Triturus Macedonicus Research Articles

Extending the known vertical distribution for the highly adaptive Triturus macedonicus (Karaman, 1922)

Current knowledge of the vertical distribution of Triturus macedonicus places the species at altitudes up to 2140 m. Herein, we report its presence in the alpine lake Gistova at 2360 m on Mt. Grammos, Greece, thus extending the highest altitudinal point for the species, as well as for the Triturus genus. This record showcases the adaptive capacity of the Macedonian newt, which allows it to occupy markedly different environments. More populations may exist at similar or even higher altitudes and their discovery could positively impact conservation efforts.

Defining Evolutionary Conservation Units in the Macedonian Crested Newt, Triturus macedonicus (Amphibia; Salamandridae), in a Biodiversity Hotspot

In this study, we used genetic approaches to assess the conservation status of a protected amphibian species, the Macedonian crested newt, Triturus macedonicus, in Northern Pindos National Park (Epirus, Greece). Mitochondrial DNA sequences and multilocus genotypes of individuals from 38 breeding sites were used to infer their phylogenetic position and to detect and measure genetic variation patterns, population genetic structure, and levels of gene flow. The examined individuals fell within two major clades of the Macedonian crested newt phylogeny, being geographically separated by the Aoos River valley and Vikos Gorge. Both groups constitute separate gene pools, bearing private haplotypes and alleles, and the groups were found to be highly differentiated in both their mitochondrial and microsatellite markers. Thus, they meet all of the criteria needed to be characterized as evolutionary significant units (ESUs) that deserve a separate conservation status. Within each ESU, the particularly high inter-population differentiation and low migration rates imply a lack of connectivity between breeding sites and local isolation in favorable habitats. Our results can inform future programs and actions towards the conservation and management of the Macedonian crested newt in the area.

Consequences of hybridization on life history and growth in postmetamorphic Triturus macedonicus

Abstract The Macedonian crested newt T. macedonicus forms a complex hybrid zone with the Balkan crested newt T. ivanbureschi in the central part of the Balkan Peninsula. In this study, we compared life history parameters (sex ratio, survival rate and age of sexual maturation) and growth parameters (body length and mass) of T. macedonicus and T. macedonicus × T. ivanbureschi hybrids over the first three postmetamorphic years. Survival rates were high and similar for both genotypes. Sexual dimorphism in body size (length and mass) was evident at the beginning of the third postmetamorphic year, after the first breeding year, for both T. macedonicus and hybrids. We did not find clear and consistent difference in any of the analysed traits suggesting that hybridization with T. ivabureschi does not affect postmetamorphic growth and survival of T. macedonicus.

Infectious disease threats to amphibians in Greece: new localities positive for Batrachochytrium dendrobatidis.

In the early 2000s, numerous cases of European amphibian population declines and mass die-offs started to emerge. Investigating those events led to the discovery that wild European amphibians were confronted with grave disease threats caused by introduced pathogens, namely the amphibian and the salamander chytrid fungi Batrachochytrium dendrobatidis (Bd) and B. salamandrivorans (Bsal) and ranaviruses. In Greece, Bd was previously documented among wild amphibian populations in 2 different locations and 3 different species. However, no disease-related mass declines or mortality events have been reported. In this work, we build upon previous findings with new, subsequently obtained data, resulting in a 225-sample dataset of 14 species from 17 different locations throughout Greece, in order to examine the occurrence status of all 3 pathogens responsible for emerging infectious diseases in European amphibians. No positive samples for Bsal or ranavirus were recorded in any location. We confirmed the presence of Bd in 4 more localities and in 4 more species, including 1 urodelan (Macedonian crested newt Triturus macedonicus) and 1 introduced anuran (American bullfrog Lithobates catesbeianus). All insular localities were negative for Bd, except for Crete, where Bd was identified in 2 different locations. Again, no mass declines or die-offs were recorded in any Bd-positive area or elsewhere. However, given the persistence of Bd across Greece over the past ~20 yr, monitoring efforts should continue, and ideally be further expanded.

