New distributional data on Haemogregarina stepanowi (Apicomplexa) and Placobdella costata (Hirudinea) parasitising the Sicilian pond turtle Emys trinacris (Testudines)

The host-parasite system “Emys trinacris – Placobdella costata – Haemogregarina stepanowi” is known for Sicily, but scarce information is available to date about the distribution of the two parasites on the island. Therefore, an extensive sampling effort through visual census and collection and analysis of blood smears of the endemic Sicilian pond turtle E. trinacris was carried out in 46 water bodies scattered throughout mainland Sicily. Our findings revealed that the distribution of both parasites is limited to the Nebrodi area, where the infection of H. stepanowi has shown a high incidence on the local turtle populations. Our data suggest no correlation between the current distribution of the two parasite species and environmental features. The current distribution of H. stepanowi and P. costata seems not to be relictual, but rather the outcome of a recent colonisation process. Considering the possible negative impact of both H. stepanowi and P. costata on their turtle host, their long-term effect on E. trinacris should be investigated.

Stand out from the Crowd: Small-Scale Genetic Structuring in the Endemic Sicilian Pond Turtle

The geographical pattern of genetic diversity was investigated in the endemic Sicilian pond turtle Emys trinacris across its entire distribution range, using 16 microsatellite loci. Overall, 245 specimens of E. trinacris were studied, showing high polymorphic microsatellite loci, with allele numbers ranging from 7 to 30. STRUCTURE and GENELAND analyses showed a noteworthy, geographically based structuring of the studied populations in five well-characterized clusters, supported by a moderate degree of genetic diversity (FST values between 0.075 and 0.160). Possible explanations for the genetic fragmentation observed are provided, where both natural and human-mediated habitat fragmentation of the Sicilian wetlands played a major role in this process. Finally, some conservation and management suggestions aimed at preventing the loss of genetic variability of the species are briefly reported, stressing the importance of considering the five detected clusters as independent Management Units.

Molecular Cytogenetic Characterization of the Sicilian Endemic Pond Turtle Emys trinacris and the Yellow-Bellied Slider Trachemys scripta scripta (Testudines, Emydidae)

Turtles, a speciose group consisting of more than 300 species, demonstrate karyotypes with diploid chromosome numbers ranging from 2n = 26 to 2n = 68. However, cytogenetic analyses have been conducted only to 1/3rd of the turtle species, often limited to conventional staining methods. In order to expand our knowledge of the karyotype evolution in turtles, we examined the topology of the (TTAGGG)n telomeric repeats and the rDNA loci by fluorescence in situ hybridization (FISH) on the karyotypes of two emydids: the Sicilian pond turtle, Emys trinacris, and the yellow-bellied slider, Trachemys scripta scripta (family Emydidae). Furthermore, AT-rich and GC-rich chromosome regions were detected by DAPI and CMA3 stains, respectively. The cytogenetic analysis revealed that telomeric sequences are restricted to the terminal ends of all chromosomes and the rDNA loci are localized in one pair of microchromosomes in both species. The karyotype of the Sicilian endemic E. trinacris with diploid number 2n = 50, consisting of 13 pairs of macrochromosomes and 12 pairs of microchromosomes, is presented here for first time. Our comparative examination revealed similar cytogenetic features in Emys trinacris and the closely related E. orbicularis, as well as to other previously studied emydid species, demonstrating a low rate of karyotype evolution, as chromosomal rearrangements are rather infrequent in this group of turtles.

