The recognition that invasive alien species (IAS) are among the greatest threats to biodiversity has stimulated a growing interest in their impacts on native amphibians. Here we describe the multifaceted consequences of biological invasions on native amphibians and identify potential mechanisms and strategies that could better enable the long-term persistence of native species. IAS can influence amphibian fitness, population size, and community structure via multiple pathways and can exert major, direct impacts through predation, competition, and hybridization. The consequences of indirect impacts, too, such as habitat alteration and the spread of emerging diseases, can be particularly severe in native populations. Native amphibians may respond to IAS by modulating aspects of their behavior, morphology, or life history. Nevertheless, it is still unclear the extent to which phenotypic plasticity and rapid evolution may help native species withstand the impacts of IAS in invaded communities. Practical management strategies focused on prevention, monitoring, and early control are the most effective approaches to allay the impacts of IAS and should be prioritized in proactive conservation plans. Eradications of IAS and mitigation approaches, should IAS become established, are feasible and can greatly improve the status of native populations.

In the last decade, eDNA and metabarcoding have opened new avenues to biodiversity studies; amphibians and reptiles are animals for which these new approaches have allowed great leaps forward. Here we review different approaches through which eDNA can be used to study amphibians, reptiles and many more organisms. eDNA is often used to evaluate the presence of target species in freshwaters; it has been particularly useful to detect invasive alien amphibians and secretive or rare species, but the metabarcoding approach is increasingly used as a cost-effective approach to assess entire communities. There is growing evidence that eDNA can be also useful to study terrestrial organisms, to evaluate the relative abundance of species, and to detect reptiles. Metabarcoding has also revolutionized studies on the microbiome associated to skin and gut, clarifying the complex relationships between pathogens, microbial diversity and environmental variation. We also identify additional aspects that have received limited attention so far, but can greatly benefit from innovative applications of eDNA, such as the study of past biodiversity, diet analysis and the reconstruction of trophic interactions. Despite impressive potential, eDNA and metabarcoding also bear substantial technical and analytical complexity; we identify laboratory and analytical strategies that can improve the robustness of results. Collaboration among field biologists, ecologist, molecular biologists, and bioinformaticians is allowing fast technical and conceptual advances; multidisciplinary studies involving eDNA analyses will greatly improve our understanding of the complex relationships between organisms, and our effectiveness in assessing and preventing the impact of human activities.

The 15th UN Convention on Biological Diversity (CBD) (COP15) will be held in Kunming, China in October 2021. Historically, CBDs and other multilateral treaties have either alluded to or entirely overlooked the subterranean biome. A multilateral effort to robustly examine, monitor, and incorporate the subterranean biome into future conservation targets will enable the CBD to further improve the ecological effectiveness of protected areas by including groundwater resources, subterranean ecosystem services, and the profoundly endemic subsurface biodiversity. To this end, we proffer a conservation roadmap that embodies five conceptual areas: (1) science gaps and data management needs; (2) anthropogenic stressors; (3) socioeconomic analysis and conflict resolution; (4) environmental education; and (5) national policies and multilateral agreements.