Testudo Graeca Research Articles

Mitochondrial phylogeography of Testudo graeca in the Western Mediterranean: Old complex divergence in North Africa and recent arrival in Europe

Abstract We investigated the mitochondrial phylogeography of spur-thighed tortoises (Testudo graeca) in the Western Mediterranean. In North Africa, four major lineages (A-D) occur that together constitute a well-supported clade corresponding to one of the six major clades within T. graeca; the North African clade is sister to a Caucasian clade representing the subspecies T. g. armeniaca. Phylogenetic relationships between the North African lineages are badly resolved. Lineage A is distributed in Tunisia and adjacent Algeria, lineage B in Algeria and northern Morocco, lineage C in the Libyan Cyrenaica Peninsula, and lineage D north of the High Atlas Mts. and in the Souss Valley (southern Morocco). Lineage B is subdivided into two subgroups, B1 (eastern Morocco and Algeria) and B2 (north-western Morocco). Italian tortoises harbour haplotypes of lineage A, Spanish tortoises of subgroup B1. Based on a relaxed molecular clock calibrated with fossil evidence, the six major mtDNA clades of T. graeca are estimated to have diverged approximately 4.2-1.8 Ma ago; the split between the clades representing the eastern subspecies T. g. ibera and T. g. terrestris is younger than the split between Western Mediterranean tortoises and T. g. armeniaca. The Western Mediterranean lineages A-D were dated to have diverged at least 1.4-1.1 Ma ago; B1 and B2 split approximately 0.7 Ma ago. Our results suggest that Italian and Spanish tortoises were either introduced or originated from trans-oceanic dispersal in historic or prehistoric times. Spur-thighed tortoises invaded North Africa probably across Near Eastern landbridges that emerged in the Late Tertiary. Their diversification in North Africa seems to be correlated with habitat aridization cycles during the Pleistocene. The ranges of the Western Mediterranean lineages largely correspond to the distribution of morphologically defined subspecies in North Africa, with exception of T. g. graeca and T. g. whitei, and of T. g. lamberti and T. g. marokkensis, which are not differentiated. We propose to lump the first two subspecies under the name of T. g. graeca and the latter under the name of T. g. marokkensis. The complex differentiation of spur-thighed tortoises in North Africa implies that the model of a bipartite east-west differentiation, as proposed for other Maghrebian amphibians and reptiles, may be too simplistic, reflecting incomplete locality sampling rather than actual phylogeographic differentiation.

International Testudo Trade

Abstract The global trade in tortoises of the genus Testudo was reviewed by using compiled import data from Convention on International Trade in Endangered Species of Wild Fauna and Flora (CITES) for 1975–2005. A total of 58 importing and 112 exporting countries were recorded as being involved with the Testudo trade. Turkey, the former Soviet Union, the former Yugoslavia, Morocco, Uzbekistan, Russia, and Ukraine accounted for 81% of the recorded exports; whereas, the United Kingdom, Germany, and the United States dominated the import market with 67% of the individuals. Testudo horsfieldi accounted for 48% of the exports, followed by Testudo graeca (37%) and Testudo hermanni (13%). A total of 2,062,289 individuals were traded internationally, and of these, 99% were live animals. Of the animals from known sources, 67% were from the wild, and 16% were allegedly captive bred; the source of 70% of all declared trade animals was unknown.

Food choice of an Algerian population of the spur‐thighed tortoise, Testudo graeca

