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Environmental DNA (eDNA) approaches to monitoring biodiversity in terrestrial environments have largely focused on sampling water bodies, potentially limiting the geographic and taxonomic scope of eDNA investigations. We assessed the performance of two strictly terrestrial eDNA sampling approaches to detect arboreal mammals, a guild with many threatened and poorly studied taxa worldwide, within two central New Jersey (USA) woodlands. We evaluated species detected with metabarcoding using two eDNA collection methods (tree bark vs. soil sampling), and compared the performance of two detection methods (qPCR vs. metabarcoding) within a single species. Our survey, which included 94 sampling events at 21 trees, detected 16 species of mammals, representing over 60% of the diversity expected in the area. More DNA was found for the 8 arboreal versus 8 non-arboreal species detected (mean: 2466 vs. 289 reads/sample). Soil samples revealed a generally similar composition, but a lower diversity, of mammal species. Detection rates for big brown bat were 3.4 × higher for qPCR over metabarcoding, illustrating the enhanced sensitivity of single-species approaches. Our results suggest that sampling eDNA from on and around trees could serve as a useful new monitoring tool for cryptic arboreal mammal communities globally.

Skin color patterns are ubiquitous in nature, impact social behavior, predator avoidance, and protection from ultraviolet irradiation. A leading model system for vertebrate skin patterning is the zebrafish; its alternating blue stripes and yellow interstripes depend on light-reflecting cells called iridophores. It was suggested that the zebrafish’s color pattern arises from a single type of iridophore migrating differentially to stripes and interstripes. However, here we find that iridophores do not migrate between stripes and interstripes but instead differentiate and proliferate in-place, based on their micro-environment. RNA-sequencing analysis further reveals that stripe and interstripe iridophores have different transcriptomic states, while cryogenic-scanning-electron-microscopy and micro-X-ray diffraction identify different crystal-arrays architectures, indicating that stripe and interstripe iridophores are different cell types. Based on these results, we present an alternative model of skin patterning in zebrafish in which distinct iridophore crystallotypes containing specialized, physiologically responsive, organelles arise in stripe and interstripe by in-situ differentiation. The skin of zebrafish is patterned by alternating blue stripes and yellow interstripes which arises from guanine crystal-containing cells called iridophores that reflect light. Here the authors track iridophores and see that they do not migrate between stripes and interstripes, but instead differentiate and proliferate in place based on their micro-environment.

Colour produced by wavelength-dependent light scattering is a key component of visual communication in nature and acts particularly strongly in visual signalling by structurally-coloured animals during courtship. Two miniature peacock spiders ( Maratus robinsoni and M. chrysomelas ) court females using tiny structured scales (~ 40 × 10 μm 2 ) that reflect the full visual spectrum. Using TEM and optical modelling, we show that the spiders’ scales have 2D nanogratings on microscale 3D convex surfaces with at least twice the resolving power of a conventional 2D diffraction grating of the same period. Whereas the long optical path lengths required for light-dispersive components to resolve individual wavelengths constrain current spectrometers to bulky sizes, our nano-3D printed prototypes demonstrate that the design principle of the peacock spiders’ scales could inspire novel, miniature light-dispersive components. Several animals and plants get their often spectacular colouration and iridescence from structural colouration. Here, Hsiung et al. show the 3-dimensional nanostructures that produce the rainbow colours on the abdomen of miniature peacock spiders.

Environmental DNA (eDNA) analyses have become invaluable for detecting and monitoring aquatic and terrestrial species and assessing site biodiversity within aquatic environments or soil. Recent studies have extended these techniques by using eDNA to identify the presence of aboveground terrestrial arthropods directly from vegetative surfaces. However, while the dynamics of eDNA state, transport, and fate (its “ecology”) have been explored within aquatic environments and soil, they have yet to be explored within aboveground terrestrial systems. Here, we explore the ecology of terrestrial eDNA deposited by fluid‐feeding arthropods on leaf surfaces. We carried out a series of experiments to evaluate the optimal filter pore size for intracellular eDNA collection, how eDNA is affected by rain events, and its degradation rate under different solar radiation conditions. We found that the captured concentration of intracellular eDNA was not significantly affected by an increase in filter pore size, suggesting a wide range of viable pore size options exist for targeting intracellular eDNA. We also found extracellular eDNA from fluid excrement degrades more rapidly than intracellular when exposed to solar radiation, indicating the latter is a more viable target for collection. Finally, we identified that rainfall or mist will remove most terrestrial eDNA present on vegetation surfaces. We provide researchers and environmental managers key insights into successfully designing and carrying out aboveground terrestrial arthropod eDNA surveys that maximize detection probability. Here, we explore the ecology of terrestrial eDNA deposited by fluid‐feeding arthropods on leaf surfaces. We carried out a series of experiments to evaluate the optimal filter pore size for intracellular eDNA collection, how eDNA is affected by rain events, and its degradation rate under different solar radiation conditions. We found that the captured concentration of intracellular eDNA was not significantly affected by an increase in filter pore size, extracellular eDNA from fluid excrement degraded more rapidly than intracellular when exposed to solar radiation, and rainfall or mist will remove most terrestrial eDNA present on vegetation surfaces.

