Next-generation monitoring of aquatic biodiversity using environmental DNA metabarcoding.

Global biodiversity in freshwater and the oceans is declining at high rates. Reliable tools for assessing and monitoring aquatic biodiversity, especially for rare and secretive species, are important for efficient and timely management. Recent advances in DNA sequencing have provided a new tool for species detection from DNA present in the environment. In this study, we tested whether an environmental DNA (eDNA) metabarcoding approach, using water samples, can be used for addressing significant questions in ecology and conservation. Two key aquatic vertebrate groups were targeted: amphibians and bony fish. The reliability of this method was cautiously validated in silico, in vitro and in situ. When compared with traditional surveys or historical data, eDNA metabarcoding showed a much better detection probability overall. For amphibians, the detection probability with eDNA metabarcoding was 0.97 (CI = 0.90-0.99) vs. 0.58 (CI = 0.50-0.63) for traditional surveys. For fish, in 89% of the studied sites, the number of taxa detected using the eDNA metabarcoding approach was higher or identical to the number detected using traditional methods. We argue that the proposed DNA-based approach has the potential to become the next-generation tool for ecological studies and standardized biodiversity monitoring in a wide range of aquatic ecosystems.

An extensive database on the traits and occurrences of amphibian species in Turkey.

Amphibians are the most endangered taxa among vertebrates, and they face many threats during their complex life cycles. The species' life history traits and occurrence database help understand species responses against ecological factors. Consequently, the species-level-trait database has gained more prominence in recent years as a useful tool for understanding the dimensions of communities, assembly processes of communities, and conserving biodiversity at the ecosystem level against environmental changes. However, in Turkey, there are deficiencies in the knowledge of the ecological traits of amphibians compared to other vertebrate taxa, as most studies have focused on their distribution or taxonomic status. Consequently, there is a need to create such a database for future research on all known extant amphibians in Turkey. We compiled a species-level data set of species traits and occurrences for all amphibians in Turkey using 436 literature sources. We completed 36 trait categories with 5611 occurrence data for 37 amphibian species in Turkey. This study provides an open, useful, and comprehensive database for macroecological and conservation studies on amphibians in Turkey.

Pesticide concentrations in a threatened freshwater turtle (Emys orbicularis): Seasonal and annual variation in the Camargue wetland, France.

Wetlands are among the most threatened ecosystems on the planet and pollution is a major factor causing the decline of wetland biodiversity. Despite the increasing use of pesticides, their fate and effects on freshwater reptiles remain largely unknown. We studied the European pond turtle (Emys orbicularis), a long-lived species at risk with a high exposure potential to pesticides. Between 2018 and 2020, we measured 29 pesticides and metabolites in 408 blood samples of turtles from two populations in the Camargue wetland (France). We were able to quantify 24 compounds and at least one pesticide or one degradation product in 62.5% of samples. Pesticide occurrences and concentrations were low, except for a herbicide widely used in rice cultivation and locally detected in water: bentazone that reached high blood concentrations in E. orbicularis. The occurrence and the concentration of pesticides in E. orbicularis blood depended mainly on the site and the sampling date in relation to pesticide application. Individual characteristics (sex, age, body condition) did not explain the occurrence or the concentration of pesticides found in turtle blood. Assessing the exposure of aquatic wildlife to a cocktail of currently-used pesticides is a first and crucial step before studying their effects at the individual and population levels.

Diverse aging rates in ectothermic tetrapods provide insights for the evolution of aging and longevity.

Comparative studies of mortality in the wild are necessary to understand the evolution of aging; yet, ectothermic tetrapods are underrepresented in this comparative landscape, despite their suitability for testing evolutionary hypotheses. We present a study of aging rates and longevity across wild tetrapod ectotherms, using data from 107 populations (77 species) of nonavian reptiles and amphibians. We test hypotheses of how thermoregulatory mode, environmental temperature, protective phenotypes, and pace of life history contribute to demographic aging. Controlling for phylogeny and body size, ectotherms display a higher diversity of aging rates compared with endotherms and include phylogenetically widespread evidence of negligible aging. Protective phenotypes and life-history strategies further explain macroevolutionary patterns of aging. Analyzing ectothermic tetrapods in a comparative context enhances our understanding of the evolution of aging.

Determinants of Legacy Persistent Organic Pollutant Levels in the European Pond Turtle (Emys orbicularis) in the Camargue Wetland, France.

Many banned persistent organic pollutants (POPs) remain for decades in the aquatic environment and can have harmful effects on long-lived predators because of their high bioaccumulation and biomagnification potentials. We investigated the occurrence and levels of 18 polychlorinated biphenyls (PCBs) and 16 organochlorine pesticides in European pond turtles (n = 174) from April to July 2018 in the Camargue wetland, France. Although the Camargue was highly contaminated in previous decades, plasma occurrence and levels of POPs were very low: we were able to quantify only 3 of the 34 compounds we analyzed in >10% of the turtles. The burdens from POPs did not differ between males and females and were uncorrelated with sampling date and body mass. We observed differences in POP burdens between turtles from the 2 sampling sites. One possible explanation is that the sampling sites were in different agricultural hydraulic systems: plasma occurrence and levels were higher for PCB-52 and hexachlorobenzene in turtles captured in drainage channels and for PCB-153 at the site that receives irrigation. Finally, the occurrence and levels of PCB-153 in turtles increased with age, likely because of bioaccumulation and much higher exposure 20 to 30 yr ago than now. Environ Toxicol Chem 2021;40:2261-2268. © 2021 SETAC.

