Acta Herpetologica 12(2): 209-214, 2017 ISSN 1827-9635 (print) © Firenze University Press ISSN 1827-9643 (online) www.fupress.com/ah DOI: 10.13128/Acta_Herpetol-20283 Who are you? The genetic identity of some insular populations of Hierophis viridiflavus s.l. from the Tyrrhenian Sea Ignazio Avella, Riccardo Castiglia, Gabriele Senczuk* Dipartimento di Biologia e Biotecnologie “Charles Darwin”, Università di Roma “La Sapienza”, 00151, Roma, Italy. *Corresponding author. E-mail: gabriele.senczuk@uniroma1.it Submitted on: 2017, 3rd February; revised on: 2017, 13th April; accepted on: 2017, 6th June Editor: Adriana Bellati Abstract. This work investigates the genetic identity of Hierophis viridiflavus s.l. specimens from insular populations, to determine which of the two previously identified species is present on each island. Here, the authors hypothesise about times and modes of colonization and discuss the faunistic value of the obtained results. This follows the recent proposal to consider the two clades as two different species. Specimens from the islands of Favignana, Lipari and Vul- cano belong to H. carbonarius and probably all belong to putative Sicilian source populations. Conversely, all individ- uals from the Pontine Islands (Ponza, Palmarola, Ventotene) should be considered to belong to H. viridiflavus. Even if genetically identical to the specimens from the Tyrrhenian Italian coast, these individuals show a darker colouration, very similar to the one usually shown by H. carbonarius specimens. Considering that the Pontine H. viridiflavus pop- ulations probably have a very recent origin, the dark livery of these individuals could be the result of a rapid morpho- logical adaptation to insular environments. Keywords. Colour pattern, Hierophis viridiflavus, islands, nd4, phylogeography. The western whip snake Hierophis viridiflavus s.l. (Lacépède, 1789) is a colubrid snake with a wide distri- bution range. It can be found in Central Europe from Eastern Spain to Central France, Luxemburg, Switzer- land, Slovenia and Croatia. Its range also includes all of the Italian Peninsula, Sicily, Sardinia and most of the smaller Italian islands, Corsica and some Croatian islands (Vanni and Nistri, 2006; Zuffi, 2007). The species is found from sea level to 1500-1800 m a.s.l., although it is extremely rare above 1500 m in the Alps (Farinello and Bonato, 2000). Interestingly, individuals of this spe- cies show two main phenotypes, one named “viridiflavus” (usually brown/blackish with yellow stripes and spots) and the other named “carbonarius”, typically melanic (completely black with blackish/grey ventral colouration) or melanotic (almost completely black livery, but paler/ yellowish head scales and ventral surface). Individu- als from some of the islands of the Tuscan Archipelago, Sardinia and Corsica show a third phenotype, a middle ground between “carbonarius” and “viridiflavus”, called “abundistic” (Zuffi, 2008). In the past, four H. viridifla- vus s.l. subspecies have been described for Italy, mainly due to the chromatic pattern of the analysed individu- als: H. v. viridiflavus (Rimpp, 1979), H. v. carbonarius (Bonaparte, 1833), H. v. kratzeri (Kramer, 1971) and H. v. antoniimanueli (Capolongo, 1984). However, based on mitochondrial and nuclear DNA evidences, they have all been recently rejected (Vanni and Zuffi, 2011). Indeed, Nagy et al. (2002) and Rato et al. (2009) showed the pres- ence of two mitochondrial distinct haplogroups: the first, roughly corresponding to the subspecies H. v. viridiflavus (clade W) includes individuals from