Development of a tetraplex digital PCR (dPCR) assay for the detection of
invasive snake species in Florida, U.S.A.
Abstract
Florida, U.S.A. is a hotspot of biological invasions with over 500
non-native species reported. Reptiles encompass the majority of
non-native wildlife with over 50 species established, many of which are
sympatric and are identified as invasive due to their impacts to the
environment, economy, and human health and safety. Reports of new
non-native reptiles occur and many established non-native reptiles
continue to expand their ranges in Florida, increasing the need for
multi-taxa detection and monitoring capabilities. Invasive constrictor
snakes are a primary focus of management efforts due to life history
traits that favor successful establishment and dispersal in Florida as
well as their impacts to native wildlife and Everglades restoration
efforts. While traditional survey methods that rely on visual detections
fail to reliably detect invasive constrictors, environmental DNA (eDNA)
has proven to be a promising method for detection of cryptic and rare
species across the landscape. To address emerging needs for
multi-species detection and monitoring in Florida we developed the first
tetraplex dPCR assay designed for detection of four species of invasive
constrictor snakes, including Burmese pythons (Python bivittatus),
northern African pythons (P. sebae), boa constrictors (Boa constrictor),
and rainbow boas (Epicrates cenchria). In this tetraplex assay, no
cross-amplification across species was documented. This assay serves as
a valuable tool for faster and more accurate monitoring efforts of these
invasive species in south Florida.