Optimized and validated protocol to the detection of the invasive bivalve Limnoperna fortunei from eDNA plankton samples

Josiane Ribolli,Flávia Lucena Zacchi,Sophia Cassol,Grasiela Fagundes Minatto Cardoso,Evoy Zaniboni Filho,Samara Hermes Silva,Jacó Joaquim Mattos,Alex Pires De Oliveira Nuñer

doi:10.1590/s2179-975x7620

Abstract

Abstract: We optimized a methodology for plankton environmental DNA detection of the invasive golden mussel and validated it in samples from a Southern Brazil reservoir. Limnoperna fortunei is a successful invasive alien species that causes significant impacts on freshwater ecosystems. We adjusted and validated the methodology to detect L. fortunei in plankton samples, with a SYBR Green assay. Based on the standard curve analysis, the observed theoretical minimal qPCR detection level was 0.0005625 ng.µL-1 (R2 = 0.99) at a PCR quantification cycle of 14.09–29.56. We also presented a practical guide to be used in monitoring and detection of L. fortunei. The optimized protocol was efficient in detecting L. fortunei and can be used to monitor already infested environments or invasions in new environments.

Highlights

Invasive alien species (IAS) are changing environments worldwide at an unprecedented rate, threatening to deplete and to homogenize the ecosystems and their respective environmental services (Mack et al, 2000; Rahel & Olden, 2008)
We optimized a methodology for plankton environmental DNA detection of the invasive golden mussel and validated it in samples from a Southern Brazil reservoir
Environment with slow‐moving waters located in the reservoir body; I4 - Lentic environment with slow‐moving waters located in one of the marginal area formed by the reservoir; and I5 - Lentic environment with still or slow‐moving waters located in the reservoir body immediately upstream of the Itá dam

Summary

Introduction

Invasive alien species (IAS) are changing environments worldwide at an unprecedented rate, threatening to deplete and to homogenize the ecosystems and their respective environmental services (Mack et al, 2000; Rahel & Olden, 2008). The use of eDNA has experienced considered growth and has being used in community and invasion ecology, because it allows you to obtain information from a bulk DNA sample (Taberlet et al, 2012). For this reason, the combination of qPCR and eDNA has been used as a technique to monitor aquatic environments (Jerde et al, 2011; Goldberg et al, 2013; Stoeckle et al, 2017), speeding up and bringing reliability to the use of non-lethal methods

Methods

Results

Conclusion