Nikolay Tzankov

Phylogeography and cryptic variation within the Lacerta viridis complex (Lacertidae, Reptilia)

It is well known that the current genetic pattern of many European species has been highly influenced by climatic changes during the Pleistocene. While there are many well known vertebrate examples, knowledge about squamate reptiles is sparse. To obtain more data, a range‐wide sampling of Lacerta viridis was conducted and phylogenetic relations within the L. viridis complex were analysed using an mtDNA fragment encompassing part of cytochrome b, the adjacent tRNA genes and the noncoding control region. Most genetic divergence was found in the south of the distribution range. The Carpathian Basin and the regions north of the Carpathians and Alps are inhabited by the same mitochondrial lineage, corresponding to Lacerta viridis viridis. Three distinct lineages occurred in the south‐eastern Balkans — corresponding to L. v. viridis, L. v. meridionalis, L. v. guentherpetersi— as well as a fourth lineage for which no subspecies name is available. This distribution pattern suggests a rapid range expansion of L. v. viridis after the Holocene warming, leading to a colonization of the northern part of the species range. An unexpected finding was that a highly distinct genetic lineage occurs along the western Balkan coast. Phylogenetic analyses (Bayesian, maximum likelihood, maximum parsimony) suggested that this west Balkan lineage could represent the sister taxon of Lacerta bilineata. Due to the morphological similarity of taxa within the L. viridis complex this cryptic taxon was previously assigned to L. v. viridis. The distribution pattern of several parapatric, in part highly, distinct genetic lineages suggested the existence of several refuges in close proximity on the southern Balkans. Within L. bilineata sensu stricto a generally similar pattern emerged, with a high genetic diversity on the Apennine peninsula, arguing for two distinct refuges there, and a low genetic diversity in the northern part of the range. Close to the south‐eastern Alps, three distinct lineages (L. b. bilineata, L. v. viridis, west Balkan taxon) occurred within close proximity. We suggest that the west Balkan lineage represents an early offshoot of L. bilineata that was isolated during a previous Pleistocene glacial from the more western L. bilineata populations, which survived in refuges on the Apennine peninsula.

Tracing glacial refugia of Triturus newts based on mitochondrial DNA phylogeography and species distribution modeling

IntroductionThe major climatic oscillations during the Quaternary Ice Age heavily influenced the distribution of species and left their mark on intraspecific genetic diversity. Past range shifts can be reconstructed with the aid of species distribution modeling and phylogeographical analyses. We test the responses of the different members of the genus Triturus (i.e. the marbled and crested newts) as the climate shifted from the previous glacial period (the Last Glacial Maximum, ~21 Ka) to the current interglacial.ResultsWe present the results of a dense mitochondrial DNA phylogeography (visualizing genetic diversity within and divergence among populations) and species distribution modeling (using two different climate simulations) for the nine Triturus species on composite maps.ConclusionsThe combined use of species distribution modeling and mitochondrial phylogeography provides insight in the glacial contraction and postglacial expansion of Triturus. The combined use of the two independent techniques yields a more complete understanding of the historical biogeography of Triturus than both approaches would on their own. Triturus newts generally conform to the ‘southern richness and northern purity’ paradigm, but we also find more intricate patterns, such as the absence of genetic variation and suitable area at the Last Glacial Maximum (T. dobrogicus), an ‘extra-Mediterranean’ refugium in the Carpathian Basin (T. cristatus), and areas where species displaced one another postglacially (e.g. T. macedonicus and western T. karelinii). We provide a biogeographical scenario for Triturus, showing the positions of glacial refugia, the regions that were postglacially colonized and the areas where species displaced one another as they shifted their ranges.

Contrasting evolutionary histories of the legless lizards slow worms (Anguis) shaped by the topography of the Balkan Peninsula

