Tail regeneration alters the digestive performance of lizards.

Tissue regeneration is a fundamental evolutionary adaptation, which is well known in lizards that can regenerate their entire tail. However, numerous parameters of this process remain poorly understood. Lizard tail serves many functions. Thus, tail autotomy comes with many disadvantages and the need for quick regeneration is imperative. To provide the required energy and materials for caudal tissue building, lizards are expected to undergo a number of physiological and biochemical adjustments. Previous research showed that tail regeneration induces changes in the digestive process. Here, we investigated if and how tail regeneration affects the digestive performance in five wall lizard species deriving from mainland and island sites and questioned whether the association of tail regeneration and digestion is affected by species relationships or environmental features, including predation pressure. We expected that lizards from high predation environments would regenerate their tail faster and modify accordingly their digestive efficiency, prioritizing the digestion of proteins; the main building blocks for tissue repair. Second, we anticipated that the general food shortage on islands would inhibit the process. Our findings showed that all species shifted their digestive efficiency, as predicted. Elongation rate was higher in sites with stronger predation regime and this was also applied to the rate with which protein digestion raised. Gut passage time increases during regeneration so as to improve the nutrient absorbance, but among the islanders, the pace was more intense. The deviations between species should be attributed to the different ecological conditions prevailing on islands rather than to their phylogenetic relationships.

Investigating the sex-related geometric variation of the human cranium.

Accurate sexing methods are of great importance in forensic anthropology since sex assessment is among the principal tasks when examining human skeletal remains. The present study explores a novel approach in assessing the most accurate metric traits of the human cranium for sex estimation based on 80 ectocranial landmarks from 176 modern individuals of known age and sex from the Athens Collection. The purpose of the study is to identify those distance and angle measurements that can be most effectively used in sex assessment. Three-dimensional landmark coordinates were digitized with a Microscribe 3DX and analyzed in GNU Octave. An iterative linear discriminant analysis of all possible combinations of landmarks was performed for each unique set of the 3160 distances and 246,480 angles. Cross-validated correct classification as well as multivariate DFA on top performing variables reported 13 craniometric distances with over 85% classification accuracy, 7 angles over 78%, as well as certain multivariate combinations yielding over 95%. Linear regression of these variables with the centroid size was used to assess their relation to the size of the cranium. In contrast to the use of generalized procrustes analysis (GPA) and principal component analysis (PCA), which constitute the common analytical work flow for such data, our method, although computational intensive, produced easily applicable discriminant functions of high accuracy, while at the same time explored the maximum of cranial variability.

Population-specificity of sexual dimorphism in cranial and pelvic traits: evaluation of existing and proposal of new functions for sex assessment in a Greek assemblage.

The current paper examines the accuracy of existing binary logistic regression equations for sex prediction based on pelvic and cranial traits in a modern Greek assemblage and proposes new equations with the aim of improving correct classification rates for Balkan material. Our results suggest that existing equations based on pelvic traits perform very well on the Greek material, which can be attributed to the fact that sexual dimorphism in the pelvis results from common evolutionary forces across populations. In contrast, equations based on cranial traits are highly dependent upon the populations based on which they were developed, stressing the need to produce more population-specific functions. Our proposed equations achieve correct sex classification in 92.59% of the females and 95.79% of the males for pelvic traits, while these percentages rise to 97.53% for females and 98.95% for males when the vertical femoral head diameter is included in the models. Our functions based on cranial traits produced correct classifications in up to 92.59% of females and 88.42% of males, and when the cranial scores where combined with the vertical femoral head diameter, the correct classification rates increased to 93.83% for females and 94.73% for males. Prior to the generalization of the use of these functions, further research examining their accuracy in other groups is required, but our results appear promising.

Tail regeneration affects the digestive performance of a Mediterranean lizard.

In caudal autotomy, lizards shed their tail to escape from an attacking predator. Since the tail serves multiple functions, caudal regeneration is of pivotal importance. However, it is a demanding procedure that requires substantial energy and nutrients. Therefore, lizards have to increase energy income to fuel the extraordinary requirements of the regenerating tail. We presumed that autotomized lizards would adjust their digestion to acquire this additional energy. To clarify the effects of tail regeneration on digestion, we compared the digestive performance before autotomy, during regeneration, and after its completion. Tail regeneration indeed increased gut passage time but did not affect digestive performance in a uniform pattern: though protein income was maximized, lipid and sugar acquisition remained stable. This divergence in proteins may be attributed to their particular role in tail reconstruction, as they are the main building blocks for tissue formation.

