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Home , Alpine newt, Ichthyosaura, Lissotriton, Moravian Gate, Newt, Salamander

Herpetology Notes, volume 12: 923-930 (2019) (published online on 23 September 2019)

An Alpine ( alpestris) population on the Baltic coast of

Joanna Jakóbik1, Paweł Janowski2, Jacek Błażuk3, Tomasz Narczyński4, and Maciej Pabijan1,*

Abstract. This study describes a new locality of the located on the Baltic coast of northern Poland, establishing the most northeastern population of this in . The population inhabits the Tricity Landscape Park (TLP, Trójmiejski Park Krajobrazowy) and is isolated from the main part of the range of this species by about 350 km. By sequencing a fragment of the mitochondrial ND4 gene and incorporating data from recent phylogenetic analyses, we restricted the potential origin of from TLP to populations carrying a specific variant of the C3 lineage present in northwestern and . More precise inference of the source population is hindered by low genetic differentiation within the C3 lineage. We suggest that a human-mediated XXth century introduction (purposeful or accidental) could have established the Alpine newt in TLP, although historical information of the release is currently unavailable. Alternatively, the TLP population may be a relict of a regressing range border due late climatic shifts and in the southeastern Baltic region.

Keywords. urodele, patchy distribution, introduction, mtDNA

Introduction Sura, 2018). Small and isolated localities occur in the highlands of south-central Poland (Pabijan et al., 2009; The Alpine newt, Ichthyosaura alpestris (Laurenti, Kurek and Święciak, 2010) and scattered lowland sites 1768), has a broad distribution encompassing most close to the continuous range in southwestern Poland of western and central Europe, the Balkan Peninsula (Rychła et al., 2002; Najbar et al., 2005). and parts of Italy and Spain (Sillero et al., 2014). Three major mitochondrial DNA (mtDNA) lineages The nominate is present in western and occur across the range of the Alpine newt (Fig.1; central parts of the continent, while genetically and Sotiroupoulos et al., 2007; Recuero et al., 2014), two morphologically distinct subspecies and lineages occur of these (western and eastern lineages) show further in the southern parts of the distribution (Roček et al., subdivisions in the southern European peninsulas. 2003; Sotiropoulos et al., 2007; Recuero et al., 2014). Western and central Europe, including Poland, are Introduced populations have been documented in Great populated by newts carrying the widespread C3 mtDNA Britain, Holland, central Spain, southern France and lineage (Sotiroupoulos et al., 2007; Recuero et al., 2014). New Zealand (Bell and Bell, 1995; Sillero et al., 2014; Despite limited mtDNA variation within C3, there is Arntzen et al., 2016; Palomar et al., 2017). In Poland, clear geographic substructure with two haplotypes this species is mostly limited to the Carpathians, differing by two substitutions in the ND4 gene (Pabijan and Holy Cross mountain ranges and their foothills, and Babik, 2006). The western variant, termed C3-Sud in the southern part of the country (Głowaciński and hereafter, occurs in the Sudetes mountains of Poland and the and westwards into Germany, France and Austria, while the eastern haplotype, hereafter termed C3-Carp, is restricted to the of Poland, Ukraine and northern Romania (Fig. 1). 1 Department of Comparative , Institute of Zoology Additional haplotypes with limited distributions, e.g. and Biomedical Research, Jagiellonian University, ul. from Hungary and France, occur locally across central Gronostajowa 9, 30-387 Kraków. and western Europe (Recuero et al., 2014; Arntzen et 2 Towarzystwo Herpetologiczne NATRIX. 3 ul. Opolska 10, 80-395 Gdańsk-Oliwa. al., 2016). The Moravian Gate, a depression situated 4 Polskie Towarzystwo Ochrony Przyrody „”. between the Carpathian and Sudetes Mountains, divides * Corresponding author: [email protected] the distributions of haplotypes C3-Sud and C3-Carp and 924 Joanna Jakóbik et al.

