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The use of pesticides is important for growing crops and protecting human health by reducing the prevalence of targeted pest species. However, less attention is given to the potential unintended effects on nontarget species, including taxonomic groups that are of current conservation concern. One issue raised in recent years is the potential for pesticides to become more lethal in the presence of predatory cues, a phenomenon observed thus far only in the laboratory. A second issue is whether pesticides can induce unintended trait changes in nontarget species, particularly trait changes that might mimic adaptive responses to natural environmental stressors. Using outdoor mesocosms, I created simple wetland communities containing leaf litter, algae, zooplankton, and three species of tadpoles (wood frogs [ Rana sylvatica or Lithobates sylvaticus ], leopard frogs [ R. pipiens or L. pipiens ], and American toads [ Bufo americanus or Anaxyrus americanus ]). I exposed the communities to a factorial combination of environmentally relevant herbicide concentrations (0, 1, 2, or 3 mg acid equivalents [a.e.]/L of Roundup Original MAX) crossed with three predator-cue treatments (no predators, adult newts [ Notophthalmus viridescens ], or larval dragonflies [ Anax junius ]). Without predator cues, mortality rates from Roundup were consistent with past studies. Combined with cues from the most risky predator (i.e., dragonflies), Roundup became less lethal (in direct contrast to past laboratory studies). This reduction in mortality was likely caused by the herbicide stratifying in the water column and predator cues scaring the tadpoles down to the benthos where herbicide concentrations were lower. Even more striking was the discovery that Roundup induced morphological changes in the tadpoles. In wood frog and leopard frog tadpoles, Roundup induced relatively deeper tails in the same direction and of the same magnitude as the adaptive changes induced by dragonfly cues. To my knowledge, this is the first study to show that a pesticide can induce morphological changes in a vertebrate. Moreover, the data suggest that the herbicide might be activating the tadpoles' developmental pathways used for antipredator responses. Collectively, these discoveries suggest that the world's most widely applied herbicide may have much further-reaching effects on nontarget species than previous considered.

Carryover effects occur when experiences early in life affect an individual's performance at a later stage. Many studies have shown carryover effects to be important for future performance. However, it is currently unclear whether variation in later environments could overwhelm factors from an earlier life stage. We were interested in whether similar patterns would emerge under the same experimental design with similar taxa. To examine this, we implemented a four-way factorial experimental design with different forestry practices on three species of anurans (each examined in different years) in the aquatic larval environment and terrestrial juvenile environment in outdoor mesocosms in central Missouri, USA. Using Cormack-Jolly-Seber mark-recapture models implemented in program MARK, we investigated whether one environment or both environments best predicted terrestrial juvenile survival. We found only limited evidence of carryover effects for one of three species in one time period. These were the effects of time to metamorphosis and body condition at metamorphosis predicting leopard frog ( Lithobates sphenocephalus ) survival. However, both effects were counterintuitive and/or very weak. For wood frogs ( L. sylvaticus ), all of the variables predicting survival had confidence intervals that included zero, but very low survival may have limited our ability to estimate parameters. The terrestrial environment was important for predicting survival in both American toads ( Anaxyrus americanus ) and southern leopard frogs. The partial harvest forest tended to increase survival relative to control forest and early-successional forest in American toads. Alternately, early-successional forest with downed wood removed increased survival for leopard frogs, but this treatment was no different from control forest for American toads. Previous studies have shown negative effects of recent clearcuts on terrestrial amphibians. It appears that vegetative regrowth after just a few years can mitigate these initial negative effects. Our study shows that variation in later environments probably can overwhelm variation from earlier environments. However, previous evidence of carryover effects suggests that more research is needed to predict when carryover effects are likely to occur.

Ranaviruses have caused die-offs of amphibians across the globe. In North America, these pathogens cause more amphibian mortality events than any other pathogen. Field observations suggest that ranavirus epizootics in amphibian communities are common during metamorphosis, presumably due to changes in immune function. However, few controlled studies have compared the relative susceptibility of amphibians to ranaviruses across life stages. Our objectives were to measure differences in mortality and infection prevalence following exposure to ranavirus at four developmental stages and determine whether the differences were consistent among seven anuran species. Based on previous studies, we hypothesized that susceptibility to ranavirus would be greatest at metamorphosis. Our results did not support this hypothesis, as four of the species were most susceptible to ranavirus during the larval or hatchling stages. The embryo stage had the lowest susceptibility among species probably due to the protective membranous layers of the egg. Our results indicate that generalizations should be made cautiously about patterns of susceptibility to ranaviruses among amphibian developmental stages and species. Further, if early developmental stages of amphibians are susceptible to ranaviruses, the impact of ranavirus epizootic events may be greater than realized due to the greater difficulty of detecting morbid hatchlings and larvae compared to metamorphs.

