SH. Soroushnia

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Acute toxicity of nano SiO2, ZnO, MCM-41 (Meso pore silica), Cu, Multi Wall Carbon Nano Tube (MWCNT), Single Wall Carbon Nano Tube (SWCNT) , Fe (Coated) to bacteria Vibrio fischeri using a homemade luminometer , was evaluated. The values of the nominal effective concentrations (EC), causing 20% and 50% inhibition of biouminescence, using two mathematical models at two times of 5 and 30 minutes were calculated. Luminometer was designed with Photomultiplier (PMT) detector. Luminol chemiluminescence reaction was carried out for the calibration graph. In the linear calibration range, the correlation coefficients and coefficient of Variation (CV) were 0.988 and 3.21% respectively which demonstrate the accuracy and reproducibility of the instrument that are suitable. The important part of this research depends on how to optimize the best condition for maximum bioluminescence. The culture of Vibrio fischeri with optimal conditions in liquid media, were stirring at 120 rpm at a temperature of 150C to 180C and were incubated for 24 to 72 hours while solid medium was held at 180C and for 48 hours. Suspension of nanoparticles ZnO, after 30 min contact time to bacteria Vibrio fischeri, showed the highest toxicity while SiO2 nanoparticles showed the lowest toxicity. After 5 min exposure time, the toxicity of ZnO was the strongest and MCM-41 was the weakest toxicant component.

Keywords: Bioluminescence, effective concentration, nanomaterials, toxicity, Vibrio fischeri.

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Authors: E. Binaeian, A.M. Rashidi, H. Attar

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A comparative evaluation of acute toxicity of synthesized nano silvers using two different procedures (biological and chemical reduction methods) and silver ions on bacteria Vibrio fischeri was investigated. The bacterial light inhibition test as a toxicological endpoint was used by applying of a homemade luminometer. To compare the toxicity effects as a quantitative parameter, a nominal effective concentrations (EC) of chemicals and a susceptibility constant (Z-value) of bacteria, after 5 min and 30 min exposure times, were calculated. After 5 and 30 min contact times, the EC50 values of two silver nanoparticles and the EC20 values were about similar. It demonstrates that toxicity of silvers was independent of their procedure. The EC values of nanoparticles were larger than those of the silver ions. The susceptibilities(Z- Values) of V.fischeri (L/mg) to the silver ions were greater than those of the nano silvers. According to the EC and Z values, the toxicity of silvers decreased in the following order: Silver ions >> silver nanoparticles from chemical reduction method ~ silver nanoparticles from biological method.

Keywords: Bioluminescence, Luminometer, silver nano particles, Toxicity, Vibrio fischeri

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Authors: N. B. Dhayanithi, T. T. Ajith Kumar, T. Balasubramanian

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Present study summarizes the control of Vibrio alginolyticus infection in hatchery reared Clownfish, Amphiprion sebae with the extract of the mangrove plant, Avicennia marina. Fishes with visible symptoms of hemorrhagic spots were chosen and the genomic DNA of the causative bacterium was isolated and sequenced based on 16S rDNA gene. The in vitro assay revealed that a fraction of A. marina leaf extract elucidated with ethyl acetate: methanol (6:4) showed a high activity (28 mm) at 125 μg/ml concentrations. About 4 % of the fraction fed along with live V. alginolyticus was significantly decreased the cumulative mortality (P<0.05) in the experimental groups than the control group. The responsible fraction was investigated by gas chromatography - mass spectroscopy and found the presence of active compounds. This is the first research in India to control vibriosis infection in marine ornamental fish with mangrove leaf extract.

Keywords: Amphiprion seabe, Avicennia marina, Gas Chromatography - Mass Spectroscopy, Vibrio alginolyticus

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Authors: Kusumawadee Thancharoen, Rungrat Nontawong, Thanawat Junsom

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Pathogenic bacterial flora was isolated from giant freshwater prawns, Macrobrachium rosenbergii. Infected shrimp samples were collected from BuaBan Aquafarm in Kalasin Province, Thailand, between June and September 2018. Bacterial species were isolated by serial dilution and plated on Thiosulfate Citrate Bile Salt Sucrose (TCBS) agar medium. A total 89 colonies were isolated and identified using the API 20E biochemical tests. Results showed the presence of genera Aeromonas, Citrobacter, Chromobacterium, Providencia, Pseudomonas, Stenotrophomonas and Vibrio. Maximum number of species was recorded in Pseudomonas (50.57%) with minimum observed in Chromobacterium and Providencia (1.12%).