Figure 1. Maximum likelihood tree (lower left) of ND4 sequences of Alpine newts depicting major mtDNA lineages (asterisks on branches denote bootstrap support >70%). Haplotype network (upper left) showing nucleotide variation within the C3 mitochondrial lineage. Map shows the range of the Alpine newt with colors depicting the major mtDNA lineages. Haplotypes C3-Sud and C3-Carp are denoted by filled circles and triangles, respectively. Grey squares indicate introduced populations of unknown provenance; question marks denote areas in which major mtDNA lineage is in need of confirmation. Map based on Sotiropoulos et al. (2007), Recuero et al. (2014), Sillero et al. (2014) and Arntzen et al. (2016).

probably constitutes a natural barrier to dispersal for this Material and Methods species since it is devoid of Alpine newt populations. Study area.—The Tricity Landscape Park is a mostly This mtDNA phylogeographic break is mirrored by the forested region in the Cassubian lakeland of north- existence of discreet evolutionary units of this species central Poland, close to the . The young inhabiting the Carpathians and Sudetes as revealed by landscape consists of undulating hills with numerous nuclear DNA variation (Pabijan and Babik, 2006; Babik lakes and small water bodies created by the Weichselian et al., 2008) as well as allozyme (Pabijan et al., 2005) and glaciation. The cool, north-facing slopes and deep skin graft rejection data (Rafinski, 1974). These studies ravines are drained by numerous coldwater streams. also show that the isolated population in the Holy Cross Beech, oak and Scots pine predominate in the forests Mountains (southcentral Poland) is genetically distinct of this region. but most closely related to the Carpathian population. In 2016 several adult Alpine newts were unexpectedly Field methods.—Field surveys of 111 potential found in the Tricity Landscape Park (TLP; breeding sites were conducted in the spring Polish: Trójmiejski Park Krajobrazowy, German: and early summer of 2015-2016 with the aim of Landschaftsschutzpark Dreistadt), a protected area in obtaining an inventory of the batrachofauna of the TLP northern Poland close to the Baltic coast, adjacent to the (P. Janowski, unpublished). Most of the sites inhabited cities of Gdańsk, Gdynia and Sopot. This Alpine newt by Alpine newts were revisited in 2017 and 2018 to population is geographically distant from other localities confirm the presence of breeding adults and/or larvae of (about 350 km straight line distance to the nearest site). this species. We noted the characteristics of Alpine newt The aim of this study was to determine the provenance breeding (confirmed by the presence of larvae) of the Alpine newts in TLP based on mtDNA variation. by recording variables describing the size, stability, We also mapped the local range of the Alpine newt and connectivity and vegetation of ponds during the time of described its breeding habitat characteristics in this surveys (April-August). area. Molecular methods.—Genomic DNA was extracted from 22 Alpine newts (Table 1), five of which originated An Alpine newt Ichthyosaura alpestris population on the Baltic coast of Poland 925 from TLP. One of the individuals from TLP was an adult using primers ND4 and Leu (Pabijan and Babik, 2006). female (MPFC2190) found dead and partly decomposed After purification, the PCR products were sequenced near site 4 (see Fig. 1). Four larvae (MPFC3220, using Big Dye chemistry (Applied Biosystems) and MPFC3230, MPFC3232, MPFC3233) from different resolved on an ABI 3130 machine (Applied Biosystems). sites (sites 4, 5 and 8, Fig. 1) were sampled by taking Newly obtained sequences were deposited in GenBank tailfin clips in 2018. All other Alpine newt samples (accession numbers MN211320-MN211341). were collected from different locations in central DNA sequence analysis.—Chromatograms were Europe over the past 15 years and stored in alcohol at edited in CodonCode Aligner (v. 2.0.6, Codon Code -20ºC in the amphibian tissue collection maintained at Corporation). The new sequences were aligned with the Department of Comparative Anatomy, Jagiellonian published sequences representing the range-wide University. DNA extraction, PCR and amplicon mtDNA diversity of this species. The final alignment sequencing were conducted according to Pabijan and included the following sequences from Pabijan and Babik (2006). DNA in the decomposed adult female Babik (2006), Recuero et al., (2014) and Arntzen et (MPFC2190) had deteriorated and necessitated the al., (2016): Ambleteuse A3 2,3; Charnohora A2; 12 application of an extraction kit dedicated to retrieving Krempna B1; 40 Beskid Wyspowy; 94, 96 Dulowa B2; trace amounts of DNA from degraded samples (Sherlock 120 Alwernia C1; 154, 177 Prudnik A1; 181, 182 Bielnik; AX, A&A Biotechnology). DNA from all other samples 184 Suchedniów; 215, 218, 249, 250 Muszkowice A1; was extracted using a phenol/chloroform protocol. We 282, 283, 285 Kąty; 315 Złoty Stok A1; 330 Lubawka amplified a fragment of NADH dehydrogenase subunit A1; 303, 312 Riu Marului; 313 Colonia Tomeasa; 268, 4 (ND4), tRNA-His, tRNA-Ser and tRNA-Leu genes 269 Baiu Mts C2; MAB301, MAB302, MAB305 -