Spain, France, Cor- sica, Sardinia and central to north-western Italy on the West side of the Apennines; the second, in part matching with the subspecies H. v. carbonarius (clade E), occurs on the other side of the Apennines, from north-eastern 210 Ignazio Avella et alii Italy to southern Italy (including Sicily). More recently, a study based on morphometric, genetic and karyologi- cal data, proposed the elevation of the two genetic groups to species status (Mezzasalma et al., 2015). In particular, the authors emphasized significant differences in sexual chromosomes, as while females from the Eastern group have a submetacentric W sex chromosome, females from the Western group have a telocentric W sex chromo- some. Thus, individuals from the Eastern clade have been recognized as Hierophis carbonarius, while individuals from the Western clade have been recognized as Hiero- phis viridiflavus. Interestingly, the relationship between colour variation and genetic repartition does not match completely (Zuffi, 2008). The brown/blackish colouration with yellow stripes and spots pattern, generally almost exclusive of the H. viridiflavus range, can be found also in H. carbonarius specimens (Rato et al., 2009). Although the distribution of the two species in the Italian Penin- sula and on the largest Mediterranean islands (Sardinia, Corsica and Sicily) has already been studied (Nagy et al., 2002; Rato et al., 2009; Mezzasalma et al., 2015), there is a lack of molecular data from smaller Italian archi- pelagos. The aim of the present work is to determine the genetic identity of individuals collected in some Tyrrhe- nian islands including: Ponza, Palmarola and Ventotene from the Pontine Archipelago; Vulcano and Lipari from the Aeolian Islands; and Favignana from the Aegadian Islands, in order to update the distribution of the two species of whip snakes. We sampled a total of seven individuals of H. viridi- flavus s.l. from six different islands between March 2014 and July 2015 (geographic locations are reported in Table 1 and showed in Fig. 1). Snakes were caught and han- dled following standard protocols (Fowler, 1978), and some ventral scales were removed and preserved in pure ethanol. In one case (RS296 from Lipari), the tissue was obtained from a shedded skin. Genomic DNA was extracted following the protocol described in Aljanabi and Martinez (1997). A fragment including the terminal portion of the NADH dehydroge- nase subunit 4 (nd4) was amplified by standard PCR pro- tocols using primers published by Arèvalo et al. (1994). Amplification conditions were the same as described by Pinho et al. (2006). The PCR products were purified with a Sure Clean (Bioline©) purification kit and the sequenc- ing reactions were run under Big-Dye TM Terminator cycling conditions by a commercial company, Macro- gen (www.macrogen.com). The electropherograms were checked using the software FinchTV (http://www.geo- spiza.com/finchtv/) to ensure the absence of double peaks and ambiguous positions. The obtained sequences were deposited to GenBank (accession numbers: KY923281- KY923287) and joined with additional 91 nd4 sequences of Hierophis viridiflavus s.l. retrieved from GenBank (accession numbers: FJ430621-FJ430660, Rato et al., 2009; LN552045-LN552095, Mezzasalma et al., 2015). Nucleotide sequences were translated into amino acids with MEGA 6.0 (Tamura et al., 2013) using the vertebrate mitochondrial genetic code in order to assess the absence of pseudogenes. One nd4 sequence of Hierophis gemonensis (Laurenti, 1768) was downloaded from GenBank (accession num- ber: AY487044, Nagy et al., 2004) and included in the analysis as outgroup, as it is considered the closest related species to H. viridiflavus (Schätti, 1988). The software jModelTest (Posada, 2008) was used to determine the most appropriate model of sequence evo- lution for the nd4 dataset. According to the Akaike infor- mation criterion (AIC), the most supported evolutionary model was the TrN + I, therefore applied in the subse- quent analysis. To reconstruct phylogenetic relationships, we used a coalescent Bayesian approach as implemented in MrBayes 3.2.6 (Ronquist et al., 2012). We run 2 million generations, with 4 Markov chains sampling every 1000 steps. After a burn-in of 10%, the remaining trees were used to compute a 50% majority rule consensus tree. In addition, a statistical parsimony network under 95% probability connection limits was constructed using TCS 1.21 (Clement et al., 2000). Number of haplotypes, nucleotide diversity (π) and haplotype diversity (H) were Table 1. individuals analysed including sampling location, group of islands, colour pattern, haplotype number and haplogroup. Sample code Locality Archipelago Colour pattern Haplotype Species RL22 Ponza Pontine “abundistic” H1 H. viridiflavus RL66 Ponza Pontine “abundistic” H1 H. viridiflavus RL79 Ventotene Pontine “abundistic” H1 H. viridiflavus RL80 Palmarola Pontine “abundistic” H1 H. viridiflavus RS85 Favignana Aegadian “carbonarius” H10 H. carbonarius RS276 Vulcano Aeolian “carbonarius” H9 H. carbonarius RS296 Lipari Aeolian “carbonarius” H9 H. carbonarius 211Genetic identity from Tyrrhenian insular populations of Hierophis viridiflavus s.l. also calculated for each group using DnaSP 5.1 (Librado and Rozas, 2009). The final nd4 alignment (568 bp) of 99 sequences returned 67 polymorphic sites and 13 haplotypes. The phylogenetic analysis confirmed the presence of two well defined mitochondrial clades (Fig. 1A), as already stated in previous works and corresponding to the two spe- cies H. viridiflavus and H. carbonarius (Rato et al., 2009; A Hv24-11 V39-15 Hv6-29 V10-79 Hv17-52 Hv32-62 Hv42-40 Hv8-30 Hv18-14 RL66-2 V4-31 Hv30-33 RS276-6 Hv26-74 V13-57 Hv49-70 Hv43-47 Hv38-72 V19-55 V30-77 V20-51 V27-42 Hv22-41 Hv25-69 Hv41-65 V26-45 V7-31 V1-46 Hv35-37 RL80-3 Hv45-67 V12-67 Hv14-28 V32-75 V25-45 V21-53 Hv13-18 V8-31 V42-43 Hv28-54 Hv5-57 Hv16-21 Hv40-36 Hv29-73 Hv46-13 Hv4-34 RS85-4 Hv9-10 V9-79 V14-57 V15-57 V38-26 Hv37-78 Hv33-56 Hv2-23 V41-66 Hv3-76 Hv34-49 Hv1-12 V11-79 Hv19-35 V3-58 V40-22 Hv48-68 V22-67 V16-57 V29-77 Hv21-32 V33-34 Hv12-9 Hv36-27 Hv31-38 RL79-1 Hv20-8 Hv51-24 RL22-2 V17-17 Hv23-63 V31-77 V35-16 V6-31 V24-44 V2-48 Hv47-7 Hv11-20 Hv15-61 V23-39 V28-64 Hv27-59 V36-21 Hierophis gemonensis Hv10-28 V18-50 Hv50-25 Hv39-71 Hv7-19 Hv44-60 RS296-5 V37-7 0,99 0,99 0,8 0,98 1 C H1 Hierophis viridiflavus Hierophis carbonarius H11 H9 H5 H13 H12 H6 H7 H8 H10 H2 H3 H4 61 56 52 41 60 57 49 59 54 47 3236 40 62 3435 3337 38 67 29 28 30 27 25 23 19 20 21 24 14 12 10 9 13 18 11 7 8 69 74 73 78 72 71 6870 63 65 76 44 55 46 58 5051 45 48 53 43 3942 31 66 26 17 16 15 2264 79 77 75 Ponza 2 Favignana 4 Vulcano 5 Ventotene 3 Palmarola 1 Lipari 6 B H10 H6 H8 H1 H4 H3 H2 H5 H9 H12 H13 H7 H11 Fig. 1. Bayesian phylogenetic tree (A) based on nd4 sequences for 98 ingroup specimens of H. viridiflavus s.l. and one outgroup (H. gem- onensis). The posterior probabilities are indicated at each node. Each label indicates the specimen code, the locality number and the rela- tive haplotype. Insular individuals are shown in bold. Geographic distribution (B) of the two mitochondrial lineages corresponding to H. viridiflavus (blue) and H. carbonarius (red). Statistical parsimony network (C) connecting haplotypes. The circle size is proportional to the sequence frequencies and each filled rectangle represent one substitution. 