Assessing spatial and temporal population dynamics of cryptic species: an example with the European pond turtle.

Within the current context of biodiversity loss a number of biodiversity indicators have been developed to help measure the state of nature and how it is changing. However, most indicators are derived from bird populations. Reptiles and amphibians could be useful ecosystem indicators, but this requires obtaining precise and unbiased population parameters. This is a particularly challenging task for these two groups of species, because individuals are extremely difficult to detect for various reasons. We illustrate the use of a sampling and analytical method that explicitly takes into account imperfect detection to assess the population dynamics of a reptile species and its temporal and spatial variation. European pond turtles (Emys orbicularis) were sampled at two different locations during a 10-year study. The two sites differed by their water management regimes and number of livestock. At each site and for each sex, the data were modeled using a robust design capture-mark-recapture framework to obtain and compare estimates of survival, temporary emigration, time-specific abundance, density, and population growth rate, while taking into account effects of individual heterogeneity, trap-response, and time on capture probabilities. Temporary emigration was higher in males (0.344 +/- 0.046) (mean +/- SE) than in females (0.228 +/- 0.071), did not differ between sites for each sex, and was mainly Markovian. Apparent adult survival was higher in females (0.966 +/- 0.031) than in males (0.864 +/- 0.024), and was negatively related to the proportion of individuals with repaired shell fractures in the population. Average density was 63.7 +/- 6.6 turtles/km2 but was higher for females (39.2 +/- 14.0 females/km2) than for males (23.5 +/- 5.7 males/km2). The population sizes of males and females decreased where long periods of artificial drought and the highest density of livestock occurred. This research highlights differences in demographic parameters according to sex and site in a turtle species, and it provides evidence that significant human-induced disturbance represents a potential risk to European pond turtle populations. The sampling and analytical approaches illustrated in this study are applicable to many other species of reptiles and amphibians, and estimated population parameters could be combined to produce population indicators useful for conservation and management.

Estimating biodiversity changes in the Camargue wetlands: An expert knowledge approach.

Mediterranean wetlands are critical strongholds for biodiversity and the provision of ecosystem functions and services; yet, they are being severely degraded by a number of socio-economic drivers and pressures, including climate change. Moreover, we still lack comprehensive understanding of the extent to which biodiversity loss in Mediterranean wetlands will accelerate change in ecosystem processes. Here, we evaluate how changes in biodiversity can alter the ecosystem of the Camargue (southern France). We collected data on species presence/absence, trends and abundance over a 40-year period by combining observations from the scholarly literature with insights derived from expert knowledge. In total, we gathered more than 1500 estimates of presence/absence, over 1400 estimates of species abundance, and about 1400 estimates of species trends for eight taxonomic groups, i.e. amphibians, reptiles, breeding birds, fish, mammals, dragonflies (odonates), orthopterans and vascular plants. Furthermore, we used information on recently arrived species and invasive species to identify compositional changes across multiple taxa. Complementing targeted literature searches with expert knowledge allowed filling important gaps regarding the status and trends of biodiversity in the Camargue. Species trend data revealed sharp population declines in amphibians, odonates and orthopterans, while birds and plants experienced an average increase in abundance between the 1970s and the 2010s. The general increasing trends of novel and invasive species is suggested as an explanation for the changing abundance of birds and plants. While the observed declines in certain taxa reflect the relative failure of the protection measures established in the Camargue, the increasing exposure to novel and invasive species reveal major changes in the community structure of the different taxonomic groups. This study is the first attempt to assess changes in biodiversity in the Camargue using an expert knowledge approach, and can help manage the uncertainties and complexities associated with rapid social-ecological change in other Mediterranean wetlands.

Determinants of mercury contamination in viperine snakes, Natrix maura, in Western Europe.

The effects of Hg contamination are presumably widespread across the components of aquatic ecosystems, but investigations have been mainly focused on freshwater fish, because this biota represents a major source of Hg for human populations. Yet, the possible bioaccumulation of Hg on other freshwater meso- and apex-predators (e.g., amphibians, reptiles) has been largely overlooked, especially in Western Europe. In this study, the determinants of Hg concentrations were assessed for the viperine snake (Natrix maura) across 6 populations (>130 individuals sampled in 2016 and 2017) in France and Spain. Specifically, body size, sex, and diet were compared with Hg concentrations measured in ventral scales. Overall, N. maura accumulated Hg in their scales. Sex did not seem to influence Hg concentrations in this species. Significant differences in Hg concentrations were observed between study sites, and these differences were likely to be mediated by site-specific diet. Frog-eating individuals were characterized not only by lower mean values of Hg (0.194±0.018µg·g-1 versus 0.386±0.032µg·g-1 for piscivorous individuals), but also by weaker slopes of the body size-Hg relationship as compared to fish-eating snakes, suggesting strong differences in accumulation rates due to food resources. Importantly, the highest slope of the body size-Hg relationship and the highest values of Hg were found in individuals foraging on trout raised by a fish farm, suggesting that fish farming may contribute to Hg contamination in inland freshwater systems. Finally, our results are compared with data on Hg concentrations in other species of aquatic snakes, in order to provide a comparative point for future studies.