Selectivity and Specificity of a Chromogenic Medium for Detecting Vibrio parahaemolyticus†

2005 ◽  
Vol 68 (7) ◽  
pp. 1454-1456 ◽  
Author(s):  
YI-CHENG SU ◽  
JINGYUN DUAN ◽  
WEN-HSIN WU
Keyword(s):  
Vibrio Parahaemolyticus ◽  
Bile Salts ◽  
Vibrio Vulnificus ◽  
Enterobacter Cloacae ◽  
Most Probable Number ◽  
Chromogenic Medium ◽  
Probable Number ◽  
Vibrio Fluvialis ◽  
Vibrio Mimicus ◽  
Vibrio Spp

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.

Effects of Flash Freezing, Followed by Frozen Storage, on Reducing Vibrio parahaemolyticus in Pacific Raw Oysters (Crassostrea gigas)

2009 ◽  
Vol 72 (1) ◽  
pp. 174-177 ◽  
Author(s):  
CHENGCHU LIU ◽  
JIANZHANG LU ◽  
YI-CHENG SU
Keyword(s):  
Crassostrea Gigas ◽  
Vibrio Parahaemolyticus ◽  
Vibrio Vulnificus ◽  
Frozen Storage ◽  
Most Probable Number ◽  
Freezing Process ◽  
Probable Number ◽  
Pacific Oysters ◽  
Subsequent Storage ◽  
Shellfish Sanitation

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.

Depuration of Oysters (Crassostrea gigas) Contaminated with Vibrio parahaemolyticus and Vibrio vulnificus with UV Light and Chlorinated Seawater

2012 ◽  
Vol 75 (8) ◽  
pp. 1501-1506 ◽  
Author(s):  
ROBERTA JULIANO RAMOS ◽  
MARÍLIA MIOTTO ◽  
FRANCISCO JOSÉ LAGREZE SQUELLA ◽  
ANDRÉIA CIROLINI ◽  
JAIME FERNANDO FERREIRA ◽  
...  
Keyword(s):  
Vibrio Parahaemolyticus ◽  
Vibrio Vulnificus ◽  
Uv Light ◽  
Control Treatment ◽  
Most Probable Number ◽  
Brazilian Coast ◽  
Uv Treatment ◽  
Probable Number ◽  
Postharvest Treatment ◽  
Small Producers

The efficacy of depuration using UV light and chlorinated seawater for decontaminating Vibrio parahaemolyticus and Vibrio vulnificus from oysters was investigated. Oysters were contaminated with a five-strain cocktail of V. parahaemolyticus or V. vulnificus to levels of 104 to 105 CFU ml−1 for bioaccumulation. The depuration was conducted in a closed system in which 350 liters of seawater was recirculated at a rate of 7 liters/min for 48 h at room temperature. Counts of V. parahaemolyticus or V. vulnificus were determined at 0, 6, 18, 24, and 48 h. Three treatments were conducted: T1, control treatment; T2, UV treatment; and T3, UV plus chlorine treatment. After 48 h of depuration of V. parahaemolyticus, T3 reduced the count by 3.1 log most probable number (MPN) g−1 and T2 reduced the count by 2.4 log MPN g−1, while T1 reduced the count by only 2.0 log MPN g−1. After 48 h of depuration of V. vulnificus, T2 and T3 were efficient, reducing the counts by 2.5 and 2.4 log MPN g−1, respectively, while T1 reduced the count by only 1.4 log MPN g−1. The UV light plus chlorine treatment was more efficient for controlling V. parahaemolyticus in oysters. Both UV light and UV light plus chlorine were efficient for V. vulnificus. The present study is the first report showing the efficacy of depuration systems for decontaminating V. parahaemolyticus and V. vulnificus in oysters cultivated on the Brazilian coast. This study provides information on processes that can contribute to controlling and preventing such microorganisms in oysters and could be used for effective postharvest treatment by restaurants and small producers of oysters on the coast of Brazil.