Extensive cytonuclear discordance in a crested newt from the Balkan Peninsula glacial refugium

Abstract Integration of multilocus data and species distribution modelling into phylogeography allows mitochondrial DNA (mtDNA)-based scenarios to be fine-tuned. We address the question of whether extensive mtDNA substructuring in the crested newt Triturus macedonicus from the Balkan Peninsula is matched in the nuclear genome. We determine the intraspecific population structure based on 52 nuclear DNA markers and project a species distribution model on climate layers for the Last Glacial Maximum. We show that T. macedonicus accumulated nuclear DNA population structure in an area predicted to have been climatically stable during the Pleistocene, with four nuclear DNA groups in the western part of the species range. The distribution of these nuclear DNA groups shows little agreement with that of mtDNA structuring, which shows three highly distinct species-specific clades and a fourth one introgressed from another crested newt species. This cytonuclear discordance conveys that historical biogeographical scenarios based on mtDNA exclusively should be interpreted with caution. Our findings further highlight the important role the Balkan Peninsula has played in the evolution and preservation of European biodiversity.

The Effect of Shelter on Oxidative Stress and Aggressive Behavior in Crested Newt Larvae (Triturus spp.).

Simple SummaryA lack of adequate refuge, even in the absence of predators, can alter the metabolism, levels of corticosteroids, growth and behavior in various groups of animals. Even though some of those processes (higher metabolic rates and activation of the stress response of the hypothalamic-pituitary-adrenal axis) in animals can result in enhanced reactive species (RS) production and oxidative stress, there is no study examining the effects of shelter on oxidative stress parameters. The results from this study showed that in crested newt larvae the absence of refuges altered their oxidative/antioxidative status and movement, but did not affect their aggressivity/injuries rate. Higher values of catalase, glutathione peroxidase, glutathione S-transferase and glutathione can indicate increased production of hydrogen peroxide in individuals without an adequate hiding place. This boosted antioxidant defense has a certain physiological cost that can be expressed in terms of the consumption of energy needed to maintain it as upregulated.Shelters are important for animal survival. Provision of adequate hiding places allow animals to express their natural sheltering behavior and it can have different positive effects on cortisol levels, physiological processes and mental performance. Although the absence of a refuge activates some stress response, its effect on oxidative stress has not been adequately examined. This study investigated whether the presence/absence of a shelter modifies the oxidative status (the antioxidant system and oxidative damage) and aggressive behavior of crested newt larvae (Triturus macedonicus and its hybrid with T. ivanbureschi). Our results show that individuals reared with shelters had lower values of the tested antioxidant parameters (catalase, glutathione peroxidase, glutathione S-transferase and glutathione), indicating a lower production of reactive species than individuals reared without shelter. The same pattern was observed in both T. macedonicus and its hybrid. Contrary to the activation of some physiological pathways, shelter availability did not significantly affect the rate of intraspecific aggressive behavior. The physiological benefits of shelter use can be manifested as a lower requirement for investment in the energy necessary for the maintenance of the upregulated antioxidant defenses, activation of repair systems and synthesis of endogenous antioxidants. This study highlights the importance of shelter provision, which may be valuable in habitat restoration and animal conservation studies.

Constrained by aliens, shifting landscape, or poor water quality? Factors affecting the persistence of amphibians in an urban pond network

Abstract Freshwater communities and especially pond‐breeding amphibians are extremely vulnerable to land‐use change, alien species introductions and the use of pesticides or other toxic chemicals, as reflected in their worldwide decline. Effective conservation and management of ponds requires a better understanding of the biotic and abiotic factors that shape diversity patterns and species distribution, especially in cases of habitat loss The present study aimed to reveal and classify which pond and landscape characteristics are the most important determinants for the occurrence patterns of amphibian species as well as for the overall amphibian species richness in an urban pond network. To achieve this aim, 17 biotic and abiotic variables were measured for 61 ponds and the dataset was analysed by means of a machine‐learning approach, suitability indices and co‐occurrence modelling The highest probability of Triturus macedonicus occurrence was found at fishless artificial and semi‐natural ponds. The persistence of Lissotriton graecus in ponds was predicted by high pond connectivity and the absence of fish reflecting the species dispersive potential. Pelophylax kurtmuelleri showed a higher probability of occurrence with increased pond connectivity and macrophyte cover. Amphibian species richness was higher in fishless ponds with well‐oxygenated waters located in sites with a low to intermediate road density network. Ponds categorized as ‘average’ in terms of newt suitability were more frequently inhabited by fish than Triturus macedonicus. Alien fish species showed negative associations with both newt species, while both newt species showed a positive association between them. The results support the view that conserving a greater number of water bodies with increased landscape connectivity and without any fish presence could provide amphibian species with alternative habitat choices, especially in sites with human pressure where pond stability is at stake owing to continuous landscape changes.

