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In Pakistan, the family Microhylidae is represented by two species: the Marbled Balloon Frog (Uperodon systoma) and Ant Frog (Microhyla nilphamariensis). Although these two frog species are listed as Least Concern (LC) in the IUCN red list of threatened species, the former is rare in Pakistan. When a species is not detected at certain locations, it can indicate that the species may be truly absent or not detected despite concerted efforts, which could pose a serious issue for conservation efforts. Here we estimate the abundance and determine if the likelihood of occurrence of U. systoma and M. nilphamariensis was influenced by the site and survey covariates using logistic regression in a subtropical scrub forest (Rawalpindi-Islamabad, Pakistan). We did not record U. systoma from the 24 sampled sites either through active searching or pitfall trapping. We recorded 37 M. nilphamariensis from eight of the 24 sampled sites. Our findings could be used to update the conservation status of the studied narrow-mouthed frog species in Pakistan. [ABSTRACT FROM AUTHOR]

Endemic anurans are particularly vulnerable to environmental changes, and are susceptible to population declines because of their restricted distribution ranges. The Murree Hills Frog Nanorana vicina and Hazara Torrent Frog Allopaa hazarensis are associated with the torrential streams and nearby clear water pools in subtropical chir pine forest and other forest types, at elevations higher than 1000 m in Pakistan. In this study, we have provided data on the extent of movement of these frog species for the first time. We installed radio transmitters on a total of 13 Murree Hills Frogs and 13 Hazara Torrent Frogs during eight consecutive days in September 2017 and 2018. Our results showed that these frogs did not move long distances along the stream or away from the stream into the forest. All the radio-tracked frogs showed movement of < 3 m. We found a significant differences only in the distance moved by Murree Hills Frogs between the two years studied. Based on our findings, we propose a movement paradigm that focuses on conservation implications for these endemic frogs. [ABSTRACT FROM AUTHOR]

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 both A. hazarensis and N. vicina than for visual encounter surveys. Detection probability using eDNA was greater for both species, particularly for A. 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. [ABSTRACT FROM AUTHOR]

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. [ABSTRACT FROM AUTHOR]

The larvae of some amphibian species grow and develop more slowly at lower temperature and attain larger body size at the end of metamorphosis, but this is not true for all amphibians. We have for the first time provided empirical data by using a combination of field and lab-based studies with a robust sample size (360 adults, 1000 tadpoles) for Hazara Torrent Frog (Allopaa hazarensis) and Murree Hills Frog (Nanorana vicina). In this study, we examined how tadpoles responded to different temperatures through metamorphosis, body size, developmental complications or deformities, fitness and survival. We found that the tadpoles of A. hazarensis and N. vicina showed the same response to elevated temperatures under laboratory conditions including faster metamorphosis, reduction in the body size, more frequent developmental complications or deformities such as edema and tail kinks, lower fitness and higher mortality at elevated temperatures (>26 °C). The comparison of thermal sensitivity between the two species showed that N. vicina was more sensitive to higher temperatures. Pakistan has been experiencing changes in climatic patterns, and the ecosystems in northern Pakistan are losing biodiversity due to increasing temperature, frequency of drought and intensity of floods. It is feared that these frogs may experience local extinction in future. Our findings contribute to filling the information gap regarding impacts of temperature increase on biodiversity of high altitude forested montane ecosystems and is an important contribution to future studies associating biodiversity and climate change. • The study provides empirical data by using combination of field and lab-based studies for Hazara Torrent Frog (Allopaa hazarensis) and Murree Hills Frog (Nanorana vicina). • The tadpoles of both species showed the same response to elevated temperatures under laboratory conditions. • Faster metamorphosis, reduction in the body size, lower fitness and higher mortality were observed in tadpoles at elevated temperatures (> 26 °C). [ABSTRACT FROM AUTHOR]