The diet of an Algerian population of spur‐thighed tortoise (Testudo graeca) was studied with the aims of exploring: (i) the variation in diet among males, females and juveniles, (ii) the relationships between consumption and relative availability of the plant species, and (iii) which plant tissues, vegetative or reproductive, are eaten by tortoises. We recorded more than forty plant species at the study area, with monocot species (n = 7) having a greater percent cover than that of dicot species (n = 31 species) or Gymnospermae (n = 2). Tortoise diet was studied by categorising 4422 plant and animal fragments in faecal pellets of 20 males, 16 females and eight juveniles. Tortoises ate a wide variety of plant species, including 13 dicots and three monocots, and occasionally invertebrates. The number of fragments for a plant species was correlated with plant species cover, and plant vegetative tissues exceeded plant reproductive tissues in the faeces. Dicots (Fabaceae, Composeae, Primulaceae, and Caryophyllaceae) accounted for over 70% of the diet (faecal fragments). The high dietary (niche) overlap, and null model analysis (RA3 algorithm with 30,000 Monte Carlo simulations), indicate that males, females and juveniles did not partition food resources; all three groups ate the same plant species.

Sexual Dimorphism in the Greek Tortoise: A Test of the Body Shape Hypothesis

In most animal species, it is expected that females should exhibit a greater abdominal volume than males to hold the progeny, when compared with females, males should exhibit more developed attributes that enhance mobility. We tested this hypothesis in the Greek tortoise. In chelonians, a reduction of the openings in the shell improves protection against predation but also constrains the abdominal volume and limits the space available to move the limbs. As expected, our data show that the shell provides a larger abdominal volume relative to tortoise size in females than in males. In males, deep notches in the shell and a reduction of several plastron plates offer more freedom to the limbs and to the tail; these characteristics presumably enhance mating success. Further studies are necessary to assess the applicability of these results in other chelonians, notably freshwater and marine turtles.

The Geographic Distribution of Turtles in Armenia and the Nagorno-Karabakh Republic (Artsakh)

The turtles of the Caucasus are poorly studied, and basic data on their geographic distribution in some Caucasian regions are poorly collated, hard to access, or entirely lacking. We present a comprehensive review of turtle localities in such a region, Armenia and Nagorno-Karabakh Republic (Artsakh), based on literature records, personal observations, and museum specimens. The compiled data for the 3 native species (Emys orbicularis, Mauremys caspica, and Testudo graeca) show that their distributions in this region are restricted to 3 areas of low elevation. In the northeast, turtles can be found in the Kura River drainage. In the south and west, turtles can be found in the Arax River drainage. The distributions of species within these areas vary, with E. orbicularis being the most restricted by elevation. We note that the 3 regions here form part of an important transect from the interior Arax drainage (in the Ararat region of western Armenia) to near its confluence with the Kura drainage (in southern Armenia and Nagorno-Karabakh) to the Kura drainage sensu stricto (in northeast Armenia). Future studies of turtles in these areas will play an important role in understanding the historical biogeography of Caucasian turtles.

The distal forelimb musculature in aquatic and terrestrial turtles: phylogeny or environmental constraints?

We compared the muscular anatomy of the distal front limb in terrestrial and aquatic chelonians to test whether observed differences between the two groups are associated with their divergent lifestyles and locomotor modes. Given the different use of the forelimb in the two environments (body support and propulsion on land vs. mainly propulsion in water) we expected that: (1) aquatic and terrestrial turtles would show differences in their muscular anatomy, with aquatic species having more individualized muscle bundlesto allow for the complex forearm movements observed during swimming, and (2) that terrestrial turtles would have more robust muscles to support their body weight against gravity. To address these questions, we examined the forelimb myology and associated tissues in six aquatic or semi-aquatic turtles (Phyrnops hilarii, Podocnemis unifilis, Trachemys scripta, Sacalia bealei, Cuora amboinensis and Mauremys caspica) and six terrestrial or semi-terrestrial turtles (Geochelone chilensis, Testudo graeca, Cuora galbinifrons, Glyptemys insculpta, Terrapene carolina and Rhinoclemmys pulcherrima). This paper describes the general structure of the forelimb musculature in all species, and quantifies muscle masses in those species with more than five specimens available (Ph. hilarii, Po. unifilis and Ge. chilensis). The general structure of the forelimb muscles in the strictly terrestrial species Ge. chilensis and Tes. graeca was found to be notably different from the pattern of the aquatic and semi-aquatic species examined, showing a distinct fusion of the different muscular bodies. Ter. carolina also show a distinctly terrestrial pattern, but a less extensive tendon development. R. pulcherrima and GI. insculpta were found to be morphologically intermediate; in the geoemydids the strictly terrestrial bauplan never appears. Quantitative differences in the robustness or mass of the distal forelimb muscles were also observed for the species investigated, supporting our prediction that the extensor muscles are more robust in terrestrial turtles. However, in contrast to our expectations, not only the extensor muscles of the distal forelimb (which are crucial in providing both body support and propulsion), but all muscles acting around the wrist were found to be heavier in terrestrial turtles.