212 Ignazio Avella et alii Mezzasalma et al., 2015). Nei’s standard genetic distance between the two species was 4.2%. Specimens from Favignana, Vulcano and Lipari, belong to the species H. carbonarius (Fig. 1A). This clade showed the presence of nine haplotypes (Fig. 1C, see Table A1 in supplementary materials for all the haplo- type references) with H = 0.623 ± 0.071, and π = 0.00389 ± 0.00057 (mean ± SD). The specimens from Vulcano (RS276) and Lipari (RS296) shared the same haplotype (H9) with individuals from localities seven and eight (Fig. 1B), corresponding to Iria and Lago Spartà (Sicily). These results may suggest a recent colonization, either human-mediated or by oversea dispersal, from Sicily to the Aeolian islands. On the other hand, the specimen from the island of Favignana (RS85) showed a new pri- vate haplotype (H10), separated by one mutational step from haplotype H9. In this case, the single fixed substitu- tion may have occurred on the island through a vicari- ant mechanism. Indeed, during the last glacial phase this island was connected to Sicily and become separated following the Last Glacial Maximum because of the sea level drop. A similar scenario has also been suggested to explain the genetic differentiation observed in other rep- tiles from Favignana (Mizan, 2015; Senczuk et al., 2017). However, due to the small sample size from Sicily, we cannot completely rule out that insular distinctiveness may have derived from a recent dispersal process of a haplotype not yet sampled in Sicily. All the individuals from the Pontine Islands (RL22, RL66, RL79, RL80) shared one single haplotype (H1) and should therefore be recognized as belonging to H. viridi- flavus (Fig. 1A). This clade is genetically less differenti- ated than H. carbonarius clade (H = 0.128 ± 0.067; π = 0.00023 ± 0.00012; mean ± SD) and is composed by four haplotypes: a single highly represented haplotype (H1; with an allele frequency of 94%) and three derived and extremely localized haplotypes (H2, H3 and H4). This result may suggest an anthropic introduction in modern times or a recent colonization of the Pontine Islands from the Tyrrhenian coast of the Italian Peninsula. Interestingly, the four specimens from the Pontine Islands showed a colour pattern which resemble the “abundistic” morph, which is in the middle between the “carbonarius” (melanic/melanotic) and the “viridiflavus” (black and yellowish) colour patterns. The “abundistic” phenotype was previously reported only in Sardinia, Cor- sica and the Tuscan Archipelago. However, Schätti and Vanni (1986) reported similarities between specimens from the Pontine Islands and the dark coloured ones from Emilia Romagna now considered belonging to H. carbonarius. In particular, the individual sampled from Ventotene (RL79, Fig. 2) had a very dark dorsal with a yellowish ventral colouration, showing a phenotype which could easily be mistaken with the one observed in many populations of H. carbonarius. This observation confirms that colour pattern alone cannot help identify- ing the species to which a specimen belongs. The four H. viridiflavus specimens from the Pontine Islands are genetically indistinguishable from the usu- ally brown-yellowish western whip snakes located on the Tyrrhenian coast of Italy, but show a darker phenotype. It has been reported in previous works that