Systematic assessments of species extinction risk at regular intervals are necessary for informing conservation action^1,2. Ongoing developments in taxonomy, threatening processes and research further underscore the need for reassessment^3,4. Here we report the findings of the second Global Amphibian Assessment, evaluating 8,011 species for the International Union for Conservation of Nature Red List of Threatened Species. We find that amphibians are the most threatened vertebrate class (40.7% of species are globally threatened). The updated Red List Index shows that the status of amphibians is deteriorating globally, particularly for salamanders and in the Neotropics. Disease and habitat loss drove 91% of status deteriorations between 1980 and 2004. Ongoing and projected climate change effects are now of increasing concern, driving 39% of status deteriorations since 2004, followed by habitat loss (37%). Although signs of species recoveries incentivize immediate conservation action, scaled-up investment is urgently needed to reverse the current trends.

The current study was focused on documentation of amphibian assemblage in North Punjab and Islamabad Capital Territory, Pakistan, by using mitochondrial gene sequences of 16S rRNA. Our study entailed 37% of the known amphibian species of the country. We provided a phylogenetic analysis based on 74 newly generated mitochondrial 16S rRNAs from nine species of genus Microlyla, Duttaphrynus, Allopaa, Nanorana, Sphaerotheca, Minervarya, Hoplobatrachus, and Euphlyctis. We employed the maximum‐likelihood inference and Bayesian analysis to assess the taxonomic status of the samples obtained from Pakistan, with respect to other congeneric species from surrounding regions. Our findings confirmed the taxonomic status of South Asian anuran species Duttaphrynus stomaticus, Duttaphrynus melanostictus, Microhyla nilphamariensis, Allopaa hazarensis, Nanorana vicina, Sphaerotheca maskeyi (synonym: S. pashchima), Minervarya pierrei, Hoplobatrachus tigerinus, and Euphlyctis kalasgramensis in Pakistan. We have reported new country records of genus Minervarya (M. pierrei). Minervarya pierrei was previously misidentified as Fejervarya limnocharis, due to dearth of genetic information. We provided the first genetic records of our endemic species N. vicina. The results revealed the taxonomic placement of N. vicina with respect to its congeners and validated the taxonomic status of N. vicina from its type locality (Murree) for the first time. The findings of the present study also indicated the paraphyletic relationship of A.‐ hazarensis with Nanorana species. So, based on our phylogenetic inferences, morphological characters, and habitat preferences, validity of generic status of A. hazarensis is undecided. As our data were not enough to resolve this issue, we suggest sequencing of additional mitochondrial and nuclear genes in the future studies to get a better resolution. We recommend carrying out extensive surveys throughout the country for proper scientific documentation of amphibians of Pakistan. Many new species, some of them might be endemic to Pakistan, are expected to be discovered, and taxonomic status of other species would be resolved.

The Indian pangolin (Manis crassicaudata) has been recently listed by the International Union for Conservation of Nature as an endangered species throughout its range, but in Pakistan it is categorized as vulnerable. Very little is known about the breeding habits of this nocturnal and fossorial animal in the wild. The present paper provides information on breeding ecology of its population in Potohar Plateau in Pakistan. A total of 13 specimens were trapped to record breeding condition of the captured animals. Additionally, a questionnaire survey was conducted in the study area to collect breeding data on the species. Our limited data show a male-to-female ratio of 1.6:1. The local population seems to breed once a year, usually from July to October, with a litter size of one to two. The juvenile pangolins were observed during the months of January, April and December.

Wildlife monitoring programs are instrumental for the assessment of species, habitat status, and for the management of factors affecting them. This is particularly important for species found in freshwater ecosystems, such as amphibians, as they have higher estimated extinction rates than terrestrial species. We developed and validated two species-specific environmental DNA (eDNA) protocols and applied them in the field to detect the Hazara Torrent Frog (Allopaa hazarensis) and Murree Hills Frog (Nanorana vicina). Additionally, we compared eDNA surveys with visual encounter surveys and estimated site occupancy. eDNA surveys resulted in higher occurrence probabilities for bothA. hazarensisandN. vicinathan for visual encounter surveys. Detection probability using eDNA was greater for both species, particularly forA. hazarensis. The top-ranked detection model for visual encounter surveys included effects of both year and temperature on both species, and the top-ranked occupancy model included effects of elevation and year. The top-ranked detection model for eDNA data was the null model, and the top-ranked occupancy model included effects of elevation, year, and wetland type. To our knowledge, this is the first time an eDNA survey has been used to monitor amphibian species in the Himalayan region.

Research on amphibians and their conservation have gained worldwide attention, as the group includes the highest number of threatened and Data Deficient species when compared to other vertebrates. However, amphibians have long been neglected in wildlife conservation, management decisions, policy making, and research agendas in Pakistan. In this paper, an annotated checklist of the 21 amphibian species of Pakistan, a key to their identification, and detailed discussions on variation in species, including the genera Minervarya and Sphaerotheca, are provided. We found a statistically significant difference in the morphometric measurements of males but non-significant difference in the females of the two forms (rusty dorsum and dotted dorsum) of S. maskeyi. Some genera, such as Microhyla, Uperodon, Minervarya, Allopaa, Chrysopaa, Euphlyctis, Nanorana, and Sphaerotheca, in Pakistan are in need of additional data for molecular and morphological comparisons with taxa in other South Asian countries. The predicaments of amphibian research in Pakistan are discussed, gaps identified, and suggestions are made. Although the occurrence of chytrid fungus in Pakistan is predicted of low likelihood, a lack of data merits studying the prevalence of the fungus, particularly in the northern regions of the country which exhibit complex and dynamic ecosystems. It is recommended that systematic and coordinated surveys are conducted throughout the country to build a database of species occurrences and distributions. Additionally, the monitoring of wild populations and threat mitigation, as well as appropriate legislation, are suggested as long-term measures. By adopting an inclusive wildlife conservation approach in Pakistan, amphibians could be integrated into wildlife conservation and management efforts.

The quantification of anuran abundance and habitat provides valuable baseline data for future monitoring in areas of planned or anticipated human activities. We carried out the present study to see if anuran abundance is associated with habitat variables (water quality, gravel size and vegetation) in Rawalpindi–Islamabad Area, Pakistan. We used area-constrained searches and quadrat method to gather data on anuran abundance and vegetation diversity, respectively, from September, 2012 to July, 2013. We recorded 28±4.83 (mean number±SE) individuals of six anuran species from the study area. We recorded Euphlyctis cyanophlyctis (10±2.39) as the most abundant anuran species while Microhyla ornata (<1±0.09) as the least abundant species. The Kernel regression revealed strong and statistically significant association between habitat variables and abundance of Hoplobatrachus tigerinus (R2=0.678) and Bufo stomaticus (R2=0.624) but weak and statistically significant association between habitat variables and abundance of E. cyanophlyctis (R2=0.482); Duttaphrynus melanostictus (R2=0. 451); M. ornata (R2=0.223) and Limnonectes limnocharis (R2=0. 006). We concluded that the common frogs and toads in our area belong to families Dicroglossidae and Bufonidae while uncommon frogs are of family Microhylidae. We suggest inclusion of monitoring of water quality (dissolved oxygen and pH) and maintenance of native wild vegetation particularly herbs, shrubs and hydrophytes of the area in the on-going and proposed development schemes of Rawalpindi–Islamabad Areas