1 Table 1. Origin of new mitochondrial ND4 sequences of Alpine newt individuals obtained in this study. Table 1. Origin of newly obtained mitochondrial ND4 sequences of Alpine newt individuals obtained. GenBank accession numbers will be submitted after manuscript acceptance.

ID Locality N E Haplotype GenBank accs. MPFC2190 Tricity Landscape Park, PL 54.383 18.531 C3-Sud MN211336 MPFC3220 Tricity Landscape Park, PL 54.383 18.531 C3-Sud MN211337 MPFC3230 Tricity Landscape Park, PL 54.385 18.531 C3-Sud MN211338 MPFC3232 Tricity Landscape Park, PL 54.395 18.516 C3-Sud MN211339 MPFC3233 Tricity Landscape Park, PL 54.395 18.516 C3-Sud MN211340 MPFC1037 Leszczawa Dolna, PL 49.665 22.442 C3-Carp MN211320 MPFC1038 Leszczawa Dolna, PL 49.665 22.442 C3-Carp MN211321 MPFC1283 Zawoja, PL 49.620 19.538 C3-Carp MN211325 MPFC2186 Tunel, PL 50.437 19.999 C3-Carp MN211332 MPFC2187 Tunel, PL 50.437 19.999 C3-Carp MN211333 MPFC2188 Pasmo Brzanki, PL 49.840 21.117 C3-Carp MN211334 MPFC2189 Pasmo Brzanki, PL 49.840 21.117 C3-Carp MN211335 MPFC705 Harz mts., DE 51.938 10.288 C3-Sud MN211341 MPFC1041 Harz mts., DE 51.906 10.333 C3-Sud MN211322 MPFC1042 Harz mts., DE 51.918 10.344 C3-Sud MN211323 MPFC1043 Harz mts., DE 51.908 10.330 C3-Sud MN211324 MPFC1854 Bradetelu, RO 46.751 25.108 C3-Carp MN211326 MPFC1872 Larga, RO 46.843 24.946 C3-Carp MN211327 MPFC1878 Lapusna, RO 46.771 25.271 C3-Carp MN211328 MPFC1919 Borzont, RO 46.731 25.360 C3-Carp MN211329 MPFC1977 Karlovice, CZ 50.098 17.433 C3-Sud MN211330 MPFC1978 Karlovice, CZ 50.098 17.433 C3-Sud MN211331 926 Joanna Jakóbik et al.