Evaluation of the Compact Dry VP Method for Screening Raw Seafood for Total Vibrio parahaemolyticus

2009 ◽  
Vol 72 (1) ◽  
pp. 169-173 ◽  
Author(s):  
HIDEMASA KODAKA ◽  
HAJIME TERAMURA ◽  
SHINGO MIZUOCHI ◽  
MIKAKO SAITO ◽  
HIDEAKI MATSUOKA
Keyword(s):  
Vibrio Parahaemolyticus ◽  
Cold Water ◽  
Correlation Coefficients ◽  
Water Soluble ◽  
Coliform Bacteria ◽  
Most Probable Number ◽  
Probable Number ◽  
Different Types ◽  
Phosphate Buffered Saline ◽  
Vibrio Spp

Compact Dry VP (CDVP) is a ready-to-use method for enumerating Vibrio parahaemolyticus in food. The presterilized plates contain a culture medium comprising peptone, NaCl, bile salts, antibiotics, chromogenic substrates, and polysaccharide gum as a cold water–soluble gelling. After diluting raw seafood samples in a phosphate-buffered saline solution, a 1-ml aliquot was inoculated onto the center of the plate and allowed to diffuse by capillary action. Blue-green colonies forming on the plates were counted after 18 to 20 h of incubation at 35°C. A total of 85 V. parahaemolyticus strains (62 tdh+ strains and 23 tdh− strains) were studied for inclusivity, 81 (95.3 %) of which produced blue-green colonies. When 97 strains (14 strains of Vibrio spp., 33 strains of coliform bacteria, and 50 strains of noncoliform bacteria) were assessed for exclusivity, 10 strains of Vibrio spp. produced non–blue-green colonies, and 87 strains failed to grow. The CDVP and U.S. Food and Drug Administration Bacteriological Analytical Manual (FDA-BAM) methods were compared with the use of four different types of raw seafood that were inoculated with four different V. parahaemolyticus strains. For raw tuna and oysters, the FDA-BAM colony lift method was used, whereas the FDA-BAM most-probable-number method was used for salmon and scallop. The linear correlation coefficients between the CDVP and FDA-BAM methods were 0.99 for fresh raw tuna, 0.95 for fresh raw oysters, 0.95 for frozen raw salmon, and 0.95 for frozen raw scallops. These results suggest that the CDVP method is useful for screening raw seafood for V. parahaemolyticus.

scholarly journals Relationship of aquatic environmental factors with the abundance of Vibrio cholerae, Vibrio parahaemolyticus, Vibrio mimicus and Vibrio vulnificus in the coastal area of Guaymas, Sonora, Mexico

2013 ◽  
Vol 11 (4) ◽  
pp. 700-712 ◽  
Author(s):  
A. León Robles ◽  
E. Acedo Félix ◽  
B. Gomez-Gil ◽  
E. I. Quiñones Ramírez ◽  
M. Nevárez-Martínez ◽  
...  
Keyword(s):  
Environmental Factors ◽  
Pathogenic Bacteria ◽  
Vibrio Vulnificus ◽  
Sampling Site ◽  
Principal Component ◽  
Most Probable Number ◽  
Probable Number ◽  
Vibrio Mimicus ◽  
Positive Correlations ◽  
Relationship Of

Members of the genus Vibrio are common in aquatic environments. Among them are V. cholerae, V. vulnificus, V. parahaemolyticus and V. mimicus. Several studies have shown that environmental factors, such as temperature, salinity, and dissolved oxygen, are involved in their epidemiology. Therefore, the main objective of this study is to determine if there is a correlation between the presence/amount of V. cholerae, V, vulnificus, V. parahaemolyticus and V. mimicus and the environmental conditions of the seawater off the coast of Guaymas, México. Quantification of all four pathogenic bacteria was performed using the most probable number method, and suspected colonies were identified by polymerase chain reaction (PCR). Correlations were found using principal component analysis. V. parahaemolyticus was the most abundant and widely distributed bacteria, followed by V. vulnificus, V. mimicus and V. cholerae. Positive correlations between V. parahaemolyticus, V. vulnificus and V. mimicus with temperature, salinity, electric conductivity, and total dissolved solids were found. The abundance of V. cholerae was mainly affected by the sampling site and not by physicochemical parameters.

Effects of Dry Storage and Resubmersion of Oysters on Total Vibrio vulnificus and Total and Pathogenic (tdh+/trh+) Vibrio parahaemolyticus Levels

2015 ◽  
Vol 78 (8) ◽  
pp. 1574-1580 ◽  
Author(s):  
THOMAS P. KINSEY ◽  
KERI A. LYDON ◽  
JOHN C. BOWERS ◽  
JESSICA L. JONES
Keyword(s):  
Vibrio Parahaemolyticus ◽  
Storage Time ◽  
Vibrio Vulnificus ◽  
Ambient Air ◽  
Most Probable Number ◽  
Probable Number ◽  
Ambient Temperatures ◽  
Dry Storage ◽  
Hemolysin Gene ◽  
Thermostable Direct Hemolysin