Reproductive characteristics of two Triturus species (Amphibia: Caudata)

During three consecutive years, we compared the reproductive characteristics and oviposition dynamics of two crested newt species, Triturus ivanbureschi and T. macedonicus. These two well-defined species are of special interest because of complex interactions at their contact zone, which include hybridization, species replacement and asymmetric mitochondrial DNA introgression. In common garden experiments, females were introduced to conspecific males, to males of other species and to hybrid males. We monitored the total number and size of the deposited eggs, as well as the dynamics and duration of oviposition during three consecutive years. The number of deposited eggs of Triturus species was much higher than previously reported. Triturus macedonicus lay more eggs that were of larger size in comparison to T. ivanbureschi. Also, the onset of T. macedonicus oviposition was considerably delayed compared to T. ivanbureschi. These are fundamental data, important for understanding complex species interactions in their contact zones.

Oxidative cost of interspecific hybridization: a case study of two Triturus species and their hybrids.

Oxidative stress has most recently been suggested as one of the possible mechanisms responsible for reduced fitness of hybrids. To explore possible oxidative cost of hybridization, we examined anti-oxidant defence system parameters (superoxide dismutase, catalase, glutathione peroxidase, glutathione S-transferase, glutathione reductase, glutathione, SH groups), their interconnectedness (index of integration) and levels of oxidative damage [concentrations of lipid peroxides, TBARS (thiobarbituric acid reactive substances)] in laboratory-reared newt species, Triturus macedonicus and T. ivanbureschi, and their hybrid. Our results showed that parental species differed in anti-oxidant defence system parameters, but not in the levels of integration of the whole system and oxidative damage. Individuals of T. ivanbureschi had higher activities of superoxide dismutase, glutathione S-transferase and concentrations of glutathione. Hybrid individuals of crested newts displayed higher levels of the anti-oxidant defence system (higher superoxide dismutase, catalase, glutathione peroxidase activities and concentrations of SH groups), and a lower overall correlation of anti-oxidant system (lower index of integration) in comparison with both parental species, suggesting that they may possess a less efficient anti-oxidant defence system and a higher investment in maintaining oxidative balance. The higher investment in the anti-oxidant system could divert limited resources away from other functions and affect further hybrid fitness. The presented findings contribute to a better understanding of the anti-oxidant defence system of crested newts and their interspecies differences, and support the hypothesis that oxidative stress is one of the costs of interspecific hybridization.

Evidence for paedomorphosis in Macedonian crested newt (Triturus macedonicus) from Montenegro

One of the most interesting heterochronic processes in the European newt (genus Triturus) is paedomorphosis, a phenomenon of attaining reproductive maturity while retaining larval features. However, paedomorphosis seems to be rare in the crested newt (Triturus cristatus superspecies). In the locality Vrba, near the town of Tuzi (Montenegro), Dr Georg Džukic found paedomorphic Macedonian crested newt (Triturus macedonicus). In this paper, new evidence for this heterochronic process in the crested newt from Montenegro is reported.

Fine scale spatial genetic structure of two syntopic newts across a network of ponds: implications for conservation

In this study we used genetic approaches to assess the influence of landscape features on the dispersal patterns and genetic structure of two newt species (Triturus macedonicus and Lissotriton vulgaris) living syntopically in a network of ponds. Multilocus genotypes were used to detect and measure genetic variation patterns, population genetic structure and levels of gene flow. We interpret results on the basis of the different dispersal properties of the two species and explored the influence of certain landscape features, such as road and channel networks, on population connectivity. We found marked differences in the spatial genetic patterns of the respective species, which can be explained by their different dispersal properties. The road network seems to act as a barrier to dispersal in the overland dispersing L. vulgaris, while the channel network maintains connectivity in the aquatic dispersing T. macedonicus. The simultaneous and comparative consideration of species in a given area offers a much better understanding of the processes that govern population dynamics and persistence, providing valuable knowledge useful in conservation and management design.

