Location: Harry K. Dupree Stuttgart National Aquaculture Research CntrTitle: Molecular detection and quantification of the fish pathogens Saprolegnia spp. using qPCR and Loop Mediated Isothermal Amplification (LAMP) from recirculating aquaculture systems
|GHOSH, SATYAKI - Bowling Green State University|
|Straus, David - Dave|
|GOOD, CHRISTOPHER - Freshwater Institute|
|PHUNTUMART, VIPAPORN - Bowling Green State University|
Submitted to: Meeting Abstract
Publication Type: Abstract Only
Publication Acceptance Date: 4/1/2019
Publication Date: 6/10/2019
Citation: Ghosh, S., Straus, D.L., Good, C., Phuntumart, V. 2019. Molecular detection and quantification of the fish pathogens Saprolegnia spp. using qPCR and Loop Mediated Isothermal Amplification (LAMP) from recirculating aquaculture systems [abstrast]. 62nd Annual Conference on the Great Lakes Research, June 10-14, 2019, Brockport, NY. p. 1.
Technical Abstract: Saprolegniasis is a disease caused by oomycete pathogens of the genus Saprolegnia, which can infect fish in both natural habitats and commercial aquaculture. It is estimated to cause economic losses of 10% in salmon production alone and 30% in fish production worldwide. In production and natural settings, a sudden drop in water temperature immunocompromises the fish host, while facilitating the virulence of the Saprolegnia spp. pathogens. The goal of this study is to establish Loop Mediated Isothermal Amplification (LAMP) as a rapid and sensitive molecular tool for on-field detection and quantification of Saprolegnia spp. from water samples, specifically focusing on detection of zoospores. This will facilitate informed decisions about the timing and extent of disease treatment. LAMP reactions are performed at a constant temperature, and results can be estimated visually within 30 minutes, making it suitable for field diagnostics. Our developed LAMP technique could detect as low as 10fg of Saprolegnia spp. DNA, and one zoospore of Saprolegnia spp. directly. Additionally, we have developed a simple strategy to filter large volumes of water through polycarbonate track etch (PCTE) membranes, followed by a simple heat-based DNA extraction strategy for LAMP. This procedure was successfully applied to detect Saprolegnia spp. in water collected from recirculating aquaculture systems.