Vibrio vulnificus (Vv) and Vibrio parahaemolyticus (Vp) are the two leading causes of bacterial illnesses associated with raw shellfish consumption. Levels of these pathogens in oysters can increase during routine antifouling aquaculture practices involving dry storage in ambient air conditions. After storage, common practice is to resubmerge these stored oysters to reduce elevated Vv and Vp levels, but evidence proving the effectiveness of this practice is lacking. This study examined the changes in Vv and in total and pathogenic (thermostable direct hemolysin gene and the tdh-related hemolysin gene, tdh+ and trh+) Vp levels in oysters after 5 or 24 h of dry storage (28 to 32°C), followed by resubmersion (27 to 32°C) for 14 days. For each trial, replicate oyster samples were collected at initial harvest, after dry storage, after 7 days, and after 14 days of resubmersion. Oysters not subjected to dry storage were collected and analyzed to determine natural undisturbed vibrio levels (background control). Vibrio levels were measured using a most-probable-number enrichment followed by real-time PCR. After storage, vibrio levels (excluding tdh+ and trh+ Vp during 5-h storage) increased significantly (P < 0.001) from initial levels. After 7 days of resubmersion, Vv and total Vp levels (excluding total Vp in oysters stored for 5 h) were not significantly different (P > 0.1) from levels in background oysters. Vv and total and pathogenic Vp levels were not significantly different (P > 0.1) from levels in background oysters after 14 days of resubmersion, regardless of dry storage time. These data demonstrate that oyster resubmersion after dry storage at elevated ambient temperatures allows vibrio levels to return to those of background control samples. These results can be used to help minimize the risk of Vv and Vp illnesses and to inform the oyster industry on the effectiveness of routine storing and resubmerging of aquaculture oysters.

Enumeration of Vibrio parahaemolyticus and Vibrio vulnificus in Various Seafoods with Two Enrichment Broths

1994 ◽  
Vol 57 (5) ◽  
pp. 403-409 ◽  
Author(s):  
CURTIS J. HAGEN' ◽  
EDNA M. SLOAN ◽  
GAYLE A. LANCETTE ◽  
JAMES T. PEELER ◽  
JOHN N. SOFOS
Keyword(s):  
Cold Stress ◽  
Vibrio Parahaemolyticus ◽  
Vibrio Vulnificus ◽  
Polymyxin B ◽  
Most Probable Number ◽  
Geometric Means ◽  
Probable Number ◽  
Alkaline Peptone Water ◽  
Peptone Water ◽  
Stressed Cells

This study compares recoveries of Vibrio parahaemolyticus and Vibrio vulnificus with salt-polymyxin B broth (SPB) and alkaline peptone water (APW) from samples of crab legs, oysters, shrimp, lobster and shark, which were inoculated at three levels (approximately 101 to 102, 102 to 103 and 104 to 105/g) with each of the pathogens. Six samples of each product were analyzed [3-tube most probable number (MPN)] with each broth. Inoculated samples of oysters and slurries of crab and lobster were also tested after cold stress (refrigerated at 2 to 4°C, 3 or 7 days, or frozen at −15°C for 21 or 28 days). For each seafood, geometric means of cells recovered with APW were significantly (P < 0.05) higher than the corresponding means of recovery with SPB. In addition, 12 of 15 calculated estimates of 50% relative detectable levels (RDL50) were lower (P < 0.05) for APW than for SPB. In these samples, the level of detection by APW was found to be 40 to 32,000 and 6- to 42-fold lower for V parahaemolyticus and V. vulnificus, respectively, than the level of detection by SPB. In cold-stored samples, overall detection of the pathogens was greatly reduced, but APW was also more efficient than SPB in recovering stressed cells.

scholarly journals Rapid Proliferation of Vibrio parahaemolyticus, Vibrio vulnificus, and Vibrio cholerae during Freshwater Flash Floods in French Mediterranean Coastal Lagoons

2015 ◽  
Vol 81 (21) ◽  
pp. 7600-7609 ◽  
Author(s):  
Kevin Esteves ◽  
Dominique Hervio-Heath ◽  
Thomas Mosser ◽  
Claire Rodier ◽  
Marie-George Tournoud ◽  
...  
Keyword(s):  
Vibrio Cholerae ◽  
Vibrio Parahaemolyticus ◽  
Vibrio Vulnificus ◽  
Coastal Lagoons ◽  
Flash Floods ◽  
Low Flow ◽  
Most Probable Number ◽  
Abrupt Decrease ◽  
Probable Number ◽  
Content Type