MAB314, MAB319 - MAB321, MAB338 - MAB340, central Germany (Harz mountains) and northern Czech MAB346, MAB348- MAB350, MAB363 - MAB365, Republic carried the C3-Sud haplotype; newts from MAB369, MAB370, MAB374 - MAB380, MAB382, southeastern and south-central Poland and northern MAB383, MAB388 - MAB390, MAB392 - MAB397, Romania had the C3-Carp haplotype. Two nucleotide MAB399 - MAB406, MAB409 - MAB416, MAB426, substitutions (p distance of 0.003) differentiate between MAB427, MAB430, MAB441, Ma1, Ma4, Ma7, the C3-Sud and C3-Carp haplotypes. As expected, the Ma13; KR107551, KR107552, KR107553, KR107554, obtained sequences clustered with other haplotypes KR107555, KR107556, KR107557. ClustalW of the C3 lineage and I. a. cyreni from northern Spain implemented in MEGA7 (Kumar et al., 2016) was (Fig. 1). The C3 + I. a. cyreni (C1) was sister to used for sequence alignment. A maximum likelihood I. a. apuana from Italy. Our topology reveals slight was constructed in MEGA7 using differences from that of Sotiroupoulos et al. (2007) the GTR+G model of nucleotide substitution; statistical and Recuero et al. (2014), the discrepancies stem from support for branches was assessed by 500 bootstrap our use of a single gene fragment as compared to the replicates. Nucleotide variation within lineage C3 was multilocus analyses of the previous authors. Network illustrated by constructing a phylogenetic network analysis revealed that most sampled newts across based on statistical parsimony in TCS v1.21 (Clement western and central Europe carry either the C3-Sud et al., 2000). (14 localities) or C3-Carp (19 localities) haplotypes, all other haplotypes have local distributions (i.e. found at a single locality). Results Our field surveys showed that Alpine newts are We obtained ND4 sequences (658-729 bp; for present in a small enclave of TLP along the Baltic coast network analysis we trimmed all sequences to 658 of Poland (Fig. 2). All breeding sites (Table 2, Fig. 3) bp) of 22 Alpine newts from 16 localities (Table 1) were small and mainly temporary water bodies, highly representing two haplotypes: all specimens from TLP, shaded, fishless, and generally overgrown by dense

Figure 2. Map of the southern part of Tricity Landscape Park showing the spatial distribution of 11 breeding sites of Alpine newts. Numbers refer to localities in Table 2. An Alpine newt Ichthyosaura alpestris population on the Baltic coast of Poland 927

Table 2. Habitat characteristics of Alpine newt breeding sites in the Tricity Landscape Park during April-August 2017. Alpine newt larvae were recorded at all sites. Area: approximate area of the pond during the survey; stability: describes whether the pond is known to dry out during periods of low rainfall; shade: describes the extent of shading of the site during the time of survey; # 2 ponds : number of ponds (with or without newts) within a 500 m perimeter of the focal site; aquatic veg.: describes the extent of submergedTable 2. Habitat vegetation characteristics in the ofpond Alpine at the newt time breeding of survey; sites in terrestrial the Tricity veg.: Landscape dominant Park duringtree species April-August in the 2017.immediate Alpine vicinity newt larvae of the were recorded at all sites. Area: siteapproximate (Aglu – area Alnus of the glutinosa pond during, Fsyl the –survey; Fagus stability sylvatica: describes, Psyl whether– Pinus the sylvestris, pond is known Sal –to Salixdry out sp.); during other periods sp.: of other low rainfall;amphibian shade species: describes the extent of shading of the site during the time of survey; # ponds: number of ponds (with or without newts) within a 500 m perimeter of the focal site; aquatic veg.: describes the extent of submerged breedingvegetation atin the pondsite (Lvulat the time– of survey; terrestrialvulgaris ,veg.: Rtemp dominant – Rana tree temporaria species in the). immediate vicinity of the site (Aglu – Alnus glutinosa, Fsyl – Fagus sylvatica, Psyl – Pinus sylvestris, Sal – Salix sp.); other sp.: other species breeding at the site (Lvul – Lissotriton vulgaris, Rtemp – Rana temporaria).

Site N E Area (m2) Stability Shade (%) # Ponds Aquatic veg. (%) Terrestrial veg. Other sp. 1 54.401 18.533 50 temporary 100 3 0 Aglu Lvul 2 54.401 18.533 50 temporary 100 3 0 Aglu Lvul 3 54.383 18.530 850 temporary 85 6 100 Fsyl, Psyl Lvul 4 54.384 18.527 250 temporary 100 6 0 Fsyl, Psyl Lvul, Rtemp 5 54.385 18.532 645 temporary 70 6 40 Fsyl, Psyl Lvul 6 54.384 18.504 850 temporary 40 0 95 Fsyl, Psyl Lvul 7 54.408 18.522 230 stable 50 0 0 Fsyl Lvul 8 54.395 18.517 1000 stable 100 4 50 Fsyl, Aglu - 9 54.395 18.516 700 temporary 70 4 100 Fsyl - 10 54.395 18.516 1600 temporary 60 4 100 Fsyl - 11 54.399 18.557 180 temporary 100 3 100 Fsyl , Sal Lvul

aquatic vegetation. The surveyed fragment of TLP has numerous ponds and water-filled depressions but is nonetheless poor in amphibian species.