Chersine Merrem, 1820 and Chersina Gray, 1831: a nomenclatural survey

In the tenth Edition of his ‘Systema Naturae’, Carolus Linnaeus presented among the species of the genus Testudo (including all the turtles and tortoises, thus corresponding to the present order of Chelonians) the species Testudo graeca (Linnaeus 1758). The reference was the description and the plate published by George Edwards (1751) of a tortoise from North Africa, more precisely from the old fort of Santa-Cruz (presently Djebel Murdjadjo or “pic de l’Aïdour” 35°42’N, 0°45’W, Oran, Algeria). Several hypotheses tried to explain Linnaeus’s choice of the name graeca, one proposing that the species is the tortoise of the old Greek authors. Nevertheless, since 1758, the identity of Testudo graeca is rather well established, with a precise type locality, although the pictured holotype is lost. Testudo graeca Linnaeus, 1758 is the type species of Testudo Linnaeus, 1758 by subsequent designation of Bell (1828).

Plasma concentrations of 25‐hydroxycholecalciferol in 22 captive tortoises (Testudo species)

The plasma concentration of 25-hydroxycholecalciferol was measured in 13 adult Hermann's tortoises (Testudo hermanni), seven adult spur-thighed tortoises (Testudo graeca) and two adult marginated tortoises (Testudo marginata) during 2004. They were healthy, of both sexes, and kept in captivity under natural unfiltered sunlight in southern England with no dietary sources of cholecalciferol. Blood samples were taken in March, June and August, and the concentration of 25-hydroxycholecalciferol did not vary significantly with the seasons. However, the concentrations in the female tortoises were always significantly lower than in the males.

Prevalence and intensity of blood apicomplexan infections in reptiles from Romania

In order to evaluate prevalence and intensity of apicomplexan hemoparasites in free-ranging reptiles from Romania, blood smears were collected from European pond turtles (Emys orbicularis), sand lizards (Lacerta agilis), and spur-thighed tortoises (Testudo graeca). All three host species were positive for blood parasites, with prevalence of infected individuals between 60.71% and 100% and variable intensity. Similarities and differences with other epidemiological data are discussed.

Educação Ambiental e para o desenvolvimento sustentável no marco da década das Nações Unidas: um caso de reorientação curricular ao nível de pós-graduação na Universidade Federal de Tocantins

In this paper, we investigate the social dimension of the conservation of species by means of social intervention for the conservation of the spur-thighed (Greek) tortoise in the south-west of the Iberian peninsula and the management of a captive population of spur-thighed tortoises.

Cryptosporidium from tortoises: Genetic characterisation, phylogeny and zoonotic implications

Cryptosporidiosis is a disease caused by apicomplexan parasites of the genus Cryptosporidium, which affect the gastrointestinal tract of humans and animals worldwide. In the present study, Cryptosporidium isolates from captive European tortoises ( Testudo graeca, Testudo hermanni and Testudo marginata) in Italy were genetically characterised using gene/s encoding Cryptosporidium oocyst wall protein (COWP) and/or the hypervariable segment internal to the SSU rDNA. Six isolates from T. graeca, T. hermanni and T. marginata revealed 100% sequence identity with the zoonotic Cryptosporidium pestis ( Cryptosporidium parvum ‘bovine genotype’), while a sequence from one T. marginata was a unique uncharacterised genotype ( Cryptosporidium sp. ex T. marginata). The new genotype is phylogenetically characterised as belonging to the intestinal cryptosporidial lineage. The present study indicates that tortoises may disseminate Cryptosporidium oocysts in the captive environment. The identification of the zoonotic genotype in the faeces of captive European tortoises indicates a potential risk for humans within the household.

