Threatened species are susceptible to irreversible population decline caused by adverse sub-lethal effects of chemical contaminant exposure. It is therefore vital to develop the necessary tools to predict and detect these effects as early as possible. Biomarkers of contaminant exposure and effect are widely applied to this end, and a significant amount of research has focused on development and validation of sensitive and diagnostic biomarkers. However, progress in the use biomarkers that can be measured using non-destructive techniques has been relatively slow and there are still many difficulties to overcome in the development of sound methods. This paper systematically quantifies and reviews studies that have aimed to develop or validate non-destructive biomarkers in wildlife, and provides an analysis of the successes of these methods based on the invasiveness of the methods, the potential for universal application, cost, and the potential for new biomarker discovery. These data are then used to infer what methods and approaches appear the most effective for successful development of non-destructive biomarkers of contaminant exposure in wildlife. This review highlights that research on non-destructive biomarkers in wildlife is severely lacking, and suggests further exploration of in vitro methods in future studies.

The Mediterranean Sea has been described as one of the most affected areas by marine litter in the world. Although effects on organisms from marine plastic litter ingestion have been investigated in several oceanic areas, there is still a lack of information from the Mediterranean Sea. The main objectives of this paper are to review current knowledge on the impact of marine litter on Mediterranean biodiversity, to define selection criteria for choosing marine organisms suitable for use as bioindicator species, and to propose a methodological approach to assessing the harm related to marine litter ingestion in several Mediterranean habitats and sub-regions. A new integrated monitoring tool that would provide the information necessary to design and implement future mitigation actions in the Mediterranean basin is proposed.According to bibliographic research and statistical analysis on current knowledge of marine litter ingestion, the area of the Mediterranean most studied, in terms of number of species and papers in the Mediterranean Sea is the western sub-area as well as demersal (32.9%) and pelagic (27.7%) amongst habitats.Applying ecological and biological criteria to the most threatened species obtained by statistical analysis, bioindicator species for different habitats and monitoring scale were selected. A threefold approach, simultaneously measuring the presence and effects of plastic, can provide the actual harm and sub-lethal effects to organisms caused by marine litter ingestion. The research revealed gaps in knowledge, and this paper suggests measures to close the gap. This and the selection of appropriate bioindicator species would represent a step forward for marine litter risk assessment, and the implementation of future actions and mitigation measures for specific Mediterranean areas, habitats and species affected by marine litter ingestion.

Wildlife lead exposure is an increasing conservation threat that is being widely investigated. However, for some areas of the world (e.g., South America) and certain species, research on this subject is still scarce or only local information is available. We analyzed the extent and intensity of lead exposure for a widely distributed threatened species, the Andean Condor (Vultur gryphus). We conducted the study at two different scales: 1) sampling of birds received for rehabilitation or necropsy in Argentina, and 2) bibliographic review and extensive survey considering exposure event for the species' distribution in South America. Wild condors from Argentina (n = 76) presented high lead levels consistent with both recent and previous exposure (up to 104 μg/dL blood level, mean 15.47 ± 21.21 μg/dL and up to 148.20 ppm bone level, mean 23.08 ± 31.39 ppm). In contrast, captive bred individuals -not exposed to lead contamination- had much lower lead levels (mean blood level 5.63 ± 3.08 μg/dL, and mean bone level 2.76 ± 3.06 ppm). Condors were exposed to lead throughout their entire range in continental Argentina, which represents almost sixty percent (>4000 km) of their geographical distribution. We also present evidence of lead exposure events in Chile, Ecuador, and Peru. Lead poisoning is a widespread major conservation threat for the Andean Condor, and probably other sympatric carnivores from South America. The high number and wide range of Andean Condors with lead values complement the results for the California Condor and other scavengers in North America suggesting lead poisoning is a continental threat. Urgent actions are needed to reduce this poison in the wild.