Partitioning thermal habitat on a vertical rock, a herculean task.

Species occurring in sympatry have to effectively segregate their niche in order to co-exist. In the case of ectotherms in particular, the very important parameter of thermal biology has to be taken into account. Here we investigated the thermoregulatory effectiveness (E) of two endemic Greek lizards (Hellenolacerta graeca and Podarcis peloponnesiacus) that live syntopically on a rocky cliff in the Peloponnese. We presumed that the two species would select different microhabitats, to avoid interspecific competition, and follow a similar thermoregulation pattern as they experience the same conditions. We also expected that E values for both species would differ depending on the season. Overall, we found that the two species had similar E values for each season but differentiated partial thermoregulatory attributes. Though they both occurred in the same types of microhabitat, H. graeca selected higher sites (average 99cm above ground) than P. peloponnesiacus (average 44cm). Also, the latter achieved higher preferred temperatures during summer and winter. Finally, the effectiveness of thermoregulation for both species varied interseasonally and received its highest values during summer, in response to the lowest thermal quality that was observed then. Similar studies stress the importance of thermal shifts for ectotherm co-existence.

Living in sympatry: The effect of habitat partitioning on the thermoregulation of three Mediterranean lizards.

The ability for effective, accurate and precise thermoregulation is of paramount importance for ectotherms. Sympatric lizards often partition their niche and select different microhabitats. These microhabitats, however, usually differ in their thermal conditions and lizards have to adapt their thermoregulation behavior accordingly. Here, we evaluated the impact of habitat partitioning on the thermal biology of three syntopic, congeneric lacertids (Podarcis peloponnesiacus, P. tauricus and P. muralis) from central Peloponnese, Greece. We assessed thermoregulation effectiveness (E) using the three standard thermal parameters: body (Tb), operative (Te) and preferred (Tpref) temperatures. We hypothesized that the microhabitats used by each species would differ in thermal quality. We also predicted that all species would effectively thermoregulate, as they inhabit a thermally challenging mountain habitat. As expected, the partition of the habitat had an effect on the thermoregulation of lizards since microhabitats had different thermal qualities. All three species were effective and accurate thermoregulators but one of them achieved smaller E values as a result of the lower Tb in the field. This discrepancy could be attributed to the cooler (but more benign) thermal microhabitats that this species occupies.

The particularities of a remote islet shape the thermoregulatory profile of an endemic Mediterranean lizard.

Environmental temperatures considerably affect the reptilian ability for thermoregulation and harsh climatic conditions may impose a highly effective body temperature regulation to lizards. Such demanding conditions are more common to extreme mainland habitats (e.g. deserts or mountains). To the contrary, islands have more benign climate conditions thanks to the thermal buffering effect of the surrounding sea. However, this favorable effect may be eliminated in small size islets where the scarcity of thermal shelters and exposure to high winds create challenging conditions. Here we investigate the impact of a tough islet habitat on the thermoregulation of Podarcis levendis, a lacertid lizard endemic to two rocky islets in the north Cretan Sea, Greece. To evaluate the thermoregulatory effectiveness of P. levendis we measured operative and body temperatures in the field and the preferred body temperatures in the lab. Analyses of the thermal data revealed an accurate, precise, and effective thermoregulator, achieving very high thermoregulation values (E =0.91, de¯-db¯ =7.6). This high effectiveness comes to compensate living in an inhospitable habitat as the operative temperatures denote (de =7.79). Our findings, together with the limited published literature, suggested the lack of a general pattern for all insular lizards and indicated a possible deviation for islet habitats.

Effects of insularity on digestion: living on islands induces shifts in physiological and morphological traits in island reptiles.

Living on islands entails numerous challenges for animals, among which resource scarcity stands out. In order to survive, animals have to optimize energy acquisition. We examined the impact of insularity on digestion comparing a series of physiological and morphological traits of adult males between insular and mainland populations of the Balkan green lizard. Island lizards had longer gastrointestinal tracts and gut passage times and higher digestive efficiencies. The dissection of the hindgut revealed an unexpected finding, the presence of cecal valves that were more frequent in island lizards. Thanks to all above islanders retain food for longer periods and thus maximize energy income and increase the amount of the extracted nutrients. That way, they secure energy income from the limited, in time and quantity, food resources of the islands.