Similarities and differences in adult tortoises : a morphological approach and its implication for reproduction and mobility between species.

Sexes in Chelonia display marked differences. Sexual size dimorphism (SSD) is important in evolutionary biology. Different sexual strategies result in species specific selection. Biometric variation in male and female tortoises of two species is studied. Eighteen biometrics were measured in 75 museum specimens (20 Testudo graeca; 55 T. hermanni). Nine of 18 parameters in T. hermanni and two of 18 in T. graeca were sexually dimorphic. Multivariate analyses (principal component analysis) highlighted two components, with bridge length the first and anal divergence the second component. The bridge length can be used to separate sexes and species. Males of both species were most different, whereas females of two species overlapped in body shape measurements. We hypothesise that female similarity could be a by-product of reproductive biology and sexual selection that optimise individual fitness.

Leptoconops bezzii (Diptera: Ceratopogonidae) parasitizing tortoises Testudo graeca (Testudines: Testudinidae) in mountain ranges of Lebanon and western Syria

Biting midges of the genus Leptoconops, subgenus Leptoconops (Diptera: Ceratopogonidae) were repeatedly found parasitizing on spur-thighed tortoises Testudo graeca in western Syria and Lebanon. Collected females were assigned to the species L. bezzii according to their morphological characteristics. Tortoises parasitized by midges were observed throughout the daytime, under mild temperatures, and slight winds, in Mediterranean or stony steppe habitats. Intensity of the infestations ranged from a few specimens up to thousands of midges per tortoise. Midges infested predominantly the tortoises' carapax, finding their approach to the capillaries in the seams between the keratinized scutes. The vector potential of ceratopogonids is discussed.

Population structure and demography of an Algerian population of the Moorish tortoise, Testudo graeca

Abstract Demography and population biology aspects of the Moorish tortoise, Testudo graeca graeca were studied in the Parc National d'El Kala, north-eastern Algeria. Density, which was assessed by capture-mark-recapture procedures in a study plot of 30 ha surface, was 3.73 individuals × ha−1. Adult sex-ratio was nearly equal (1.06 : 1). Sexual maturity was reached at 7-8 years of age in the males and at 9-11 years in the females. The mean age of tortoises was 13.8 ± 6.4 years (n = 88). Maximum longevity was over 20 years in both sexes. The females reached larger body size than the males. The growth was rapid during the early years of age in both sexes, and tended to stop completely by the 9-12 years of life in the males and by the 8-10 years in the females. The instantaneous growth of adults which were recaptured in multiple years indicated an average growth rate of 11.34 mm/year in the males, and of 26.30 mm/year in the females. Mortality was estimated at 3.94%. The main mortality cause for the tortoises of this population was the occurrence of fires spreading throughout the study area. These data are of interest because demographic patterns of Testudo graeca seem to vary not only because of their relative genetical variability, but also because of their relative environmental stress.

Effects of Storage and Sample Type on Ionized Calcium, Sodium and Potassium Levels in Captive Tortoises, Testudo spp.

