Molecular Methods to Study Vibrio parahaemolyticus and Vibrio vulnificus From Atypical Environments

Author(s):  
Indrani Karunasagar ◽  
Biswajit Maiti ◽  
Ballamoole Krishna Kumar
2008 ◽  
Vol 15 (10) ◽  
pp. 1541-1546 ◽  
Author(s):  
S. Datta ◽  
M. E. Janes ◽  
J. G. Simonson

ABSTRACT Mice were immunized by injection of Vibrio parahaemolyticus ATCC 17802 polar flagellin in order to produce monoclonal antibodies (mAbs). mAbs were analyzed by anti-H enzyme-linked immunosorbent assay using V. parahaemolyticus polar flagellar cores. The mAb exhibiting the highest anti-H titer was coated onto Cowan I Staphylococcus aureus cells at a concentration of 75 μg/ml cell suspension and used for slide coagglutination. Of 41 isolates identified genetically as V. parahaemolyticus, 100% coagglutinated with the anti-H mAb within 30 s, and the mAb did not react with 30 isolates identified as Vibrio vulnificus. A strong coagglutination reaction with V. parahaemolyticus ATCC 17802 was still observed when the S. aureus cells were armed with as little as 15 μg of mAb/ml S. aureus cell suspension. At this concentration, the mAb cross-reacted with three other Vibrio species, suggesting that they share an identical H antigen or antigens. The anti-H mAb was then used to optimize an immunomagnetic separation protocol which exhibited from 35% to about 45% binding of 102 to 103 V. parahaemolyticus cells in phosphate-buffered saline. The mAb would be useful for the rapid and selective isolation, concentration, and detection of V. parahaemolyticus cells from environmental sources.


2021 ◽  
Author(s):  
Oluwakemi Victoria Ayodele ◽  
Anthony Ifeanyi Okoh

Abstract Background: The use of antibiotics globally has helped reduce mortality and morbidity rate due to its ability to effectively treat bacterial infections in both humans and animals. However, the menace of antimicrobial resistance has become a challenge to public health due to its increased mortality and morbidity rate. This study determined the antibiogram pattern of non-cholera causing Vibrio species against a panel of 11 antibiotics that are wildly used for treatment. Multiple antibiotic resistance phenotype, multiple antibiotic resistant indices and minimum inhibitory concentration (MIC) of test antibiotics were also determined.Results: Polymerase chain reaction (PCR) was used to confirm 100 isolates of Vibrio parahaemolyticus, 82 and 46 isolates of Vibrio vulnificus and Vibrio fluvialis respectively, collected from the culture collections of the Applied and Environmental Microbiology Research Group (AEMREG), University of Fort Hare. Thereafter, disc diffusion method was used to determine the antibiogram pattern of target non-cholera causing Vibrio species against a panel of 11 antibiotics that are of clinical importance. The highest rate of Vibrio parahaemolyticus resistance was observed against tetracycline (22 %) and nalidixic acid (16 %). Vibrio fluvialis also displayed highest rate of resistance against tetracycline (28 %) and nalidixic acid (28 %), while Vibrio vulnificus isolates exhibited highest rate resistance against imipenem (40 %) and tetracycline (22 %). A total of 38 MARP patterns were observed and the MAR indices ranged between 0.3 and 0.8. Against the resistant Vibrio parahaemolyticus and Vibrio fluvialis isolates, minimum inhibitory concentration ranged from 16 µg/ml to 2048 µg/ml for both tetracycline and nalidixic acid, while against Vibrio vulnificus isolates, minimum inhibitory concentration ranged from 8 µg/ml to 256 µg/ml for both imipenem and nalidixic acid. Conclusions: Results obtained from this study is an indication that antibiotic resistant bacteria that could pose as threat to health of humans and animals are present in the environment.


2005 ◽  
Vol 68 (7) ◽  
pp. 1454-1456 ◽  
Author(s):  
YI-CHENG SU ◽  
JINGYUN DUAN ◽  
WEN-HSIN WU

The thiosulfate–citrate–bile salts–sucrose agar (TCBS) used in the most-probable-number method for detecting Vibrio parahaemolyticus cannot differentiate growth of V. parahaemolyticus from Vibrio vulnificus or Vibrio mimicus. This study examined the selectivity and specificity of Bio-Chrome Vibrio medium (BCVM), a chromogenic medium that detects V. parahaemolyticus on the basis of the formation of distinct purple colonies on the medium. A panel consisting of 221 strains of bacteria, including 179 Vibrio spp. and 42 non-Vibrio spp., were examined for their ability to grow and produce colored colonies on BCVM. Growth of Salmonella, Shigella, Escherichia coli, Enterobacter cloacae, Yersinia enterocolitica, and Aeromonas was inhibited by both BCVM and TCBS. All 148 strains of V. parahaemolyticus grew on BCVM, and 145 of them produced purple colonies. The remaining 31 Vibrio spp., except one strain of Vibrio fluvialis, were either unable to grow or produced blue-green or white colonies on BCVM. Bio-Chrome Vibrio medium was capable of differentiating V. parahaemolyticus from other species, including V. vulnificus and V. mimicus. Further studies are needed to evaluate the sensitivity and specificity of BCVM for detecting V. parahaemolyticus in foods.