ABSTRACTVibrio parahaemolyticus,Vibrio vulnificus, andVibrio choleraeof the non-O1/non-O139 serotype are present in coastal lagoons of southern France. In these Mediterranean regions, the rivers have long low-flow periods followed by short-duration or flash floods during and after heavy intense rainstorms, particularly at the end of the summer and in autumn. These floods bring large volumes of freshwater into the lagoons, reducing their salinity. Water temperatures recorded during sampling (15 to 24°C) were favorable for the presence and multiplication of vibrios. In autumn 2011, before heavy rainfalls and flash floods, salinities ranged from 31.4 to 36.1‰ and concentrations ofV. parahaemolyticus,V. vulnificus, andV. choleraevaried from 0 to 1.5 × 103most probable number (MPN)/liter, 0.7 to 2.1 × 103MPN/liter, and 0 to 93 MPN/liter, respectively. Following heavy rainstorms that generated severe flash flooding and heavy discharge of freshwater, salinity decreased, reaching 2.2 to 16.4‰ within 15 days, depending on the site, with a concomitant increase inVibrioconcentration to ca. 104MPN/liter. The highest concentrations were reached with salinities between 10 and 20‰ forV. parahaemolyticus, 10 and 15‰ forV. vulnificus, and 5 and 12‰ forV. cholerae. Thus, an abrupt decrease in salinity caused by heavy rainfall and major flooding favored growth of human-pathogenicVibriospp. and their proliferation in the Languedocian lagoons. Based on these results, it is recommended that temperature and salinity monitoring be done to predict the presence of theseVibriospp. in shellfish-harvesting areas of the lagoons.

Effects of Electrolyzed Oxidizing Water Treatment on Reducing Vibrio parahaemolyticus and Vibrio vulnificus in Raw Oysters

2006 ◽  
Vol 69 (8) ◽  
pp. 1829-1834 ◽  
Author(s):  
TINGTING REN ◽  
YI-CHENG SU
Keyword(s):  
Water Treatment ◽  
Vibrio Parahaemolyticus ◽  
Vibrio Vulnificus ◽  
Safety Concern ◽  
Most Probable Number ◽  
Probable Number ◽  
Pacific Oysters ◽  
Oxidation Reduction ◽  
Oxidation Reduction Potential ◽  
Pure Cultures

Contamination of Vibrio parahaemolyticus and Vibrio vulnificus in oysters is a food safety concern. This study investigated effects of electrolyzed oxidizing (EO) water treatment on reducing V. parahaemolyticus and V. vulnificus in laboratory-contaminated oysters. EO water exhibited strong antibacterial activity against V. parahaemolyticus and V. vulnificus in pure cultures. Populations of V. parahaemolyticus (8.74 × 107 CFU/ml) and V. vulnificus (8.69 × 107 CFU/ml) decreased quickly in EO water containing 0.5% NaCl to nondetectable levels (>6.6 log reductions) within 15 s. Freshly harvested Pacific oysters were inoculated with a five-strain cocktail of V. parahaemolyticus or V. vulnificus at levels of 104 and 106 most probable number (MPN)/g and treated with EO water (chlorine, 30 ppm; pH 2.82; oxidation-reduction potential, 1131 mV) containing 1% NaCl at room temperature. Reductions of V. parahaemolyticus and V. vulnificus in oysters were determined at 0 (before treatment), 2, 4, 6, and8hof treatment. Holding oysters inoculated with V. parahaemolyticus or V. vulnificus in the EO water containing 1% NaCl for 4 to 6 h resulted in significant (P < 0.05) reductions of V. parahaemolyticus and V. vulnificus by 1.13 and 1.05 log MPN/g, respectively. Extended exposure (>12 h) of oysters in EO water containing high levels of chlorine (>30 ppm) was found to be detrimental to oysters. EO water could be used as a postharvest treatment to reduce Vibrio contamination in oysters. However, treatment should be limited to 4 to6hto avoid death of oysters. Further studies are needed to determine effects of EO water treatment on sensory characteristics of oysters.