ABSTRACT Blood samples were obtained from Hermann's tortoises, Testudo hermanni, spur-thighed tortoises, Testudo graeca, and Horsfield tortoises, Testudo horsfieldi, by jugular venipuncture and were collected into heparin. Ionized calcium, sodium and potassium analyses were performed using the ion selective electrode principle. The effect of sample type (heparinized whole blood vs. heparinized plasma) and storage were evaluated. No statistically significant differences were found between ionized calcium levels determined from heparinized plasma samples stored for 48 hr at 5°C (23°F) and fresh heparinized whole blood. Mean ionized calcium levels determined from the heparinized plasma were: males 3.72 mEq/L (1.86 mmol/L), females 3.86 mEq/L (1.93 mmol/L) and 3.80 mEq/L (1.90 mmol/L) for the combined data. Sodium and potassium levels were also determined and there were statistically significantly lower sodium levels and higher potassium levels in the stored heparinized whole blood compared to the fresh hepar...

Phenotypic plasticity leads to incongruence between morphology-based taxonomy and genetic differentiation in western Palaearctic tortoises (Testudo graeca complex; Testudines, Testudinidae)

Abstract Tortoises of the Testudo graeca complex inhabit a patchy range that covers part of three continents (Africa, Europe, Asia). It extends approximately 6500 km in an east-west direction from eastern Iran to the Moroccan Atlantic coast and about 1600 km in a north-south direction from the Danube Delta to the Libyan Cyrenaica Peninsula. Recent years have seen a rapid increase of recognized taxa. Based on morphological investigations, it was suggested that this group consists of as many as 20 distinct species and is paraphyletic with respect to T. kleinmanni sensu lato and T. marginata. Based on samples from representative localities of the entire range, we sequenced the mitochondrial cytochrome b gene and conducted nuclear genomic fingerprinting with ISSR PCR. The T. graeca complex is monophyletic and sister to a taxon consisting of T. kleinmanni sensu lato and T. marginata. The T. graeca complex comprises six well-supported mtDNA clades (A-F). Highest diversity is found in the Caucasian Region, where four clades occur in close neighbourhood. This suggests, in agreement with the fossil record, the Caucasian Region as a radiation centre. Clade A corresponds to haplotypes from the East Caucasus. It is the sister group of another clade (B) from North Africa and western Mediterranean islands. Clade C includes haplotypes from western Asia Minor, the southeastern Balkans and the western and central Caucasus Region. Its sister group is a fourth, widely distributed clade (D) from southern and eastern Asia Minor and the Levantine Region (Near East). Two further clades are distributed in Iran (E, northwestern and central Iran; F, eastern Iran). Distinctness of these six clades and sister group relationships of (A + B) and (C + D) are well-supported; however, the phylogeny of the resulting four clades (A + B), (C + D), E and F is poorly resolved. While in a previous study (Fritz et al., 2005a) all traditionally recognized Testudo species were highly distinct using mtDNA sequences and ISSR fingerprints, we detected within the T. graeca complex no nuclear genomic differentiation paralleling mtDNA clades. We conclude that all studied populations of the T. graeca complex are conspecific under the Biological Species Concept. There is major incongruence between mtDNA clades and morphologically defined taxa. Morphologically well-defined taxa, like T. g. armeniaca or T. g. floweri, nest within clades comprising also geographically neighbouring, but morphologically distinctive populations of other taxa (clade A: T. g. armeniaca, T. g. ibera, T. g. pallasi ; clade D: T. g. anamurensis, T. g. antakyensis, T. g. floweri, T. g. ibera, T. g. terrestris), while sequences of morphologically similar tortoises of the same subspecies (T. g. ibera sensu stricto or T. g. ibera sensu lato) scatter over two or three genetically distinct clades (A, C or A, C, D, respectively). This implies that pronounced morphological plasticity, resulting in phenotypes shaped by environmental pressure, masks genetic differentiation. To achieve a more realistic taxonomic arrangement reflecting mtDNA clades, we propose reducing the number of T. graeca subspecies considerably and regard in the eastern part of the range five subspecies as valid (T. g. armeniaca, T. g. buxtoni, T. g. ibera, T. g. terrestris, T. g. zarudnyi). As not all North African taxa were included in the present study, we refrain from synonymizing North African taxa with T. g. graeca (mtDNA clade B) that represents a further valid subspecies.