BackgroundGenetic architecture of a species is a result of historical changes in population size and extent of distribution related to climatic and environmental factors and contemporary processes of dispersal and gene flow. Population-size and range contractions, expansions and shifts have a substantial effect on genetic diversity and intraspecific divergence, which is further shaped by gene-flow limiting barriers. The Balkans, as one of the most important sources of European biodiversity, is a region where many temperate species persisted during the Pleistocene glaciations and where high topographic heterogeneity offers suitable conditions for local adaptations of populations. In this study, we investigated the phylogeographical patterns and demographic histories of four species of semifossorial slow-worm lizards (genus Anguis) present in the Balkan Peninsula, and tested the relationship between genetic diversity and topographic heterogeneity of the inhabited ranges.ResultsWe inferred phylogenetic relationships, compared genetic structure and historical demography of slow worms using nucleotide sequence variation of mitochondrial DNA. Four Anguis species with mostly parapatric distributions occur in the Balkan Peninsula. They show different levels of genetic diversity. A signature of population growth was detected in all four species but with various courses in particular populations. We found a strong correlation between genetic diversity of slow-worm populations and topographic ruggedness of the ranges (mountain systems) they inhabit. Areas with more rugged terrain harbour higher genetic diversity.ConclusionsPhylogeographical pattern of the genus Anguis in the Balkans is concordant with the refugia-within-refugia model previously proposed for both several other taxa in the region and other main European Peninsulas. While slow-worm populations from the southern refugia mostly have restricted distributions and have not dispersed much from their refugial areas, populations from the extra-Mediterranean refugia in northern parts of the Balkans have colonized vast areas of eastern, central, and western Europe. Besides climatic historical events, the heterogeneous topography of the Balkans has also played an important role in shaping genetic diversity of slow worms.

Envenoming following bites by the Balkan adder Vipera berus bosniensis – First documented case series from Bulgaria

We report the first detailed accounts of bites by the Balkan adder, Vipera berus bosniensis from Bulgaria. Documentation of bites by this subspecies is very rare in the literature and most available accounts are from the northern limit of its distribution. V. berus bosniensis is considered to possess neurotoxic venom but little evidence has hitherto been available to support this supposition. In this case series symptoms typical of adder bites developed including oedema, nausea, dizziness, lymphangitis, vomiting, and diarrhoea together with aberrant symptoms such as diplopia and ptosis that confirm the presence of neurotoxic venom in Balkan adders. In addition, unusual and atypical symptoms of adder bites such as painless bites and muscle cramps appeared. The inadequate treatment in hospital and the remote habitats in which this species is encountered are potential sources of complication.

Feeding behaviour in a ‘basal’ tortoise provides insights on the transitional feeding mode at the dawn of modern land turtle evolution

Almost all extant testudinids are highly associated with terrestrial habitats and the few tortoises with high affinity to aquatic environments are found within the genus Manouria. Manouria belongs to a clade which forms a sister taxon to all remaining tortoises and is suitable as a model for studying evolutionary transitions within modern turtles. We analysed the feeding behaviour of Manouria emys and due to its phylogenetic position, we hypothesise that the species might have retained some ancestral features associated with an aquatic lifestyle. We tested whether M. emys is able to feed both in aquatic and terrestrial environments. In fact, M. emys repetitively tried to reach submerged food items in water, but always failed to grasp them—no suction feeding mechanism was applied. When feeding on land, M. emys showed another peculiar behaviour; it grasped food items by its jaws—a behaviour typical for aquatic or semiaquatic turtles—and not by the tongue as generally accepted as the typical feeding mode in all tortoises studied so far. In M. emys, the hyolingual complex remained retracted during all food uptake sequences, but the food transport was entirely lingual based. The kinematical profiles significantly differed from those described for other tortoises and from those proposed from the general models on the function of the feeding systems in lower tetrapods. We conclude that the feeding behaviour of M. emys might reflect a remnant of the primordial condition expected in the aquatic ancestor of the tortoises.

A comparative study of the mating call of Pelophylax ridibundus and Pelophylax kurtmuelleri (Anura: Ranidae) from syntopic and allotopic populations

The mating call of Pelophylax ridibundus and Pelophylax kurtmuelleri from three syntopic and five allotopic populations was studied. All recordings were made during the mating season. We tested the hypothesis that the call of P. ridibundus from allotopic populations would differ from that in syntopic populations with P. kurtmuelleri. For the comparative analysis we used univariate and multivariate statistical methods and the following characteristics: call duration, pulse group duration, pulse group interval, pulse group period, pulse group number, minimum, maximum, fundamental and dominant frequencies and pulse energy. Analyses reveal a trait divergence in most of the studied characteristics of the signals, with stronger differentiation between P. kurtmuelleri and P. ridibundus in syntopic populations than between the two species in allotopic ones. The possible evolutionary implications of this divergence are discussed.