colour variation in reptiles can be associated to adaptive processes (Nor- ris and Lowe, 1964; Rosenblum et al., 2004). For exam- ple, darker or melanotic colouration may give a benefit in terms of thermoregulation (Trullas et al., 2007; Broenni- mann et al., 2014) and reproduction (Capula and Luiselli, 1994), and similar conclusion had been already drawn by Rato et al. (2009) and Zuffi (2007), as they consider the colour types in Hierophis viridiflavus s.l. a by-product of different environmental conditions. Therefore, the dark colouration of the snakes from the Pontine Islands could be the result of adaptive morphological evolution which occurred in a very short time, a phenomenon already observed in other insular reptile populations (Losos et al., 1997; Herrel et al., 2008). Finally, despite changes in colour polymorphism might also be the outcome of non- adaptive processes (King, 1988; Lorioux et al., 2008), the independent recurrence of the “abundistic” chromatism in all the northern Tyrrhenian Islands suggests a prominent role of adaptive forces acting in similar insular environ- mental conditions, which would deserve further studies. ACKNOWLEDGEMENTS Thanks to Dario D’Eustacchio, Emanuela De Simone, Marco Basile and Mattia Menchetti for providing sam- ples, to Laura Gramolini for helping in laboratory and Fig. 2. The specimen from Ventotene island (RL79). The individual was found stuck in a mist net trap and later released. 213Genetic identity from Tyrrhenian insular populations of Hierophis viridiflavus s.l. to Brinna Barlow for editorial assistance. We thanks Sara Riello for providing photos of the Ventotene specimen. All the tissue samples used in this work were collected with permission of the Italian Environment Ministry for the Environment, Land and Sea to RC. (Prot. 00017879/ PNM del 09/09/2012) and no animals were killed. SUPPLEMENTARY MATERIALS Supplementary material associated with this article can be found at Manuscript number 20283. REFERENCES Aljanabi, S.M., Martinez, I. (1997): Universal and rapid salt-extraction of high quality genomic DNA for PCR- based techniques. Nucleic Acids Res. 25: 4692-4693. Arèvalo, E., Davis, S.K., Sites, J.W. (1994): Mitochondrial DNA sequence divergence and phylogenetic relation- ships among eight chromosome races of the Scelopo- rus grammicus complex (Phrynosomatidae) in central Mexico. Syst. Biol. 43: 387-418. Bonaparte, C.L. (1833): Iconografia della fauna italica per le quattro classi di Animali Vertebrati. II. Anfibi. Salviucci, Roma. Broennimann, O., Ursenbacher, S., Meyer, A., Golay, P., Monney, J.C., Schmocker, H., Guisan, A., Dubey, S. (2014): Influence of climate on the presence of colour polymorphism in two montane reptile species. Biol. Lett. 10: 2014.0638. Capolongo, D. (1984): Note sull’erpetofauna Pugliese. Att. Soc. Ital. Sci. Nat. Mus. Civ. St. Nat. Mil. 125: 189-200. Capula, M., Luiselli, L. (1994): Reproductive strategies in alpine adders, Vipera berus: the black females bear more often. Acta Oecol. 15: 207-214. Clement, M., Posada, D.C.K.A., Crandall, K.A. (2000): TCS: a computer program to estimate gene genealo- gies. Mol. Ecol. 9: 1657-1659. Farinello, F., Bonato, R. (2000): Biacco, Hierophis vir- idif lavus (Lacépède, 1789); Coluber viridif lavus (Lacépède, 1789). In: Atlante degli Anfibi e dei Ret- tili della provincia di Vicenza, pp. 157-160. Gruppo Nisoria, Mus. Nat. Vic., Eds, Padovan Editore, Vice- nza. Fowler, M.E. (1978): Restraint and handling of wild and domestic animals. Iowa State University Press, Ames, Iowa. Herrel, A., Huyghe, K., Vanhooydonck, B., Backeljau, T., Breugelmans, K., Grbac, I., Van