Triturus Vulgaris Research Articles

Assessing the Use of Artificial Hibernacula by the Great Crested Newt (Triturus cristatus) and Smooth Newt (Lissotriton vulgaris) in Cold Climate in Southeast Norway

Construction of artificial overwintering habitats, hibernacula, or newt hotels, is an important mitigation measure for newt populations in urban and agricultural areas. We have monitored the use of four artificial hotels built in September 2011 close to a 6000 m2 breeding pond in Norway. The four hotels ranged from 1.6 to 12.4 m3 and were located from 5 to 40 m from the breeding pond. In 2013–2015, 57 Great Crested Newts (Triturus cristatus) and 413 Smooth Newts (Lissotriton vulgaris) spent the winter in the hotels. The proportions of juveniles were 75% and 62%, respectively, and the hotels may be important to secure recruitment. Knowledge on emigration routes and habitat quality for summer use and winter hibernation is important to find good locations for newt hotels. The study documented that newts may survive a minimum temperature of −6.7 °C. We recommend that newt hotels in areas with harsh climate are dug into the ground in slopes to reduce low-temperature exposure during winter.

An improved microsatellite panel to assess genetic variability of the Italian smooth newt (Lissotriton vulgaris meridionalis)

Habitat loss and fragmentation are threatening amphibians by increasing population isolation. However, artificial waterbodies created for livestockmay contrast this phenomenon by providing surrogate habitats for amphibians. Here, we performed a genetic study on an amphibian species, Lissotriton vulgaris meridionalis, in a rural area in central Italy where natural wetlands are disappearing and drinking troughs for cattle watering are widespread. Specifically, we tested a panel of microsatellite markers to identify a suitable tool for addressing conservation genetic issues of this species that is undergoing severe local decline. Twelve of the 20 tested loci produced reliable amplifications and were polymorphic. Three distinct units with a low level of gene flow were distinguished and the population genetic structuring overlapped with geographic distribution. Such loci will be useful to assess the genetic diversity of the species across multiscale levels for its management and conservation.

Egg-laying environment modulates offspring responses to predation risk in an amphibian.

Predator-induced plasticity has been in the focus of evolutionary ecological research in the last decades, but the consequences of temporal variation in the presence of cues predicting offspring environment have remained controversial. This is partly due to the fact that the role of early environmental effects has scarcely been scrutinized in this context while also controlling for potential maternal effects. In this study, we investigated how past environmental conditions, that is different combinations of risky or safe adult (prenatal) and oviposition (early post-natal) environments, affected offspring's plastic responses in hatching time and locomotor activity to predation risk during development in the smooth newt (Lissotriton vulgaris). We found that females did not adjust their reproductive investment to the perceived level of risk in the adult environment, and this prenatal environment had generally negligible effect on offspring phenotype. However, when predator cues were absent during oviposition, larvae raised in the presence of predator cues delayed their hatching and exhibited a decreased activity compared to control larvae developing without predator cues, which responses are advantageous when predators pose a threat to hatched larvae. In the presence of predator cues during oviposition, the difference in hatching time persisted, but the difference in general locomotor activity disappeared between risk-exposed and control larvae. Our findings provide clear experimental evidence that fine-scale temporal variation in a predictive cue during and after egg-laying interactively affects offspring phenotype, and highlight the importance of the early post-natal environment, which may exert a substantial influence on progeny's phenotype also under natural conditions.

Major biological activities and protein profiles of skin secretions of Lissotriton vulgaris and Triturus ivanbureschi

Abstract Objective The aim of this study was to determine the total protein amounts, protein profiles, in vitro cytotoxicities, antimicrobial activities and hemolytic effects of skin secretions of the Lissotriton vulgaris and Triturus ivanbureschi. Methods Skin secretions were obtained, clarified, supernatants snap-frozen then lyophilized. Total protein amounts were determined by BCA assay kit. Protein profiles were revealed by the SDS-PAGE. The cytotoxicity and antimicrobial activity were determined by using MTT assay and minimum inhibitory concentration (MIC) method. Hemolytic effects were measured on rabbit red blood cells. Results Lissotriton vulgaris and T. ivanbureschi skin secretions have totally 18 and 20 protein fractions. IC50 values were detected between 1.40 and 40.28 μg/mL. The MIC results were found between 7.8 and 250 μg/mL. Lissotriton vulgaris skin secretion showed low hemolytic effect while T. ivanbureschi skin secretion showed high hemolytic effect. Conclusion This study is the first report showing the potential of L. vulgaris and T. ivanbureschi skin secretions for cytotoxicity, antimicrobial and hemolytic activity as an alternative therapeutic approach for traditional uses. Further studies need to focus on purification of the active components from these skin secretions and mode of action on cancer cell lines and microorganisms as anti-agents.

