Improved recovery of pathogenic Vibrio parahaemolyticus from oysters using colony hybridization following enrichment

2003 ◽  
Vol 52 (2) ◽  
pp. 273-277 ◽  
Author(s):  
Jessica L Nordstrom ◽  
Angelo DePaola
Keyword(s):  
Vibrio Parahaemolyticus ◽  
Colony Hybridization ◽  
Pathogenic Vibrio

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 Seasonal Variation in Abundance of Total and Pathogenic Vibrio parahaemolyticus Bacteria in Oysters along the Southwest Coast of India

2005 ◽  
Vol 71 (7) ◽  
pp. 3575-3580 ◽  
Author(s):  
A. Deepanjali ◽  
H. Sanath Kumar ◽  
I. Karunasagar ◽  
I. Karunasagar
Keyword(s):  
Vibrio Parahaemolyticus ◽  
Pcr Amplification ◽  
Seasonal Abundance ◽  
Southwest Coast Of India ◽  
Colony Hybridization ◽  
Southwest Coast ◽  
Alkaline Peptone Water ◽  
Pathogenic Vibrio ◽  
Hemolysin Gene ◽  
Peptone Water

ABSTRACT The seasonal abundance of Vibrio parahaemolyticus in oysters from two estuaries along the southwest coast of India was studied by colony hybridization using nonradioactive labeled oligonucleotide probes. The density of total V. parahaemolyticus bacteria was determined using a probe binding to the tlh (thermolabile hemolysin) gene, and the density of pathogenic V. parahaemolyticus bacteria was determined by using a probe binding to the tdh (thermostable direct hemolysin) gene. Furthermore, the prevalence of V. parahaemolyticus was studied by PCR amplification of the toxR, tdh, and trh genes. PCR was performed directly with oyster homogenates and also following enrichment in alkaline peptone water for 6 and 18 h. V. parahaemolyticus was detected in 93.87% of the samples, and the densities ranged from <10 to 104 organisms per g. Pathogenic V. parahaemolyticus could be detected in 5 of 49 samples (10.2%) by colony hybridization using the tdh probe and in 3 of 49 samples (6.1%) by PCR. Isolates from one of the samples belonged to the pandemic serotype O3:K6. Twenty-nine of the 49 samples analyzed (59.3%) were positive as determined by PCR for the presence of the trh gene in the enrichment broth media. trh-positive V. parahaemolyticus was frequently found in oysters from India.

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 Longitudinal Study of Total and Pathogenic Vibrio parahaemolyticus (tdh+ and/or trh+) in Two Natural Extraction Areas of Mytilus chilensis in Southern Chile

2021 ◽  
Vol 12 ◽  
Author(s):  
Cristina Bacian ◽  
Cristobal Verdugo ◽  
Katherine García ◽  
Josu Perez-Larruscain ◽  
Ignacio de Blas ◽  
...  
Keyword(s):  
Water Temperature ◽  
Vibrio Parahaemolyticus ◽  
River Estuary ◽  
Environmental Parameters ◽  
Economic Losses ◽  
Pcr Analysis ◽  
Potential Hazard ◽  
Detection Rates ◽  
Pathogenic Vibrio ◽  
Interaction Factor

Vibrio parahaemolyticus is the leading cause of seafood-associated bacterial gastroenteritis worldwide. Although different studies have focused on its pattern of variation over time, knowledge about the environmental factors driving the dynamics of this pathogen, within the Chilean territory, is still lacking. This study determined the prevalence of total and pathogenic V. parahaemolyticus strains (tdh and/or trh genes) in mussels (Mytilus chilensis) collected from two natural growing areas between 2017 and 2018, using selective agar and PCR analysis. V. parahaemolyticus was detected in 45.6% (93/204) of pooled samples from the Valdivia River Estuary. The pathogenic strains carrying the tdh and/or trh gene were detected in 11.8% (24/204): tdh in 9.8% (20/204), trh in 0.5% (1/204), and 1.5% (3/204) presented both genes. In Reloncaví Fjord, V. parahaemolyticus was detected in 14.4% (30/209) of the samples, pathogenic V. parahaemolyticus carrying the trh gene was detected in 0.5% (1/209) of the samples, while the tdh gene was not detected in the samples from this area. The total count of mauve-purple colonies typical of V. parahaemolyticus on CHROMagar was positively associated by multivariate analysis with area, water temperature, and salinity. Similarly, V. parahaemolyticus detection rates by PCR had a positive correlation with the area and water temperature. The chances of detecting total V. parahaemolyticus in the Valdivia River Estuary are significantly higher than in the Reloncaví Fjord, but inversely, during spring-summer months, the interaction factor between the area and temperature indicated that the chances of detecting V. parahaemolyticus are higher in the Reloncaví Fjord. Interestingly, this period coincides with the season when commercial and natural-growing shellfish are harvested. On the other hand, pathogenic V. parahaemolyticus tdh+ was significantly correlated with an increase of water temperature. These environmental parameters could be used to trigger a warning on potential hazard, which would influence human health and economic losses in aquaculture systems.