Keywords: Biochemical test, giant freshwater prawn, isolation, salt tolerance, shrimp diseases.

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Authors: Uun Yanuhar

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The purpose of research was to know the role of immunogenic protein of 49 kDa from V.alginolyticus which capable to initiate molecule expression of MHC Class II in receptor of Cromileptes altivelis. The method used was in vivo experimental research through testing of immunogenic protein 49 kDa from V.alginolyticus at Cromileptes altivelis (size of 250 - 300 grams) using 3 times booster by injecting an immunogenic protein in a intramuscular manner. Response of expressed MHC molecule was shown using immunocytochemistry method and SEM. Results indicated that adhesin V.alginolyticus 49 kDa which have immunogenic character could trigger expression of MHC class II on receptor of grouper and has been proven by staining using immunocytochemistry and SEM with labeling using antibody anti MHC (anti mouse). This visible expression based on binding between epitopes antigen and antibody anti MHC in the receptor. Using immunocytochemistry, intracellular response of MHC to in vivo induction of immunogenic adhesin from V.alginolyticus was shown.

Keywords: C.altivelis, immunogenic, MHC, V.alginolyticus.

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Authors: Nelson R. Osungbemiro, Rafiu O. Sanni, Rotimi F. Olaniyan, Abayomi O. Olajuyigbe

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Bacteria flora of Clarias gariepinus collected from two natural habitats namely Owena River (freshwater) and Igbokoda lagoon (brackish water) were examined using standard microbiological procedures. Thirteen bacterial species were identified. The result indicated that from the identified bacteria isolated, Vibrio sp, Proteus sp. Shigella sp. and E. coli were present in both habitats (fresh and brackish waters). Others were habitat-selective such as Salmonella sp., Pseudomonas sp, Enterococcus sp, Staphylococcus sp. that were found only in freshwater habitat. While Branhamella sp, Streptococcus sp. and Micrococcus sp. were found in brackish water habitat. Bacteria load from Owena river (freshwater) was found to be the highest load recorded at 6.21 x 10⁴cfu. T-test analysis also revealed that there was a marked significant difference between bacterial load in guts of sampled Clarias from fresh water and brackish water habitats.

Keywords: Bacteria flora, gut, Clarias gariepinus, Owena river

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Authors: Joan Iliopoulou-Georgudaki, Chris Theodoropoulos, Danae Venieri, Maria Lagkadinou

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Monitoring of microbial flora in aquacultured sea bream, in relation to the physicochemical parameters of the rearing seawater, ended to a model describing the influence of the last to the quality of the fisheries. Fishes were sampled during eight months from four aqua farms in Western Greece and analyzed for psychrotrophic, H2S producing bacteria, Salmonella sp., heterotrophic plate count (PCA), with simultaneous physical evaluation. Temperature, dissolved oxygen, pH, conductivity, TDS, salinity, NO3 - and NH4 + ions were recorded. Temperature, dissolved oxygen and conductivity were correlated, respectively, to PCA, Pseudomonas sp. and Shewanella sp. counts. These parameters were the inputs of the model, which was driving, as outputs, to the prediction of PCA, Vibrio sp., Pseudomonas sp. and Shewanella sp. counts, and fish microbiological quality. The present study provides, for the first time, a ready-to-use predictive model of fisheries hygiene, leading to an effective management system for the optimization of aquaculture fisheries quality.

Keywords: Microbiological, model, physicochemical, Seabream.

