Evaluation of an Alkaline Phosphatase–Labeled Oligonucleotide Probe for the Detection and Enumeration of the Thermostable-Related Hemolysin (trh) Gene of Vibrio parahaemolyticus

2006 ◽  
Vol 69 (11) ◽  
pp. 2770-2772 ◽  
Author(s):  
JESSICA L. NORDSTROM ◽  
RACHEL RANGDALE ◽  
MICHAEL C. L. VICKERY ◽  
ANDREA M. B. PHILLIPS ◽  
SHELLEY L. MURRAY ◽  
...  
Keyword(s):  
Alkaline Phosphatase ◽  
Vibrio Parahaemolyticus ◽  
Oligonucleotide Probe ◽  
The United States ◽  
Bacterial Strains ◽  
Specific Detection ◽  
Colony Hybridization ◽  
The United Kingdom ◽  
Pathogenic Vibrio ◽  
Reliable Methods

Reliable methods are needed to detect total and pathogenic Vibrio parahaemolyticus. One marker of V. parahaemolyticus virulence is the thermostable-related hemolysin. We developed an alkaline phosphatase–labeled DNA probe method for the specific detection and enumeration of trh-positive V. parahaemolyticus by colony hybridization. The probe was tested against a panel of 200 bacterial strains and determined to be specific for trh-positive V. parahaemolyticus. Additionally, the trh alkaline phosphatase probe colony hybridization was successfully used to detect and enumerate trh-positive V. parahaemolyticus in seafood and water samples collected from the United States and the United Kingdom.

scholarly journals Isolation and selection of bacteria against shrimp pathogenic Vibrio parahaemolyticus from shrimp pond water on Duyen Hai district, Tra Vinh province

2020 ◽  
Vol 10 (1) ◽  
pp. 43-52
Author(s):  
Tran Vu Phuong ◽  
Dang Thi Ngoc Thanh ◽  
Cao Ngoc Diep
Keyword(s):  
Vibrio Parahaemolyticus ◽  
Pathogenic Bacteria ◽  
Pond Water ◽  
Shrimp Farming ◽  
Diffusion Method ◽  
Resistant Bacteria ◽  
Bacterial Strains ◽  
Shrimp Pond ◽  
Pathogenic Vibrio ◽  
The Status

Antibiotic has frequently been used in the shrimp-farming process in Vietnam. This leads to the status that antibiotic-resistant bacteria and products do not receive in the market. Bacteria had the resistant ability to pathogenic bacteria in water, and they have an important role in sustainable aquaculture. This study aimed to isolate and select good bacterial strains against Vibrio parahaemolyticus, pathogenic bacteria, on shrimp from 8 samples of shrimp pond water at 3 villages Ngu Lac, Phuoc An and Long Toan of Duyen Hai district, Tra Vinh province on NB agar medium. As a result, fifty-nine bacterial isolates were isolated and 10/59 isolates (16.95%) were identified as resistant to Vibrio parahaemolyticus by the well diffusion method. In 10 isolates, there were 7 isolates had good resistance to select for PCR technique and sequencing. The result indicated that these seven strains, including DH1m, DH2f, DH4d, DH8i, DH8m, DH8n, belonged to Bacilli and DH1n strain belonged to Streptomyces sp.

scholarly journals Seasonal Abundance of Total and Pathogenic Vibrio parahaemolyticus in Alabama Oysters

2003 ◽  
Vol 69 (3) ◽  
pp. 1521-1526 ◽  
Author(s):  
Angelo DePaola ◽  
Jessica L. Nordstrom ◽  
John C. Bowers ◽  
Joy G. Wells ◽  
David W. Cook
Keyword(s):  
United States ◽  
Vibrio Parahaemolyticus ◽  
Inverse Relationship ◽  
The United States ◽  
Seasonal Abundance ◽  
Focused Attention ◽  
Mobile Bay ◽  
Pathogenic Vibrio ◽  
Thermostable Direct Hemolysin ◽  
Efficient Screening

