Israel Journal of Ecology & Evolution, 2017 http://dx.doi.org/10.1163/22244662-06303001 Compassionate approaches for the conservation and protection of fire salamanders Leon Blausteina,*, Ori Segeva,**, Valentina Rovellia, Shirli Bar-Davida,b, Lior Blanka,c, Antonina Polevikova, Nadav Pezaroa, Tamar Krugmana, Simona Showstacka, Avi Koplovicha, Lital Ozeria and Alan R. Templetona,d aInstitute of Evolution and Department of Evolutionary and Environmental Biology, University of Haifa, Haifa 3498838 Israel bMitrani Department of Desert Ecology, Blaustein Institutes for Desert Research, Ben Gurion University of the Negev, Sede Boqer Campus, Sede Boker 84990 Israel cDepartment of Plant Pathology and Weed Research, ARO, Volcani Center, Bet-Dagan 50250, Israel dDepartment of Biology and Division of Statistical Genomics, Washington University, St. Louis, MO 63130-4899 USA **Current Address: Faculty of Aerospace Engineering, Technion, Haifa, Israel Abstract The Near Eastern fire salamander,Salamandra infraimmaculata, is considered an endangered species in Israel and is near-threatened regionally. For 25 years, our laboratory has sought ethical sampling methods to protect individuals and populations of Salamandra. To “mark” individuals for estimating dispersal and population size, we use non-invasive individual-specific markings from photographs of larvae and adults. We demonstrated through mesocosm experiments (which are less mortality-driven than in nature) that exotic Gambusia affinis have extreme negative mortality effects on Salamandra larvae. From a compassionate conservation aspect, G. affinis should not be killed and placed in habitats where amphibians are not in danger and mosquitoes can be controlled. We identified breeding-site characteristics demonstrating that permanent breeding sites support larger adult populations than temporary breeding sites. For population genetics studies, we take minimal sized tail tips from adults (which have no adverse effects) for microsatellite data. For gene expression studies, rather than sacrifice entire bodies, we demonstrated that by taking only small larval tail tips, we could follow gene expression. We additionally demonstrated that tail tip removal does not affect survival, time to or size at metamorphosis. We documented high road kill rates at a specific breeding site. To prevent potential disease spread, we sterilize boots and sampling gear. We use results for implementing or recommending conservation of individuals and populations – e.g., identifying: movement corridors for breeding site dispersal; roadkill hotspots for under-road tunnels; suitable habitat for pool construction for more effective conservation; utilizing population genetics for recommending management units; information on demography and genetic diversity to identify hotspots for conservation; removal of Gambusia for amphibian protection. Keywords animal ethics; compassionate conservation; Gambusia affinis; gene expression; Salamandra infraimmaculata Introduction at least 190 m above sea level (asl) (Blank and Blaustein, Compassionate conservation is a discipline that not only 2012, 2014). In Syria also, they are also not found below acts to prevent extinctions of populations but also protects 200 m asl (Bogaerts et al., 2013). Salamandra infraim- individuals from unnecessary harm and death (Ben-Ami maculata retreat into deep crevices and underground caves et al., 2014; Ramp and Beckoff, 2015, Wallach et al., 2015). when rains stop, where they remain relatively dormant The Near Eastern fire salamander, Salamandra infraim- throughout the hot and dry season (approximately early maculata, is considered an endangered species in Israel May through mid-October). Adults resume activity and (Dolev and Perevolotsky, 2004) and near-threatened re- emerge from estivation sites once rains begin in October gionally (IUCN Red List, 2017). For 2.5 decades, our or November. Gravid females emerge from their over- laboratory has sought ethical sampling methods to protect summering sites, carrying a full complement of larvae (as individuals and populations of S. infraimmaculata while many as 193 larvae has been deposited by a single clutch performing research to further our knowledge and under- from a single mother although S. infraimmaculata females standing of their life history, ecology, population genetics from Tel Dan are smaller and deposit far fewer larvae [De- and ecological genomics, which in turn, can be used to gani, 1996]). Adults are typically active on rainy nights and help preserve individuals and populations. females larviposit in a variety of water bodies including temporary lentic ponds, permanent lentic ponds, tempo- rary ponds in ephemeral wadis and in permanent streams Natural History of Salamandra