Mitochondrial phylogeography of the European pond turtle, Emys orbicularis (Linnaeus 1758)

The phylogeny and phylogeography of Emys orbicularis was inferred from mitochondrial nucleotide sequences of the cytochrome b gene analysed by DNA sequencing and RNA heteroduplex analysis. Within the family Emydidae the monotypic genus Emys is affiliated with the nearctic taxa Emydoidea blandingii and Clemmys marmorata. The analysis of 423 individuals of E. orbicularis, originating throughout its distribution range, revealed a remarkable intraspecific differentiation in 20 different haplotypes with distinct geographical ranges. Maximum parsimony analysis produced a star-like phylogeny with seven main lineages which may reflect separations in the late Pliocene. The haplotype distribution examined by partial Mantel tests and analysis of molecular variance revealed a substantial effect of glacial periods. This historical perspective suggests the existence of multiple glacial refugia and considerable Holocene range expansion which was modulated by climatic traits. Further support is gained for the occurrence of long-term parapatry in glacial refugia.     

Characterization of microsatellite loci in the European pond turtle Emys orbicularis

A set of eight highly polymorphic microsatellite markers was isolated and characterized from a genomic library enriched for dinucleotide repeats in the European pond turtle, Emys orbicularis. The markers were tested for polymorphism in a total of 33 turtles sampled in two natural ponds in the nature reserve of Kerkini, northern Greece. Number of alleles varied from 10 to 18, and expected heterozygosity ranged between 0.738 and 0.921. This novel set of loci will be particularly useful to assess fine-scale population structure and for parentage analysis in E. orbicularis.     

Factors affecting the predation of otter (Lutra lutra) on European pond turtle (Emys orbicularis)

In this case study, the ecological background of an unusual hunting behaviour was investigated, when otters Lutra lutra preyed upon European pond turtles Emys orbicularis in a Hungarian fish pond system during an 18-month period. Predation on turtle was found only during cold periods (established by spraint analysis and also by the collection of 182 turtle carcasses in 2003). The relationship was not close between fish availability and turtle consumption ( r _P =−0.325, P =0.19). The crude protein content of the turtle head and leg was higher than that of fish, frog and turtle body, whereas the energy content of the samples was similar. The mean body weight of the killed turtles (460 g) fell within the range of the optimal prey size of the otter. Turtles were used as cache foods by otters during extreme environmental conditions (as in the long winter), but occurred only rarely as buffer foods during moderate winter. In fish ponds, the conservation of the coexistent otter and turtle depends on pond management. The maintenance of a higher fish availability in ponds during winter makes it possible to avoid the need to acquire a proper hunting technique on turtle, indicated by the scarcity of primary fish food.     

The morphometrics and colouration of the European pond turtle Emys orbicularis in Lubuskie province (West Poland)

The European pond turtle Emys orbicularis, is an endangered species in Poland. The region of the Ilanka River, in the western part of the country (Lubuskie province), is the most densely inhabited area by the species. Several populous sites are dispersed over the whole area of the province. Over the past 10 years, 279 turtles were captured at 11 sites. In 218 individuals, those with a straight carapace length (SCL) between 2.10–12.10 cm, sex was not determined; the remaining 61 with SCL 13.33–19.85 cm were sexed. Females had bigger bodies and were heavier than males. The SCL of females was 17.10 ± 1.7 cm (average ± SD, n = 33), their body mass (BM) was 790.6 ± 228.7 g (n = 27), and in the case of males SCL was 15.44 ± 1.5 cm and BM was 554.2 ± 138.8 g (n = 28). SCL of the smallest egg laying female was 15.30 cm, and SCL of the smallest mating male was 13.33 cm. The colouration of the turtles which undergoes change with age can be varied.     

Reproductive strategies and body shape in the European pond turtle (Emys orbicularis) from contrasting habitats in Italy

Comparisons within and among populations offer important insights into variation in life-history traits and possible adaptive patterns to environmental conditions. We present the results of observed differences in body size, body shape and patterns of reproduction in four separate populations of the European pond turtle Emys orbicularis in central and southern Italy – coastal ( n =3) and mountainous ( n =1) sites and pond ( n =2) and canal ( n= 2) habitats – to determine whether phenotypic plasticity affects reproductive output. Although we did not find any significant latitudinal variation in body size, we observed significant differences in body shape between canal (rounded body shape) and pond (elongated body shape) systems and smaller size with rounded shape in the mountainous population. Reproductive output is similar among populations (median=5 eggs per clutch), whereas reproductive investment (relative clutch mass to maternal body mass) is higher in the mountain population (one clutch per year) than in coastal populations (two clutches per year), suggesting differential trade-offs between geographic locality, elevation and habitat type. Turtle shell shape and geographic location together affect reproductive output in E. orbicularis in Italy.     

