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Use of environmental DNA for wildlife species detection is a field of research that has seen rapid growth in recent years, however, the majority of research to date has been focused on aquatic species. Here, we propose and test a novel source for the detection of terrestrial species with environmental DNA: drinking water from watering holes and wildlife water developments. We hypothesized that when terrestrial animals drink from a water source, DNA from saliva and buccal cells is shed and can be isolated for species identification. We tested this hypothesis in a pilot study by filtering drinking water supplied to coyotes (Canis latrans) at a captive coyote research facility. DNA was successfully extracted from filters, amplified by the polymerase chain reaction, and sequenced, and sequences were positively identified as belonging to coyotes. We believe this environmental DNA based approach holds great promise for the detection of terrestrial species of conservation concern.

Kin selection theory predicts that under certain conditions animals will tolerate related individuals in their home ranges. We examined the relationship between spatiotemporal overlap and genetic relatedness in ocelots (Leopardus pardalis) to determine if kin selection plays a role in structuring ocelot populations. We used 3 years of camera trapping to examine the spatial organization of an ocelot population on Barro Colorado Island in Panama. We also placed camera traps on ocelot latrines to match photographs of individual ocelots with microsatellite genotypes from feces. Strengths of spatiotemporal overlap between individual ocelots were calculated using a half-weight association index based on how often individuals were photographed at the same camera within 30 days of one another. We calculated relatedness between individuals based on 11 variable microsatellite loci. Male ocelots overlapped with ≤ 11 females, and females overlapped with ≤ 7 males. We detected no clear evidence of strict intersexual territoriality in either sex. Mean overlap among males was more than 5 times greater than overlap among females; however, spatiotemporal overlap was strong between some female pairs. Overall, overlapping individuals were more related to one another than was the sample population as a whole, consistent with the hypothesis that kin selection influences ocelot spatial organization. This finding was driven by relatedness among overlapping females, and by relatedness among overlapping individuals of opposite sex, but not by overlapping males.

Native freshwater mussels have seen dramatic declines in western North America, but effective monitoring and management of these organisms can be difficult due to their cryptic nature. We used environmental DNA sampling—a sensitive, noninvasive genetic technique—to examine the contemporary distribution of 2 native freshwater mussel species, Anodonta nuttalliana and Margaritifera falcata, in Utah and western Wyoming. We sampled water bodies with historical presence, as well as additional water bodies and locations with promising habitat for the species. We detected A. nuttalliana from 39% of the 31 water bodies sampled and M. falcata from 16% of the 25 water bodies sampled. We demonstrated that environmental DNA technology is an efficient method for determining the distributions of freshwater mussels. Mejillones de agua dulce nativos han visto reducciones dramáticas en el oeste norteamericano; sin embargo, pueden ser difícil de detectar debido a su comportamiento críptico. Usamos el ADN del ambiente, una técnica genética sensitiva y no invasiva, para examinar la distribución contemporánea de 2 especies de mejillones de agua dulce, Anodonta nuttalliana y Margaritifera falcata, en los estados de Utah y el Oeste de Wyoming. Muestreamos masas de agua con presencias históricas, así como masas de agua adicionales con hábitat para las especies. Detectamos a A. nuttalliana en 39% de 31 masas de agua muestreadas, y M. falcata de 16% de 25 masas de agua muestreadas. Demostramos que la tecnología que implica al ADN del ambiente es un método efectivo de determinar las distribuciones de los mejillones de agua dulce.

Non-invasive genetic techniques utilising DNA extracted from faeces hold great promise for felid conservation research. These methods can be used to establish species distributions, model habitat requirements, analyse diet, estimate abundance and population density, and form the basis for population, landscape and conservation genetic analyses. Due to the elusive nature of most felid species, non-invasive genetic methods have the potential to provide valuable data that cannot be obtained with traditional observational or capture techniques. Thus, these methods are particularly valuable for research and conservation of endangered felid species. Here, we review recent studies that use non-invasive faecal genetic techniques to survey or study wild felids; provide an overview of field, laboratory and analysis techniques; and offer suggestions on how future non-invasive genetic studies can be expanded or improved to more effectively support conservation.