The reintroduction of a species into its historic range is a critical component of conservation programmes designed to restore extirpated metapopulations. However, many reintroduction efforts fail, and the lack of rigorous monitoring programmes and statistical models have prevented a general understanding of the factors affecting metapopulation viability following reintroduction. Spatially explicit metapopulation theory provides the basis for understanding the dynamics of fragmented populations linked by dispersal, but the theory has rarely been used to guide reintroduction programmes because most spatial metapopulation models require presence–absence data from every site in the network, and they do not allow for observation error such as imperfect detection. We develop a spatial occupancy model that relaxes these restrictive assumptions and allows for inference about metapopulation extinction risk and connectivity. We demonstrate the utility of the model using six years of data on the Chiricahua leopard frog Lithobates chiricahuensis, a threatened desert‐breeding amphibian that was reintroduced to a network of sites in Arizona USA in 2003. Our results indicate that the model can generate precise predictions of extinction risk and produce connectivity maps that can guide conservation efforts following reintroduction. In the case of L. chiricahuensis, many sites were functionally isolated, and 82% of sites were characterized by intermittent water availability and high local extinction probabilities (0·84, 95% CI: 0·64–0·99). However, under the current hydrological conditions and spatial arrangement of sites, the risk of metapopulation extinction is estimated to be <3% over a 50‐year time horizon. Low metapopulation extinction risk appears to result from the high dispersal capability of the species, the high density of sites in the region and the existence of predator‐free permanent wetlands with low local extinction probabilities. Should management be required, extinction risk can be reduced by either increasing the hydroperiod of existing sites or by creating new sites to increase connectivity. Synthesis and applications. This work demonstrates how spatio‐temporal statistical models based on ecological theory can be applied to forecast the outcomes of conservation actions such as reintroduction. Our spatial occupancy model should be particularly useful when management agencies lack the funds to collect intensive individual‐level data.

Understanding how natural and anthropogenic processes affect population dynamics of species with patchy distributions is critical to predicting their responses to environmental changes. Despite considerable evidence that demographic rates and dispersal patterns vary temporally in response to an array of biotic and abiotic processes, few applications of metapopulation theory have sought to explore factors that explain spatiotemporal variation in extinction or colonization rates. To facilitate exploring these factors, we extended a spatially explicit model of metapopulation dynamics to create a framework that requires only binary presence–absence data, makes few assumptions about the dispersal process, and accounts for imperfect detection. We apply this framework to 22 yr of biannual survey data for lowland leopard frogs, Lithobates yavapaiensis, an amphibian that inhabits arid stream systems in the southwestern United States and northern Mexico. Our results highlight the importance of accounting for factors that govern temporal variation in transition probabilities, as both extinction and colonization rates varied with hydrologic conditions. Specifically, local extinctions were more frequent during drought periods, particularly at sites without reliable surface water. Colonization rates increased when larval and dispersal periods were wetter than normal, which increased the probability that potential emigrants metamorphosed and reached neighboring sites. Extirpation of frogs from all sites in one watershed during a period of severe drought demonstrated the influence of site-level features, as frogs persisted only in areas where most sites held water consistently and where the amount of sediment deposited from high-elevation wildfires was low. Application of our model provided novel insights into how climate-related processes affected the distribution and population dynamics of an arid-land amphibian. The approach we describe has application to a wide array of species that inhabit patchy environments, can improve our understanding of factors that govern metapopulation dynamics, and can inform strategies for conservation of imperiled species.