Postglacial species displacement in Triturus newts deduced from asymmetrically introgressed mitochondrial DNA and ecological niche models.

BackgroundIf the geographical displacement of one species by another is accompanied by hybridization, mitochondrial DNA can introgress asymmetrically, from the outcompeted species into the invading species, over a large area. We explore this phenomenon using the two parapatric crested newt species, Triturus macedonicus and T. karelinii, distributed on the Balkan Peninsula in south-eastern Europe, as a model.ResultsWe first delimit a ca. 54,000 km2 area in which T. macedonicus contains T. karelinii mitochondrial DNA. This introgression zone bisects the range of T. karelinii, cutting off a T. karelinii enclave. The high similarity of introgressed mitochondrial DNA haplotypes with those found in T. karelinii suggests a recent transfer across the species boundary. We then use ecological niche modeling to explore habitat suitability of the location of the present day introgression zone under current, mid-Holocene and Last Glacial Maximum conditions. This area was inhospitable during the Last Glacial Maximum for both species, but would have been habitable at the mid-Holocene. Since the mid-Holocene, habitat suitability generally increased for T. macedonicus, whereas it decreased for T. karelinii.ConclusionThe presence of a T. karelinii enclave suggests that T. karelinii was the first to colonize the area where the present day introgression zone is positioned after the Last Glacial Maximum. Subsequently, we propose T. karelinii was outcompeted by T. macedonicus, which captured T. karelinii mitochondrial DNA via introgressive hybridization in the process. Ecological niche modeling suggests that this replacement was likely facilitated by a shift in climate since the mid-Holocene. We suggest that the northwestern part of the current introgression zone was probably never inhabited by T. karelinii itself, and that T. karelinii mitochondrial DNA spread there through T. macedonicus exclusively. Considering the spatial distribution of the introgressed mitochondrial DNA and the signal derived from ecological niche modeling, we do not favor the hypothesis that foreign mitochondrial DNA was pulled into the T. macedonicus range by natural selection.

The evolution of the adult body form of the crested newt (Triturus cristatussuperspecies, Caudata, Salamandridae)

We characterized the adult body form of the crested newt (Triturus cristatus superspecies) and explored its evolution. From seven morphometric traits, we determined that body size, interlimb distance and head width define the body form. None of the morphometric traits showed a phylogenetic signal. Three body-shape morphotypes (Triturus dobrogicus + T. cristatus, Triturus carnifex + Triturus macedonicus and Triturus karelinii + Triturus arntzeni) and three body-size morphotypes (T. dobrogicus, T. cristatus and all other crested newts) could be recognized. The ancestral phenotype (a large body with a short trunk and a wide head) characterized T. karelinii and T. arntzeni. Triturus carnifex and T. macedonicus had a somewhat different phenotype (large body and wide head, accompanied by mild body elongation). The most derived phenotype included body size reduction and more pronounced body elongation in T. cristatus and, especially, in T. dobrogicus. Body elongation occurred by trunk lengthening but not head and tail lengthening. Additionally, contrary to other tetrapods, evolutionary axis elongation in crested newts was followed by a decrease in body size. We advocate the hypothesis that ecology drives the evolution of body form in crested newts.

A multi-scale approach to facultative paedomorphosis of European newts (Salamandridae) in the Montenegrin karst: Distribution pattern, environmental variables, and conservation

Facultative paedomorphosis, a process in which newt larvae can opt for reproduction before or after metamorphosis, is geographically heterogeneous. Despite numerous ecological studies and recent evidence of declines in paedomorphic populations, however, no attempt to model environmental variables that explain the presence of paedomorphs has been made at a multi-scale level. Our aim was to fill this gap in studying three newt species ( Lissotriton vulgaris, Mesotriton alpestris, and Triturus macedonicus) of the Montenegrin karst as model species. To this end, we used multivariate analysis on three scales of habitat: the breeding pond, the land use and the climatologic features. Results show that the study area is both an important hotspot for paedomorphosis and where intraspecific diversity is quickly disappearing (20–47% extirpation) because of fish introductions. Other habitat variables (water permanency, pH or the habitat origin) were shown to act on paedomorphosis but not consistently across species, confirming complexity of the evolutionary and ecological processes. This study appeals for more long-term and detailed landscape studies of polyphenisms, a neglected but promising topic, to better understand and protect alternative modes of development. Particularly, measures should be taken to identify hotspots of intraspecific diversity at a global scale and stop fish introductions before we reach a point of no-return.

