Urbanization is a severe form of habitat fragmentation that can cause many species to be locally extirpated and many others to become trapped and isolated within an urban matrix. The role of drift in reducing genetic diversity and increasing genetic differentiation is well recognized in urban populations. However, explicit incorporation and analysis of the demographic and temporal factors promoting drift in urban environments are poorly studied. Here, we genotyped 15 microsatellites in 320 fire salamanders from the historical city of Oviedo (Est. 8th century) to assess the effects of time since isolation, demographic history (historical effective population size; N ) and patch size on genetic diversity, population structure and contemporary N . Our results indicate that urban populations of fire salamanders are highly differentiated, most likely due to the recent N declines, as calculated in coalescence analyses, concomitant with the urban development of Oviedo. However, urbanization only caused a small loss of genetic diversity. Regression modelling showed that patch size was positively associated with contemporary N , while we found only moderate support for the effects of demographic history when excluding populations with unresolved history. This highlights the interplay between different factors in determining current genetic diversity and structure. Overall, the results of our study on urban populations of fire salamanders provide some of the very first insights into the mechanisms affecting changes in genetic diversity and population differentiation via drift in urban environments, a crucial subject in a world where increasing urbanization is forecasted.

a b s t r a c tAlthough locating wildlife roadkill hotspots is essential to mitigate road impacts, the influence of study design on hotspot identification remains uncertain. We evaluated how sampling frequency affects the accuracy of hotspot identification, using a dataset of vertebrate roadkills (n ¼ 4427) recorded over a year of daily surveys along 37 km of roads. "True" hotspots were identified using this baseline dataset, as the 500-m segments where the number of road-killed vertebrates exceeded the upper 95% confidence limit of the mean, assuming a Poisson distribution of road-kills per segment. "Estimated" hotspots were identified likewise, using datasets representing progressively lower sampling frequencies, which were produced by extracting data from the baseline dataset at appropriate time intervals (1e30 days). Overall, 24.3% of segments were "true" hotspots, concentrating 40.4% of roadkills. For different groups, "true" hotspots accounted from 6.8% (bats) to 29.7% (small birds) of road segments, concentrating from <40% (frogs and toads, snakes) to >60% (lizards, lagomorphs, carnivores) of roadkills. Spatial congruence between "true" and "estimated" hotspots declined rapidly with increasing time interval between surveys, due primarily to increasing false negatives (i.e., missing "true" hotspots). There were also false positives (i.e., wrong "estimated" hotspots), particularly at low sampling frequencies. Spatial accuracy decay with increasing time interval between surveys was higher for smaller-bodied (amphibians, reptiles, small birds, small mammals) than for larger-bodied species (birds of prey, hedgehogs, lagomorphs, carnivores). Results suggest that widely used surveys at weekly or longer intervals may produce poor estimates of roadkill hotspots, particularly for small-bodied species. Surveying daily or at two-day intervals may be required to achieve high accuracy in hotspot identification for multiple species.

The existence of two or more distinctly coloured phenotypes among individuals of an interbreeding population is known as colour polymorphism. In amphibians, this phenomenon is pervasive among anurans, but rare or absent among salamanders and caecilians, respectively. Here, we examine whether various distinct phenotypes of Salamandra salamandra in North Spain, used as a basis to describe the subspecies S. s. bernardezi and S. s. alfredschmidti, indeed warrant separate taxonomic status or that these co-occur and belong to a single taxon. Based on a sample of 1147 individuals from 27 local populations, six phenotype classes were designated. Although two phenotypes that are attributable to S. s. alfredschmidti show some degree of geographical restriction, these co-occur with those representing typical S. s. bernardezi. A fifth phenotype class could not be unambiguously attributed to either subspecies due to an overlap in previously suggested diagnostic characteristics. Mitochondrial (cytochrome b) and nuclear (b-fibrinogen) DNA analyses revealed S. s. alfredschmidti to be nested within several subclades of S. s. bernardezi, without displaying unique lineages. Furthermore, no significant divergence was recovered by means of niche overlap analyses. As a result, we revoke the subspecies status of S. s. alfredschmidti, which should be regarded as a junior synonym of S. s. bernardezi. The current findings confirm the existence of colour polymorphism in S. salamandra and the family Salamandridae, which provides exciting possibilities for future research.