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Authors: Jackson H. O. Onyuka, Rose Kakai, David M. Onyango, Peter F. Arama, John Gichuki, Ayub V.O. Ofulla

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A cross sectional study design and standard microbiological procedures were used to determine the prevalence and antimicrobial susceptibility patterns of Escherichia coli, Salmonella enterica serovar typhimurium and Vibrio cholerae O1 isolated from water and two fish species Rastrineobola argentea and Oreochromis niloticus collected from fish landing beaches and markets in the Lake Victoria Basin of western Kenya. Out of 162 samples analyzed, 133 (82.1%) were contaminated, with S. typhimurium as the most prevalent (49.6%), followed by E. coli (46.6%), and lastly V. cholerae (2.8%). All the bacteria isolates were sensitive to ciprofloxacin. E. coli isolates were resistant to ampicillin, tetracycline, cotrimoxazole, chloramphenical and gentamicin while S. typhimurium isolates exhibited resistance to ampicillin, tetracycline, and cotrimoxazole. The V. cholerae O1 isolates were resistant to tetracycline and ampicillin. The high prevalence of drug resistant enteric bacteria in water and fish from the study region needs public health intervention from the local government.

Keywords: Aquatic environments, Antimicrobial resistance, Enteric bacteria, Lake Victoria Basin

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Authors: Mauro Giacomini, Stefania Bertone, Federico Caneva Soumetz, Carmelina Ruggiero

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Environmental micro-organisms include a large number of taxa and some species that are generally considered nonpathogenic, but can represent a risk in certain conditions, especially for elderly people and immunocompromised individuals. Chemotaxonomic identification techniques are powerful tools for environmental micro-organisms, and cellular fatty acid methyl esters (FAME) content is a powerful fingerprinting identification technique. A system based on an unsupervised artificial neural network (ANN) was set up using the fatty acid profiles of standard bacterial strains, obtained by gas-chromatography, used as learning data. We analysed 45 certified strains belonging to Acinetobacter, Aeromonas, Alcaligenes, Aquaspirillum, Arthrobacter, Bacillus, Brevundimonas, Enterobacter, Flavobacterium, Micrococcus, Pseudomonas, Serratia, Shewanella and Vibrio genera. A set of 79 bacteria isolated from a drinking water line (AMGA, the major water supply system in Genoa) were used as an example for identification compared to standard MIDI method. The resulting ANN output map was found to be a very powerful tool to identify these fresh isolates.

Keywords: Cellular fatty acid methyl esters, environmental bacteria, gas-chromatography, unsupervised ANN.

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Authors: M.D. Murcia, M. Gomez, E. Gomez, J.L. Gomez, N. Christofi

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The removal efficiency of 4-chlorophenol with different advanced oxidation processes have been studied. Oxidation experiments were carried out using two 4-chlorophenol concentrations: 100 mg L-1 and 250 mg L-1 and UV generated from a KrCl excilamp with (molar ratio H2O2: 4-chlorophenol = 25:1) and without H2O2, and, with Fenton process (molar ratio H2O2:4- chlorophenol of 25:1 and Fe2+ concentration of 5 mg L-1). The results show that there is no significant difference in the 4- chlorophenol conversion when using one of the three assayed methods. However, significant concentrations of the photoproductos still remained in the media when the chosen treatment involves UV without hydrogen peroxide. Fenton process removed all the intermediate photoproducts except for the hydroquinone and the 1,2,4-trihydroxybenzene. In the case of UV and hydrogen peroxide all the intermediate photoproducts are removed. Microbial bioassays were carried out utilising the naturally luminescent bacterium Vibrio fischeri and a genetically modified Pseudomonas putida isolated from a waste treatment plant receiving phenolic waste. The results using V. fischeri show that with samples after degradation, only the UV treatment showed toxicity (IC50 =38) whereas with H2O2 and Fenton reactions the samples exhibited no toxicity after treatment in the range of concentrations studied. Using the Pseudomonas putida biosensor no toxicity could be detected for all the samples following treatment due to the higher tolerance of the organism to phenol concentrations encountered.

Keywords: 4-chlorophenol, Fenton, photodegradation, UV, excilamp.