ABSTRACT Recent Vibrio parahaemolyticus outbreaks associated with consumption of raw shellfish in the United States focused attention on the occurrence of this organism in shellfish. From March 1999 through September 2000, paired oyster samples were collected biweekly from two shellfish-growing areas in Mobile Bay, Ala. The presence and densities of V. parahaemolyticus were determined by using DNA probes targeting the thermolabile hemolysin (tlh) and thermostable direct hemolysin (tdh) genes for confirmation of total and pathogenic V. parahaemolyticus, respectively. V. parahaemolyticus was detected in all samples with densities ranging from <10 to 12,000 g−1. Higher V. parahaemolyticus densities were associated with higher water temperatures. Pathogenic strains were detected in 34 (21.8%) of 156 samples by direct plating or enrichment. Forty-six of 6,018 and 31 of 6,992 V. parahaemolyticus isolates from enrichments and direct plates, respectively, hybridized with the tdh probe. There was an apparent inverse relationship between water temperature and the prevalence of pathogenic strains. Pathogenic strains were of diverse serotypes, and 97% produced urease and possessed a tdh-related hemolysin (trh) gene. The O3:K6 serotype associated with pandemic spread and recent outbreaks in the United States was not detected. The efficient screening of numerous isolates by colony lift and DNA probe procedures may account for the higher prevalence of samples with tdh + V. parahaemolyticus than previously reported.

scholarly journals Detection and Enumeration of Vibrio vulnificus in Oysters from Two Estuaries along the Southwest Coast of India, Using Molecular Methods

2004 ◽  
Vol 70 (11) ◽  
pp. 6909-6913 ◽  
Author(s):  
Ammini Parvathi ◽  
H. Sanath Kumar ◽  
Indrani Karunasagar ◽  
Iddya Karunasagar
Keyword(s):  
Alkaline Phosphatase ◽  
Environmental Factors ◽  
Vibrio Vulnificus ◽  
Oligonucleotide Probe ◽  
Monsoon Season ◽  
Seasonal Distribution ◽  
Southwest Coast Of India ◽  
Colony Hybridization ◽  
Alkaline Peptone Water ◽  
Peptone Water

ABSTRACT This study was conducted to understand the seasonal distribution of Vibrio vulnificus in oysters from two estuaries and the effect of environmental factors on the abundance of V. vulnificus in tropical waters. V. vulnificus was detected in 56.6% of the samples tested by colony hybridization with an alkaline phosphatase-labeled oligonucleotide probe (VV-AP), and the counts ranged from <10/g during the summer months to 103/g in the monsoon season at both sites. The density of V. vulnificus appeared to be controlled more by salinity than by temperature. A nested PCR used in this study detected V. vulnificus in 85% of the samples following 18 h of enrichment in alkaline peptone water.

Oyster-to-Oyster Variability in Levels of Vibrio parahaemolyticus

2003 ◽  
Vol 66 (1) ◽  
pp. 125-129 ◽  
Author(s):  
G. E. KAUFMAN ◽  
A. K. BEJ ◽  
J. BOWERS ◽  
A. DePAOLA
Keyword(s):  
Alkaline Phosphatase ◽  
Vibrio Parahaemolyticus ◽  
Mobile Bay ◽  
Pathogenic Vibrio ◽  
Sampling Protocols ◽  
Thermostable Direct Hemolysin ◽  
The Mean ◽  
June July ◽  
A Site

This study examined the variability in the levels of total and pathogenic Vibrio parahaemolyticus in individual oysters. Twenty oysters were collected on three occasions (in June, July, and September 2001) from a site near Mobile Bay, Ala. Ten of these oysters were tested immediately, and 10 were tested after 24 h of storage at 26°C. Levels of total and pathogenic V. parahaemolyticus were determined by alkaline phosphatase–labeled DNA probe procedures targeting the thermolabile hemolysin and thermostable direct hemolysin genes, respectively. Similar V. parahaemolyticus levels (200 to 2,000 CFU/g) were found in nearly 90% of the oysters (for all sampling occasions) prior to storage. The log-transformed densities (means ± standard deviations) of V. parahaemolyticus in oysters immediately after harvest were 2.90 ± 0.91, 2.88 ± 0.36, and 2.47 ± 0.26 log10 CFU/g for June, July, and September, respectively. After storage for 24 h at 26°C, the mean V. parahaemolyticus densities increased approximately 13- to 26-fold. Before storage, pathogenic V. parahaemolyticus was detected in 40% (10 to 20 CFU/g) of the oysters collected in June and July but was not detected in any oysters collected in September. After storage, pathogenic V. parahaemolyticus was detected in some oysters at levels of &gt;100 CFU/g. These data should aid in the development of sampling protocols for oyster monitoring programs and in the determination of exposure distributions associated with raw oyster consumption.

