A multi-method approach for assessing the distribution of a rare, burrowing North American crayfish species.

Primary burrowing crayfishes face high extinction risk, but are challenging to study, manage, and conserve due to their difficult-to-sample habitat (i.e., terrestrial burrows) and low population densities. We apply here a variety of methods to characterize the distribution, habitat associations, and conservation status of the Boston Mountains Crayfish Cambarus causeyi (Reimer, 1966), an endemic burrowing crayfish found only in the Ozark Mountains of Arkansas, United States. We used species distribution modeling (SDM) on historic occurrence records to characterize the distribution and macro-scale habitat associations of this species. We then ground-truthed SDM predictions with conventional sampling, modeled fine-scale habitat associations with generalized linear models (GLM), and lastly developed and tested an environmental DNA (eDNA) assay for this species in comparison to conventional sampling. This represents, to our knowledge, the first successful eDNA assay for a terrestrial burrowing crayfish. Our MaxEnt-derived SDM found a strong effect of average annual precipitation on the historic distribution of C. causeyi, which occurred most frequently at locations with moderately high average annual precipitation (140-150 cm/yr) within our study region. Cambarus causeyi was difficult to detect by conventional sampling in 2019 and 2020, found at only 9 of 51 sites (17.6%) sampled by searching for and manually excavating crayfish burrows. Surprisingly, habitat suitability predicted from our MaxEnt models was not associated with contemporary C. causeyi occurrences per GLMs. Instead, C. causeyi presence was negatively associated with both sandy soils and the presence of other burrowing crayfish species. Poor SDM performance in this instance was likely caused by the omission of high resolution fine-scale habitat data (e.g., soils) and biotic interactions from MaxEnt models. Finally, our eDNA assay detected C. causeyi from six of 25 sites (24.0%) sampled in 2020, out-performing conventional surveys by burrow excavation for this species. Given the difficulty of studying primary burrowing crayfishes and their high conservation need, we propose that eDNA may become an increasingly important monitoring tool for C. causeyi and similar species.

Increased Temperature Influenced Growth and Development of Lithobates pipiens Tadpoles Exposed to Leachates of the Invasive Plant European Buckthorn (Rhamnus cathartica) and a Triclopyr Herbicide.

Multiple factors including habitat loss, pollutants, invasive species, and disease have contributed to the global decline of amphibians, and further declines can be expected as a result of climate change. Warming temperatures may allow for range expansion of invasive plants, and because herbicides are the primary method to control invasive plants, chemical use may increase. A laboratory experiment was performed to examine the individual and combined effects of leachates from the invasive plant European buckthorn (Rhamnus cathartica, L.) and a triclopyr herbicide (Renovate® 3; 0.21 mg/L), which is commonly used to manage R. cathartica, on northern leopard frog (Lithobates pipiens, Schreber) tadpoles at 2 temperature regimes (20 and 25 °C). We measured tadpole growth weekly and body and intestine morphology at the conclusion of the experiment after 8 wk. In the presence of R. cathartica leachates, tadpole growth increased at 25 °C, but only during the first 3 to 4 wk of the experiment. From week 5 until the end of the experiment, tadpoles were significantly smaller at 25 °C compared with 20 °C, but had more developed limb buds at the end of the experiment (except in the triclopyr treatment). Triclopyr had minimal effects on tadpole growth at the low dose used in this study. These results encourage further examination of potential effects of global climate changes in combination with other environmental factors that may impact amphibian populations. Environ Toxicol Chem 2021;40:2547-2558. © 2021 SETAC.

No evidence that crayfish carcasses produce detectable environmental DNA (eDNA) in a stream enclosure experiment.