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Authors: Sarkar M. A. Kawsar, Sarkar M. A. Mamun, Md S. Rahman, Hidetaro Yasumitsu, Yasuhiro Ozeki

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Lectins have a good scope in current clinical microbiology research. In the present study evaluated the antimicrobial activities of a D-galactose binding lectin (PnL) was purified from the annelid, Perinereis nuntia (polychaeta) by affinity chromatography. The molecular mass of the lectin was determined to be 32 kDa as a single polypeptide by SDS-PAGE under both reducing and non-reducing conditions. The hemagglutinating activity of the PnL showed against trypsinized and glutaraldehyde-fixed human erythrocytes was specifically inhibited by D-Gal, GalNAc, Galβ1-4Glc and Galα1-6Glc. PnL was evaluated for in vitro antibacterial screening studies against 11 gram-positive and gram-negative microorganisms. From the screening results, it was revealed that PnL exhibited significant antibacterial activity against gram-positive bacteria. Bacillus megaterium showed the highest growth inhibition by the lectin (250 μg/disc). However, PnL did not inhibit the growth of gram-negative bacteria such as Vibrio cholerae and Pseudomonas sp. PnL was also examined for in vitro antifungal activity against six fungal phytopathogens. PnL (100 μg/mL) inhibited the mycelial growth of Alternaria alternata (24.4%). These results indicate that future findings of lectin applications obtained from annelids may be of importance to life sciences.

Keywords: Perinereis nuntia, Lectin, Inhibition zone, Mycelial growth.

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Authors: T. Citarasu, M. Michaelbabu V. N. Vakharia

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The rhizome of Java grass, Cyperus rotundus was extracted different organic polar and non-polar solvents and performed the in vitro antiviral and immunostimulant activities against White Spot Syndrome Virus (WSSV) and Vibrio harveyi respectively. Based on the initial screening the ethyl acetate extract of C. rotundus was strong activities and further it was purified through silica column chromatography and the fractions were screened again for antiviral and immunostimulant activity. Among the different fractions screened against the WSSV and V. harveyi, the fractions, FIII to FV had strong activities. In order to study the in vivo influence of C. rotundus, the fractions (F-III to FV) were pooled and delivered to the F. indicus through artificial feed for 30 days. After the feeding trail the experimental and control diet fed F. indicus were challenged with virulent WSSV and studied the survival, molecular diagnosis, biochemical, haematological, and immunological parameters. Surprisingly, the pooled fractions (F-IV to FVI) incorporated diets helped to significantly (P<0.01) suppressed viral multiplication, showed significant (P<0.01) differences in protein and glucose levels, improved total haemocyte count (THC), coagulase activity, significantly increased (P <= 0.001) prophenol oxidase and intracellular superoxide anion production compared to the control shrimps. Based on the results, C. rotundus extracts effectively suppressed WSSV multiplication and improve the immune system in F. indicus against WSSV infection and this knowledge will helps to develop novel drugs from C. rotundus against WSSV.

Keywords: Antiviral drugs, Cyperus rotundus, Fenneropenaeus indicus, WSSV.

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Authors: Benjamin Osei-Tutu, Henrietta Awewole Kolson

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Background: Acute gastroenteritis is one of the frequently reported Out-Patient Department (OPD) cases. However, the causative pathogens of these cases are rarely identified at the OPD due to delay in laboratory results or failure to obtain specimens before antibiotics is administered. Method: A retrospective review of surveillance data from the Adentan Municipality, Accra, Ghana that were recorded in the National foodborne disease surveillance system of Ghana, was conducted with the main aim of describing the epidemiology and food practice of cases reported from the Adentan Municipality. The study involved a retrospective review of surveillance data kept on patients who visited health facilities that are involved in foodborne disease surveillance in Ghana, from January 2015 to December 2016. Results: A total of 375 cases were reviewed and these were classified as viral hepatitis (hepatitis A and E), cholera (Vibrio cholerae), dysentery (Shigella sp.), typhoid fever (Salmonella sp.) or gastroenteritis. Cases recorded were all suspected case and the average cases recorded per week was 3. Typhoid fever and dysentery were the two main clinically diagnosed foodborne illnesses. The highest number of cases were observed during the late dry season (Feb to April), which marks the end of the dry season and the beginning of the rainy season. Relatively high number of cases was also observed during the late wet seasons (Jul to Oct) when the rainfall is the heaviest. Home-made food and street vended food were the major sources of suspected etiological food, recording 49.01% and 34.87% of the cases respectively. Conclusion: Majority of cases recorded were classified as gastroenteritis due to the absence of laboratory confirmation. Few cases were classified as typhoid fever and dysentery based on clinical symptoms presented. Patients reporting with foodborne diseases were found to consume home meal and street vended foods as their predominant source of food.

Keywords: Accra, etiologic food, food poisoning, gastroenteritis, illness, surveillance.

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