Pathogenic Vibrio Species in Dutch Shellfish Destined for Direct Human Consumption

2010 ◽  
Vol 73 (4) ◽  
pp. 734-738 ◽  
Author(s):  
FRANCISKA M. SCHETS ◽  
HAROLD H. J. L. van den BERG ◽  
SASKIA A. RUTJES ◽  
ANA MARIA de RODA HUSMAN

Vibrio parahaemolyticus is a common cause of shellfish-related gastroenteritis all over the world. V. parahaemolyticus and Vibrio alginolyticus have previously been detected in water samples from the Oosterschelde, a large inlet on the North Sea, which is used for both recreational purposes and shellfish production. In 2006, oysters (Crassostrea gigas) from a noncommercial oyster bed in the Oosterschelde and oysters bought in Dutch fish shops were tested for the presence of pathogenic Vibrio species; in 2007 and 2008, oysters (C. gigas) and mussels (Mytilus edulis) from Oosterschelde production areas were examined. Total Vibrio numbers were related to water temperatures to study joint patterns. Vibrio was found in oysters and mussels from the production areas, and levels ranged from 6 to 622 most probable number (MPN) per g in oysters and 6 to 62 MPN/g in mussels. Vibrio levels in oysters from fish shops were 231 to >333 MPN/g, whereas levels in noncommercial oysters ranged from 231 to >2,398 MPN/g. About 80% of the isolated strains were V. alginolyticus, and approximately 10% were identified as V. parahaemolyticus. Vibrio counts in shellfish samples increased with increasing water temperature and declined when water temperatures dropped; Vibrio was not detected when water temperatures declined to <13.5°C. Based on the obtained results and the known high V. parahaemolyticus dose (<104 cells per serving of oysters) required for infection, it is concluded that the risk of gastrointestinal infections with V. parahaemolyticus through consumption of shellfish from the Oosterschelde production sites is presumably low.

2015 ◽  
Vol 81 (7) ◽  
pp. 2320-2327 ◽  
Author(s):  
C. D. Cruz ◽  
D. Hedderley ◽  
G. C. Fletcher

ABSTRACTThe food-borne pathogenVibrio parahaemolyticushas been reported as being present in New Zealand (NZ) seawaters, but there have been no reported outbreaks of food-borne infection from commercially grown NZ seafood. Our study determined the current incidence ofV. parahaemolyticusin NZ oysters and Greenshell mussels and the prevalence ofV. parahaemolyticustdhandtrhstrains. Pacific (235) and dredge (21) oyster samples and mussel samples (55) were obtained from commercial shellfish-growing areas between December 2009 and June 2012. TotalV. parahaemolyticusnumbers and the presence of pathogenic genestdhandtrhwere determined using the FDA most-probable-number (MPN) method and confirmed using PCR analysis. In samples from the North Island of NZ,V. parahaemolyticuswas detected in 81% of Pacific oysters and 34% of mussel samples, while the numbers ofV. parahaemolyticustdhandtrhstrains were low, with just 3/215 Pacific oyster samples carrying thetdhgene.V. parahaemolyticusorganisms carryingtdhandtrhwere not detected in South Island samples, andV. parahaemolyticuswas detected in just 1/21 dredge oyster and 2/16 mussel samples. Numbers ofV. parahaemolyticusorganisms increased when seawater temperatures were high, the season when most commercial shellfish-growing areas are not harvested. The numbers ofV. parahaemolyticusorganisms in samples exceeded 1,000 MPN/g only when the seawater temperatures exceeded 19°C, so this environmental parameter could be used as a trigger warning of potential hazard. There is some evidence that the totalV. parahaemolyticusnumbers increased compared with those reported from a previous 1981 to 1984 study, but the analytical methods differed significantly.


Author(s):  
Qingyao Wang ◽  
Songzhe Fu ◽  
Qian Yang ◽  
Jingwei Hao ◽  
Can Zhou ◽  
...  