Evaluating the efficiency of different recording protocols for entheseal changes in regards to expressing activity patterns using archival data and cross-sectional geometric properties.

OBJECTIVES: In the past decade there have been extensive discussions on the potential utility of entheseal changes (EC) as activity markers. Nevertheless, no study to date has compared different EC recording protocols with respect to their correlation to activity patterns. MATERIALS AND METHODS: This article records changes on fibrocartilaginous entheses of the upper limbs of 90 male skeletons from the documented Athens Collection using the Hawkey and Merbs (Int J Osteoarchaeol 5 (1995) 324-338), Mariotti et al. (Collegium Antropol 28 (2004) 145-159), and Villotte et al. (Am J Phys Anthropol 142 (2010) 224-234) recording schemes in order to determine which one exhibits the highest correlation with activity. Activity is assessed by means of the recorded profession of each individual, as well as employing cross-sectional geometric properties. Generalized Linear Models are used to explore the impact of age, body mass, and activity on EC expression. RESULTS: Our results agree with previous studies that age is the primary factor determining EC, whereas body mass is the second most influential factor. In contrast, activity in the form of profession or cross-sectional geometry rarely showed a significant correlation to EC expression and no clear pattern could be discerned irrespective of the recording technique. However, bilateral differences in the impact of age and body mass in EC expression were traced and may relate to activity patterns. CONCLUSIONS: The differences found in the bilateral impact of age and body mass highlight the fact that the activity patterns of the individuals under examination must play an underlying role to EC expression, though current recording schemes for EC do not capture this, rendering further work in the direction of developing more elaborate recording standards imperative.

Molecular systematics and historical biogeography of the green lizards (Lacerta) in Greece: insights from mitochondrial and nuclear DNA.

The green lizards of the genus Lacerta (Sauria, Lacertidae) comprise nine recognized species, which in Europe are mainly restricted to the southern peninsulas. Four of them (L. trilineata, L. viridis, L. bilineata and L. agilis) occur in Greece. The uncertainty of morphological diversification renders the taxonomic assignment into species and subspecies problematic. In this study sequence data derived from two mitochondrial (cytochrome b and 16S rRNA) genes and one nuclear (NKTR) gene were used to (a) evaluate the taxonomic status of the genus Lacerta in Greece with emphasis on L. trilineata group and (b) investigate the evolutionary history of the genus through the application of phylogenetic and phylogeographic analyses, using Gallotia and Timon as outgroups. The phylogenetic analyses revealed the existence of four major clades. The first clade corresponds to L. trilineata group, the second to L. media, the third to L. agilis and the fourth to a complex of L. viridis and L. bilineata. However, the produced phylogenetic relationships are not congruent with the current taxonomy, especially in the first clade in which L. trilineata appeared to be paraphyletic in regard to L. pamphylica. Six distinct lineages were inferred within L. trilineata, despite the current recognition of nine morphological subspecies, the genetic differentiation of which exceeds that of other Lacerta species, imposing a thorough taxonomic revision of the species. Our results suggested a rapid diversification of L. trilineata group during the late Miocene. We believe that the present distribution of the genus in Greece is the result of several dispersal and vicariant events that took place during the late Miocene and early Pliocene.

Predation Risk, and Not Shelter or Food Availability, as the Main Determinant of Reproduction Investment in Island Lizards.

Reproductive investment, including the number of offspring produced, is one of the fundamental characteristics of a species. It is particularly important for island vertebrates, which face a disproportionate number of threats to their survival, because it predicts, among other things, a species' resilience to environmental disruption. Taxa producing more offspring recover more quickly from environmental perturbations and survive environmental change better. However, ecologists do not understand which primary drivers shape a species' reproductive investment well. Here, we compare the reproductive efforts of 14 island populations of the Aegean Wall Lizard (Podarcis erhardii), which lives across widely diverging environmental conditions. We test three hypotheses, namely that reproductive investment (measured as clutch size, clutch volume) is (1) positively associated with predation risk ['Predation Risk Hypothesis']; (2) positively associated with the presence of reliable vegetation cover that provides shelter ['Gravid Female Protection Hypothesis']; and (3) limited by (and hence positively correlated with) food availability ['Food Limitation Hypothesis']. Although field data are somewhat consistent with all three hypotheses, statistical analyses provide strong support for the Predation Risk Hypothesis. The results not only shed light on which fundamental forces shape reproductive investment in island vertebrates, but can also help shape conservation priorities.