New polymorphic microsatellite loci for the Macedonian crested newt, Triturus macedonicus, and cross‐priming testing in four other crested newt species

Eleven microsatellites have been characterized for Triturus macedonicus. Nine loci showed different variation patterns in a sample of 40 individuals from a single breeding pond in Zagori province (Greece), with an average number of 4.2 alleles per locus and an expected heterozygosity ranging from 0.1199 to 0.8079. Distinct cross-priming amplification rates were recovered on four additional crested newt species. Two monomorphic T. macedonicus loci were polymorphic in other Triturus species. The microsatellites developed herein could be a useful intraspecific genetic tool to undertake fine-scale population genetic analyses as well as in the study of contact zones between crested newt species.

Sexual size and shape evolution in European newts (Amphibia: Caudata: Salamandridae) on the Balkan Peninsula

We used a phylogenetic perspective in an examination of the direction and extent of sexual dimorphism in body size and body shape in European newts from the Balkan Peninsula (alpine newts, Mesotriton alpestris; crested newts, Triturus cristatus superspecies; smooth newts, Lissotriton vulgaris). We found a strong, female-biased sexual size dimorphism (SSD) in the analysed clades of alpine newt, whereas within crested newts we found a less stringent female-biased SSD in Triturus carnifex, Triturus macedonicus and Triturus karelinii, and no significant SSD in T. cristatus or Triturus dobrogicus. Among the smooth newts, we found male-biased SSD in Lissotriton vulgaris vularis and Lissotriton vulgaris greacus and no SSD in Lissotriton vulgaris meridionalis. Most of these newts also exhibit a significant sexual dimorphism in body shape, which varied more randomly than body size, regardless of SSD level. Female and male body size as well as the degree of SSD displayed statistically significant phylogenetic signal, while sexual dimorphism in body shape was phylogenetically independent. The relationship between independent contrast data for female size and male size indicated that SSD in European newts could be driven by a disproportionate increase in female size as increase in female size was not accompanied by a proportional increase in male size.

Distribution patterns and environmental determinants of European newts in the Montenegrin karst area

The distribution patterns of three European newt species (Triturus macedonicus, Lissotriton vulgaris, and Mesotriton alpestris) was determined in the Montenegrin karst region and the influence of breeding ponds, the adjacent landscape, and climate variables on distributions within the framework of allotopy vs. syntopy occurrences was tested. After surveying 145 newt breeding sites in a 10,080 km2 area, the ranges of T. macedonicus and M. alpestris were found not come into contact, while L. vulgaris, the most widely distributed species, covers the entire range of T. macedonicus and the lower part of the range of M. alpestris. Using Akaike’s Information Criterion (AIC) to evaluate the effectiveness of 70 a priori models in predicting the presence of the three newt species, it was found that: (1) elevation is the most important variable in predicting newt presence; (2) only a few elevation-dependent variables can replace elevation in the building of a good descriptive model; (3) precipitation in combination with elevation contributes significantly to the building of a better descriptive model; and (4) in comparison with allotopy, syntopy mostly occurs at intermediate elevation ranges of species in sympatry.

Breeding site traits of European newts (Triturus macedonicus, Lissotriton vulgaris, and Mesotriton alpestris: Salamandridae) in the Montenegrin karst region

We recorded the occurrence of three European newt species - the smooth newt (Lissotriton vulgaris), the eastern alpine crested newt (Triturus macedonicus), and the alpine newt (Mesotriton alpestris) - in the Montenegrin karst, as well as their breeding site characteristics. In terms of long-lasting breeding site numbers and occupation rate, the most common species is the smooth newt, followed by the alpine newt and the crested newt. The examined water bodies with?out newts showed no significant differences of aquatic habitat characteristics compared to water bodies with newts. The factors that explained most of the observed variation in newt breeding site traits were the habitat category and habitat origin. The alpine newt primarily inhabits natural lakes, while the crested newt inhabits artificial breeding sites such as lithotelma and ubao. The smooth newt is less choosy and occurs in different types of natural and artificial habitats. The aquatic requirements of Montenegrin newt species do not differ substantially in many respects from requirements of the core species range populations.