Genetic assays for guano-based identification of species and sex in bats of the United States and Canada

Abstract

Bat guano is a noninvasive, data-rich genetic resource. However, the constituent bat DNA is relatively scant, degraded, and complexed with polymerase chain reaction inhibitors. It also is comingled with a rich pool of nontarget DNA from microbes, parasites, and dietary items. We designed and tested new DNA assays for bat species identification (COX1-Bat) and sex identification (XGXYC) for use with guano and other challenging samples. We reviewed previously published assays that can be used with guano samples to obtain the same species and sex data, and attempted to validate these assays for species in which they had not previously been tested. Our results demonstrate that guano-derived DNA can be used successfully to 1) identify nearly all US and Canadian bats at the species level, or to one of three Myotis species clusters, and 2) identify the sex of at least 23 US and Canadian bat species. Our newly developed assays, and validation of previously published assays, for guano-based identification of species and sex in bats, significantly enhance the power of noninvasive sampling and genetic analysis for bat studies, management, and conservation.

Multifaceted DNA metabarcoding of guano to uncover multiple classes of ecological data in two different bat communities

“…Two ASVs were assigned to Eptesicus fuscus Palisot de Beauvois (Big Brown Bat), both with sequence matches of 100% to E. fuscus 16S rRNA sequence. Although M. lucifugus sequences for this locus maybe identical or highly similar to homologous sequence in some other Myotis species (Guan et al, 2020 ), no such species occurs in the sampled region.…”

“…Two, noninvasive sample collection does not require specialized training. Three, scat may contain DNA evidence for many different key organismal attributes or states, which can be uncovered using different “universal” assays (Guan et al, 2020 , 2021 ; Swift et al, 2018 ; Walker et al, 2016 ; Zeale et al, 2011 ). Further, some of these attributes, such as the presence of endoparasites and diet, are very difficult to quantify using traditional approaches (involving detailed necropsies or microscopic analyses of scat), and the use of DNA metabarcoding may vastly improve the ease and efficiency of collecting these data (Edwards et al, 2019 ; Swift et al, 2018 ).…”

Sampling methodology influences habitat suitability modeling for chiropteran species

“…While manual vetting may reduce false‐positive rates, subjectivity among various call ID software and specialists remains (Fritsch & Bruckner, 2014 ; Russo & Voigt, 2016 ). This extends beyond acoustics to include misidentification of feces, incorrect track identification, and blurry camera trap photos, where genetic analyses provide confidence for some detection methods (Clare et al, 2017 ; Guan et al, 2020 ; Louvrier et al, 2019 ). For species with high false identification rates, occupancy models have begun incorporating false‐positive parameterizations, thus improving model estimation (Clement et al, 2014 ; Miller et al, 2011 ; Rojas et al, 2019 ).…”

Influencing Activity of Bats by Dimly Lighting Wind Turbine Surfaces with Ultraviolet Light

“…To date, the most common metric of efficacy for fatality reduction tests has been to compare the number of dead bats found beneath wind turbines following nights with and without applied treatments (e.g., curtailment, acoustic deterrents [ 20 , 21 , 25 , 26 , 27 , 109 , 110 ]). This gold standard of counting carcasses to judge if a method works is fully justified and supported by robust statistical measures and laboratory techniques to enhance the quality, precision, and reliability of such data [ 111 , 112 , 113 , 114 , 115 , 116 , 117 , 118 , 119 , 120 , 121 ]. Had we relied on fatality ground searches as a metric of bat response to dim-UV treatment, we would not have been able to statistically test for treatment effects, by either attraction or deterrence, simply (and fortunately) due to the low sample size.…”