Unexpected early extinction of the European pond turtle (Emys orbicularis) in Sweden and climatic impact on its Holocene range

Using ancient DNA sequences of subfossil European pond turtles ( Emys orbicularis ) from Britain, Central and North Europe and accelerator mass spectrometry radiocarbon dating for turtle remains from most Swedish sites, we provide evidence for a Holocene range expansion of the pond turtle from the southeastern Balkans into Britain, Central Europe and Scandinavia, according to the 'grasshopper pattern' of Hewitt. Northeastern Europe and adjacent Asia were colonized from another refuge located further east. With increasing annual mean temperatures, pond turtles reached southern Sweden approximately 9800 years ago. Until approximately 5500 years ago, rising temperatures facilitated a further range expansion up to Östergötland, Sweden (approximately 58°30'N). However, around 5500 years ago pond turtle records suddenly terminate in Sweden, some 1500 years before the Holocene thermal maximum ended in Scandinavia and distinctly earlier than previously thought. This extinction coincides with a temporary cooling oscillation during the Holocene thermal maximum and is likely related to lower summer temperatures deteriorating reproductive success. Although climatic conditions improved later again, recolonization of Sweden from southern source populations was prevented by the Holocene submergence of the previous land connection via the Danish Straits that occurred approximately 8500 years ago.     

Oxygen consumption as related to the development of the extraembryonic membranes and cardiovascular system in the European pond turtle (Emys orbicularis) embryogenesis

The rate of oxygen consumption (V(O2)), embryo mass, distribution and mass of the chorioallantoic membrane (CAM), heart rate (HR), heart mass, and amnion rhythmic contractions (ARC) were studied in eggs of the European pond turtle (Emys orbicularis) incubated at 28 degrees C for 62.5+/-0.3 days. The V(O2) rapidly increased beginning from incubation day 19 (D19) to a maximum on D50 and then decreased until pipping. The rapid V(O2) rise was correlated with an increase in the CAM surface and mass, heart mass, and ARC amplitude, whereas the functional parameters such as HR and ARC frequency remained unchanged. The drop in V(O2) before pipping was accompanied by a decrease in HR, while the heart and CAM masses were almost constant. In the cases of short-term temperature deviations of +/-3 degrees C from 28 degrees C, changes in (O2) were significant until D50 and nonsignificant after that, the changes in ARC frequency and HR being significant at all stages studied. Thus, the developmental V(O2) changes were contributed mainly by the slow morphogenetic processes during D19-D50, whereas changes in functional parameters began to play a role at later stages. The response to temperature fluctuations was mediated by a rapid change in functional parameters at all these stages.     

Absence of purinoceptors in the heart of the turtle (Emys orbicularis)

The effects of adenosine, adenosine 5'-triphosphate (ATP), a slowly degradable ATP analogue beta,gamma-methylene ATP (APPCP) and a degradation resistant ATP analogue alpha,beta-methylene ATP (APCPP) were examined on the turtle heart. Adenosine, ATP, APPCP and APCPP had no effect on the rate or force of contraction of either the atrium or ventricle. The effects of acetylcholine and noradrenaline were also examined on the turtle heart. Acetylcholine decreased the force and rate of contraction of turtle atria in a concentration-dependent manner. Noradrenaline increased the rate of contraction but caused a slight decrease in the force of contraction of the atrium. Neither acetylcholine nor noradrenaline produced an inotropic effect on the ventricle.     

Rapid response of a long-lived species to improved water and grazing management: The case of the European pond turtle (Emys orbicularis) in the Camargue,France

Among human activities, the effect of habitat management by grazing on population viability is ambiguous. Indeed, beneficial effects of grazing are expected by maintaining open meadows, but overgrazing is supposed to increase mortality by trampling. Grazing has been shown to negatively impact the survival of European pond turtle (Emys orbicularis) in the Camargue. Consequently, a new management plan was defined. We investigated the consequences of this management using capture–recapture methods to estimate variations of population sizes in this managed site and a control site over a 17 years period. Results show an increase of the number of adults and juveniles on the managed site after the management change. Our results suggest that improved water management with flooding in autumn provided better hibernation conditions, and that reduced grazing intensity in autumn/winter likely decreased the risk of trampling. Population size significantly increased in less than 4 years following the management change, probably by the relaxation of density-dependence. It is an original result for a long lived-species supposed to have an important time of resilience to perturbations.     