Threatened or endangered reptiles, such as sea turtles, are generally understudied within the field of wildlife toxicology, with even fewer studies on how contaminants affect threatened species reproduction. This paper aimed to better inform threatened species conservation by systematically and quantitatively reviewing available research on the reproductive toxicology of all reptiles, threatened and non-threatened. This review found 178 studies that matched our search criteria. These papers were categorized into location conducted, taxa studied, species studied, effects found, and chemicals investigated. The most studied taxa were turtles (n = 87 studies, 49%), alligators/crocodiles (n = 54, 30%), and lizards (n = 37, 21%). Maternal transfer, sex steroid alterations, sex reversal, altered sexual development, developmental abnormalities, and egg contamination were the most common effects found across all reptile taxa, providing guidance for avenues of research into threatened species. Maternal transfer of contaminants was found across all taxa, and taking into account the foraging behavior of sea turtles, could help elucidate differences in maternal transfer seen at nesting beaches. Sex steroid alterations were a common effect found with contaminant exposure, indicating the potential to use sex steroids as biomarkers along with traditional biomarkers such as vitellogenin. Sex reversal through chemical exposure was commonly found among species that exhibit temperature dependent sex determination, indicating the potential for both environmental pollution and climate change to disrupt population dynamics of many reptile species, including sea turtles. Few studies used in vitro, DNA, or molecular methodologies, indicating the need for more research using high-throughput, non-invasive, and cost-effective tools for threatened species research. The prevalence of developmental abnormalities and altered sexual development and function indicates the need to further study how anthropogenic pollutants affect reproductive output in threatened reptiles.

To assess ozone sensitivity of subtropical broadleaved tree species and explore possible underlying mechanisms, six evergreen and two deciduous native species were exposed to either charcoal-filtered air or elevated O₃ (E-O₃, ∼150ppb) for one growing season. Initial visible symptoms in deciduous species appeared much earlier than those in evergreen species. The species which first showed visible symptoms also had the largest reductions in biomass. E-O₃ induced significant decreases in photosynthesis rate, chlorophyll content and antioxidant capacity but a significant increase in malondialdehyde content in two deciduous species and two evergreen species (Cinnamomum camphora and Cyclobalanopsis glauca). Except C. glauca, however, E-O₃ had no significant effects on stomatal conductance (gₛ), total phenols and ascorbate contents. Difference in O₃ sensitivity among all species was strongly attributed to specific leaf mass rather than gₛ. It suggests that some subtropical tree species will be threatened by rising O₃ concentrations in the near future.

Here we review mechanisms and factors influencing contaminant exposure among terrestrial vertebrate wildlife. There exists a complex mixture of biotic and abiotic factors that dictate potential for contaminant exposure among terrestrial and semi-terrestrial vertebrates. Chemical fate and transport in the environment determine contaminant bioaccessibility. Species-specific natural history characteristics and behavioral traits then play significant roles in the likelihood that exposure pathways, from source to receptor, are complete. Detailed knowledge of natural history traits of receptors considered in conjunction with the knowledge of contaminant behavior and distribution on a site are critical when assessing and quantifying exposure. We review limitations in our understanding of elements of exposure and the unique aspects of exposure associated with terrestrial and semi-terrestrial taxa. We provide insight on taxa-specific traits that contribute, or limit exposure to, transport phenomenon that influence exposure throughout terrestrial systems, novel contaminants, bioavailability, exposure data analysis, and uncertainty associated with exposure in wildlife risk assessments. Lastly, we identify areas related to exposure among terrestrial and semi-terrestrial organisms that warrant additional research. Both biotic and abiotic factors determine chemical exposure for terrestrial vertebrates.