Recent methodological advances permit surveys for terrestrial insects from the direct collection of environmental DNA (eDNA) deposited on vegetation or other surfaces. However, in contrast to well‐studied aquatic applications, little is known about how detection rates for such terrestrial eDNA‐based surveys compare with conventional survey methods. Lycorma delicatula, the spotted lanternfly, is an emerging invasive insect in eastern North America, and a significant ecological and economic pest of forested and agricultural systems, especially grapes. During fall 2019, we conducted two rounds of paired eDNA and visual surveys for spotted lanternflies within 48 plots at 12 vineyards in New Jersey, USA. We compared detection probabilities within a multimethod occupancy modeling framework and used the results to extrapolate and inform survey design. The probability of detecting spotted lanternflies given presence in a plot was over two times higher for eDNA (84%) versus visual surveys (36%). In mid‐September, lanternfly eDNA was detected at five plots in three vineyards, while visual surveys revealed only a single individual in one plot. In early October, after dispersal of lanternflies into vineyards, lanternfly eDNA was detected in 12 plots within six vineyards compared with visual detections in six plots in two vineyards. Extrapolations based on detection and local‐scale occupancy rates indicate that only five and 12 plots would have been needed to positively detect lanternfly presence with 95% confidence using eDNA in contrast to 14 and 29 plots with visual surveys alone, respective to survey rounds. Log‐linear models revealed that visual counts of lanternflies were positively related to eDNA concentrations (R2 = 71%). We provide some of the first quantitative evidence to support the enhanced sensitivity of terrestrial eDNA approaches compared with conventional methods. Such methods can augment efforts to combat invasive species through improved ability to delimit invasion fronts, identify satellite populations, and confirm local eradications. We field‐tested a recently‐developed protocol for detecting insect eDNA on vegetation surfaces. Lycorma delicatula, the spotted lanternfly, is an emerging invasive insect in the USA that is highly destructive to wine grapes among other crops. Paired surveys in vineyard plots and multimethod occupancy modeling revealed nearly three times higher detection probability for eDNA‐based methods compared with standard visual surveys.

Environmental DNA surveys have revolutionized monitoring of rare or cryptic species and species inhabiting areas where conventional sampling is difficult or dangerous. Recent advancements within terrestrial environments include the capture of eDNA deposited by animals on surfaces such as tree bark and foliage, hereafter “surface eDNA.” Notably, a technique which uses commercial paint rollers to aggregate surface eDNA has been deployed with success to detect the presence of forest insect pests providing a potentially powerful new management tool. However, before widespread adoption is feasible, the efficiency and logistics of roller sample collection and study design, especially relative to realistic survey conditions, must be evaluated. We compared the performance of two DNA preservation treatments—cold and ethanol—on their ability to reduce the loss of captured eDNA on rollers over time. Additionally, we evaluated how the detection probability of our target species, the spotted lanternfly (Lycorma delicatula), varied with sampling effort (time spent rolling per sample) and the initial quantity of eDNA present. Finally, we evaluated how the number of trees sampled per roller influenced the final concentrations of lanternfly eDNA remaining on the roller. We found storing rollers with ethanol or cold temperatures resulted in 3–10‐fold greater concentrations of experimentally controlled eDNA relative to no treatment after 24 h. Detection probability declined as the amount of lanternfly eDNA decreased, but did not change in response to sampling effort over sample time (10–80 s/tree). Finally, recovered lanternfly eDNA decreased as more trees were sampled by a single roller—a 91% reduction after 7 trees—potentially due to captured DNA being transferred back from the roller onto the bark. Our results provide improved guidance for deploying roller surface eDNA methods for spotted lanternfly surveys, and for invasive insect pest surveillance and monitoring programs generally. Surveying plant surfaces for eDNA is relatively novel. We present several experiments to improve on a recent technique used for surveying bark and branches of trees for insect DNA. Cold temperatures can preserve DNA on paint rollers, used to sample trees, while sampling more than a few trees can lead to DNA loss.

