Two novel qPCR assays to enhance black rail (Laterallus jamaicensis) eDNA surveys in the United States.

Black rails (Laterallus jamaicensis) are a rare and protected bird species. They are also notoriously difficult to monitor. eDNA surveys have proven beneficial for the detection of this elusive species, but additional developments are needed to enhance eDNA survey utility (including reliability of outcomes) in light of black rail status and associated high-profile conservation actions. The use of multiple assays functions to increase eDNA detection confidence by increasing the probability of amplification in low concentration, highly degraded samples (i.e., reducing the risk of false negatives) and by providing a multiplicative test for ruling out contamination as the source of qPCR amplification (i.e., reducing the risk of false positives). The two eDNA assays developed, optimized, and validated in this study will prove valuable for the continued surveillance of the secretive black rail, offering data to measure its distribution and occurrence over time and as influenced by anticipated climate-change-induced disturbances and/or habitat restoration/creation initiatives. The assays are applicable to both Eastern black rails and California black rails, and do not exhibit qPCR amplification in non-target, co-distributed taxa (including other Rallidae species) in the United States. [ABSTRACT FROM AUTHOR]