No abstract

Evolutionary changes in reproductive mode may affect co‐evolving traits, such as dispersal, although this subject remains largely underexplored. The shift from aquatic oviparous or larviparous reproduction to terrestrial viviparous reproduction in some amphibians entails skipping the aquatic larval stage and, thus, greater independence from water. Accordingly, amphibians exhibiting terrestrial viviparous reproduction may potentially disperse across a wider variety of suboptimal habitats and increase population connectivity in fragmented landscapes compared to aquatic‐breeding species. We investigated this hypothesis in the fire salamander (Salamandra salamandra), which exhibits both aquatic‐ (larviparity) and terrestrial‐breeding (viviparity) strategies. We genotyped 426 larviparous and 360 viviparous adult salamanders for 13 microsatellite loci and sequenced a mitochondrial marker for 133 larviparous and 119 viviparous individuals to compare population connectivity and landscape resistance to gene flow within a landscape genetics framework. Contrary to our predictions, viviparous populations exhibited greater differentiation and reduced genetic connectivity compared to larviparous populations. Landscape genetic analyses indicate viviparity may be partially responsible for this pattern, as water courses comprised a significant barrier only in viviparous salamanders, probably due to their fully terrestrial life cycle. Agricultural areas and, to a lesser extent, topography also decreased genetic connectivity in both larviparous and viviparous populations. This study is one of very few to explicitly demonstrate the evolution of a derived reproductive mode affects patterns of genetic connectivity. Our findings open avenues for future research to better understand the eco‐evolutionary implications underlying the emergence of terrestrial reproduction in amphibians.

Amphibians are the most threatened vertebrates on Earth, and one of the main factors involved in their decline is the loss and fragmentation of their natural habitats. Contemporary urban development is a major cause of habitat fragmentation, and populations trapped within urban environments offer a unique opportunity to study effects of fragmentation. Here, we compared, for the first time in fire salamanders (Salamandra salamandra), estimates of census (N) and effective population size (N e ) in a small urban population in the city of Oviedo (Spain). We performed a 4 year capture-mark-recapture study and used three single-sample N e estimators based on 58 individuals genotyped for 15 polymorphic microsatellite loci. Our study showed a small (N ¼ 113 salamanders; 95 % CI 100-142) but dense population (mean 0.45 individuals per m 2 ), while single-sample estimators provided congruent N e estimates. A high N e /N population size ratio (range 0.50-0.84) obtained in this small and isolated population suggests the existence of mechanisms of genetic compensation (low reproductive variance and multiple paternity) in fire salamanders.

No abstract

Examining historical and contemporary processes underlying current patterns of genetic variation is key to reconstruct the evolutionary history of species and implement conservation measures promoting their long-term persistence. Combining phylogeographic and landscape genetic approaches can provide valuable insights, especially in regions harboring high levels of biodiversity that are currently threatened by climate and land cover changes, like southern Iberia. We used genetic (mtDNA and microsatellites) and spatial data (climate and land cover) to infer the evolutionary history and contemporary genetic connectivity in a short-range endemic salamander subspecies, Salamandra salamandra longirostris, using a combination of ecological niche modelling, phylogeographic, and landscape genetic analyses. Ecological-based analyses support a role of the Guadalquivir River Basin as a major vicariant agent in this taxon. The lower genetic diversity and greater differentiation of peripheral populations, together with analyses of climatically stable areas throughout time, suggest the persistence of a population in the central part of the current range since the Last Inter Glacial [LIG; ~120,000 -140,000 years BP], and a micro refugium in the eastern end of the range. Habitat heterogeneity plays a major role in shaping patterns of genetic differentiation in S. s. longirostris, with forests representing key areas for its long-term persistence under scenarios of environmental change. Our study stresses the importance of maintaining population genetic connectivity in low-dispersal organisms under rapidly changing environments, and will inform management plans for the long-term survival of this evolutionarily distinct Mediterranean endemic.