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.     

Isolation of microsatellite markers in the Emys orbicularis complex and development of multiplex PCR amplification

We report the development of 15 new microsatellite markers for Emys orbicularis and Emys trinacris. A survey of 20 individuals showed that all loci are highly polymorphic with 3–14 alleles per locus. Additionally, 22 Glyptemys muhlenbergii primers were checked for cross-species amplification, with 14 being amplified successfully and polymorphic (2–14 alleles). A set of eight markers was selected and combined into two multiplex PCRs. Levels of genetic diversity were assessed in 648 individuals covering the complete distribution area. The number of alleles ranged from 13 to 24 and observed heterozygosity varied between 0.515 and 0.852.     

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.     

<Emphasis Type="Italic">Msx</Emphasis> genes are expressed in the carapacial ridge of turtle shell: a study of the European pond turtle,<Emphasis Type="Italic"> Emys orbicularis</Emphasis>

The turtle shell forms by extensive ossification of dermis ventrally and dorsally. The carapacial ridge (CR) controls early dorsal shell formation and is thought to play a similar role in shell growth as the apical ectodermal ridge during limb development. However, the molecular mechanisms underlying carapace development are still unknown. Msx genes are involved in the development of limb mesenchyme and of various skeletal structures. In particular, precocious Msx expression is recorded in skeletal precursors that develop close to the ectoderm, such as vertebral spinous processes or skull. Here, we have studied the embryonic expression of Msx genes in the European pond turtle, Emys orbicularis. The overall Msx expression in head, limb, and trunk is similar to what is observed in other vertebrates. We have focused on the CR area and pre-skeletal shell condensations. The CR expresses Msx genes transiently, in a pattern similar to that of fgf10. In the future carapace domain, the dermis located dorsal to the spinal cord expresses Msx genes, as in other vertebrates, but we did not see expansion of this expression in the dermis located more laterally, on top of the dermomyotomes. In the ventral plastron, although the dermal osseous condensations form in the embryonic Msx-positive somatopleura, we did not observe enhanced Msx expression around these elements. These observations may indicate that common mechanisms participate in limb bud and CR early development, but that pre-differentiation steps differ between shell and other skeletal structures and involve other gene activities than that of Msx genes.Edited by D.A. Weisblat     

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.     

Patterns of nuclear and mitochondrial DNA variation in Iberian populations of <Emphasis Type="Italic">Emys orbicularis</Emphasis> (Emydidae): conservation implications

The European pond turtle (Emys orbicularis) is threatened and in decline in several regions of its natural range, due to habitat loss combined with population fragmentation. In this work, we have focused our efforts on studying the genetic diversity and structure of Iberian populations with a fine-scale sampling (254 turtles in 10 populations) and a representation from North Africa and Balearic island populations. Using both nuclear and mitochondrial markers (seven microsatellites, ∼1048 bp nDNA and ∼1500 bp mtDNA) we have carried out phylogenetic and demographic analyses. Our results show low values of genetic diversity at the mitochondrial level although our microsatellite dataset revealed relatively high levels of genetic variability with a latitudinal genetic trend decreasing from southern to northern populations. A moderate degree of genetic differentiation was estimated for Iberian populations (genetic distances, F _ST values and clusters in the Bayesian analysis). The results in this study combining mtDNA and nDNA, provide the most comprehensive population genetic data for E. orbicularis in the Iberian Peninsula. Our results suggest that Iberian populations within the Iberian–Moroccan lineage should be considered as a single subspecies with five management units, and emphasize the importance of habitat management rather than population reinforcement (i.e. captive breeding and reintroduction) in this long-lived species.     

Blood parasite diversity (Apicomplexa: Haemogregarinidae) within the western populations of the European pond turtle Emys orbicularis

Systematic Parasitology - Molecular tools have revolutionized assessments of blood parasites in freshwater turtles. In the Iberian Peninsula and North Africa, two native species of terrapins occur,...     

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.     