Salmonella in free living terrestrial and aquatic turtles

Detection of Salmonella in pet turtles has been the focus of extensive research, but its incidence in free living turtles is not well known. The aim of this study was to evaluate the incidence of Salmonella in terrestrial and aquatic species of chelonians inhabiting a National Park in southwestern Spain. Individuals of the terrestrial tortoise Testudo graeca ( n = 16) and the aquatic turtles Emys orbicularis ( n = 26) and Mauremys leprosa ( n = 50) were investigated. Maximum incidence of Salmonella was recorded in the terrestrial species (100%). In contrast, the incidence of infected animals was low in the aquatic species, 12% in M. leprosa and 15.4% in E. orbicularis. Five serotypes of Salmonella belonging to subspecies enterica (I) and salamae (II) were identified. All serotypes were found in the terrestrial species, and three in the aquatic ones, suggesting that wild terrestrial chelonians are important reservoirs of Salmonella in our study area. Cloacal transmission during mating is the most probable mode of transmission among individuals.

Chemical Composition of Turtles and Tortoises1–3

Thirty-two aquatic turtles (Sternotherus odoratus; juvenile, semi-adult, adult; body weight 46–238g), 55 tortoises (29 Testudo hermanni,12 Testudo horsfieldii, 11 Geochelone pardalis, 3 Testudo graeca;hatchlings, juvenile, semiadult, adult; body weight 17–1104 g), and5 semiaquatic turtles (Rhinoclemmis pulcherrima; juvenile, semiadult;body weight 31–155 g) were available for the total body analysis. Theywere donated by commercial animal traders and private persons whenthey died from various causes.Crude nutrients were determined by the Weende method (1). Formineralanalysiswetdigestionofsampleswascarriedoutinamicrowavedigestion unit. Calcium was determined by flame emission spectrogra-phy (2), and phosphorus photometrically with ammonium molybdateand ammonium vanadate in HNO

Habitat Selection by the Spur-thighed Tortoise Testudo graeca in a Multisuccessional Landscape: Implications for Habitat Management

As a result of human activities, natural Mediterranean landscapes (including agro-ecosystems) are characterised by a mosaic-like structure with habitat-patches at different successional stages. These systems have high biodiversity levels and are home to a large number of species protected by European laws whose habitats should be adequately managed. In the present work, we study habitat use from an applied point of view in the spur-thighed tortoise Testudo graeca, an endangered reptile present in semi-arid Mediterranean agro-ecosystems. Results show that, at a landscape scale, the species selects simplified vegetation structures and includes in its home range re-colonisation shrubland and small non-irrigated fields. Within the home range, habitat selection patterns vary and areas with higher vegetation cover and complexity are selected. Detected patterns are discussed in terms of the ecological requirements of the species and with a hierarchical view of resources and conditions. The implications of our findings for habitat management aimed at the conservation of the species are also discussed.

Factors determining the distribution of the spur‐thighed tortoiseTestudo graecain south‐east spain: a hierarchical approach

This work represents the first attempt to model the habitat‐species relationships of a species of terrestrial tortoise on a large scale. We applied hierarchical variance partition methodology to Generalised Lineal Models (GLMs), with the presence of the tortoise in 1 km2cells as the response variable. We posited the existence of a hierarchical scheme of factors (including climate, relief and lithology, and land‐use) that determine the distribution ofTestugo graecain southeastern Spain. We also identified the environmental variables within each factor with the greatest explanatory power and decoupled local vs landscape effects. Climate, followed by relief and lithology, and then land‐use, turned out to be the most important factor shaping the distribution ofT. graecain south‐east Spain as well as determining the presence of the species within its range. Univariate models showed that the main climate constraints were related to rainfall and extreme minimum temperatures, two factors which could be related to constraints imposed by the length of the annual activity period and productivity. Finally, multi‐scale decomposition suggested that neighbouring habitat and local dynamics may also be important in the distribution of the species at the landscape scale.