A SNAIL OUT OF WATER: HITTING THE TARGET ON PRIMER OPTIMIZATION FOR APPLE SNAIL DETECTION
Conservation efforts increasingly rely on environmental DNA (eDNA) detection to sound an alarm for non-native invasive species. However, successful invaders often include morphologically similar species. Detecting species with eDNA relies on understanding how amplification success varies across temperatures and how barcoding uses specific targets to identify species. To test for eDNA in water samples (250 mL), we developed primers designed to amplify a non-native apple snail, Pomacea maculata. We used known sequences [Folmer region of cytochrome c oxidase subunit I (COI)] as the primer template to identify four candidate assays of varied amplicon length and location. All four initially worked on P. maculata, so we sought to confirm their specificity through experiments with tissue-derived DNA from other Pomacea species and other non-native aquatic snails. We optimized our qPCR process by increasing annealing temperature in 2° increments so that our primers only amplified P. maculata at higher temperatures, but other non-native snails at lower temperatures. Our work provides the first successful eDNA detection of apple snails and could help confirm presence of one species (P. maculata) that often gets confused for a better known invader that co-occurs across Asia (P. canaliculata).
Madison Granier (Co-Presenter/Co-Author), Southwestern University, email@example.com;
Romi Burks (Co-Presenter/Co-Author), Department of Biology, Southwestern University, Georgetown, TX, USA, firstname.lastname@example.org;
Matthew Barnes (Co-Presenter/Co-Author), Texas Tech University, email@example.com;
Lauren Muskara (Primary Presenter/Author), Southwestern University, firstname.lastname@example.org;
Shellsea Miller (Co-Presenter/Co-Author), Southwestern University, email@example.com;