Discussion Amphibian field surveys conducted in 2016 and 2017 in the Tricity Landscape Park revealed the presence of a disjunct population of Alpine newts (Fig. 1), demarcating the most northeastern occurrence of this species in Europe. The TLP locality is composed of at least 11 demes at breeding sites in close proximity to each other (Fig. 2): the local range of the species probably does not exceed 15 km2. Despite ongoing amphibian inventory and monitoring, Alpine newts have not been found in seemingly suitable habitat in other parts of the TLP, and in general have not been reported from northern Poland over the last century (Glowaciński and Sura, 2018). Nonetheless it is conceivable that other populations exist in the Pomeranian region of northern Poland, as many potential amphibian remain unsurveyed. It is noteworthy that the TLP population is adjacent to a large city harbouring several academic institutions, but has nonetheless remained undocumented until recently. The terrestrial habitat of Alpine newts in TLP is heavily Figure 3. Representative breeding habitat of the Alpine newt forested with a predominance of Fagus sylvatica and in TLP (top: site 4, photo M. Pabijan; bottom: site 5, photo P. Pinus sylvestris, complemented by pockets of Alnus Janowski). glutinosa in wetter areas. The packed forest stands of 928 Joanna Jakóbik et al. this area are generally over 100 years of age (Polish originated west of the Moravian Gate, a depression Forest Data Bank, https://www.bdl.lasy.gov.pl/portal/ dividing the distributions of these haplotypes. Potential mapy), and in most places form a closed canopy. Newt sources include southwestern Poland, Germany, Czech breeding sites in TLP are therefore heavily shaded and Republic, Austria or France. We are unaware of any are generally small, shallow and temporary (Table 2, intentional introductions of Alpine newts in TLP, Fig. 3). Most are probably natural depressions in the however, a zoological park (ZOO Gdańsk) occurs in clayey soil, although some may have been excavated by the immediate vicinity of the Alpine newt habitat in people. Many of these vernal pools dry out during late TLP, from which it could have been released into the summer, and may lack water until spring rains replenish wild. Because the Alpine newt occupies an area of at them (usually March). These characteristics limit the least 15 km2 and is by far the most abundant amphibian number of regularly breeding amphibian species in this species in the vernal pools it inhabits in the TLP, we forest fragment: besides Alpine newts, we only recorded think that it has persisted in this region for at least smooth newts and common at these sites. However, several generations, being possibly introduced in the they also limit the occurrence of predatory fish, with XXth century. none being noted in any of the newt breeding sites. The Alternatively, the Alpine newt population in TLP may rather dense network of potential breeding sites and lack be a relict of a previous, more widespread distribution of of dispersal barriers within the forest suggest that the lineage C3-Sud in the Holocene. Although this scenario Alpine newts exhibit metapopulation dynamics in this seems less likely due to the large gap between the TLP area. However, densely populated urban areas to the population and its continuous range to the south, climatic east, west and south of this forest fragment, including changes as well as deforestation could have fragmented a busy four lane highway demarcating the western and extirpated the intervening local populations in boundary, probably constitute an insurmountable barrier western and central Poland, leading to the disjunct to the dispersal of this species. distribution of today. The majority of Alpine newt populations at the northern margin of its range occur Provenance of Alpine newts from TLP.—The Alpine in beech or deciduous forest (Juszczyk, 1987; Bringsøe newt population in TLP is without doubt isolated from and Mikkelsen, 1997; Babik and Rafiński, 2001; Pabijan the nearest populations of the species located about et al., 2009; Głowaciński and Sura, 2018), including 350 km southwards, close to the city of Zielona Góra the population in TLP. Climatic and geomorphological (Glowaciński and Sura, 2018). Alpine newt populations conditions conducive to the development of beech from southwestern Poland are peripheral and tend to forest can therefore be considered as a proxy for the be small, forming a patchwork of more or less isolated presence of optimal habitat for the Alpine newt in this demes (Pabijan et al., 2005; Pabijan and Babik, 2006); area. Palaeoecological reconstructions based on cores larger populations, presumably more continuous, from peat deposits close to TLP at Stążki bog (ca. 30 occur to the south in the Sudetes mountain range. The km, Lamentowicz et al., 2008; Gałka et al., 2013) and disjunction exhibited by the population from TLP can Kusowskie Bagno bog (ca.150 km, Lamentowicz et be explained by two alternative historical scenarios: al., 2015) show that beech in the western Pomeranian (1) newts could have been introduced intentionally or region appeared between 200 BC – 100 AD and was nonintentionally by people or (2) the TLP population a dominant forest component until about 1900 AD, may be an autochthonous relict from a former, more with an abrupt but short-term decline around 1500 widespread distribution of this species. AD. These studies suggest that optimum conditions Our sequence data firmly place the TLP population in for the colonization of the southern Baltic coast by the C3 lineage that had colonized most of central and Alpine newts occurred relatively recently (i.e. in the western Europe after the last ice age, when climatic last two millennia) and that deforestation, first in the conditions had fragmented the range of the species to Middle Ages, and then in the XIX century, may have enclaves in southern Europe (Sotiropolous et al., 2007; caused the fragmentation and decline of populations in Recuero et al., 2014).Thus our diagnosis refutes a northern and central Poland. Although speculative, this human-mediated introduction of newts from any of the hypothesis finds backing in rather numerous documents southern subspecies or lineages. Moreover, because our from the XIX century (Bujack, 1837; Rathke 1846; mtDNA fragment differentiates between haplotypes C3- Conwentz, 1884; Treichel, 1889) recording Alpine Sud and C3-Carp (as well as other locally distributed C3 newt populations from the Pomeranian and Masurian haplotypes), the potential source population must have Lakelands of northern Poland (then the provinces of An Alpine newt Ichthyosaura alpestris population on the Baltic coast of Poland 929

