Naslov
Development of Saprolegnia parasitica PCR detection-assay

Autori
Pavić, Dora ; Vujović, Tamara ; Miljanović, Anđela ; Bielen, Ana

Vrsta, podvrsta i kategorija rada
Sažeci sa skupova, sažetak, znanstveni

Izvornik
8th Congress of European Microbiologists (FEMS2019) Abstract Book / - , 2019, 1260-1260

Skup
8th Congress of European Microbiologists (FEMS 2019)

Mjesto i datum
Glasgow, Ujedinjeno Kraljevstvo, 07.07.2019. - 11.07.2019

Vrsta sudjelovanja
Poster

Vrsta recenzije
Međunarodna recenzija

Ključne riječi
Saprolegnia parasitica ; PCR assay ; Monitoring

Sažetak
Background: Some oomycetes from the genus Saprolegnia are causing saprolegniosis, a disease of salmonid fish important in aquaculture. Saprolegnia species cause significant economic losses in freshwater aquaculture, with Saprolegnia parasitica as the most destructive pathogen. Despite its negative effects, no monitoring protocol of S. parasitica has been established to date. Objectives: We aimed to develop PCR-detection assay for S. parasitica in order to enable its monitoring in freshwater aquaculture facilities. Methods: We constructed multiple sequence alignment of 958 internal transcribed spacer (ITS) sequences belonging to various oomycetes. We have identified S. parasitica specific regions and created three primer pairs, designated A, B and C. Next, we used gradient PCR and S. parasitica (CBS 233.65) genomic DNA as a template to determine optimal annealing temperature for each primer pair. Results: Primer pairs A and B have amplified unspecific bands that could not be eliminated, but primer pair C amplified a single band of expected size. The sensitivity of the assay with primer pair C was high: lower detection limit was < 1 pg. Also, we have demonstrated the specificity of the assay using a range of Saprolegnia species and other, more distantly related Oomycetes. In conclusion, intense fish farming in large- scale aquaculture can lead to a significant increase in saprolegniosis. S. parasitica detection assay developed here will enable early detection of S. parasitica in fish farms and therefore help in improving poor farming conditions.

Izvorni jezik
Engleski

Znanstvena područja
Biologija, Biotehnologija

POVEZANOST RADA