Environmental DNA (eDNA) is an emerging tool for monitoring invasive and imperiled species, particularly at low densities. However, the factors that control eDNA production, transport, and persistence in aquatic systems remain poorly understood. For example, the extent to which carcasses produce detectable eDNA is unknown. If positive detections are associated with dead organisms, this could confound monitoring for imperiled or invasive species. Here, we present results from one of the first studies to examine carcass eDNA in situ by deploying carcasses of the invasive red swamp crayfish (Procambarus clarkii) in a stream enclosure experiment for 28 days. We predicted that carcasses would initially produce eDNA that would decline over time as carcasses decayed. Unsurprisingly, crayfish carcasses lost biomass over time, but at the conclusion of our experiment much of the carapace and chelae remained. However, no eDNA of P. clarkii was detected in any of our samples at the crayfish density (15 P. clarkii carcasses at ∼615 g of biomass initially), stream flow (520-20,319 L/s), or temperature (∼14-25 °C) at our site. Subsequent analyses demonstrated that these results were not the consequence of PCR inhibition in our field samples, poor performance of the eDNA assay for intraspecific genetic diversity within P. clarkii, or due to the preservation and extraction procedure used. Therefore, our results suggest that when crayfish are relatively rare, such as in cases of new invasive populations or endangered species, carcasses may not produce detectable eDNA. In such scenarios, positive detections from field studies may be more confidently attributed to the presence of live organisms. We recommend that future studies should explore how biomass, flow, and differences in system (lentic vs. lotic) influence the ability to detect eDNA from carcasses.

Effects of temperature, salinity, and sediment organic carbon on methylmercury bioaccumulation in an estuarine amphipod.

Mercury (Hg) is a global contaminant that poses a human health risk in its organic form, methylmercury (MeHg), through consumption of fish and fishery products. Bioaccumulation of Hg in the aquatic environment is controlled by a number of factors expected to be altered by climate change. We examined the individual and combined effects of temperature, sediment organic carbon, and salinity on the bioaccumulation of MeHg in an estuarine amphipod, Leptocheirus plumulosus, when exposed to sediment from two locations in the Gulf of Maine (Kittery and Bass Harbor) that contained different levels of MeHg and organic carbon. Higher temperatures and lower organic carbon levels individually increased uptake of MeHg by L. plumulosus as measured by the biota-sediment accumulation factor (BSAF), while the effect of salinity on BSAF differed by sediment source. Multi-factor statistical modeling using all data revealed a significant interaction between temperature and organic carbon for both sediments, in which increased temperature had a negative effect on BSAF at the lowest carbon levels and a positive effect at higher levels. Our results suggest that increased temperature and carbon loading, of a magnitude expected as a result from climate change, could be associated with a net decrease in amphipod BSAF of 50 to 71%, depending on sediment characteristics. While these are only first-order projections, our results indicate that the future fate of MeHg in marine food webs is likely to depend on a number of factors beyond Hg loading.

Bioaccumulation of methylmercury in wood frogs and spotted salamanders in Vermont vernal pools.

Mercury (Hg) has accumulated in forested landscapes in the Northeastern U.S., and hotspots with enhanced deposition have been identified throughout the region. Due to a variety of favorable landscape characteristics, including relatively high dissolved organic carbon (DOC), fluctuating water levels, and low pH and dissolved oxygen, vernal pools provide ideal conditions for the conversion of Hg to its more toxic and bioavailable form, methylmercury (MeHg). Yet little is known about the concentrations, speciation, and bioavailability of Hg in vernal pools, or its bioaccumulation in vernal pool fauna and potential export into terrestrial systems. We investigated the role of forest cover type on the bioaccumulation of MeHg in wood frog (Lithobates sylvatica) and spotted salamander (Ambystoma maculatum) eggs, larvae, and adults, and investigated relationships among MeHg and water chemistry (pH, DOC). Water samples from pools located in coniferous stands had greater concentrations of THg and MeHg compared to deciduous pool water, and showed significant positive correlation to DOC (r = 0.683, P < 0.001) and correlated negatively with pH (r = -0.613, P < 0.001). Methylmercury levels in amphibian embryos were similar between the two species (L. sylvatica mean = 5.4 ng/g dw; A. maculatum mean = 3.5 ng/g dw). Concentrations of MeHg increased substantially in larvae, and were significantly greater in A. maculatum (mean = 237.6 ng/g ± 18.5 SE) than L. sylvatica larvae (62.5 ng/g ± 5.7 SE). Forest cover type did not explain variation in MeHg concentration among amphibian embryos or larvae. Methylmercury levels in adult tissue samples were significantly greater in A. maculatum (mean = 79.9 ng/g ± 8.9 SE) compared to L. sylvatica (mean = 47.7 ng/g ± 9.7 SE). This research demonstrates that vernal pools are important hotspots where amphibians bioaccumulate MeHg, which may then be transferred to terrestrial ecosystems. The abundance of amphibian larvae suggests they could be important bioindicators for monitoring MeHg loading and bioavailability.

