Unveiling a hotspot of genetic diversity in southern Italy for the endangered Hermann's tortoise Testudo hermanni.

BACKGROUND: Hotspots of intraspecific genetic diversity represent invaluable resources for species to cope with environmental changes, and their identification is increasingly recognized as a major goal of conservation ecology research. However, even for iconic and endangered species, conservation strategies are often planned without thorough information on the geographic patterns of genetic variation. Here, we investigated the spatial patterns of genetic variation of the endangered Hermann's tortoise Testudo hermanni in the Italian Peninsula by genotyping 174 individuals at 7 microsatellite loci, with the aim to contribute to planning effective conservation strategies. RESULTS: Ordination-based and Bayesian clustering analyses consistently identified three main genetic clusters, one spread in the central and northern part of the peninsula, and two restricted to southern Italy and Sicily, respectively. The highest levels of genetic diversity were found in populations of the southern cluster and, in particular, at the northern edges of its distribution (He > 0.6, Ar > 2.8 ), that correspond to areas of putative secondary contact and admixture between distinct lineages. Our results clearly identify a hotspot of genetic diversity for the Hermann's tortoise in southern Italy. CONCLUSION: We inferred the evolutionary history and the spatial patterns of genetic variation of the Hermann's tortoise in the Italian Peninsula. We identified three main genetic clusters along the peninsula and a hotspot of intraspecific diversity in southern Italy. Our results underline the urgent need for conservation actions to warrant the long-term persistence of viable tortoise populations in this area. Furthrmore, these data add further evidence to the role of southern Italy as a biodiversity hotspot for temperate fauna, claiming for higher consideration of this area in large scale conservation programs.

The Trophic Niche of Two Sympatric Species of Salamanders (Plethodontidae and Salamandridae) from Italy.

The trophic niche of a species is one of the fundamental traits of species biology. The ideal trophic niche of a species is realized in the absence of interspecific competition, targeting the most profitable and easy-to-handle food resources. However, when a competitor is present, species adopt different strategies to reduce competition and promote coexistence. In this study, we assessed the potential mechanisms that allow the coexistence of two generalist salamanders: the Italian cave salamander (Speleomantes italicus) and the fire salamander (Salamandra salamandra). We surveyed, in April 2021, a forested area of Emilia-Romagna (Italy) during rainy nights. Analyzing the stomach contents of the captured individuals, we obtained information on the trophic niche of these two sympatric populations. Comparing our results with those of previous studies, we found that the two species did not modify their trophic niche, but that alternative mechanisms allowed their coexistence. Specifically, different prey preferences and predator metabolisms were likely the major factors allowing reduced competition between these two generalist predators.

Capture-mark-recapture data on the strictly protected Speleomantes italicus.

This data set collects capture-mark-recapture data, biometric data, and stomach contents of seven populations of the Italian cave salamander (Speleomantes italicus), one of the strictly protected European plethodontid species endemic to mainland Italy. We monitored six subterranean populations inside caves, and one fully epigean population living in a forest, surveying a total area of >5200 m2 . Data collection was performed 24 times throughout a year for each of the subterranean populations, and seven times in late winter-early spring for the epigean population. Salamanders were individually identified using two different marking methods: subcutaneous injection of visual implant elastomers (VIE) and photographic recognition of the dorsal pattern. Overall, the data set contains information on 1283 captured salamanders, corresponding to 783 different individuals and 500 recapture events. This type of data can be used to assess the species detection probability and to estimate the size of the populations, which are fundamental parameters for the assessment of its conservation status. Captured salamanders were weighed using a digital scale and photographed next to a reference ruler to perform post hoc measurements. This allows to assess the potential variation of the body condition of individuals through the time, and the potential divergences between conspecific populations. Furthermore, repeated measurements of recaptured individuals can allow to evaluate the seasonal growth rates of Speleomantes. Before their release, the salamanders underwent stomach flushing, a non-invasive technique that allows us to investigate the food residues in the salamanders' stomach. In 951 salamanders, we were able to recognize a total of 7077 consumed prey items belonging to 37 different prey categories (i.e., order level or lower), completing the information on the consumed prey for the entire Speleomantes genus. Data on consumed prey can be used to assess potential divergences between populations or between individuals of different ages/sexes, but also to assess the potential trophic specialization of individuals. The distinctiveness of this data set is that, by combining the capture-mark-recapture data with those on the diet of individuals, it allows to perform detailed studies on the consistency of individuals' food preference over time, an analysis that has never been performed on these salamanders. We release the data set into the public domain under Creative Commons Attribution 4.0 International license (CC-BY). When you use this data in your publication, we request that you cite this data paper; if you are using the whole data set related to the entire project, please cite all the related papers. If this data set will be an important part of the data analyzed in your study, you should consider discussing a collaboration with the data set contact person. If you plan to use data from the European plethodontid salamanders trophic niche project, please contact the contact person to find out if similar analyses are already underway or if unpublished updates are available.

Assessing Stress Response in Lizards from Agroecosystems with Different Management Practices.

Despite the importance of reptiles in agroecosystems, little is known about the effects of agricultural intensification and pesticide use on these animals. We compared antioxidant and haematological biomarkers in the wild Italian wall lizards Podarcis siculus from three olive groves representing a gradient of management intensity. Lizards from the conventional grove showed induced antioxidant defences relative to those from the organic field. However, this induction did not avoid the occurrence of oxidative stress in males from intensively managed olive groves, who showed TBARS levels 58%-133% higher than males from the other sites. Haematological responses also suggested increased stress in females from the intensively managed olive groves, with a heterophil-to-lymphocyte ratio 5.3 to 14.8-fold higher than in the other sites. The observed stress responses of lizards along the studied gradient of agricultural management suggest their potential usefulness as non-destructive biomarkers to environmental stressors associated with agricultural intensification.

Mapping the geographic origin of captive and confiscated Hermann's tortoises: A genetic toolkit for conservation and forensic analyses.

The illegal trade has been threatening tortoise populations worldwide for decades. Nowadays, however, DNA typing and forensic genetic approaches allow us to investigate the geographic origin of confiscated animals and to relocate them into the wild, providing that suitable molecular tools and reference data are available. Here we assess the suitability of a small panel of microsatellite markers to investigate patterns of illegal translocations and to assist forensic genetic applications in the endangered Mediterranean land tortoise Testudo hermanni hermanni. Specific allelic ladders were created for each locus and tested on several reference samples. We used the microsatellite panel to (i) increase our understanding of the population genetic structure in wild populations with new data from previously unsampled geographic areas (overall 461 wild individuals from 28 sampling sites); (ii) detect the presence of non-native individuals in wild populations; and (iii) identify the most likely geographic area of origin of 458 confiscated individuals hosted in Italian seizure and recovery centers. Our analysis initially identified six major genetic clusters corresponding to different geographic macro-areas along the Mediterranean range. Long-distance migrants among wild populations, due to translocations, were found and removed from the reference database. Assignment tests allowed us to allocate approximately 70 % of confiscated individuals of unknown origin to one of the six Mediterranean macro-areas. Most of the assigned tortoises belonged to the genetic cluster corresponding to the area where the respective captivity center was located. However, we also found evidence of long-distance origins of confiscated individuals, especially in centers along the Adriatic coast and facing the Balkan regions, a well-known source of illegally traded individuals. Our results clearly show that the microsatellite panel and the reference dataset can play a beneficial role in reintroduction and repatriation projects when confiscated individuals need to be re-assigned to their respective macro-area of origin before release, and can assist future forensic genetic applications in detecting the illegal trade and possession of Testudo hermanni individuals.