Are species genetically more sensitive to habitat fragmentation on the periphery of their range compared to the core? A case study on the sand lizard (Lacerta agilis)

ContextSpecies show different sensitivity to habitat loss and fragmentation depending on their specialization. Populations of a species at the range margin are generally assumed to be more stenoecious than populations at the core of the distribution and should therefore be more sensitive to habitat fragmentation.ObjectivesWe evaluated the hypothesis that fragmentation effects species more strongly at the range periphery of their range compared to the core, resulting in lower genetic variability in comparable patch sizes and lower gene flow among populations.MethodsWe compared the genetic diversity and structure of five sand lizard (Lacerta agilis) populations at the margin of its range in Bulgaria and of 11 populations at the core of its distribution in Germany. We based the analysis on microsatellites, comprising 15 loci in Bulgaria and 12 in Germany.ResultsAll diversity indices declined with patch size. For medium-sized patches all diversity indices were lower at the range periphery compared to the core, with two of them being significant. AICc based model selection showed strong support for core/periphery and patch size effects for observed and expected heterozygosity but only a patch size effect for allelic richness. There was no isolation-by-distance and each sampled population was allocated to a separate cluster with high probability for both countries, indicating that all populations are (almost) completely isolated.ConclusionOur study indicates an increased sensitivity of a species to fragmentation at the periphery compared to the core of its distribution. This differential sensitivity should be accounted for when prioritizing species based on their fragmentation sensitivity in landscape management.

Contributions to the functional morphology of caudate skulls: kinetic and akinetic forms

A strongly ossified and rigid skull roof, which prevents parietal kinesis, has been reported for the adults of all amphibian clades. Our μ-CT investigations revealed that the Buresch’s newt (Triturus ivanbureschi) possess a peculiar cranial construction. In addition to the typical amphibian pleurokinetic articulation between skull roof and palatoquadrate associated structures, we found flexible connections between nasals and frontals (prokinesis), vomer and parasphenoid (palatokinesis), and between frontals and parietals (mesokinesis). This is the first description of mesokinesis in urodelans. The construction of the skull in the Buresch’s newts also indicates the presence of an articulation between parietals and the exocipitals, discussed as a possible kind of metakinesis. The specific combination of pleuro-, pro-, meso-, palato-, and metakinetic skull articulations indicate to a new kind of kinetic systems unknown for urodelans to this date. We discuss the possible neotenic origin of the skull kinesis and pose the hypothesis that the kinesis in T. ivanbureschi increases the efficiency of fast jaw closure. For that, we compared the construction of the skull in T. ivanbureschi to the akinetic skull of the Common fire salamander Salamandra salamandra. We hypothesize that the design of the skull in the purely terrestrial living salamander shows a similar degree of intracranial mobility. However, this mobility is permitted by elasticity of some bones and not by true articulation between them. We comment on the possible relation between the skull construction and the form of prey shaking mechanism that the species apply to immobilize their victims.

On the amphibious food uptake and prey manipulation behavior in the Balkan-Anatolian crested newt (Triturus ivanbureschi, Arntzen and Wielstra, 2013).

Feeding behavior in salamanders undergoing seasonal habitat shifts poses substantial challenges caused by differences in the physical properties of air and water. Adapting to these specific environments, urodelans use suction feeding predominantly under water as opposed to lingual food prehension on land. This study aims to determine the functionality of aquatic and terrestrial feeding behavior in the Balkan-Anatolian crested newt (Triturus ivanbureschi) in its terrestrial stage. During the terrestrial stage, these newts feed frequently in water where they use hydrodynamic mechanisms for prey capture. On land, prey apprehension is accomplished mainly by lingual prehension, while jaw prehension seems to be the exception (16.67%) in all terrestrial prey capture events. In jaw prehension events there was no detectable depression of the hyo-lingual complex. The success of terrestrial prey capture was significantly higher when T. ivanbureschi used lingual prehension. In addition to prey capture, we studied the mechanisms involved in the subduction of prey. In both media, the newts frequently used a shaking behavior to immobilize the captured earthworms. Apparently, prey shaking constitutes a significant element in the feeding behavior of T. ivanbureschi. Prey immobilization was applied more frequently during underwater feeding, which necessitates a discussion of the influence of the feeding media on food manipulation. We also investigated the osteology of the cranio-cervical complex in T. ivanbureschi to compare it to that of the predominantly terrestrial salamandrid Salamandra salamandra.

Sexual Size Dimorphism in Genus Triturus Rafinesque, 1815 (Amphibia: Salamandridae) in Bulgaria—Preliminary Results

ABSTRACT We studied a sexual size dimorphism (SSD) in three species from the genus Triturus. We used fifteen body measures for calculating the SSD. We performed the multivariate SSD index to calculate the dimorphism in overall body size. We found a well express female-biased SSD in T. karelinii, and male-biased in T. macedonicus. In T. cristatus we also found a male-biased SSD but weakly pronounced. In general SSD possess similar values but is express by different traits. The tendencies of displaying the sexual dimorphism in all the species are also similar. It is still not clear which factors affect the sexual dimorphism in newts.