East and West Prussia). These reports were later ruled as References misidentifications (see Berger et al., 1969 and references Arntzen, J.W., King, T.M., Denöel, M., Martínez-Solano, I., Wallis, therein), but we note that the authors questioning their G.P. (2016): Provenance of Ichthyosaura alpestris (: th validity worked in the early and mid XX century, ) introductions to France and New Zealand i.e. after deforestation and the potential demise of the assessed by mitochondrial DNA analysis. Herpetological Alpine newt in this area. In line with this idea, in a Journal 26: 49–56. checklist for East and West Prussia, Rathke Babik, W., Rafinski, J. (2001): Amphibian breeding site (1846) commented that this species was already “very characteristics in the , Poland. Herpetological Journal 11: 41–51. rare” (“sehr selten”) in the first half of the XIX century. Babik, W., Pabijan, M., Radwan, J. (2008): Contrasting patterns of th In light of the present results, we suggest that the XIX variation in MHC loci in the Alpine newt. Molecular century records require reconsideration. Interestingly, 17: 2339–2355. numerous sightings of the fire Salamandra Bell, B.D., Bell, A.P. (1995): Distribution of the introduced alpine salamandra exist for the southeastern part of the Baltic newt alpestris and of native Triturus species in north region (Litvinchuk 1996; Głowaciński and Sura, 2018), Shropshire, England. Australian Journal of Ecology 20: 367– far-removed from the continuous populations of this 375. Berger, L., Jaskowska, J., Młynarski, M. (1969): Amphibia et species in Central European mountain ranges. Similarly Reptilia, Catalogus Faunae Poloniae, XXXIX. Warszawa, to the Alpine newt, fire inhabit cool and Poland, Państwowe Wydawnictwo Naukowe. damp conditions associated with beech forest, and it has Böhme, M.U., Schneeweiss, N., Fritz, U., Moravec, J., Majláth, I., been suggested that since the Holocene Majláthová, V., Berendonk, T.U. (2006): Genetic differentiation climatic optimum could have led to the demise of a once and diversity of Lacerta viridis viridis (Laurenti, 1768) within the widespread and autochthonous salamander population northern part of its range: an investigation using mitochondrial in this area (Litvinchuk 1996). haplotypes. Salamandra, 42: 29–40. Bringsøe, H., Mikkelsen, U.S. (1997): Newt in progress: Status In conclusion, it is not yet clear whether the disjunct for Triturus alpestris in Denmark. Memoranda Societas Fauna locality of Alpine newts in TLP originated from a human- Flora Fennica, 73: 105–108. mediated introduction or is a relictual population from Bujack, J.G. (1837): Naturgeschichte der höheren Thiere mit a wider range during the late Holocene. In this respect, besonderer Berücksichtingung der Fauna Prussica, ein Handbuch our