Microplastics (MPs) have been reported to threaten a wide variety of terrestrial, marine, and freshwater organisms. However, knowledge about the effects of MPs on anuran amphibians, one of the most threatened taxa worldwide, is still limited. To assess the effects of MPs on the growth and survival of the Italian agile frog (Rana latastei) and green toad (Bufotes balearicus), we exposed tadpoles to three different concentrations (1, 7, and 50 mg L⁻¹) of an environmental relevant mixture of microplastics (HPDE, PVC, PS and PES), recording data on their activity level, weight and mortality rates. While the effects of MPs on green toad tadpoles were negligible, Italian agile frog tadpoles were severely affected both in terms of growth and activity level, with high mortality rates even at the lowest MP density (1 mg L⁻¹). Our results suggest that MP contamination of freshwater habitats may contribute to the ongoing decline of anuran amphibians.

The use of bird feathers to assess environmental contamination has steadily increased in ecotoxicological monitoring programs over the past decade. The Olrog’s Gull (Larus atlanticus) is a species endemic to the Atlantic coast of southern South America, constituting one of the three threatened gull species listed in the entire American continent. The aim of this study was to assess the exposure to Persistent Organic Pollutants (POPs) and chlorpyrifos in the Near Threatened Olrog’s Gull through the analysis of body feathers sampled at the Mar Chiquita coastal lagoon, the main wintering area of the species in Argentina, controlling for sex and age class. Chlorpyrifos showed the highest concentrations among all contaminants and groups of individuals (X¯ = 263 ng g⁻¹), while among POPs the concentration of organochlorine pesticides was higher than polychlorinated biphenyls and polybrominated diphenyl ethers, likely indicating the current use of these agricultural contaminant in the region. The highest values of total POP concentrations (males X¯ = 280 ng g⁻¹, females X¯ = 301 ng g⁻¹) were found in juvenile gulls, likely as a consequence of the incorporation of pollutants during the breeding season. Subadult and adult birds showed difference between sexes in the concentration of contaminants, with higher levels in males than females. The results highlight the need to include birds of different sex and age classes in order to better understand the variation in pollutants loads. The present study provides relevant information to improve the conservation status of the Olrog’s Gull and new insights about the environmental health of the Mar Chiquita coastal lagoon, Argentina, a MAB-UNESCO World Biosphere Reserve. However, there is a continued need for long-term monitoring programs focusing on this threatened species to understand the effects of pollutants on its population.

Stabled livestock reared in housed conditions are often subjected to intensive treatments with veterinary drug, which residues may be present in livestock meat ingested by scavengers, but nothing is known about their presence in eggs of wild birds and their potential detrimental effects on breeding success. We searched for residues of veterinary drugs and other toxicants in infertile and embryonated unhatched eggs of griffon vultures (Gyps fulvus) and red kites (Milvus milvus), two threatened avian scavengers. Quinolones (ciprofloxacin and enrofloxacin) were found in most unhatched eggs of both scavenger species clearly associated with severe alterations in the development of embryo cartilage and bones that could preclude embryo movements and subsequently normal development, pre-hatch position and successful hatching. The detrimental effects on developing eggs of veterinary drugs from livestock operations may help to explain reduced breeding success of avian scavengers. Fluoroquinolones used in livestock farming and found in eggs of avian scavenger caused severe alterations in embryo cartilage and bone development.

Methods to assess the effects of contaminants on marine organisms typically involve euthanasia to obtain samples, but less invasive techniques may be more appropriate for working with threatened species. In this study, were assessed the biological responses of crabs exposed to microplastics and contaminants of emerging concern. Biochemical and cellular effects (lipid peroxidation, DNA damage, cholinesterase activity, and lysosomal membrane stability) in hemolymph were analyzed in a kinetic study, at 3 and 7 days, in U. cordatus exposed to microplastics spiked with Triclosan (TCS) or 17α-Ethynylestradiol (EE2). The results showed that the contaminants were produced toxic effects in the crabs exposed either to the microplastics alone (oxidative stress, genotoxicity, and neurotoxicity), or to microplastics with TCS or EE2 adsorbed (neurotoxic and cytotoxic). The present study showed the responsiveness of non-lethal analyzes to understanding the biological effects of combined exposure to microplastics and chemical pollution.