Tropical shrimp aquaculture farms harbour pathogenic Vibrio parahaemolyticus with high genetic diversity and Carbapenam resistance

2020 ◽  
Vol 160 ◽  
pp. 111551
Author(s):  
Sreejith V. Narayanan ◽  
Toms C. Joseph ◽  
Shaheer Peeralil ◽  
Reshmi Koombankallil ◽  
Murugadas Vaiyapuri ◽  
...  
Keyword(s):  
Genetic Diversity ◽  
Vibrio Parahaemolyticus ◽  
Shrimp Aquaculture ◽  
High Genetic Diversity ◽  
Pathogenic Vibrio ◽  
Tropical Shrimp

Real-Time PCR Quantification of Vibrio parahaemolyticus in Oysters Using an Alternative Matrix

2004 ◽  
Vol 67 (11) ◽  
pp. 2424-2429 ◽  
Author(s):  
G. E. KAUFMAN ◽  
G. M. BLACKSTONE ◽  
M. C. L. VICKERY ◽  
A. K. BEJ ◽  
J. BOWERS ◽  
...  
Keyword(s):  
Real Time ◽  
Real Time Pcr ◽  
Vibrio Parahaemolyticus ◽  
Pcr Amplification ◽  
Oyster Tissue ◽  
Pcr Assay ◽  
Colony Hybridization ◽  
The Real ◽  
Mantle Fluid ◽  
June July

This study examined the relationship between levels of total Vibrio parahaemolyticus found in oyster tissues and mantle fluid with the goal of using mantle fluid as a template matrix in a new quantitative real-time PCR assay targeting the thermolabile hemolysin (tlh) gene for the enumeration of total V. parahaemolyticus in oysters. Oysters were collected near Mobile Bay, Ala., in June, July, and September and tested immediately after collection and storage at 26°C for 24 h. Initial experiments using DNA colony hybridization targeting tlh demonstrated that natural V. parahaemolyticus levels in the mantle fluid of individual oysters were strongly correlated (r = 0.85, P &lt; 0.05) with the levels found in their tissues. When known quantities of cultured V. parahaemolyticus cells were added to real-time PCR reactions that contained mantle fluid and oyster tissue matrices separately pooled from multiple oysters, a strong linear correlation was observed between the real-time PCR cycle threshold and the log concentration of cells inoculated into each PCR reaction (mantle fluid: r = 0.98, P &lt; 0.05; and oyster: r = 0.99, P &lt; 0.05). However, the mantle fluid exhibited less inhibition of the PCR amplification than the homogenized oyster tissue. Analysis of natural V. parahaemolyticus populations in mantle fluids using both colony hybridization and real-time PCR demonstrated a significant (P &lt; 0.05) but reduced correlation (r =−0.48) between the two methods. Reductions in the efficiency of the real-time PCR that resulted from low population densities of V. parahaemolyticus and PCR inhibitors present in the mantle fluid of some oysters (with significant oyster-to-oyster variation) contributed to the reduction in correlation between the methods that was observed when testing natural V. parahaemolyticus populations. The V. parahaemolyticus–specific real-time PCR assay used for this study could estimate elevated V. parahaemolyticus levels in oyster mantle fluid within 1 h from sampling time.

Seasonal distribution of total and pathogenic Vibrio parahaemolyticus in Chesapeake Bay oysters and waters

2008 ◽  
Vol 128 (2) ◽  
pp. 354-361 ◽  
Author(s):  
Salina Parveen ◽  
Kumidini A. Hettiarachchi ◽  
John C. Bowers ◽  
Jessica L. Jones ◽  
Mark L. Tamplin ◽  
...  
Keyword(s):  
Chesapeake Bay ◽  
Vibrio Parahaemolyticus ◽  
Seasonal Distribution ◽  
Pathogenic Vibrio

Effect of Ploidy on Vibrio parahaemolyticus and Vibrio vulnificus Levels in Cultured Oysters

2020 ◽  
Vol 83 (11) ◽  
pp. 2014-2017
Author(s):  
JESSICA L. JONES ◽  
KERI A. LYDON ◽  
WILLIAM C. WALTON
Keyword(s):  
Crassostrea Virginica ◽  
Vibrio Parahaemolyticus ◽  
Pathogenic Bacteria ◽  
Vibrio Vulnificus ◽  
High Quality ◽  
Colony Hybridization ◽  
Naturally Occurring ◽  
Rapid Growth Rate ◽  
Vibrio Spp ◽  
Half Sibling

ABSTRACT Vibrio parahaemolyticus and Vibrio vulnificus are naturally occurring human pathogenic bacteria commonly found in estuarine environments where oysters are cultured. The use of triploid oysters has increased due to their rapid growth rate and because they maintain a high quality throughout the year. Previous work suggested levels of Vibrio spp. may be lower in triploid oysters than diploid oysters. Therefore, this study aimed to determine whether there is a difference in the abundances of V. parahaemolyticus and V. vulnificus between half-sibling diploid and triploid American oysters (Crassostrea virginica). In four trials, 100 individual oysters (either iced or temperature abused) were analyzed for V. parahaemolyticus and V. vulnificus by using direct plating followed by colony hybridization. Mean levels of V. parahaemolyticus in iced and abused diploid oysters were 3.55 and 4.21 log CFU/g, respectively. Mean levels in iced and abused triploid oysters were 3.49 and 4.27 log CFU/g, respectively. Mean levels of V. vulnificus in iced and abused diploid oysters were 3.53 and 4.56 log CFU/g, respectively. Mean levels in iced and abused triploid oysters were 3.54 and 4.55 log CFU/g, respectively. The differences in Vibrio spp. abundances between diploid and triploid oysters was not significant (P &gt; 0.05). However, the differences across treatments were significant (P &lt; 0.05), with the exception of V. parahaemolyticus levels in trial 3 (P = 0.83). Variation between individual oysters was also observed, with 12 of 808 measurements being outside of the 95th percentile. This phenomenon of occasional statistical outliers (“hot” or “cold” oysters) has been previously described and supports the appropriateness of composite sampling to account for inherent animal variability. In summary, the data indicate that abundances of V. parahaemolyticus and V. vulnificus are not dependent on the ploidy of cultured oysters but vary with the type of handling. HIGHLIGHTS

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