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

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.

2006 ◽  
Vol 69 (8) ◽  
pp. 1829-1834 ◽  
Author(s):  
TINGTING REN ◽  
YI-CHENG SU

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.


2010 ◽  
Vol 73 (6) ◽  
pp. 1111-1115 ◽  
Author(s):  
YI-CHENG SU ◽  
QIANRU YANG ◽  
CLAUDIA HÄSE

The efficacy of refrigerated-seawater depuration for reducing Vibrio parahaemolyticus levels in Pacific oyster (Crassostrea gigas) was investigated. Raw Pacific oysters were inoculated with a mixed culture of five clinical strains of V. parahaemolyticus (105 to 106 most probable number [MPN] per g) and depurated with refrigerated seawater (5°C) in a laboratory-scale recirculation system equipped with a 15-W gamma UV sterilizer. Depuration with refrigerated seawater for 96 h reduced V. parahaemolyticus populations by >3.0 log MPN/g in oysters harvested in the winter. However, 144 h of depuration at 5°C was required to achieve a 3-log reduction in oysters harvested in the summer. Depuration with refrigerated seawater at 5°C for up to 144 h caused no significant fatality in the Pacific oyster and could be applied as a postharvest treatment to reduce V. parahaemolyticus contamination in Pacific oysters. Further studies are needed to validate the efficacy of the depuration process for reducing naturally accumulated V. parahaemolyticus in oysters.


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.


2012 ◽  
Vol 75 (8) ◽  
pp. 1501-1506 ◽  
Author(s):  
ROBERTA JULIANO RAMOS ◽  
MARÍLIA MIOTTO ◽  
FRANCISCO JOSÉ LAGREZE SQUELLA ◽  
ANDRÉIA CIROLINI ◽  
JAIME FERNANDO FERREIRA ◽  
...  

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.


1994 ◽  
Vol 57 (7) ◽  
pp. 604-606 ◽  
Author(s):  
ROGER W. PARKER ◽  
ELLEN M. MAURER ◽  
A. BILL CHILDERS ◽  
DONALD H. LEWISI

Vibrio vulnificus contamination of raw oysters is a serious public health hazard, therefore, it is necessary to investigate the persistence of V. vulnificus in harvested and stored oysters. For this study, triplicate oyster samples were split into four treatment groups: control, normal-packaged; control, vacuum-packaged; inoculated, normal-packaged; and inoculated, vacuum-packaged. Oysters in the inoculated groups were individually injected with V. vulnificus to a level of approximately 1 × 106 CFU/g. Control oysters were already naturally contaminated to a level of approximately 1 × 104 CFU/g. Oysters were then packaged, frozen and stored at −20°C. On day 0 and days 7, 14, 30 and 70 post-freezing, concentrations of total aerobic bacteria and V. vulnificus were determined using a 3-tube most probable number (MPN) estimation from enrichment Alkaline Peptone Water tubes with subsequent presumptive V. vulnificus growth on modified Cellobiose-Polymyxin B-Colistin agar. Length of frozen storage had a significant effect on decreasing total aerobic bacteria (from approximately 106 CFU/g to approximately 102.5 CFU/g) and V. vulnificus (from approximately 105 CFU/g to approximately 101 CFU/g). Also, vacuum-packaged samples showed significantly lower concentrations of V. vulnificus over the length of the study than did the normal-sealed samples.


2015 ◽  
Vol 78 (8) ◽  
pp. 1574-1580 ◽  
Author(s):  
THOMAS P. KINSEY ◽  
KERI A. LYDON ◽  
JOHN C. BOWERS ◽  
JESSICA L. JONES

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.


1994 ◽  
Vol 57 (5) ◽  
pp. 403-409 ◽  
Author(s):  
CURTIS J. HAGEN' ◽  
EDNA M. SLOAN ◽  
GAYLE A. LANCETTE ◽  
JAMES T. PEELER ◽  
JOHN N. SOFOS

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.


2015 ◽  
Vol 81 (21) ◽  
pp. 7600-7609 ◽  
Author(s):  
Kevin Esteves ◽  
Dominique Hervio-Heath ◽  
Thomas Mosser ◽  
Claire Rodier ◽  
Marie-George Tournoud ◽  
...  

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.


2002 ◽  
Vol 65 (1) ◽  
pp. 79-87 ◽  
Author(s):  
DAVID W. COOK ◽  
PAUL O'LEARY ◽  
JEFF C. HUNSUCKER ◽  
EDNA M. SLOAN ◽  
JOHN C. BOWERS ◽  
...  

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.


2016 ◽  
Vol 82 (15) ◽  
pp. 4517-4522 ◽  
Author(s):  
J. L. Jones ◽  
T. P. Kinsey ◽  
L. W. Johnson ◽  
R. Porso ◽  
B. Friedman ◽  
...  

ABSTRACTVibrio parahaemolyticusandVibrio vulnificuscan grow rapidly in shellfish subjected to ambient air conditions, such as during intertidal exposure. In this study, levels of total and pathogenic (tdh+and/ortrh+)V. parahaemolyticusand totalV. vulnificuswere determined in oysters collected from two study locations where intertidal harvest practices are common. Samples were collected directly off intertidal flats, after exposure (ambient air [Washington State] or refrigerated [New Jersey]), and after reimmersion by natural tidal cycles. Samples were processed using a most-probable-number (MPN) real-time PCR method for total and pathogenicV. parahaemolyticusorV. vulnificus. In Washington State, the mean levels ofV. parahaemolyticusincreased 1.38 log MPN/g following intertidal exposure and dropped 1.41 log MPN/g after reimmersion for 1 day, but the levels were dependent upon the container type utilized. PathogenicV. parahaemolyticuslevels followed a similar trend. However,V. vulnificuslevels increased 0.10 log MPN/g during intertidal exposure in Washington but decreased by >1 log MPN/g after reimmersion. In New Jersey, initial levels of all vibrios studied were not significantly altered during the refrigerated sorting and containerizing process. However, there was an increase in levels after the first day of reimmersion by 0.79, 0.72, 0.92, and 0.71 log MPN/g for total,tdh+andtrh+V. parahaemolyticus, andV. vulnificus, respectively. The levels of all targets decreased to those similar to background after a second day of reimmersion. These data indicate that the intertidal harvest and handling practices for oysters that were studied in Washington and New Jersey do not increase the risk of illness fromV. parahaemolyticusorV. vulnificus.IMPORTANCEVibrio parahaemolyticusandVibrio vulnificusare the leading causes of seafood-associated infectious morbidity and mortality in the United States.Vibriospp. can grow rapidly in shellfish subjected to ambient air conditions, such as during periods of intertidal exposure. When oysters are submersed with the incoming tide, the vibrios can be purged. However, data on the rates of increase and purging during intertidal harvest are scarce, which limits the accuracy of risk assessments. The objective of this study was to help fill these data gaps by determining the levels of total and pathogenic (tdh+and/ortrh+)V. parahaemolyticusandV. vulnificusin oysters from two locations where intertidal harvest practices are common, using the current industry practices. The data generated provide insight into the responses ofVibriospp. to relevant practices of the industry and public health, which can be incorporated into risk management decisions.


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