Coupling GIS spatial analysis and Ensemble Niche Modelling to investigate climate change-related threats to the Sicilian pond turtle Emys trinacris, an endangered species from the Mediterranean

The pond turtle Emys trinacris is an endangered endemic species of Sicily showing a fragmented distribution throughout the main island. In this study, we applied “Ensemble Niche Modelling”, combining more classical statistical techniques as Generalized Linear Models and Multivariate Adaptive Regression Splines with machine-learning approaches as Boosted Regression Trees and Maxent, to model the potential distribution of the species under current and future climatic conditions. Moreover, a “gap analysis” performed on both the species’ presence sites and the predictions from the Ensemble Models is proposed to integrate outputs from these models, in order to assess the conservation status of this threatened species in the context of biodiversity management. For this aim, four “Representative Concentration Pathways”, corresponding to different greenhouse gases emissions trajectories were considered to project the obtained models to both 2050 and 2070. Areas lost, gained or remaining stable for the target species in the projected models were calculated. E. trinacris’ potential distribution resulted to be significantly dependent upon precipitation-linked variables, mainly precipitation of wettest and coldest quarter. Future negative effects for the conservation of this species, because of more unstable precipitation patterns and extreme meteorological events, emerged from our analyses. Further, the sites currently inhabited by E. trinacris are, for more than a half, out of the Protected Areas network, highlighting an inadequate management of the species by the authorities responsible for its protection. Our results, therefore, suggest that in the next future the Sicilian pond turtle will need the utmost attention by the scientific community to avoid the imminent risk of extinction. Finally, the gap analysis performed in GIS environment resulted to be a very informative post-modeling technique, potentially applicable to the management of species at risk and to Protected Areas’ planning in many contexts.

Mitochondrial phylogeography of European pond turtles (Emys orbicularis, Emys trinacris) – an update

Based on more than 1100 samples of Emys orbicularis and E. trinacris, data on mtDNA diversity and distribution of haplotypes are provided, including for the first time data for Armenia, Georgia, Iran, and the Volga, Ural and Turgay River Basins of Russia and Kazakhstan. Eight mitochondrial lineages comprising 51 individual haplotypes occur in E. orbicularis, a ninth lineage with five haplotypes corresponds to E. trinacris. A high diversity of distinct mtDNA lineages and haplotypes occurs in the south, in the regions where putative glacial refuges were located. More northerly parts of Europe and adjacent Asia, which were recolonized by E. orbicularis in the Holocene, display distinctly less variation; most refuges did not contribute to northern recolonizations. Also in certain southern European lineages a decrease of haplotype diversity is observed with increasing latitude, suggestive of Holocene range expansions on a smaller scale.

Variation of Sicilian pond turtles, Emys trinacris – What makes a species cryptic?

Variation of the Sicilian pond turtle, Emys trinacris, is described, based on morphological data of more than 200 adult and immature turtles and mtDNA sequences of 31 new known-locality specimens. Emys trinacris is morphologically more variable than thought before. There exist pronounced population-specific differences. Adults are barely distinguishable from Emys orbicularis galloitalica. Hatchlings of E. trinacris are, however, significantly different patterned, allowing immediate species determination. Moreover, hatchlings of E. trinacris seem to be on average smaller and lighter than in E. orbicularis. This could be related with different reproductive strategies. Coloration polymorphism of adult Sicilian pond turtles comprises fair part of variation known in E. orbicularis. Thus, adult E. trinacris might have preserved the whole array of variation of the last common ancestor with E. orbicularis. We hypothesize that similarity of certain southern E. orbicularis subspecies and E. trinacris on one hand and pronounced differences in size, coloration and pattern of northern E. orbicularis subspecies on the other result from stabilising selection in the north. While dark coloration and large body-size seem to be beneficial in the north, light coloration and small size could be simply not disadvantageous in the south, allowing coloration polymorphisms in E. trinacris and southern E. orbicularis subspecies. Further, the term 'cryptic species' and its meaning in regard to species concepts and bar-coding is discussed. It is concluded that species delineation based on mtDNA barcoding requires application of a Phylogenetic Species Concept. Under the Biological Species Concept, animal mtDNA is not always an ideal tool for delineating species boundaries because taxa with monophyletic mitochondrial gene trees are neither necessarily genetically isolated nor must represent the same Biological Species. Cryptic species are nothing special in nature because difficulties with their identification are due to deficits in cognitive abilities of man. This is illustrated by the fact that distinct live stages of 'cryptic species' may differ to various degrees from similar species, as is the case in hatchlings and adults of E. trinacris and E. o. galloitalica.