Damme, R., Irschick, D.J. (2008): Rapid large-scale evolutionary divergence in morphology and performance associated with exploitation of a different dietary resource. Proc. Natl. Acad. Sci. 105: 4792-4795. King, R.B. (1988): Polymorphic populations of the garter snake Thamnophis sirtalis near Lake Erie. Herpetolog- ica 44: 451-458. Kramer, E. (1971): Revalidierte und neue Rassen der europäischen Schlangenfauna. Biogeographia 1: 667- 676. Librado, P., Rozas, J. (2009): DnaSP v5: a software for comprehensive analysis of DNA polymorphism data. Bioinformatics 25: 1451-1452. Lorioux, S., Bonnet, X., Brischoux, F., De Crignis, M. (2008): Is melanism adaptive in seakraits? Amphibia- Reptilia 29: 1-5. Losos, J.B., Warheit, K.I., Schoener, T.W. (1997): Adaptive differentiation following experimental island coloniza- tion in Anolis lizards. Nature 387: 70-73. Mizan, V.L. (2015): Geographic patterns of genetic and morphological variation of the Sicilian wall lizard, Podarcis wagleriana. Unpublished MSc dissertation. University of Porto, Porto. Mezzasalma, M., Dall’Asta, A., Loy, A., Cheylan, M., Lymberakis, P., Zuffi, M.A.L., Tomovic, L., Odierna, G., Guarino, F.M. (2015): A sisters’ story: comparative phylogeography and taxonomy of Hierophis viridifla- vus and H. gemonensis (Serpentes, Colubridae). Zool. Scr. 44: 495-508. Nagy, Z.T., Joger, U., Guicking, D., Wink, M. (2002): Phylogeography of the European whip snake Coluber (Hierophis) viridiflavus as inferred from nucleotide sequences of the mitochondrial cytochrome b gene and ISSR genomic fingerprinting. Biota 3: 109-118. Norris, K.S., Lowe, C.H. (1964): An Analysis of Back- ground Color‐Matching in Amphibians and Reptiles. Ecology 45: 565-580. Nagy, Z.T., Lawson, R., Joger, U., Wink, M. (2004): Molecular systematics of racers, whipsnakes and rela- tives (Reptilia: Colubridae) using mitochondrial and nuclear markers. J. Zool. Syst. Evol. Res. 42: 223-233. Pinho, C., Ferrand, N., Harris, D.J. (2006): Reexamination of the Iberian and North African Podarcis (Squamata: Lacertidae) phylogeny based on increased mitochon- drial DNA sequencing. Mol. Phylogenet. Evol. 38: 266-273. Posada, D. (2008): jModelTest: phylogenetic model aver- aging. Mol. Biol. Evol. 25: 1253-1256. Rato, C., Zuffi, M.A.L., Corti, C., Fornasiero, S., Gentilli, A., Razzetti, E., Scali, S., Carretero, M.A., Harris, D.J. (2009): Phylogeography of the European Whip Snake, Hierophis viridiflavus (Colubridae), using mtDNA and 214 Ignazio Avella et alii nuclear DNA sequences. Amphibia-Reptilia 30: 283- 289. Rimpp, K. (1979): Herpetologische Skizzen aus Sardinien. Herpetofauna 1: 24-27. Ronquist, F., Teslenko, M., Van der Mark, P., Ayres, D.L., Darling, A., Höhna, S., Huelsenbeck, J.P. (2012): MrBayes 3.2: efficient Bayesian phylogenetic inference and model choice across a large model space. Syst. Biol. 61:539-542. Rosenblum, E.B., Hoekstra, H.E., Nachman, M.W. (2004): Adaptive reptile color variation and the evolution of the MC1R gene. Evolution 58: 1794-1808. Schätti, B., Vanni, S. (1986): Intraspecific variation in Coluber viridiflavus Lacépède, 1789, and validity of its subspecies (Reptilia, Serpentes, Colubridae). Rev. Sui- sse Zool. 93: 219-232. Schätti, B. (1988): Systematik und Evolution der Schla- gengattung Hierophis Fitzinger, 1843 (Reptilia, Ser- pentes). Unpublished doctoral dissertation. University of Zürich, Zürich. Senczuk, G., Colangelo, P., De Simone, E., Aloise, G., Castiglia, R. (2017): A combination of long term frag- mentation and glacial persistence drove the evolution- ary