Sexual dimorphism of the European Pond Turtle,Emys orbicularis (Linnaeus, 1758), in Anzali Lagoon,Iran (Reptilia: Emydidae)

The morphometric characteristics of the European Pond Turtle, Emys orbicularis, were studied at Anzali lagoon on the southern coast of the Caspian Sea. Males were on average (N=249) with 272.0?g significantly lighter than females with 447.0?g, and average carapace length was significantly smaller (123.0?mm in males, 139.0 in females). Females exceeded males also in all other studied characters (carapace width, plastron length, plastron width, and scute height). The sex ratio of male to female was 1 to 1.03, and the comparison of total tail length and cloaca-tail tip length revealed a difference between the position of the cloaca in both sexes, a character useful for sex determination in this species.     

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.     

Basic morphological data of native Czech Emys orbicularis revealed by subfossil finds

Holocene Czech Emys orbicularis are rare in museum collections because the species disappeared from this region centuries ago. Fifteen reconstructed shells of subfossil Czech specimens of E. orbicularis (ranging in age from 3800–3500 BC to 1300 AD) from archaeological sites were used to assess basic morphological data. Our results provide evidence that the extinct Czech turtles fall into the size range of extant Central European E. orbicularis populations from the Hungarian Lowland. Maximum male and female straight-line carapacial lengths of the Czech specimens reached at least 160 and 175 mm, respectively.     

Nuclear gene phylogeography reveals the historical legacy of an ancient inland sea on lineages of the western pond turtle,Emys marmorata in California

The historical biogeography of California’s taxa has been the focus of extensive research effort. The western pond turtle (Emys marmorata) is an example of a wide‐ranging taxon that spans several well‐known California diversity hotspots. Using a dataset comprised of one mitochondrial and five nuclear loci, we elucidate the major biogeographic patterns of the western pond turtle across the California landscape. By employing a combination of phylogenetic and network‐based approaches, we recovered a relatively ancient (c. 2–8 Ma) north/south split among populations of E. marmorata and find an area of intergradation centred in the Central Coast Ranges of California. In addition, discordant mitochondrial/nuclear genetic patterns suggest subsequent gene flow from northern populations and from San Joaquin Valley populations into the Central Coast Ranges after the Pliocene‐Pleistocene marine embayment of the Great Central Valley subsided. Our results emphasize the utility of nuclear DNA phylogeography for recovering the impact of relatively ancient biogeographic events, and suggest that the Central Coast Ranges of California have played a major role in the geographic structuring of the western pond turtle, and possibly other co‐distributed taxa.     

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.     

The breeding colouration of male three-spined sticklebacks (Gasterosteus aculeatus) as an indicator of energy investment in vigour

Summary A necessary condition of most models of intersexual selection requires that secondary sexual traits are costly so that cheating is prevented. If the conspicuous breeding colouration of male three-spined sticklebacks (Gasterosteus aculeatus L.) is such a handicap, it must involve costs. I examined the energetic costs of the breeding colouration by varying the energy contents of the daily food supply among five groups of sticklebacks over a 10 week period. The nutritional carotenoid level, i.e. the colour pigment used in the breeding colouration, was constant for all fish. Both the increase of their condition factor and the condition level they finally achieved correlated positively with the food ration of the groups. Individuals whose condition increased during the experiment developed a more intensive red colouration. However, a direct correlation between food quantity and the red breeding colouration reached at the end of the experiment did not exist. Nevertheless, given the limitation of pigment availability, there was still variation in the breeding colouration and the costs for the metabolism of the colouration were sufficient to render it an honest signal: a female stickleback can assess a male's condition and condition change over the past few weeks by the intensity of the colour of his blue eyes (which is not based on carotenoids and whose pigments were therefore not controlled in the food) and his red jaw, respectively. How much an individual male fish invests in increase of length and increase of condition (which correlate negatively with each other) seems to be, at least partly, his own strategic decision, which could have important consequences in the competition for female mates. It is eventually this decision that a male stickleback seems to signal with his red jaw.