Breeding-migration patterns and reproductive dynamics of two syntopic newt species (Amphibia, Salamandridae) at a temporary pond in southern Greece

Amphibian breeding-migrations are strongly influenced by environmental conditions. Furthermore, an individual’s breeding-migration pattern might be influenced by their sex, size, physiological state, and/or fecundity. From 2012 to 2014, we collected mark–recapture data for two syntopic alpine (Ichthyosaura alpestris) and smooth (Lissotriton vulgaris) newt populations in southern Greece (the southern limit of their distribution), as well as morphometric data for individuals, and data on environmental factors. Using these data, we studied the reproductive dynamics of these populations and assessed whether individual traits and environmental covariates influence their breeding-migration patterns. Both species followed a similar temporal migration pattern, regardless of individual traits. Migration to the breeding site was influenced by water availability, but not by ground temperature, rainfall or photoperiod. Migration from the breeding site was not influenced by water availability, but was triggered by increasing water temperatures. Our findings corroborate the few available studies of European newt populations occurring near the southern edge of their distribution. However, compared to numerous studies of northern European newt populations, our results suggest distinct breeding-migration patterns, as well as contrasting effects of environmental factors on these migration patterns. Based on current climate change projections, the breeding period of southern newt populations breeding in temporary ponds could be severely reduced.

Helminth endoparasites of the smooth newt Lissotriton vulgaris: linking morphological identification and molecular data.

The helminth endoparasites of many European amphibian species are often known exclusively from morphological descriptions. A molecular library of DNA sequence data linked to morphological identifications is still in its infancy. In this paper, we aim to contribute to such a library on the smooth newt Lissotriton vulgaris, the intermediate and definitive host of 31 helminth parasites, according to evidence published so far. Newts (n = 69) were collected at two study sites in western Germany and examined for the presence of helminths. A total of five helminth species were detected in 56 (81%) of the newts, but only one or two species infected a single host. Four out of five helminth species were identified morphologically and based on DNA sequences as Parastrigea robusta (metacercariae), Oswaldocruzia filiformis, Megalobatrachonema terdentatum (adults and larvae) and Cosmocerca longicauda, and the corresponding sequences were provided subsequently. Oswaldocruzia molgeta was confirmed to be a junior synonym of O. filiformis. Molecular data on a fifth species (a cosmocercid nematode) that could not be identified at species level were added to GenBank. These findings increased the molecular library on morphologically identified smooth newt parasites significantly, from 12 to 15 entries.

The distributions of the six species constituting the smooth newt species complex (Lissotriton vulgaris sensu lato and L. montandoni) – an addition to the New Atlas of Amphibians and Reptiles of Europe

Abstract The ‘smooth newt’, the taxon traditionally referred to as Lissotriton vulgaris, consists of multiple morphologically distinct taxa. Given the uncertainty concerning the validity and rank of these taxa, L. vulgaris sensu lato has often been treated as a single, polytypic species. A recent study, driven by genetic data, proposed to recognize five species, L. graecus, L. kosswigi, L. lantzi, L. schmidtleri and a more restricted L. vulgaris. The Carpathian newt L. montandoni was confirmed to be a closely related sister species. We propose to refer to this collective of six Lissotriton species as the smooth newt or Lissotriton vulgaris species complex. Guided by comprehensive genomic data from throughout the range of the smooth newt species complex we 1) delineate the distribution ranges, 2) provide a distribution database, and 3) produce distribution maps according to the format of the New Atlas of Amphibians and Reptiles of Europe, for the six constituent species. This allows us to 4) highlight regions where more research is needed to determine the position of contact zones.

