<< Return to News

Identifying Pathogenic Potential – Aquaculture America Conference: Feb 9-12th

Sona Son, Technical Manager at Microbial Discovery Group, presents a new MDG Technology Development at the Aquaculture America Conference Feb 9-12th, 2014.  See poster abstract below.  If you’re dealing with problems associated with Vibrio aquaculture pathogens, we look forward to discussing our work with you.  Please contact us at info@mdgbio.com.

COMBINING THE USE OF WHATMAN FTA CARDS AND DIAGNOSTIC PCR ASSAYS TO DETERMINE THE PATHOGENIC POTENTIAL OF SHRIMP WATER SAMPLES SHIPPED OVERSEAS

Sona Son, Amy Lange, Heather Behn and Michael King
Microbial Discovery Group and JBS United

SonaSon_NewsArticle

Pathogenic Vibrio spp. are a major economic concern in the shrimp industry causing global losses of approximately $3 billion USD annually. To find potential Vibrio minimizing strategies, a thorough and regular study of virulent Vibrio species collected from shrimp ponds is required. Studies of virulent Vibrio from the field is impeded by changes in the microbial community during sample transport and current field testing methods that do not accurately assess Vibrio virulence. In this study, the use of Whatman FTA (fast technology for analysis of nucleic acids) cards to preserve Vibrio community DNA during transport was combined with a Vibrio harveyi and Vibrio campbellii haemolysin virulence gene based multiplex PCR assay1. Successful protocols will later give rise to quantitative PCR (qPCR) methods that can be used to quantify pond sample virulence potential.

Serial dilution of Vibrio cultures exhibiting virulence genes and end point multiplex PCR protocols were used to obtain semiquantiative data for evaluation. Closely related Vibrio harveyi and Vibrio campbellii culture DNA extracted from FTA cards was compared to DNA extracted with Qiagen DNA isolation columns. FTA card DNA extractions were also used to detect differences between pure cultures and cultures diluted in shrimp pond water. Further, detection differences were tested between freshly inoculated FTA cards and cards exposed to an overseas shipment simulation (21 days heat incubation at 25ºC and 32ºC). FTA card virulence gene PCR amplification protocols allowed for the detection of virulence genes from as few as 100-600 Vibrio cells per milliliter using the endpoint PCR protocol. The sensitivity of the virulence gene assay using pure culture template DNA from FTA cards was similar to results obtained using Qiagen DNA isolation columns, with a detection limit of approximately 100 cells/ml. The presence of PCR inhibitors in shrimp pond water reduced detection 100-fold compared to the detection of pure cultures. The effect of the PCR inhibitors appeared to have been removed by subsequent Qiagen column DNA purification after the FTA card, allowing virulence gene detection comparable to pure culture. Little to no difference was observed between the PCR amplifications of freshly inoculated FTA cards and inoculated cards exposed to the overseas transport simulation. These results suggest that these methods could be applicable to Vibrio virulence potential studies as applied to FTA cards for transporting overseas samples. Further studies receiving actual samples from overseas shipments are warranted.

 

1. Haldar, S. et al. (2010) Development of a haemolysin gene-based multiplex PCR for simultaneous detection of Vibrio campbellii, Vibrio harveyi, and Vibrio parahaemolyticus. Letters in Applied Microbiology 50:146-152.

Search Insights
Recently Posted Insights

Upcoming Webinar: Creating Quality Microbial Products

Producing high-quality microbial products for industrial, institutional, and consumer applications is essential for our partners. In our upcoming webinar, we will explore the critical aspects of microbial product development and how we ensure quality products for your...

Proven Success Treating Sludge with Biotifx®

If your customers are working in the biological wastewater treatment industry, chances are they’re dealing with the challenges of organic sludge. Sludge, the waste components left over after the wastewater has been treated, is a constant expense for facilities, and...

Sludge Tanks 101: Treating the Waste

Throughout the wastewater treatment process, certain components are left over that get separated from the treated water. This collection of leftovers is called sludge, which is then further treated in tanks. At Microbial Discovery Group (MDG), we have explored the...

Lagoons 101: Identification and Long-Term Treatment Approach

Lagoons and ponds have been used for treating wastewater for over 3,000 years. Today, there are over 8,000 wastewater treatment lagoons operating in the United States. One appeal to using lagoons is that they generally require less energy than other treatment systems...

Proven Success in Wastewater Seasonality

In the world of wastewater treatment, seasonal challenges can disrupt operations and affect the efficiency of treatment plants. As the industry begins to face the difficulties of this year’s spring, MDG emerges as a trusted partner with a track record of proven...

Battling Restroom Odors: Debunking Ice in Urinals

Ever since the invention of the urinal back in 1886, facilities have been battling the unwanted odors that come with them. To combat the odors, businesses have tried many different treatments… some more beneficial than others.   One of the most used techniques for...

Under The Microscope – Sierra Garcia

At MDG, we succeed by creating an environment to attract, develop, and retain the best and the brightest people who embrace our values. Learn all about Sierra Garcia, our Engagement Coordinator.   What is your role at Microbial Discovery Group (MDG) and when did...

2023 EXPANSION: CONTINUED GROWTH

In 2021, our growth journey took a significant leap as we secured an 80,000 ft2 facility in Oak Creek, WI. This expansive space now accommodates our production, blending, packaging, warehouse, three laboratories, and supplementary office areas. In addition, an...

Related News