history of the Italian wall lizard Podarcis siculus. BMC Evol. Biol. 17: 6. Tamura, K., Stecher, G., Peterson, D., Filipski, A., Kumar, S. (2013): MEGA6: molecular evolutionary genetics analysis version 6.0. Mol. Biol. Evol. 30: 2725-2729. Trullas, S.C., van Wyk, J.H.,Spotila, J.R. (2007): Ther- mal melanism in ectotherms. J. Therm. Biol. 32: 235-245. Vanni, S., Nistri, A. (2006): Hierophis viridiflavus (Lacépède, 1789). In: Atlante degli Anfibi e dei Rettili d’Italia, pp. 544-547. Sindaco, R., Doria, G., Razzetti, E., Bernini, F., Eds, Societas Herpetologica Italica, Edizioni Polistampa, Firenze. Vanni, S., Zuffi, M.A.L. (2011): Hierophis viridiflavus (Lacepede, 1789). In: Fauna d’Italia, vol. 45, Rep- tilia, pp. 509-516. Corti, C., Capula, M., Luiselli, L., Razzetti, E., Sindaco, R., Eds, Edizioni Calderini, Bologna. Zuffi, M.A.L. (2007): Patterns of phenotypic variation in the European Whip snake, Hierophis viridiflavus (Lacépède, 1789). Unpublished doctoral dissertation. Università di Pisa, Pisa. Zuffi, M.A.L. (2008): Colour pattern variation in popula- tions of the European Whip snake, Hierophis viridifla- vus: does geography explain everything? Amphibia- Reptilia 29: 229-233. Acta Herpetologica Vol. 12, n. 2 - December 2017 Firenze University Press Meristic and morphometric characters of Leptopelis natalensis tadpoles (Amphibia: Anura: Arthroleptidae) from Entumeni Forest reveal variation and inconsistencies with previous descriptions Susan Schweiger1, James Harvey2, Theresa S. Otremba1, Janina Weber1, Hendrik Müller1,* Brown anole (Anolis sagrei) adhesive forces remain unaffected by partial claw clipping Austin M. Garner*, Stephanie M. Lopez, Peter H. Niewiarowski Species and sex comparisons of karyotype and genome size in two Kurixalus tree frogs (Anura, Rhacophoridae) Shun-Ping Chang1,2, Gwo-Chin Ma2,3,4, Ming Chen2,5,6,7,8,*, Sheng-Hai Wu1,* Non-native turtles in a peri-urban park in northern Milan (Lombardy, Italy): species diversity and population structure Claudio Foglini1, Roberta Salvi2,* Species composition and richness of anurans in Cerrado urban forests from central Brazil Cláudia Márcia Marily Ferreira1,*, Augusto Cesar de Aquino Ribas2, Franco Leandro de Souza3 The life-history traits in a breeding population of Darevskia valentini from Turkey Muammer Kurnaz, Alı İhsan Eroğlu, Ufuk Bülbül*, Halıme Koç, Bılal Kutrup Influence of desiccation threat on the metamorphic traits of the Asian common toad, Duttaphrynus melanostictus (Anura) Santosh Mogali*, Srinivas Saidapur, Bhagyashri Shanbhag Predation of common wall lizards: experiences from a study using scentless plasticine lizards Jenő J. Purger*, Zsófia Lanszki, Dávid Szép, Renáta Bocz Reproductive timing and fecundity in the Neotropical lizard Enyalius perditus (Squamata: Leiosauridae) Serena Najara Migliore1,2,*, Henrique Bartolomeu Braz2,3, André Felipe Barreto-Lima4, Selma Maria Almeida-Santos1,2 Observations on the intraspecific variation in tadpole morphology in natural ponds Eudald Pujol-Buxó1,2,*, Albert Montori1, Roser Campeny3 and Gustavo A. Llorente1,2 Reliable proxies for glandular secretion production in lacertid lizards Simon Baeckens Diet of juveniles of the venomous frog Aparasphenodon brunoi (Amphibia: Hylidae) in southeastern Brazil Rogério L. Teixeira1, Ricardo Lourenço-de-Moraes2, Débora C. Medeiros3, Charles Duca3, Rogério C. Britto4, Luiz C. P. Bissoli5, Rodrigo B. Ferreira3,* Who are you? The genetic identity of some insular populations of Hierophis viridiflavus s.l. from the Tyrrhenian Sea Ignazio Avella, Riccardo Castiglia, Gabriele Senczuk*