Unraveling climate influences on the distribution of the parapatric newts Lissotriton vulgaris meridionalis and L. italicus

BackgroundClimate is often considered as a key ecological factor limiting the capability of expansion of most species and the extent of suitable habitats. In this contribution, we implement Species Distribution Models (SDMs) to study two parapatric amphibians, Lissotriton vulgaris meridionalis and L. italicus, investigating if and how climate has influenced their present and past (Last Glacial Maximum and Holocene) distributions. A database of 901 GPS presence records was generated for the two newts. SDMs were built through Boosted Regression Trees and Maxent, using the Worldclim bioclimatic variables as predictors.ResultsPrecipitation-linked variables and the temperature annual range strongly influence the current occurrence patterns of the two Lissotriton species analyzed. The two newts show opposite responses to the most contributing variables, such as BIO7 (temperature annual range), BIO12 (annual precipitation), BIO17 (precipitation of the driest quarter) and BIO19 (precipitation of the coldest quarter). The hypothesis of climate influencing the distributions of these species is also supported by the fact that the co-occurrences within the sympatric area fall in localities characterized by intermediate values of these predictors. Projections to the Last Glacial Maximum and Holocene scenarios provided a coherent representation of climate influences on the past distributions of the target species. Computation of pairwise variables interactions and the discriminant analysis allowed a deeper interpretation of SDMs’ outputs. Further, we propose a multivariate environmental dissimilarity index (MEDI), derived through a transformation of the multivariate environmental similarity surface (MESS), to deal with extrapolation-linked uncertainties in model projections to past climate. Finally, the niche equivalency and niche similarity tests confirmed the link between SDMs outputs and actual differences in the ecological niches of the two species.ConclusionsThe different responses of the two species to climatic factors have significantly contributed to shape their current distribution, through contractions, expansions and shifts over time, allowing to maintain two wide allopatric areas with an area of sympatry in Central Italy. Moreover, our SDMs hindcasting shows many concordances with previous phylogeographic studies carried out on the same species, thus corroborating the scenarios of potential distribution during the Last Glacial Maximum and the Holocene emerging from the models obtained.

Amphibian recovery after a decrease in acidic precipitation.

We here report the first sign of amphibian recovery after a strong decline due to acidic precipitation over many decades and peaking around 1980-90. In 2010, the pH level of ponds and small lakes in two heavily acidified areas in southwestern Scandinavia (Aust-Agder and Østfold in Norway) had risen significantly at an (arithmetic) average of 0.14 since 1988-89. Parallel with the general rise in pH, amphibians (Rana temporaria, R. arvalis, Bufo bufo, Lissotriton vulgaris, and Triturus cristatus) had become significantly more common: the frequency of amphibian localities rose from 33% to 49% (n=115), and the average number of amphibian species per locality had risen from 0.51 to 0.88. In two other (reference) areas, one with better buffering capacity (Telemark, n=21) and the other with much less input of acidic precipitation (Nord-Trøndelag, n=106), there were no significant changes in pH or amphibians.

Vomeropterygopalatina in larval Ichthyosaura alpestris apuanus (Amphibia: Urodela) and comments on the formation of the definite vomer in the Salamandridae

We studied the mouth roof of some larval stages of the Alpine newt Ichthyosaura alpestris apuanus, collected in summer 1977 in the field, using total preparations stained with Alizarinred, X-ray computed microtomography, and scanning electron microscopy. In all specimens we found traits indicating a delayed metamorphosis, i.e. a temporarily slowed down or even halted development of traits dependent on thyroid hormones (TH), and a slight further growth of probably TH-independent traits. Especially noteworthy is the temporary partial fusion of vomer and palatine forming a vomeropterygopalatinum. Presence of vomeropterygopalatina was recently shown in overwintering larvae of the related species Lissotriton vulgaris. On the basis of these findings we suggest that vomeropterygopalatina appear to be a quite frequent side effect of delayed metamorphosis. Further, we hypothesize that large parts of the larval vomer become incorporated into the definite (transformed) vomer. Three larvae in the series showed developmental stages of the vomerine bar, a posterior outgrowth of the vomer, typical for salamandrids, which is formed around metamorphosis. Development of the vomerine bar begins on the front end of the larval vomer and not (as generally assumed) on its posterior inner edge. These findings support our previous view that the formation of the vomerine bar in L. vulgaris and I. a. apuanus is more complex than literature assumes. However, due to some shortcomings regarding number and preservation of the available larvae a deeper study is needed using standardized conditions and a more complete developmental series of these (protected) or related species to substantiate the observations reported here.