Bioaccumulation of mercury and other metal contaminants in invasive lionfish (Pterois volitans/miles) from Curaçao.

A wide range of ecological and environmental factors influence metal bioaccumulation in fish. Studies of mercury and other metal contaminants in invasive Indo-Pacific lionfish are limited, yet consumption of the invasive predator is increasingly utilized as a management strategy. In this study, we examined the effects of body size, body condition, sex, trophic level, carbon source, diet, depth and capture location on mercury concentrations in lionfish collected from Curaçao. In addition, we examined whether or not a local petroleum refinery is the source of metal contamination in lionfish. Mercury concentrations ranged from 0.008 to 0.106 mg/kg and we found no effect of the petroleum refinery on metal bioaccumulation in lionfish. Low concentrations of metal contaminants indicate lionfish from Curaçao are safe for human consumption.

Influence of sample preparation on estuarine macrofauna stable isotope signatures in the context of contaminant bioaccumulation studies.

The ratios of stable isotopes of carbon and nitrogen provide important information on food sources of aquatic organisms and trophic structure of aquatic food webs. For many studies, trophic position and food source are linked to bioaccumulation and trophic transfer of contaminants from prey to predators. In these cases, it is useful to use measurements on whole organisms to make direct comparisons of contaminant bioaccumulation and food web attributes. There is a great deal of variation in methods used for stable isotope analysis, particularly in the selection of tissue type and sample preparation prior to stable isotope analysis. While there have been aquatic studies that examined methodological differences, few have focused on estuarine organisms. In this study, the effects of depuration and tissue dissection on the stable isotope enrichment of common estuarine invertebrates and fish were examined. Homogenized tissues of non-depurated whole organisms were compared to dissected muscle tissue or depurated whole organisms. A 24 h depuration did not change the mean δ15N and δ13C values for most species examined. Additionally, as expected, significant differences in carbon and nitrogen signatures were found when muscle tissues were compared to whole organisms. However, differences were small enough that food source as inferred by δ13C or trophic level as inferred from δ15N would not be inaccurately represented (differences of <1.9‰ for δ13C and <1.2‰ for δ15N). The results of this study suggest that for these common estuarine fish and macroinvertebrates, stable isotopes ratios of samples can be analyzed without depuration in the same way as samples for contaminant analysis, but differences in tissue types must be taken into account when combining data from different sources.

Effects of chemical management for invasive plants on the performance of Lithobates pipiens tadpoles.

Invasive plants impact amphibians by altering habitat, altering species interactions, and releasing potentially toxic secondary chemicals. Despite being costly and having the potential to affect nontarget wildlife, chemical management is commonly used to control invasive plants. Prior research has indicated that individual effects of invasive plants or herbicides can be harmful to aquatic organisms; however, information is lacking on the combined effect of these factors on amphibians. A laboratory experiment was performed to assess the impact of leachates of the invasive plants Eurasian watermilfoil (Myriophyllum spicatum) and European buckthorn (Rhamnus cathartica), the herbicide Renovate® 3 (triclopyr [3, 5, 6-trichloro-2-pyridinyloxyacetic acid]), and the combined effects of each plant leachate and the herbicide on the growth, morphology, and survival of northern leopard frog (Lithobates pipiens) tadpoles. No effects of treatment on survival were observed. Tadpole exposure to M. spicatum reduced body mass by 17%, exposure to R. cathartica increased body mass by 36%, and exposure to R. cathartica + low herbicide increased body mass by 38% (although only early in the experiment). Exposure to Renovate 3 induced a 16% and 29% decrease in tadpole size in lower (0.22 mg triclopyr active ingredient [a.i.]/L) and higher (0.92 mg triclopyr a.i./L) concentration treatments, respectively. Results from the present study highlight the importance of considering both individual and combined effects of invasive plants and herbicides because they may have different outcomes for tadpole growth and development. Environ Toxicol Chem 2017;36:2958-2964. © 2017 SETAC.