Evaluation of Ice Slurries as a Control for Postharvest Growth of Vibrio spp. in Oysters and Potential for Filth Contamination

2015 ◽  
Vol 78 (7) ◽  
pp. 1375-1379 ◽  
Author(s):  
KERI ANN LYDON ◽  
MELISSA FARRELL-EVANS ◽  
JESSICA L. JONES
Keyword(s):  
Vibrio Vulnificus ◽  
The United States ◽  
Fecal Coliforms ◽  
Coliform Bacteria ◽  
Most Probable Number ◽  
Internal Cooling ◽  
Probable Number ◽  
Route Of Exposure ◽  
C 50 ◽  
Vibrio Spp

Raw oyster consumption is the most common route of exposure for Vibrio spp. infections in humans. Vibriosis has been increasing steadily in the United States despite efforts to reduce the incidence of the disease. Research has demonstrated that ice is effective in reducing postharvest Vibrio spp. growth in oysters but has raised concerns of possible contamination of oyster meat by filth (as indicated by the presence of fecal coliform bacteria or Clostridium perfringens). This study examined the use of ice slurries (<4.5°C) to reduce Vibrio growth. Ice slurries showed rapid internal cooling of oysters, from 23.9°C (75°F) to 10°C (50°F) within 12 min. The initial bacterial loads in the ice slurry waters were near the limits of detection. Following repeated dipping of oysters into ice slurries, water samples exhibited significant (P < 0.05) increases in median levels of fecal coliforms (9.5 most probable number [MPN]/100 ml), C. perfringens (280 MPN/100 ml), Vibrio vulnificus (11,250 MPN/ml), and total Vibrio parahaemolyticus (3,900 MPN/ml). The microbial load in oyster meat, however, was unchanged after 15 min of submergence, with no significant differences (P < 0.05) in levels of filth indicator (range, 250 to 720 MPN/100 g) or Vibrio spp. (range, 9,000 to 20,000 MPN/g) bacteria. These results support the use of ice slurries as a postharvest application for rapid cooling of oysters to minimize Vibrio growth.

Vibrio vulnificus and Vibrio parahaemolyticus in U.S. Retail Shell Oysters: A National Survey from June 1998 to July 1999

2002 ◽  
Vol 65 (1) ◽  
pp. 79-87 ◽  
Author(s):  
DAVID W. COOK ◽  
PAUL O'LEARY ◽  
JEFF C. HUNSUCKER ◽  
EDNA M. SLOAN ◽  
JOHN C. BOWERS ◽  
...  
Keyword(s):  
United States ◽  
North Atlantic ◽  
Vibrio Parahaemolyticus ◽  
Vibrio Vulnificus ◽  
Gulf Coast ◽  
The United States ◽  
Most Probable Number ◽  
Probable Number ◽  
Hemolysin Gene ◽  
Thermostable Direct Hemolysin

From June 1998 to July 1999, 370 lots of oysters in the shell were sampled at 275 different establishments (71%, restaurants or oyster bars; 27%, retail seafood markets; and 2%, wholesale seafood markets) in coastal and inland markets throughout the United States. The oysters were harvested from the Gulf (49%), Pacific (14%), Mid-Atlantic (18%), and North Atlantic (11%) Coasts of the United States and from Canada (8%). Densities of Vibrio vulnificus and Vibrio parahaemolyticus were determined using a modification of the most probable number (MPN) techniques described in the Food and Drug Administration's Bacteriological Analytical Manual. DNA probes and enzyme immunoassay were used to identify suspect isolates and to determine the presence of the thermostable direct hemolysin gene associated with pathogenicity of V. parahaemolyticus. Densities of both V. vulnificus and V. parahaemolyticus in market oysters from all harvest regions followed a seasonal distribution, with highest densities in the summer. Highest densities of both organisms were observed in oysters harvested from the Gulf Coast, where densities often exceeded 10,000 MPN/g. The majority (78%) of lots harvested in the North Atlantic, Pacific, and Canadian Coasts had V. vulnificus densities below the detectable level of 0.2 MPN/g; none exceeded 100 MPN/g. V. parahaemolyticus densities were greater than those of V. vulnificus in lots from these same areas, with some lots exceeding 1,000 MPN/g for V. parahaemolyticus. Some lots from the Mid-Atlantic states exceeded 10,000 MPN/g for both V. vulnificus and V. parahaemolyticus. Overall, there was a significant correlation between V. vulnificus and V. parahaemolyticus densities (r = 0.72, n = 202, P < 0.0001), but neither density correlated with salinity. Storage time significantly affected the V. vulnificus (10% decrease per day) and V. parahaemolyticus (7% decrease per day) densities in market oysters. The thermostable direct hemolysin gene associated with V. parahaemolyticus virulence was detected in 9 of 3,429 (0.3%) V. parahaemolyticus cultures and in 8 of 198 (4.0%) lots of oysters. These data can be used to estimate the exposure of raw oyster consumers to V. vulnificus and V. parahaemolyticus.

Sign in / Sign up

Export Citation Format

Share Document