Isolation and gene flow in a speciation continuum in newts

Because reproductive isolation often evolves gradually, differentiating lineages may retain the potential for genetic exchange for prolonged periods, providing an opportunity to quantify and to understand the fundamental role of gene flow during speciation. Here we delimit evolutionary lineages, reconstruct the phylogeny and infer gene flow in newts of the Lissotriton vulgaris species complex based on 74 nuclear markers sampled from 127 localities. We demonstrate that distinct lineages along the speciation continuum in newts exchange nontrivial amounts of genes, affecting their evolutionary trajectories. By integrating a wide array of methods, we delimit nine evolutionary lineages and show that two principal factors have driven their genetic differentiation: time since the last common ancestor determining levels of shared ancestral polymorphism, and shifts in geographic distributions determining the extent of secondary contact. Post-divergence gene flow, indicative of evolutionary non-independence, has been most extensive in Central Europe, while four southern European lineages have acquired the population-genetic hallmarks of independent species (L. graecus, L. kosswigi, L. lantzi, L. schmidtleri). We obtained strong statistical support for widespread mtDNA introgression following secondary contact, previously suggested by discordance between mtDNA phylogeny and morphology. Our study reveals long-term evolutionary persistence of evolutionary lineages that may periodically exchange genes with one another: although some of these lineages may become extinct or fuse, others will acquire complete reproductive isolation and will carry signatures of this complex history in their genomes.

Linkage Map of Lissotriton Newts Provides Insight into the Genetic Basis of Reproductive Isolation.

Linkage maps are widely used to investigate structure, function, and evolution of genomes. In speciation research, maps facilitate the study of the genetic architecture of reproductive isolation by allowing identification of genomic regions underlying reduced fitness of hybrids. Here we present a linkage map for European newts of the Lissotriton vulgaris species complex, constructed using two families of F2 L. montandoni × L. vulgaris hybrids. The map consists of 1146 protein-coding genes on 12 linkage groups, equal to the haploid chromosome number, with a total length of 1484 cM (1.29 cM per marker). It is notably shorter than two other maps available for salamanders, but the differences in map length are consistent with cytogenetic estimates of the number of chiasmata per chromosomal arm. Thus, large salamander genomes do not necessarily translate into long linkage maps, as previously suggested. Consequently, salamanders are an excellent model to study evolutionary consequences of recombination rate variation in taxa with large genomes and a similar number of chromosomes. A complex pattern of transmission ratio distortion (TRD) was detected: TRD occurred mostly in one family, in one breeding season, and was clustered in two genomic segments. This is consistent with environment-dependent mortality of individuals carrying L. montandoni alleles in these two segments and suggests a role of TRD blocks in reproductive isolation. The reported linkage map will empower studies on the genomic architecture of divergence and interactions between the genomes of hybridizing newts.

Habitat suitability assessment of constructed wetlands for the smooth newt (Lissotriton vulgaris [Linnaeus, 1758]): A comparison with natural wetlands

Given the current decline of natural wetlands worldwide and the consequent negative impacts on amphibians, wetlands constructed for the treatment of wastewaters have the potential to play a role in the protection of these animals. However, there is a paucity of information regarding the value of constructed wetlands (CWs) to amphibians, particularly relating to the terrestrial phase of their life-cycle. This study compares the terrestrial habitats of natural wetlands (NWs) and CWs as refuges for the smooth newt (Lissotriton vulgaris, [L., 1758]) with the aim of developing recommendations for CWs (both new and existing) to enhance their usefulness as newt-friendly habitats. Terrestrial habitats surrounding NWs and CWs were mapped using ArcGIS. Potential barriers to newt movement in addition to the presence of features such as wood or stone which could act as potential newt refuges were also mapped. Natural wetlands had significantly more terrestrial habitat types than CWs and while woodlands at both wetland types were most likely to contain features of benefit to newts, terrestrial habitats of NWs contained more features compared to those of CWs. The application of a Habitat Suitability Index, which assesses the likelihood of the presence of newts, resulted in seven of eight NWs compared to only two of eight CWs receiving “good” scores, the lower scores for CWs being due primarily to the presence of a barrier to newt movement. Recommendations for enhancing the design and management of CWs for smooth newts include less intensive ground maintenance, reduction of barriers to newt movement, judicious planting of suitable trees or shrubs and the provision of additional refuges such as wood or stone.