The North Slope of Alaska contains arctic fish populations that are important for subsistence of local human populations, and are under threat from natural resource extraction and climate change. We designed and evaluated four quantitative PCR assays for the detection of environmental DNA from five Alaskan fish species present on the North Slope of Alaska: burbot ( Lota lota ), arctic char ( Salvelinus alpinus ), Dolly Varden ( Salvelinus malma ), arctic grayling ( Thymallus arcticus ), and slimy sculpin ( Cottus cognatus ). All assays were designed and tested for species specificity and sensitivity, and all assays detected target species from filtered water samples collected from the field. These assays will enable efficient and economical detection and monitoring of these species in lakes and rivers. This in turn will provide managers with improved knowledge of current distributions and future range shifts associated with climate and development threats, enabling more timely management.

We report a record of the endangered Central American little spotted cat (Leopardus tigrinus oncilla) from Volcán Barú National Park in the state of Chiriquí, Panama, from a scat identified through noninvasive genetics. This record, from just below the summit of Panama's highest peak, represents the highest-elevation record for the species in Panama. Additionally, we designed new DNA primers for identification of L. tigrinus oncilla from scat samples.

We report a record of the endangered Central American little spotted cat (Leopardus tigrinus oncilla) from Volcan Bard National Park in the state of Chiriqui, Panama, from a scat identified through noninvasive genetics. This record, from just below the summit of Panama's highest peak, represents the highest-elevation record for the species in Panama. Additionally, we designed new DNA primers for identification of L. tigrinus oncilla from scat samples.

Understandably, given the fast pace of biodiversity loss, there is much interest in using Earth observation technology to track biodiversity, ecosystem functions and ecosystem services. However, because most biodiversity is invisible to Earth observation, indicators based on Earth observation could be misleading and reduce the effectiveness of nature conservation and even unintentionally decrease conservation effort. We describe an approach that combines automated recording devices, high-throughput DNA sequencing and modern ecological modelling to extract much more of the information available in Earth observation data. This approach is achievable now, offering efficient and near-real-time monitoring of management impacts on biodiversity and its functions and services. A new approach is outlined for capturing multiple facets of biodiversity in near real-time by combining the latest advances in automated Earth observation recording, high-throughput sequencing and ecological modelling.

Environmental DNA (eDNA) offers a sensitive tool for detecting aquatic species, including those at low population densities. However, effective use of eDNA in conservation and monitoring efforts requires an understanding of detection probabilities and optimal replication effort necessary to minimize false negatives. This study assessed the probability of detecting Anodonta nuttalliana, a native freshwater mussel, in two Utah populations with contrasting densities (medium and low). We conducted traditional visual sampling alongside highly replicated eDNA sampling at both sites. In the medium-density population, eDNA provided high detection probabilities with minimal replication. However, in the low-density population, substantially greater replication was needed to achieve similar detection probabilities. Despite the increased eDNA replication effort needed to detect the low-density population, eDNA outperformed traditional visual sampling, which failed to detect live mussels at the low-density site. Our findings highlight the critical balance between sampling effort and detection success, offering guidance for optimizing eDNA sampling strategies in monitoring efforts, especially for species which occur at low densities.Competing Interest StatementThe authors have declared no competing interest.

We developed a species-specific, quantitative PCR assay multiplex for the detection of two threatened and endangered anuran species, arroyo toad (Anaxyrus californicus) and western spadefoot (Spea hammondii), from environmental DNA (eDNA). Both species have experienced dramatic declines over the last century throughout their ranges, mostly as a result of habitat loss. Given these declines and their cryptic life histories, improved tools for detecting and monitoring both species are needed. Species-specificity and sensitivity of assays were empirically tested in the lab, and the multiplexed assays were validated with field-collected eDNA samples. Both assays were species-specific, sensitive, and effectively detected the target species from streams, rivers and ponds, although detection was imperfect. These assays provide an efficient and economical tool that will aid in the detection, monitoring, management, and conservation of these at-risk species.Competing Interest StatementThe authors have declared no competing interest.Footnotes* New figure and table added