findings for the Alpine newt are similar to results für Lehrer der Jugend, Oekonomen, angehende Forstmänner for other ectothermic from isolated Central und Freunde der Natur. Königsberg 1837. European populations (e.g. , Böhme et al., 2006; Clement, M., Posada, D., Crandall, K. (2000): TCS: a computer program to estimate gene genealogies. Molecular Ecology 9: Jablonski et al., 2018) in which it was impossible to 1657–1660. distinguish between an autochthonous origin or human- Conwentz, H. (1884): Die einheimische Wierbelthier-Fauna. mediated introduction from geographically closest Schriften der Naturforschenden Gesellschaft in Danzig 6: 9–13. populations. Field studies dedicated to detecting Alpine Gałka, M., Miotk-Szpiganowicz, G., Goslar, T., Jęśko, M., van der newts in northern Poland, particularly in the remaining Knaap, W.O., Lamentowicz, M. (2013): Palaeohydrology, fires old stands of beech forest, could help in discerning and vegetation succession in the southern Baltic during the last between these two scenarios. A multilocus genetic 7500 years reconstructed from a raised bog based on multi-proxy data. Palaeogeography, Palaeoclimatology, Palaeoecology 370: analysis using highly variable markers (e.g. Palomar et 209–221. al., 2017) may be able to more precisely pinpoint the Głowaciński, Z., Sura P. (2018): Atlas of the Amphibians and potential origin of the newts from TLP. Reptiles of Poland. Status, Distribution, Conservation. Warszawa, Poland, Wydawnictwo Naukowe PWN SA. Acknowledgments. We thank the staff of the Tricity Landscape Jablonski, D., Gvoždík, V., Choleva, L., Jandzik, D., Moravec, J., Park for logistical support. This study was partly funded by Mačát, Z., Veselý, M. (2018): Tracing the maternal origin of the Jagiellonian University, Kraków (K/ZDS/007345). The handling common wall (Podarcis muralis) on the northern range and sampling of amphibians was done with permission from margin in Central Europe. Mitochondrion 46: 149-157. the General Directorate for Environmental Protection, permit Juszczyk, W. (1987): Płazy i gady krajowe, cz. 1-3. Wydawnictwo number DZP-WG. 6401.02.2.2018.kp.2, and the Regional Naukowe PWN, Warszawa. Directorate for Environmental Protection in Gdansk, permit Kumar, S., Stecher, G., Tamura, K. (2016): MEGA7: molecular number RDOŚ-Gd-PNII.6401.27.2016.AOL.1. in 2016, evolutionary genetics analysis version 7.0 for bigger datasets. RDOŚ-Gd-WZG.6401.26.2017.AOL.1. in 2017, RDOŚ-Gd- Molecular Biology and Evolution 33: 1870–1874. WZG.6401.33.2018.AOL.1. in 2018. Kurek, P., Święciak, T. (2010): The occurrence of the alpine newt Mesotriton alpestris on the Silesian Upland. Chrońmy Przyrodę Ojczystą 66: 57–60. 930 Joanna Jakóbik et al.

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Accepted by Spartak Litvinchuk