Small Island Effects on the Thermal Biology of the Endemic Mediterranean Lizard Podarcis gaigeae.

Ectotherms are vastly affected by climatic conditions as they rely on external sources of heat to regulate their body temperature, and changes in their habitat thermal quality could seriously affect their overall biology. To overcome the problems of a thermally unfavorable habitat, lizards need to either adjust their thermoregulatory behavior or respond to directional selection and shift their preferred body temperatures. To assess the impact of habitat thermal quality on the thermoregulatory profile, we studied multiple islet and 'mainland' populations of the Skyros wall lizard Podarcis gaigeae, an endemic lacertid to Skyros Archipelago, Greece. We evaluated the effectiveness of thermoregulation (E) using the three main thermal parameters: body (Tb), operative (Te), and preferred (Tpref) temperatures. We first hypothesized that the spatial homogeneity, the scarcity of thermal shelters, and the exposure to higher winds on islets would result in more demanding climate conditions. Second, we anticipated that islet lizards would achieve higher E in response to the lower thermal quality therein. As hypothesized, thermal parameters differed between populations but not in the expected manner. Skyros 'mainland' habitats reached higher temperatures, had more intense fluctuations, and were of lower thermal quality. As a result, lizards showed higher accuracy, precision, and effectiveness of thermoregulation. Noteworthy, we found that lizards from different populations have shifted their thermal profile and preferred body temperatures to cope with the particular conditions prevailing in their habitats. The latter supports the labile view on the evolution of thermoregulation.

Mitochondrial phylogeny and biogeographic history of the Greek endemic land-snail genus Codringtonia Kobelt 1898 (Gastropoda, Pulmonata, Helicidae).

The aim of this work was to infer the phylogeny of the Greek endemic land-snail genus Codringtonia Kobelt 1898, estimate the time frame of the radiation of the genus, and propose a biogeographic scenario that could explain the contemporary distribution of Codringtonia lineages. The study took place in the districts of Peloponnese, Central Greece and Epirus of mainland Greece. Sequence data originating from three mtDNA genes (COI, COII, and 16S rDNA) were used to infer the phylogeny of the eight nominal Codringtonia species. Furthermore, the radiation time-frame of extant Codringtonia species was estimated using a relaxed molecular clock analysis and mtDNA substitution rates of land snails. The phylogenetic analysis supported the existence of six Codringtonia lineages in Greece and indicated that one nominal species (Codringtonia neocrassa) might belong to a separate genus distantly related to Codringtonia. The time frame of differentiation of Codringtonia species was placed in the Late Miocene-Pleistocene epoch. The dispersal-vicariance analysis performed indicated that most probably Codringtonia exhibited a north-to-south spread with the ancestral area being that of central Greek mainland, accompanied with duplication (speciation) and vicariance events.

Distribution and accumulation of metals in tadpoles inhabiting the metalliferous streams of eastern Chalkidiki, northeast Greece.

The present study investigates the accumulation of heavy metals [copper (Cu), lead (Pb), zinc (Zn), magnesium (Mn), cadmium (Cd), nickel (Ni), and chromium (Cr)] in tadpoles inhabiting the metalliferous streams flowing within the Asprolakkas River basin (northeast Chalkidiki peninsula, Greece) and the effect of potentially harmful elements in stream water and sediment on the corresponding levels in their tissue. Animals were collected from six sampling sites influenced by a wide range of surface water and stream sediment trace element concentrations. The results of the chemical analyses showed that tadpoles accumulated significant levels of all of the examined metals. The range of whole-body mean measured concentrations were (in dry mass) as follows: Cu (46-182 mg/kg), Pb (103-4,490 mg/kg), Zn (494-11,460 mg/kg), Mn (1,620-13,310 mg/kg), Cd (1.2-82 mg/kg), Ni (57-163 mg/kg), and Cr (38-272 mg/kg). The mean concentrations of Pb, Zn, Mn, Ni, Cr, and Cd in Kokkinolakkas stream, which drains a currently active mining area, were the highest ever reported in tadpoles. Our results indicate that whole-body levels of Pb, Zn, Cu, and Cd increase with stream sediment concentrations and that these organisms tend to accumulate metals bound to Fe and Mn oxides. In addition, high dissolved concentrations and significant concentrations associated with more labile geochemical phases of sediments for specific metals were contributing factors determining whole-body levels. Given the observed bioconcentration factors, as well as the correlation with sediment concentrations, it is proposed that these organisms could be considered as bioindicators of environmental contamination and may be used for monitoring purposes within this metal-rich zone and, perhaps, within other rivers affected by metal mining.