The estuary is the ecological niche of pathogenic Vibrio spp. as it provides abundant organic and inorganic nutrients from seawater and rivers. However, little is known about the ecology of these Vibrio species in the inland brackish water area. In this study, their co-occurrence and relationships to key environmental constraints (salinity and temperature) in the Hun-Tai River of China were examined using the most probable number polymerase chain reaction (MPN-PCR) approach. We hereby report 2-year continuous surveillance based on six water indices of the Hun-Tai River. The results showed that seawater intrusion maximally reached inland as far as 26.5 km for the Hun-Tai River. Pathogenic Vibrio spp. were detected in 21.9% of the water samples. In particular, V. cholerae, V. parahaemolyticus, and V. vulnificus were isolated in 10 (10.4%), 20 (20.8.5%), and 2 (2.08%) samples, respectively. All V. parahaemolyticus strains were tdh gene negative, 10% were positive for the trh gene. Multi-locus sequence typing (MLST) divided V. parahaemolyticus strains into 12 sequence types (STs) for the Hun-Tai River. Five STs were respectively present in various locations along the Hun-Tai River. The PCR assay for detecting six virulence genes and Vibrio seventh pandemic island I and II revealed three genotypes in 12 V. cholerae isolates. The results of our study showed that seawater intrusion and salinity have profound effects on the distribution of pathogenic Vibrio spp. in the inland river, suggesting a potential health risk associated with the waters of the Hun-Tai River used for irrigation and drinking.


2012 ◽  
Vol 75 (1) ◽  
pp. 150-153 ◽  
Author(s):  
S. COPIN ◽  
A. ROBERT-PILLOT ◽  
P. MALLE ◽  
M. L. QUILICI ◽  
M. GAY

The most-probable-number (MPN) method is often time-consuming for the isolation, detection, and quantification of Vibrio parahaemolyticus from natural sources. MPN counting of V. parahaemolyticus bacteria usually involves the isolation of typical V. parahaemolyticus colonies on selective medium, with subsequent confirmation by biochemical identification. In this study, we evaluated the use of a PCR on MPN enrichment cultures (MPN-PCR) for the direct detection of total and pathogenic V. parahaemolyticus cells in frozen shrimp. This reaction targeted the R72H, tdh, and trh sequences. An internal amplification control was added to the samples before R72H amplification. There was an excellent correlation between the results of the two methods for artificially inoculated and natural shrimp samples. Of 36 natural samples, 28 tested positive for the presence of V. parahaemolyticus, with an MPN value of 2 × 10−1 to 9.2 × 101 per g. No pathogenic V. parahaemolyticus cells were detected. The test had a detection limit of one V. parahaemolyticus organism per g and was completed within two working days. These results support the use of the combination of PCR with MPN for the detection of total or potentially pathogenic V. parahaemolyticus cells in frozen shrimp.


2015 ◽  
Vol 78 (11) ◽  
pp. 1973-1979 ◽  
Author(s):  
HIN-CHUNG WONG ◽  
HUAI-YU JIANG ◽  
HSU-YANG LIN ◽  
YU-TING WANG

Seafood is often associated with foodborne illnesses, and Vibrio parahaemolyticus is the most common pathogen implicated in outbreaks in Taiwan. In this study, the microbiological quality of 300 raw or mixed ready-to-eat (RTE) and other cooking-needed seafood samples was examined. The total aerobic and coliform counts of the RTE samples were significantly higher than those of other cooking-needed samples. On average, 55.8 and 29.7% of the RTE samples failed to meet the local microbiological standards for total aerobic (5 log CFU/g) and coliform (3 log most probable number [MPN] per g), counts respectively; the corresponding percentages for the RTE samples from Taipei City were 9.1 and 18.2%, respectively. The total aerobic and coliform counts in the RTE samples from supermarkets and chain restaurants were significantly lower than those from traditional restaurants. The Vibrio species were more frequently identified in the cooking-needed samples than in RTE samples. Low incidences of V. parahaemolyticus (1.4%), V. vulnificus (1.9%), and V. cholerae (0%) were detected in most RTE samples. High densities of V. parahaemolyticus and V. vulnificus (1,200 MPN/g) were detected in a few RTE samples, only one of which contained toxigenic (tdh+) V. parahaemolyticus. The results of this investigation reveal that better hygiene of seafood providers such as chain restaurants, supermarkets, and traditional restaurants in Taipei City would effectively improve the microbiological quality of the seafood. The results will facilitate the establishment of measures for controlling the risks associated with seafood in Taiwan.