Amphibian skin microbiota exhibits temporal variation in community structure but stability of predicted Bd-inhibitory function.

Host-associated microbiomes are increasingly recognized to contribute to host disease resistance; the temporal dynamics of their community structure and function, however, are poorly understood. We investigated the cutaneous bacterial communities of three newt species, Ichthyosaura alpestris, Lissotriton vulgaris and Triturus cristatus, at approximately weekly intervals for 3 months using 16S ribosomal RNA amplicon sequencing. We hypothesized cutaneous microbiota would vary across time, and that such variation would be linked to changes in predicted fungal-inhibitory function. We observed significant temporal variation within the aquatic phase, and also between aquatic and terrestrial phase newts. By keeping T. cristatus in mesocosms, we demonstrated that structural changes occurred similarly across individuals, highlighting the non-stochastic nature of the bacterial community succession. Temporal changes were mainly associated with fluctuations in relative abundance rather than full turnover of bacterial operational taxonomic units (OTUs). Newt skin microbe fluctuations were not correlated with that of pond microbiota; however, a portion of community variation was explained by environmental temperature. Using a database of amphibian skin bacteria that inhibit the pathogen Batrachochytrium dendrobatidis (Bd), we found that the proportion of reads associated with 'potentially' Bd-inhibitory OTUs did not vary temporally for two of three newt species, suggesting that protective function may be maintained despite temporal variation in community structure.

Effects of a glyphosate-based herbicide and predation threat on the behaviour of agile frog tadpoles

The widespread application of pesticides emphasises the importance of understanding the impacts of these chemicals on natural communities. The most commonly applied broad-spectrum herbicides in the world are glyphosate-based herbicides, which have been suggested to induce significant behavioural changes in non-target organisms even at low environmental concentrations. To scrutinize the behavioural effects of herbicide-exposure we exposed agile frog (Rana dalmatina) tadpoles in an outdoor mesocosm experiment to three concentrations of a glyphosate-based herbicide (0, 2 and 6.5mg acid equivalent (a.e.) / L). To assess whether anti-predator behaviour is affected by the pesticide, we combined all levels of herbicide-exposure with three predator treatments (no predator, caged Aeshna cyanea dragonfly larvae or Lissotriton vulgaris newt adults) in a full factorial design. We observed hiding, activity, proximity to the predator cage and vertical position of tadpoles. We found that at the higher herbicide concentration tadpoles decreased their activity and more tadpoles were hiding, and at least at the lower concentration their vertical position was closer to the water surface than in tadpoles of the control treatment. Tadpoles also decreased their activity in the presence of dragonfly larvae, but did not hide more in response to either predator, nor did tadpoles avoid predators spatially. Further, exposure to the herbicide did not significantly influence behavioural responses to predation threat. Our study documents a definite influence of glyphosate-based herbicides on the behaviour of agile frog tadpoles and indicates that some of these changes are similar to those induced by dangerous predators. This may suggest that the underlying physiological mechanisms or the adaptive value of behavioural changes may similar.

Taxonomic status of a newly described island population of the smooth newtLissotriton vulgaris (Linnaeus, 1758) from Bozcaada (Çanakkale, Turkey)

The taxonomic status and phylogenetic position of the recently recorded smooth newt (Lissotriton vulgaris) population from the island of Bozcaada (Canakkale, Turkey) is clarified on the basis of morphological and molecular phylogenetic analyses. The L. vulgaris population from Bozcaada presents body proportions and morphological features of subsp. schmidtlerorum, such as small body length, absence of the tail filament, and dorsal crest with pointed free margin. Similarly, phylogenetic analyses of mitochondrial DNA sequences (ND4, 16S rRNA) place the Bozcaada population within Clade E of the recent L. vulgaris phylogeny, which consists of L. v. schmidtlerorum populations.

Models of amphibian myogenesis - the case of Bombina variegata.

