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.     

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.     

A new cryptic species of pond turtle from southern Italy, the hottest spot in the range of the genus Emys (Reptilia, Testudines, Emydidae)

Geographic variation in the mtDNA haplotypes (cytochrome b gene) of 127 European pond turtles from Italy was investigated. Thirty‐eight of the Italian samples were also studied by nuclear fingerprinting (ISSR PCR) and compared with samples from other parts of the range representing all nine currently known mtDNA lineages of Emys orbicularis. Our genetic findings were compared against morphological data sets (measurements, colour pattern) for 109 adult turtles from southern Italy. Italy is displaying on a small geographical scale the most complicated variation known over the entire distributional area of Emys (North Africa over Europe and Asia Minor to the Caspian and Aral Seas). The Tyrrhenic coast of the Apennine Peninsula, the Mt. Pollino area and Basilicata are inhabited by Emys orbicularis galloitalica, a subspecies harbouring a distinct mtDNA lineage. The same lineage is also found in Sardinia. Along the Adriatic coast of Italy and on the Salentine Peninsula (Apulia, southern Italy), another morphologically distinctive subspecies (Emys orbicularis hellenica) occurs, which also bears a different mtDNA lineage. A higher diversity of mtDNA haplotypes in the south of the Apennine Peninsula suggests that the glacial refugia of E. o. galloitalica and E. o. hellenica were located here. A further refuge of E. o. hellenica probably existed in the southern Balkans. The west coasts of the Balkans and Corfu have probably been colonized from Italy and not from the geographically closer southern Balkanic refuge. In Sicily, a third mtDNA lineage is distributed, which is sister to all other known lineages of Emys. Morphologically, Sicilian pond turtles resemble E. o. galloitalica. However, nuclear fingerprinting revealed a clear distinctiveness of the Sicilian taxon, whereas no significant divergence was detected between representatives of the other eight mtDNA lineages of Emys. Furthermore, nuclear fingerprinting provided no evidence for current or past gene flow between the Sicilian taxon and the mainland subspecies of E. orbicularis. Therefore, Sicilian pond turtles are described here as a species new to science. Some populations in Calabria and on the Salentine Peninsula comprise individuals of different mtDNA lineages. We interpret this as a natural contact. However, we cannot exclude that these syntopic occurrences are the result of human activity. For example, in other parts of Italy, the natural distribution pattern of Emys is obscured by allochthonous turtles. This could also be true for southern Italy. The discovery of the complex taxonomic differentiation in southern Italy requires reconsidering conservation strategies.     

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.     

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.     

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,...     

A skeletochronological study of age,growth and longevity in two freshwater turtles,Emys orbicularis and Mauremys rivulata,from Mediterranean Turkey (Reptilia: Testudines)

We estimated growth parameters of Emys orbicularis and Mauremys rivulata in Mediterranean Turkey with the skeletochronological method, using specimens drowned in fyke nets. In adult E. orbicularis, the median age was 8 years in males and 10 years in females. The median age of adult M. rivulata was 10 years for both sexes. Both species reach sexual maturity at an age of 5–7 years. No difference was found in age composition between the sexes.     

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.     

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.     

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.     

A new Eimeria (Apicomplexa: Eimeriidae), possessing mitra-shaped oocysts, from the Neotropical chelid turtle Batrachemys heliostemma (Testudines: Chelidae), and its comparison with Eimeria mitraria (Laveran & Mesnil 1902)

Eimeria jirkamoraveci sp. n. is described from faeces of two specimens of the toad-headed, side-necked turtle Batrachemys heliostemma collected at Iquitos in Peru. Oocysts are ovoid to almost spherical, 10.6 (8-12) x 8.9 (7-10) microm, without micropyle, polar granule and oocyst residuum. One conically stretched end and three blunt conical tubercles at the opposite end of oocyst give it mitra-like appearance. Sporocysts are elongated, ellipsoidal, 7.2 (6-8) x 4.1 (4-4.5) microm, with a small, knob-like Stieda body and sporocyst residuum composed of fine granules. To avoid possible conspecificity, the described new species is thoroughly compared with the most similar coccidium, E. mitraria, collected from its type host, Chinemys reevesii.     

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.     

Eimeria ornata N. Sp. (Apicomplexa: Eimeriidae) from the Ornate Box Turtle, Terrapene ornata ornata (Reptilia: Testudines), in Texas

Eimeria ornata n. sp. is described from the feces of 6/16 (37.5%) ornate box turtles, Terrapene ornata ornata , in northcentral Texas. Endogenously sporulated oocysts are ellipsoid 17.9 × 15.7(16-21 × 14-18) μm, with a thin, single-layered wall; shape index 1.14 (1.0-1.3). A micropyle is absent but a polar granule was present in one third of the oocysts. An oocyst residuum was present, consisting of numerous small globules situated either in a distinct mass or scattered within the oocyst. The sporocysts are elongate, 11.1 × 5.4 (9-13 × 5-6) μm, with an indistinct Stieda body at 1 pole. A sporocyst residuum is present, consisting either as a compact mass or as scattered granules. The sporozoites are elongate, 9.5 × 2.0 (8-12 × 2) μm, in situ, with spherical anterior and posterior refractile bodies. The new species is distinguished from the similar Eimeria carri Ernst & Forrester, 1973, from eastern box turtles, T. Carolina , by slight differences in oocyst morphology and endogenous sporulation.     

Campbellenchinae (Nematoda: Tylodoridae), a new subfamily from campbell Island,with a description of two new species

The first paragraph of the Introduction serves as an Abstract.

Specimens of an undescribed subfamily of Tylodoridae Paramonov, 1967 were found in each of 50 soil samples collected since 1951 from around the roots of native plants on Campbell Island, New Zealand. This taxon, Campbellenchinae n.subfam., its type‐genus, Campbellenchus n.gen., and 2 new species, C. poae and C. filicauda, are described. The diagnostic characters of the new subfamily include a lip region without a labial disc, a stylet of moderate length, a characteristic zone of cuticular annulation anterior to the stylet knobs, a slit‐like amphidial opening, a single ovary, and a filiform tail.     

Mitochondrial phylogeography of the Japanese pond turtle,Mauremys japonica (Testudines,Geoemydidae)

The phylogeography of the Japanese pond turtle, Mauremys japonica (Temminck and Schlegel, 1835), which is an endemic species in Japan, was studied by analyzing the variation in two mitochondrial DNA sequences, the cytochrome b gene and the control region. These analyses suggest that M. japonica comprises two major groups. The first one was found in the more eastern region, eastern Honshu Island and Shikoku Island, while the second was found in a western region, Kyushu Island and the Chugoku District (the westernmost part of Honshu Island). The boundary between the two groups is located in the Chugoku District. The nucleotide and haplotype diversities were very low, and these low diversities seem to have been caused by a bottleneck in the last glacial age. These results suggest that this species survived the last glacial period in two refugia, one in the central part of Honshu Island and the other one in Kyushu Island. Subsequently, population expansion took place in the postglacial period, and the groups from the two refugia extended their distribution ranges to the present boundary in the Chugoku District which represents a secondary contact zone.