2019 ◽  
Vol 82 (12) ◽  
pp. 2094-2099 ◽  
Author(s):  
CLAUDIA VILLICAÑA ◽  
LUIS AMARILLAS ◽  
LILIANA SOTO-CASTRO ◽  
BRUNO GÓMEZ-GIL ◽  
MARCIAL LEONARDO LIZÁRRAGA-PARTIDA ◽  
...  

ABSTRACT Seafood has frequently been associated with foodborne illness because pathogens are easily introduced during seafood cultivation, handling, and processing. Vibrio parahaemolyticus and Vibrio cholerae are human pathogens that cause gastroenteritis and cholera, respectively, and Vibrio vulnificus can cause fatal wound infections and septicemia. However, information about the occurrence of these pathogens in oysters from the Pacific coast of Mexico is limited to V. parahaemolyticus. In the present study, we evaluated the presence and abundance of these three Vibrio species in 68 raw oysters (Crassostrea corteziensis) obtained from retail seafood markets in Sinaloa, Mexico. The most probable number (MPN)–PCR assay was used for amplification of the tlh (thermolabile hemolysin), ompW (outer membrane protein), and vvhA (hemolytic cytolysin) genes that are specific to V. parahaemolyticus, V. cholerae, and V. vulnificus, respectively. All oyster samples were positive for at least one Vibrio species. V. parahaemolyticus, V. cholerae, and V. vulnificus prevalences were 77.9, 8.8, and 32.3% overall, respectively, and most species were present in all sample periods with increased prevalence in period 3. The tdh (thermostable direct hemolysin) gene was detected in 30.1%, trh (TDH-related hemolysin) was detected in 3.7%, and tdh/trh was detected in 7.5% of the total tlh-positive samples (53 of 68), whereas the pandemic serotype O3:K6 (orf8 positive) was detected in only 1 sample (1.8%). The total prevalence of tdh and/or trh was 41.5%. In none of the samples positive for V. cholerae were the cholera toxin (ctxA) and cholix (chxA) toxigenic genes or the rfb gene encoding the O1 and O139 antigens amplified, suggesting the presence of non-O1 non-O139 V. cholerae strains. Our results clearly indicated a high prevalence of pathogenic Vibrio species in raw oysters from retail seafood markets in Mexico. Consumption of these raw oysters carries the potential risk of foodborne illness, which can be limited by cooking. HIGHLIGHTS


Author(s):  
Bing Wu ◽  
Hongxia Gong ◽  
Hui Zhang ◽  
Jiabei Chen ◽  
Hongling Wang

This study aimed to investigate the prevalence of total and pathogenic Vibrio parahaemolyticus in Anadara subcrenata sampled from aquafarms and retail markets in the Zhoushan islands during June 2013 to March 2015, using the most probable number-polymerase chain reaction (MPN-PCR) method. Total V. parahaemolyticus was detected in 265 (83.86%) samples with the density 0.3 to 2400 MPN/g. In total, 30.70% and 17.41% of the samples exceeded 100 MPN/g and 1,000 MPN/g, respectively. Both highest positive rate (98.99%) and highest prevalence (median = 210.0 MPN/g) were recorded in summer. Samples from aquafarms had a higher positive rate and median than those from retail markets. Pathogenic V. parahaemolyticus was detected both in aquafarms and retail markets in all seasons but not in winter. Among the 265 tlh-positive samples, 20 (7.55%) of the samples harbored tdh, and 5 (1.89%) of the samples harbored both tdh and trh. These results indicate that the Zhoushan archipelago is severely contaminated with V. parahaemolyticus in Anadara subcrenata; these results are applicable in risk assessment and to control the risk of food-borne disease caused by V. parahaemolyticus.