Several different models of myogenesis describing early stages of amphibian paraxial myotomal myogenesis are known. Myoblasts of Xenopus laevis and Hymenochirus boettgeri change their position from perpendicular to parallel, in relation to axial organs, and differentiate into mononucleate myotubes. In Bombina variegate the myotomal myoblasts change their shape from round to elongate and then differentiate into mononuclear, morphologically mature myotubes. In Pelobates fuscus and Triturus vulgaris, myoblasts fuse into multinuclear myotubes. Mono- and multinucleate myotubes achieve morphological maturity during the differentiation process. During myogenesis of B. variegata, the nuclei of mononucleate, differentiating myotubes contain a tetraploid quantity of DNA (4C DNA). The stable quantity of DNA is confirmed by lack of 3H-thimidine incorporation into myotube nuclei. This outcome is a proof that myoblasts withdraw from the cell cycle in the G2 phase. The further development of myotomal myotubes involves myoblasts of mesenchymal origin. These myoblasts fuse with myotubes in X. laevis and B. variegate in the G1 phase. Secondary muscle fibres in amphibian myotomes have only mesenchymal origin. Mesenchymal myoblasts fuse into multinucleated myotubes. Myofibril development in the differentiating myotube and lack of DNA replication confirm the classical paradigm of myogenesis. Mesenchymal myoblasts are taking part in the myogenesis of musculus rectus abdominis and limb muscles. The mesenchymal cells in the myogenesis process show one model of myogenesis, which is a classical model of myogenesis. The mesenchymal cells probably come from dermatome.

Heavy traffic, low mortality - tram tracks as terrestrial habitat of newts

Amphibian mortality caused by rail traffic has not attracted much attention in comparison to road mortality. Density of railways in landscape, as well as traffic intensity, is usually much lower than in case of roads. As a consequence, their overall effect on amphibian populations is tacitly assumed to be less negative. To test whether very intensive rail traffic can cause substantial mortality in population of a small amphibian, we investigated a Smooth newt Lissotriton vulgaris population located in the city of Poznan, W Poland, where tram tracks border isolated breeding ponds. We performed controls during the peak of autumn migratory activity along the tracks. Less than 1% of all individuals found during the survey were killed by rail traffic. Observed mortality was very low despite large number of individuals present on the track and intensive tram traffic. As negative effects of traffic are low, rail or tram embankments can provide an important terrestrial habitat for small European newts.

Genetic characterization of a second novel picornavirus from an amphibian host, smooth newt (Lissotriton vulgaris).

In this study, a novel picornavirus was identified in faecal samples from smooth newts (Lissotriton vulgaris). The complete genome of picornavirus strain newt/II-5-Pilis/2014/HUN (KX463670) is 7755 nt long with type-IV IRES and has 39.6% aa sequence identity in the protein P1 to the corresponding protein of bat picornavirus (KJ641686, unassigned) and 42.7% and 53.5% aa sequence identity in the 2C and 3CD protein, respectively, to oscivirus (GU182410, genus Oscivirus). Interestingly, the L-protein of newt/II-5-Pilis/2014/HUN has conserved aa motifs that are similar to those found in phosphatase-1 catalytic (PP1C) subunit binding region (pfam10488) proteins. This second amphibian-origin picornavirus could represent a novel species and could be a founding member of a potential novel picornavirus genus.

Effect of different restoration approaches on two species of newts (Amphibia: Caudata) in Central European lignite spoil heaps

Post-mining sites often offer secondary habitats for various amphibians endangered in common agricultural landscapes. On the other hand, there is a general lack of quantitative studies on the effects of the common restoration practices on colonisation of such artificial freshwater habitats by amphibians. Here, we focus on two newts decreasing in Central Europe, Lissotriton vulgaris and Triturus cristatus, in pools within lignite (brown coal) spoil heaps in the western Czech Republic. We compared their abundances in pools established by technical reclamation, spontaneous succession, and their combination. In spring 2016, we sampled 29 freshwater pools in five spoil heaps and 10 fishponds in the surrounding agricultural landscape by funnel trapping. We captured 52 L. vulgaris and 138 T. cristatus in all the studied habitats together. As only two L. vulgaris and no T. cristatus were caught in fishponds, the high potential of pools in post-mining landscapes was confirmed. Both newt species generally avoided the artificially established pools, but the spontaneously established pools were equally suitable for them in both technically reclaimed and spontaneously developed heaps. L. vulgaris preferred more transparent water, higher cover of cattails and the pool placed in the forest, but with fewer trees on the pool banks, whilst T. cristatus preferred narrower zone of littoral vegetation, more trees on the pool banks, smaller water bodies and absence of fish. Such habitat preferences fully corroborate with natural habitats. The technical reclamation still prevails in restoration of post-mining sites in many regions. We reveal it is an unsuitable practice for amphibians, if not combined with natural succession processes.