infraimmaculata (Degani, 1996; Blank and Blaustein, 2012). Females have The Near Eastern fire salamander, S. infraimmaculata, ex- been shown to adjust the number of larvae they deposit in a ists in Iran, Iraq, Syria, Turkey, Lebanon, and Israel (IUCN particular water body depending on the water volume (Se- Red List, 2017). In Israel, S. infraimmaculata occurs in gev et al., 2011) or preferentially select structurally com- the northern, mountainous and more rainy regions that are plex habitats when older cohorts of conspecific individuals *Corresponding author. E-mail: [email protected] Downloaded from Brill.com09/26/2021 06:51:20AM © Koninklijke Brill NV, Leiden 2018 via free access <UN> 44 L. Blaustein et al. are already present, presumably to avoid cannibalism (Sa- wadis (Segev and Blaustein, 2014). Using the tailfin iden- deh et al., 2009). Larvae are key predators that have a large tification, we were able to document that some Salaman- impact on community structure, including negative effects dra larvae did not remain in their rock pools, but dispersed on many predators and herbivores that lead to trophic cas- downstream or were forced downstream to another rock cades (Blaustein et al., 1996, Eitam et al., 2005, Segev and pool. Also by using non-invasive photographs of adults for Blaustein, 2007). They are also predators on mosquito lar- mark-recapture, we have been able to assess how different vae and egg rafts (Blaustein et al., 2014). Some crustacean breeding habitats support different population sizes. Our species also avoid hatching in response to chemical cues research group has also recently been testing the use of Vis- of Salamandra larvae, apparently to avoid being consumed ible Implant Elastomer (VIE) tags as a marking technique (Blaustein, 1997; Spencer and Blaustein, 2001). The larvae for identifying Salamandra larvae in both field and labora- are also cannibalistic (Degani, 1993; Markman et al., 2009; tory experiments. The VIE tags consist of a silicon-based Sadeh, 2012). Once larvae metamorphose and exit the wa- liquid polymer that is injected subdermally (under anesthe- ter body, it takes about 3–5 years to become reproductively sia) and cures rapidly into a biocompatible and pliable sol- mature (Warburg, 1994) and they may live to be 25 years id (Northwest Marine Technology, Inc.). The multi-color or more (Warburg, 1994). Here, we report on our research tagging options of the VIE provides minimally invasive activities, taking compassionate conservation into account. and harmless means to mark large numbers of larvae when pattern recognition methods are unavailable (Low, 2003; Grant, 2008; Dodd, 2010). Larvae marked using VIE in Estimating S. infraimmaculata population size and our lab have also shown no indication of suffering adverse dispersal by mark-recapture effects in either survival or behaviouralchanges (Segev In assessing population size and dispersal using capture- et al., 2015; Pezaro et al., unpublished results). mark-recapture methods, many species are tracked by cut- ting off different toes for assessing individual identification. In many cases, it is not known whether cut-off toes can Is assessing impacts of introduced predators of affect mobility, survival or dispersal of particular species Salamandra larvae compassionate conservation? (Perry et al., 2011). Toe-clipping is a common technique In 2003, we found that Salamandra larvae in ponds in the that has been widely used in studies of mammals, reptiles, presence of high densities of the mosquitofish,Gambusia birds and amphibians but both the efficacy and humane affinis (which were apparently introduced for mosquito nature of the technique are subject to compassionate criti- control), had chewed-off tail fins and other appendages cism (Clark, 1972; May, 2004; McCarthy and Paris, 2004). while most Salamandra larvae in ponds without Gambusia Moreover, the technique may not be effective, especially for had normal intact tail fins and other appendages although long-term studies, if toes are lost naturally or for animals Salamandra may bite off other individuals’ tails due to can- that have the ability to regenerate their appendages (Ferner, nibalism (Degani 1993; Sadeh et al., 2009; Sadeh 2012). We 1979). In the case of estimating dispersal and population found that at the ponds of Kaukab Springs (Lower Galilee size of the fire salamander, we (Bar-David et al., 2007; Mountains), with high densities of Gambusia, no salaman- Segev et al., 2010; Goedbloed et al., 2017. I. Sinai et al., der larvae survived to metamorphosis. We submitted this unpublished results) and others (Warburg, 1994) have used work to the Israel Journal of Zoology as a likely warning photography for specific dorsal spot patterns in S. infraim- that Gambusia