Conservation of at-risk species is aided by reliable forecasts of the consequences of environmental change and management actions on population viability. Forecasts from conventional population viability analysis (PVA) are made using a two-step procedure in which parameters are estimated, or elicited from expert opinion, and then plugged into a stochastic population model without accounting for parameter uncertainty. Recently developed statistical PVAs differ because forecasts are made conditional on models fitted to empirical data. The statistical forecasting approach allows for uncertainty about parameters, but it has rarely been applied in metapopulation contexts where spatially explicit inference is needed about colonization and extinction dynamics and other forms of stochasticity that influence metapopulation viability. We conducted a statistical metapopulation viability analysis (MPVA) using 11 yr of data on the federally threatened Chiricahua leopard frog (Lithobates chiricahuensis) to forecast responses to landscape heterogeneity, drought, environmental stochasticity, and management. We evaluated several future environmental scenarios and pond restoration options designed to reduce extinction risk. Forecasts over a 50-yr time horizon indicated that metapopulation extinction risk was <4% for all scenarios, but uncertainty was high. Without pond restoration, extinction risk is forecasted to be 3.9% (95% CI 0–37%) by year 2066. Restoring six ponds by increasing their hydroperiod reduced extinction risk to <1% and greatly reduced uncertainty (95% CI 0–2%). Our results suggest that managers can mitigate the impacts of drought and environmental stochasticity on metapopulation viability by maintaining ponds that hold water throughout the year and keeping them free of invasive predators. Our study illustrates the utility of the spatially explicit statistical forecasting approach to MPVA in conservation planning efforts.

The amphibian fungal pathogenBatrachochytrium dendrobatidis(Bd) has received considerable attention due to its role in amphibian population declines worldwide. Although many amphibian species appear to be affected by Bd, there is little information on species-specific differences in susceptibility to this pathogen. We used a comparative experimental approach to examine Bd susceptibility in 6 amphibian species from the United States. We exposed postmetamorphic animals to Bd for 30 days and monitored mortality, feeding rates, and infection levels. In all species tested, Bd-exposed animals had higher rates of mortality than unexposed (control) animals. However, we found differences in mortality rates among species even though the amount of Bd detected on the different species' bodies did not differ. Of the species tested, southern toads (Anaxyrus terrestris) and wood frogs (Lithobates sylvaticus) had the highest rates of Bd-related mortality. Within species, we detected lower levels of Bd on individuals that survived longer and found that the relationship between body size and infection levels differed among species. Our results indicate that, even under identical conditions, amphibian species differ in susceptibility to Bd. This study represents a step toward identifying and understanding species variation in disease susceptibility, which can be used to optimize conservation strategies.Original Abstract: Resumen:El patogeno fungico de anfibiosBatrachochytrium dendrobatidis (Bd) ha recibido considerable atencion debido a su papel en la declinacion de poblaciones de anfibios en todo el mundo. Aunque parece que muchas especies de anfibios son afectadas por Bd, existe poca informacion sobre diferencias especificas en la susceptibilidad a este patogeno. Utilizamos un metodo experimental comparativo para examinar la susceptibilidad a Bd en 6 especies anfibios de los Estados Unidos. Expusimos a animales postmetamorficos a Bd durante 30 dias y monitoreamos las tasas de mortalidad y de alimentacion, asi como los niveles de infeccion. En todas las especies probadas, los animales expuestos a Bd tuvieron mayores tasas de mortalidad entre especies aunque la cantidad de Bd detectada sobre los cuerpos de las diferentes especies no difirio. De las especies probadas,Anaxyrus terrestrisyLithobates sylvaticustuvieron las mayores tasas de mortalidad relacionada con Bd. Entre especies, detectamos niveles menores sobre indviduos que sobrevivieron mas tiempo y encontramos que la relacion entre el tamano del cuerpo y los niveles de infeccion difirio entre especies. Nuestros resultados indican que, aunque bajo condiciones identicas, las especies de anfibios difieren en susceptibilidad a Bd y representan un paso hacia la identificacion y comprension de la variacion entre especies de la susceptibilidad a enfermedades, que pueden ser utilizados para optimizar estrategias de conservacion.