2018 ◽  
Vol 81 (4) ◽  
pp. 593-597 ◽  
Author(s):  
THI THU TRA VU ◽  
THOMAS ALTER ◽  
STEPHAN HUEHN

ABSTRACT This study was conducted to determine the prevalence of Vibrio spp. in retail seafood in Berlin, Germany. A total of 160 raw seafood samples from supermarkets and seafood shops, consisting of shrimp (n = 80) and bivalves (n = 80), were investigated for the presence of Vibrio spp. using the International Organization for Standardization ISO/TS 21872 method and a multiplex PCR. The overall prevalence of Vibrio spp. in retail seafood was 55% (95% CI: 47.2 to 62.8%). The prevalence of Vibrio spp. in shrimp was slightly higher than in bivalves (57.5 versus 52.5%); however, the difference was not statistically significant. Vibrio alginolyticus was the most prevalent species (35.6%), followed by Vibrio parahaemolyticus (27.5%), Vibrio cholerae (6.3%), and Vibrio vulnificus (0.6%). None of the V. parahaemolyticus (n = 110) isolates encoded tdh/trh genes, whereas all V. cholerae isolates (n = 27) were lacking ctxA. Among the chilled samples (n = 105), the prevalence of Vibrio spp. in unpacked samples was significantly higher than in packed samples (P = 0.006). Among the packed samples (n = 55), no significant difference in the prevalence of Vibrio spp. was observed between chilled or frozen products. The results of this study indicated a high prevalence of Vibrio spp. in retail seafood in Germany; positive samples were detected in all types of seafood investigated. The detection of tdh/trh-negative V. parahaemolyticus isolates should not be neglected because of previous findings on pathogenic strains lacking these virulence markers. Even though thorough cooking might limit the risk of foodborne illness caused by Vibrio, potential cross-contamination during preparation or consumption of raw and undercooked seafood might represent a risk of Vibrio infections.


2012 ◽  
Vol 5 (5) ◽  
pp. 607-620 ◽  
Author(s):  
Karleigh Huff ◽  
Amornrat Aroonnual ◽  
Amy E. Fleishman Littlejohn ◽  
Bartek Rajwa ◽  
Euiwon Bae ◽  
...  

2009 ◽  
Vol 72 (1) ◽  
pp. 174-177 ◽  
Author(s):  
CHENGCHU LIU ◽  
JIANZHANG LU ◽  
YI-CHENG SU

This study investigated the effects of flash freezing, followed by frozen storage, on reducing Vibrio parahaemolyticus in Pacific raw oysters. Raw Pacific oysters were inoculated with a five-strain cocktail of V. parahaemolyticus at a total level of approximately 3.5 × 105 most probable number (MPN) per gram. Inoculated oysters were subjected to an ultralow flash-freezing process (−95.5°C for 12 min) and stored at −10, −20, and −30°C for 6 months. Populations of V. parahaemolyticus in the oysters declined slightly by 0.22 log MPN/g after the freezing process. Subsequent storage of frozen oysters at −10, −20, and −30°C resulted in considerable reductions of V. parahaemolyticus in the oysters. Storing oysters at −10°C was more effective in inactivating V. parahaemolyticus than was storage at −20 or −30°C. Populations of V. parahaemolyticus in the oysters declined by 2.45, 1.71, and 1.45 log MPN/g after 1 month of storage at −10, −20, and −30°C, respectively, and continued to decline during the storage. The levels of V. parahaemolyticus in oysters were reduced by 4.55, 4.13, and 2.53 log MPN/g after 6 months of storage at −10, −20, and −30°C, respectively. Three process validations, each separated by 1 week and conducted according to the National Shellfish Sanitation Program's postharvest processing validation–verification interim guidance for Vibrio vulnificus and Vibrio parahaemolyticus, confirmed that a process of flash freezing, followed by storage at −21 ± 2°C for 5 months, was capable of achieving greater than 3.52-log (MPN/g) reductions of V. parahaemolyticus in half-shell Pacific oysters.


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