2020 ◽  
Vol 86 (23) ◽  
Author(s):  
Salina Parveen ◽  
John Jacobs ◽  
Gulnihal Ozbay ◽  
Lathadevi K. Chintapenta ◽  
Esam Almuhaideb ◽  
...  

ABSTRACT Oyster and seawater samples were collected from five sites in the Chesapeake Bay, MD, and three sites in the Delaware Bay, DE, from May to October 2016 and 2017. Abundances and detection frequencies for total and pathogenic Vibrio parahaemolyticus and Vibrio vulnificus were compared using the standard most-probable-number–PCR (MPN-PCR) assay and a direct-plating (DP) method on CHROMagar Vibrio for total (tlh+) and pathogenic (tdh+ and trh+) V. parahaemolyticus genes and total (vvhA) and pathogenic (vcgC) V. vulnificus genes. The colony overlay procedure for peptidases (COPP) assay was evaluated for total Vibrionaceae. DP had high false-negative rates (14 to 77%) for most PCR targets and was deemed unsatisfactory. Logistic regression models of the COPP assay showed high concordances with MPN-PCR for tdh+ and trh+ V. parahaemolyticus and vvhA+ V. vulnificus in oysters (85.7 to 90.9%) and seawater (81.1 to 92.7%) when seawater temperature and salinity were factored into the model, suggesting that the COPP assay could potentially serve as a more rapid method to detect vibrios in oysters and seawater. Differences in total Vibrionaceae and pathogenic Vibrio abundances between state sampling sites over different collection years were contrasted for oysters and seawater by MPN-PCR. Abundances of tdh+ and trh+ V. parahaemolyticus were ∼8-fold higher in Delaware oysters than in Maryland oysters, whereas abundances of vcgC+ V. vulnificus were nearly identical. For Delaware oysters, 93.5% were both tdh+ and trh+, compared to only 19.2% in Maryland. These results indicate that pathogenic V. parahaemolyticus was more prevalent in the Delaware Bay than in the Chesapeake Bay. IMPORTANCE While V. parahaemolyticus and V. vulnificus cause shellfish-associated morbidity and mortality among shellfish consumers, current regulatory assays for vibrios are complex, time-consuming, labor-intensive, and relatively expensive. In this study, the rapid, simple, and inexpensive COPP assay was identified as a possible alternative to MPN-PCR for shellfish monitoring. This paper shows differences in total Vibrionaceae and pathogenic vibrios found in seawater and oysters from the commercially important Delaware and Chesapeake Bays. Vibrio parahaemolyticus isolates from the Delaware Bay were more likely to contain commonly recognized pathogenicity genes than those from the Chesapeake Bay.


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.


Author(s):  
Felipe Tiago do Nascimento ◽  
Carlos Augusto do Nascimento ◽  
Fernando Rosado Spilki ◽  
Rodrigo Staggemeier ◽  
Cláudio Marcos Lauer Júnior

Natural water distillation can destroy and/or inactivate microorganisms that are sensitive to heat and ultraviolet radiation (UV). This method is currently used to provide fresh water in ships and in the desalination of brackish water. For the development of this research, a pilot-scale solar still was built and installed in the southern region of Brazil, in order to assess its efficiency in water disinfection, which was based on the most probable number (MPN) of total coliforms and  Escherichia coli, in addition to the DNA copy number of human adenovirus type 5 (HAdV-5) in raw, undistilled samples and in treated distilled water. Results showed that the distillation process removed 100% of total coliform and Escherichia coli and 4.5 log (99.997%) of HAdV-5, which meets the microbiological standards for drinking water according to national Brazilian regulations, as well as USEPA and HEALTH CANADA requirements.


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