Invasive wood-boring insects are a major economic and ecological concern worldwide as they impact native woody plant populations. These pest species are increasing in prevalence, with devastating impact, as global trade leads to higher rates of introduction and establishment. The emerald ash borer ( Agrilus planipennis ; EAB) is one such species, which has caused widespread damage across much of the United States and is now spreading across Europe. Non-indigenous woodborers such as EAB are difficult to detect at early stages of invasion, which is when management and eradication efforts are most effective and cost efficient. Environmental DNA (eDNA) surveys have demonstrated power in detecting invasive species when rare in the landscape due to their ability to detect trace amounts of DNA and identify to species. Here, we trialled a novel eDNA method for collecting environmental samples within host trees where invasive pest larvae are feeding, using EAB as a case study. We extracted tree cores approximately 1 cm in length using an increment hammer to assess detectability of eDNA from larvae feeding under the bark. In trees visibly infested with EAB, we observed a seasonal peak in EAB DNA detection probability (~ 64%; towards the end of the growing season), indicating a potential impact of ash tree phenology or EAB phenology on detection. When we trialled the method in a site with ash trees of low or uncertain EAB abundance, we did not record positive EAB eDNA detections. This outcome may have resulted from differing EAB phenology at the northern latitude of this survey site or because larval galleries were less numerous causing EAB DNA to be scarcer within the tree. Our results, however, provide preliminary evidence that increment hammer tree cores can be used to detect eDNA of EAB and, perhaps, other wood-boring pests. Further work is needed to clarify false negative survey detections at ash trees showing little to no signs or symptoms of infestation, as well as investigating the deposition, transport and persistence dynamics of EAB eDNA within trees.

Environmental DNA (eDNA) has recently emerged as an effective tool for invasive species biosecurity. We explored the use of eDNA for the detection of khapra beetle (Trogoderma granarium, Everts 1898), an invasive insect of cereal grains and other food products that has a high global economic impact. We developed a novel method for aggregating khapra beetle eDNA deposited in stored grain that entails washing a sample of rice, filtering the sample, and detecting trace beetle DNA using a standard qPCR workflow. To explore the performance of this method, we raised 500 khapra beetle larvae within 500 g of rice over a 14‐day period and then removed them. We then used this “spiked” rice to create a range of simulated densities of khapra beetle larvae. This lab approach mimics conditions that are comparable to field densities of ~1.4 to 180 beetles per 50 kg of rice (1/8 to 16 spiked rice grains per 100 g sample of clean rice), assuming DNA is uniformly distributed within the rice. We detected khapra beetle eDNA from all density levels tested. Logistic models revealed that eDNA amounts equivalent to what is left by ~1 khapra beetle larva in a 50 kg container of rice can be detected with 85% to >97% certainty, depending on the number of qPCR technical replicates run per sample. Based on this model, we estimated that for one 50 kg container of rice where beetle DNA is uniformly distributed, a single sample of 100 g with six technical replicates would be sufficient to be >99% certain that the container was free of khapra beetle eDNA (95% credible intervals: 97.7%–100%). Our results suggest that eDNA surveys may be useful as a cost‐effective, first‐step detection of khapra beetle in stored grain and provide a means to map the relative magnitude of khapra beetle transport pathways, informing allocation of conventional biosecurity inspection efforts. Khapra beetle is a agricultural pest posing high global biosecurity risk. We show eDNA sampling of stored grain can detect ~1 khapra beetle larva in a 50 kg container of rice with 85% to >97% certainty (dashed line is 95% certainty level) depending on whether 1 (red) or 3 (blue) qPCR technical replicates are performed. This result suggests, with further exploration, eDNA surveys can become cost‐effective first‐step biosecurity protocol for khapra beetle, or a means to map the relative magnitude of khapra beetle transport pathways.

Migration is a prevalent strategy among birds used to track seasonal resources throughout the year. Individual and population‐level migratory movements provide insight to life‐history variation, carry‐over effects, and impacts of climate change. Our understanding of how geographic variation in a species' breeding or wintering grounds can impact migration distances is limited. However, changes in migration distances can have important fitness consequences for individuals and conservation implications for populations, particularly if migratory connectivity is altered during the annual cycle. In this study, we use three decades of data from the United States Geological Survey Bird Banding Laboratory for six migratory species of Arctic and subarctic breeding geese. We employ a Bayesian hierarchical framework to test if the distance between breeding and wintering locations has changed over time, while accounting for the latitude of the breeding grounds. A model that included only a temporal trend estimated the average rate of change in migration distance, across all six species, at −3.0 km/year over the period 1990–2019. Five of the six species showed a significant decrease in migration distances. Including an interaction effect with breeding latitude revealed that the reduction in migration distance was strongest in the southernmost populations for four of the six species. For those species, migration distance in northern populations were all either relatively unchanged or increasing. This indicates that southern breeding populations of geese had a stronger association with the observed spatiotemporal changes in wintering ranges, potentially influenced by a combination of climatic and biotic factors (e.g. resource availability or competitive interactions) that uniquely impact these populations. Abundant, long‐term banding data shows promise for use in illuminating changes in migratory patterns under climate change, leading to improved management and conservation outcomes, from regional to continental scales.