Dual Detection of the Chytrid Fungi <i>Batrachochytrium</i> spp. with an Enhanced Environmental DNA Approach

oleh: David Lastra González, Vojtech Baláž, Jiří Vojar, Petr Chajma

Format: Article
Diterbitkan: MDPI AG 2021-03-01

Deskripsi

Environmental DNA (eDNA) is becoming an indispensable tool in biodiversity monitoring, including the monitoring of invasive species and pathogens. Aquatic chytrid fungi <i>Batrachochytrium dendrobatidis</i> (<i>Bd</i>) and <i>B. salamandrivorans</i> (<i>Bsal</i>) are major threats to amphibians. However, the use of eDNA for detecting these pathogens has not yet become widespread, due to technological and economic obstacles. Using the enhanced eDNA approach (a simple and cheap sampling protocol) and the universally accepted qPCR assay, we confirmed the presence of <i>Bsal</i> and <i>Bd</i> in previously identified sites in Spain, including four sites that were new for <i>Bsal</i>. The new approach was successfully tested in laboratory conditions using manufactured gene fragments (gBlocks) of the targeted DNA sequence. A comparison of storage methods showed that samples kept in ethanol had the best DNA yield. Our results showed that the number of DNA copies in the Internal Transcribed Spacer region was 120 copies per <i>Bsal</i> cell. Eradication of emerging diseases requires quick and cost-effective solutions. We therefore performed cost-efficiency analyses of standard animal swabbing, a previous eDNA approach, and our own approach. The procedure presented here was evaluated as the most cost-efficient. Our findings will help to disseminate information about efforts to prevent the spread of chytrid fungi.