Eimeria species from the European pond turtle, Emys orbicularis (Reptilia: Testudines), in Galicia (NW Spain), with description of two new species

Parasitological examination of feces from 44 Emys orbicularis from Galicia (NW Spain) revealed the presence of 2 new eimerian species, Eimeria gallaeciaensis sp. n. and E. emydis sp. n., as well as E. mitraria (Laveran and Mesnil, 1902) Doflein, 1909. Oocysts of E. gallaeciaensis n. sp. were found in 20 of 44 (45.4%) turtles and are subspherical to lightly ovoid-ellipsoid, 19.3 x 16.0 (17-22 x 15-18) microm, shape index 1.2 (1.1-1.3), with a smooth, single-layered wall. Micropyle and polar granule are absent, but an oocyst residuum is present. Sporocysts are ellipsoid, 9.7 x 5.1 (9-10 x 5-6) microm, shape index 1.9 (1.7-2.0), each with a sporocyst residuum and a conical Stieda body usually bearing 1-4 short and thin projections. Oocysts of E. emydis n. sp. were found in the feces of 5 of 44 (11.4%) turtles and are ovoid, rarely pear-shaped, 22.6 x 17.0 (20-25 x 15.5-18) microm, shape index 1.3 (1.2-1.5), with a smooth, single-layered wall with a slight thinning at the pointed end. Micropyle and polar granule are absent, and an oocyst residuum is present. Sporocysts are ellipsoid, 11.4 x 6.0 (9-13 x 5-7) microm, shape index 1.9 (1.6-2.2), each with sporocyst residuum and a prominent Stieda body bearing 3-5 club-shaped projections. In addition to the new species described, this is the first report of E. mitraria parasitizing E. orbicularis.     

Range-wide molecular analysis of the western pond turtle (Emys marmorata): cryptic variation, isolation by distance, and their conservation implications

We analysed phylogeography and population genetic variation across the range of the western pond turtle (Emys marmorata) using rapidly evolving mitochondrial and nuclear DNA sequence data. Nuclear DNA sequences from two unlinked introns displayed extremely low levels of variation, but phylogenetic analyses based on mtDNA recovered four well-supported and geographically coherent clades. These included a large Northern clade composed of populations from Washington south to San Luis Obispo County, California, west of the Coast Ranges; a San Joaquin Valley clade from the southern Great Central Valley; a geographically restricted Santa Barbara clade from a limited region in Santa Barbara and Ventura counties; and a Southern clade that occurs south of the Tehachapi Mountains and west of the Transverse Range south to Baja California, Mexico. An analysis of molecular variance (amova) based on regional hydrographic units revealed that populations from the Sacramento Valley north to Washington were virtually invariant, with no evidence of population substructure among northern river drainage basins. In other areas, E. marmorata contains considerable unrecognized variation, particularly in central and southern California and in northern Baja California, Mexico. Our northern clade is congruent with the distribution of the subspecies Emys marmorata marmorata (Washington-central California). However, no clade is congruent with the distribution of the southern subspecies Emys marmorata pallida from central California-Baja. Thus, recognition of the current subspecies split is not warranted, based on the available genetic evidence. Our amova and phylogenetic results, in conjunction with a growing comparative database for other codistributed aquatic taxa, confirm the occurrence of genetic breaks across the Tehachapi Mountains and Transverse Range bounding the southern end of the Great Central Valley, and point to southern California as a rich source of cryptic genetic variation.     

Molecular characterization of the gastrula in the turtle Emys orbicularis: an evolutionary perspective on gastrulation

Due to the presence of a blastopore as in amphibians, the turtle has been suggested to exemplify a transition form from an amphibian- to an avian-type gastrulation pattern. In order to test this hypothesis and gain insight into the emergence of the unique characteristics of amniotes during gastrulation, we have performed the first molecular characterization of the gastrula in a reptile, the turtle Emys orbicularis. The study of Brachyury, Lim1, Otx2 and Otx5 expression patterns points to a highly conserved dynamic of expression with amniote model organisms and makes it possible to identify the site of mesoderm internalization, which is a long-standing issue in reptiles. Analysis of Brachyury expression also highlights the presence of two distinct phases, less easily recognizable in model organisms and respectively characterized by an early ring-shaped and a later bilateral symmetrical territory. Systematic comparisons with tetrapod model organisms lead to new insights into the relationships of the blastopore/blastoporal plate system shared by all reptiles, with the blastopore of amphibians and the primitive streak of birds and mammals. The biphasic Brachyury expression pattern is also consistent with recent models of emergence of bilateral symmetry, which raises the question of its evolutionary significance.