Evaluation of DNA Colony Hybridization and Real-Time PCR for Detection of Vibrio parahaemolyticus and Vibrio vulnificus in Postharvest-Processed Oysters

2009 ◽  
Vol 72 (10) ◽  
pp. 2106-2109 ◽  
Author(s):  
JESSICA L. JONES ◽  
KATHY E. NOE ◽  
ROBIN BYARS ◽  
ANGELO DePAOLA
Keyword(s):  
Real Time ◽  
Real Time Pcr ◽  
Vibrio Parahaemolyticus ◽  
Vibrio Vulnificus ◽  
The United States ◽  
Most Probable Number ◽  
Probable Number ◽  
Colony Hybridization ◽  
Alkaline Peptone Water ◽  
Peptone Water

The applicability of real-time PCR was examined for detection of vibrios from postharvest-processed (PHP) oysters to allow for a more rapid assay and higher sample throughput than currently used. During June to October 2004, 68 PHP oyster samples were collected directly from PHP firms or from retail markets across the United States. PHP oysters were examined to determine the effectiveness of treatments in the reduction of vibrio levels and to compare the analytical methods utilized. The latter is the focus of the data presented here. Each sample was analyzed for Vibrio parahaemolyticus and V. vulnificus by using a 2-dilution, three-tube most-probable-number (MPN) and a 25-g presence/absence enrichment in alkaline peptone water. Following 6-h and overnight enrichment, aliquots from each MPN tube and the 25-g sample were streaked onto selective media and tested by real-time PCR. Colonies from the selective agar were confirmed as V. parahaemolyticus or V. vulnificus by DNA colony hybridization. DNA hybridization and real-time PCR results for each MPN tube and the 25-g enrichment at both time points were analyzed individually for each organism. The methods were in agreement for 857 (95%) of 901 and for 882 (98%) of 903 tubes for detection of V. parahaemolyticus and V. vulnificus, respectively. Overall, there was 96% agreement between real-time and DNA colony hybridization. The results obtained by real-time PCR were comparable to those from DNA colony hybridization, but analysis time was significantly reduced for the detection of vibrios in PHP-treated oysters.

scholarly journals Vibrio parahaemolyticus: a review on the pathogenicity, antibiotic resistance, foodborne outbreaks, and detection methods

Food Research ◽  
2021 ◽  
Vol 5 (3) ◽  
pp. 1-11
Author(s):  
A. Naziahsalam Kehinde ◽  
J.Y.H. Tang ◽  
Y. Nakaguchi
Keyword(s):  
Antibiotic Resistance ◽  
Vibrio Parahaemolyticus ◽  
Foodborne Pathogen ◽  
Virulence Gene ◽  
The United States ◽  
Detection Methods ◽  
Marine Habitat ◽  
Pathogenic Vibrio ◽  
Thermostable Direct Hemolysin ◽  
Foodborne Outbreaks

Vibrio parahaemolyticus is a Gram-negative bacterium that is a natural inhabitant of the marine habitat. V. parahaemolyticus is a human foodborne pathogen linked to the consumption of contaminated raw and undercooked seafood. V. parahaemolyticus pathogenicity has been linked to the presence of two virulence gene that is thermostable direct hemolysin (tdh) and TDH-related hemolysin (trh). The emergence of antibiotic resistant strain of V. parahaemolyticus is a menace to public health. V. parahaemolyticus is linked to several foodborne diseases in Asian countries including Japan, China and Taiwan and has been acknowledged as the major cause of human gastroenteritis in the United States. The emergence of pathogenic Vibrio species in shellfish in Malaysia requires persistent monitoring and public enlightenment on food safety. Several detection methods based on its virulence factors are used in detecting V. parahaemolyticus. This review will provide an insight on V. parahaemolyticus, its pathogenicity, antibiotic resistance, foodborne outbreaks and detection methods.

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