Lab protocol for investigating impacts of climate change on frogs.

We developed a method to investigate impacts of temperature (elevated) on breeding, growth and development in endemic frogs under laboratory conditions. The method provides details on housing and rearing of larvae, taking various important measurements and observing developmental deformities. The method could also be applied to rescue approach or head-start program for amphibian species experiencing climate change elsewhere in the world. • Rearing of larvae to investigate effects of temperature on larvae • Measurement of data on growth and development • Rescue/ head-start program.

Movement Paradigm for Hazara Torrent Frog Allopaahazarensis and Murree Hills Frog Nanoranavicina (Anura: Dicroglossidae).

Endemic anurans are particularly vulnerable to environmental changes, and are susceptible to population declines because of their restricted distribution ranges. The Murree Hills Frog Nanoranavicina and Hazara Torrent Frog Allopaahazarensis 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.

Rise in temperature causes decreased fitness and higher extinction risks in endemic frogs at high altitude forested wetlands in northern Pakistan.

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 tadpoles of Murree Hills Frog Nanorana vicina (Anura: Dicroglossidae).

The genus Nanorana, also called Yunnan Slow Frogs (Frank Ramus 1995), belongs to the family Dicroglossidae. Murree Hills Frog N. vicina, is an endemic species of Southeast Asian uplands in Pakistan and India and was first reported by Stoliczka (1872) from Murree, Punjab Province, Pakistan. Later Rais et al. (2014) recaptured it from its type locality and described morphology of one juvenile female and one adult male. The species is listed as Least Concern in IUCN Red List of Threatened Species while the EDGE (Evolutionarily Distinct and Globally Endangered) score is 2.64. In Pakistan, it is reported from brook water springs and streams of Murree, Galiat (North Punjab), Ayubia (Khyber Pakhtunkhwa), and parts of Azad Jammu and Kashmir (Rais et al. 2014). Although the adult form has been characterized (Khan 2006; Rais et al. 2014), the morphology of the tadpoles have not yet been described. The current study aimed to describe external morphology and morphometry of tadpoles of N. vicina.