Aim Ecological niche models (ENMs) are widely used to address urgent real‐world problems such as climate change effects or invasive species; however, the generality of models when projected through space and/or time, that is transferability, remains a key challenge. Here, we explored the effects of complex predictors and feature selection on ENM transferability in a widely employed algorithm, Maxent, using five globally invasive freshwater species as case studies. Location Global. Methods We modelled the global distributions of five notorious freshwater invasive species (African sharptooth catfish Clarias gariepinus, Mozambique tilapia Oreochromis mossambicus, American bullfrog Lithobates catesbeianus, red swamp crayfish Procambarus clarkii, and Australian redclaw crayfish Cherax quadricarinatus), using three predictor datasets of varying complexities derived from two commonly used climatic data sources (WorldClim and IPCC) and three methods of model tuning that differentially incorporated feature selection. Spatially explicit transferability assessments were then conducted using a suite of evaluation metrics previously used to quantify Maxent model performance. Results We show that in the absence of detailed biological knowledge of focal species, simpler predictor datasets produce models that are more accurate than those calibrated using comprehensive “bioclimatic” datasets. Additionally, we find that tuning models for both optimal regularization parameters as well as feature‐class combinations led to the greatest increases in transferability and geographic niche conservatism. Results indicate a tenuous link between model transferability and Akaike's information criterion corrected for small sample sizes (AICc), suggesting that the indiscriminate use of AICc as an estimate of model parsimony may lead to erratic model performance. Main conclusions Our findings demonstrate that methodological considerations can drastically affect the reliability of spatial and possibly temporal projections, which has severe implications when ENMs are used to infer species’ niches, and quantify ecological or evolutionary change across impacted landscapes.

The effects of allicin and antioxidant of bamboo leaves (AOB) on the quality of bullfrogs (Lithobates catesbeiana) during refrigerated storage (4 °C) were investigated. The quality changes in samples treated with deionized water (CK), allicin solution (All), antioxidant of bamboo leaves (AOB), and allicin solution combined with AOB solution (AA) in microbiological, physicochemical, and sensory evaluation were analyzed, respectively. The results demonstrated that combination treatment inhibited the increase in total viable counts, delayed the decrease in amino acid content, and retarded the sensory deterioration. Preservative treatment has an inhibitory effect on the early storage of PBC, which can reduce PBC by about 1.0 log CFU/g. The reduction in thiobarbituric acid (TBA) content and total volatile basic nitrogen (TVB-N) content indicated that combination treatment could better restrain the lipid oxidation and degradation of protein than the CK group and single-treatment group. In addition, the TVB-N content in the AA group still did not exceed the threshold on the 14th day. As a consequence, combination treatment prolonged the shelf life of bullfrogs for another six days. Therefore, allicin and AOB with excellent antioxidant and antimicrobial activity could be an effective approach to delay the biochemical reaction of refrigerated bullfrogs. This study has provided a potential approach for increasing the shelf life of bullfrogs and preserving their quality during refrigerated storage.

The amphibian response to climate change has been generally described as phenological shifts toward earlier breeding periods, with animals exhibiting smaller body sizes and male frogs producing breeding calls higher in pitch and shorter in duration. However, with >8000 species of amphibians now described, and the effects of climate change intensifying, the amphibian response to variable climates is likely to be broader and more nuanced than scientists have so far observed. For example, our previous work described a dramatic acceleration in the breeding season from spring to the previous fall, associated with warming, in Southern Leopard Frogs (Lithohates sphenocephalus); a correlation between precipitation levels and body mass indices in Crawfish Frogs (Lithohates areolatus); and the expression of a variety of daily activity patterns dependent on seasonal variations in temperature and humidity in crawfish Frogs. Here, we add to this literature by documenting a shift in wetland breeding habitats from seasonal/semipermanent wetlands to permanent wetlands in response to decadal-long hydrologic cycles by Northern Leopard Frogs (Lithohates pipiens) and Eastern Tiger Salamanders (Amhystoma tigrinum). Three implications follow from this result. First, when challenged by climate variability, conserving amphibian diversity requires preserving wetland diversity. Second, amphibian occupancy of any particular wetland basin shifts over time, complicating conclusions drawn from short-term survey data. Third, the wetland shifts we report in response to natural climate variability might provide amphibians the flexibility to successfully respond to future climate challenges. Our insights herein derive from field studies, and we worry that the well-documented trend toward de-emphasizing fieldwork will limit scientists' ability to accurately assess threats to species from climate challenges. As the legendary writer Jim Harrison retorted when asked why he had never accepted any of the comfortable academic jobs he had been offered, \"Somebody's got to stay outside.\"