Island biology and morphological divergence of the Skyros wall lizard Podarcis gaigeae: a combined role for local selection and genetic drift on color morph frequency divergence?

BACKGROUND: Patterns of spatial variation in discrete phenotypic traits can be used to draw inferences about the adaptive significance of traits and evolutionary processes, especially when compared to patterns of neutral genetic variation. Population divergence in adaptive traits such as color morphs can be influenced by both local ecology and stochastic factors such as genetic drift or founder events. Here, we use quantitative color measurements of males and females of Skyros wall lizard, Podarcis gaigeae, to demonstrate that this species is polymorphic with respect to throat color, and the morphs form discrete phenotypic clusters with limited overlap between categories. We use divergence in throat color morph frequencies and compare that to neutral genetic variation to infer the evolutionary processes acting on islet- and mainland populations. RESULTS: Geographically close islet- and mainland populations of the Skyros wall lizard exhibit strong divergence in throat color morph frequencies. Population variation in throat color morph frequencies between islets was higher than that between mainland populations, and the effective population sizes on the islets were small (Ne:s < 100). Population divergence (FST) for throat color morph frequencies fell within the neutral FST-distribution estimated from microsatellite markers, and genetic drift could thus not be rejected as an explanation for the pattern. Moreover, for both comparisons among mainland-mainland population pairs and between mainland-islet population pairs, morph frequency divergence was significantly correlated with neutral divergence, further pointing to some role for genetic drift in divergence also at the phenotypic level of throat color morphs. CONCLUSIONS: Genetic drift could not be rejected as an explanation for the pattern of population divergence in morph frequencies. In spite of an expected stabilising selection, throat color frequencies diverged in the islet populations. These results suggest that there is an interaction between selection and genetic drift causing divergence even at a phenotypic level in these small, subdivided populations.

Intraspecific competition and high food availability are associated with insular gigantism in a lizard.

Resource availability, competition, and predation commonly drive body size evolution. We assess the impact of high food availability and the consequent increased intraspecific competition, as expressed by tail injuries and cannibalism, on body size in Skyros wall lizards (Podarcis gaigeae). Lizard populations on islets surrounding Skyros (Aegean Sea) all have fewer predators and competitors than on Skyros but differ in the numbers of nesting seabirds. We predicted the following: (1) the presence of breeding seabirds (providing nutrients) will increase lizard population densities; (2) dense lizard populations will experience stronger intraspecific competition; and (3) such aggression, will be associated with larger average body size. We found a positive correlation between seabird and lizard densities. Cannibalism and tail injuries were considerably higher in dense populations. Increases in cannibalism and tail loss were associated with large body sizes. Adult cannibalism on juveniles may select for rapid growth, fuelled by high food abundance, setting thus the stage for the evolution of gigantism.

Assessing the accuracy of cranial and pelvic ageing methods on human skeletal remains from a modern Greek assemblage.

The present study tests the accuracy of commonly adopted ageing methods based on the morphology of the pubic symphysis, auricular surface and cranial sutures. These methods are examined both in their traditional form as well as in the context of transition analysis using the ADBOU software in a modern Greek documented collection consisting of 140 individuals who lived mainly in the second half of the twentieth century and come from cemeteries in the area of Athens. The auricular surface overall produced the most accurate age estimates in our material, with different methods based on this anatomical area showing varying degrees of success for different age groups. The pubic symphysis produced accurate results primarily for young adults and the same applied to cranial sutures but the latter appeared completely inappropriate for older individuals. The use of transition analysis through the ADBOU software provided less accurate results than the corresponding traditional ageing methods in our sample. Our results are in agreement with those obtained from validation studies based on material from across the world, but certain differences identified with other studies on Greek material highlight the importance of taking into account intra- and inter-population variability in age estimation.

Combining immunological and molecular data to assess phylogenetic relations of some Greek Podarcis species.

Most recent molecular studies revealed the phylogeny of Greek Podarcis species, which for years remained elusive, due to discordant data produced from various chromosomal, complement fixation and protein studies. In this report, we analyzed cellular immune responses of spleen-derived lymphocytes from six allopatric Podarcis species encountered in Greece, by assessing two-way mixed lymphocyte reaction (MLR)-induced proliferation. On the basis of stimulation indices (S.I.) as determined from cultures set up from xenogeneic splenocytes coincubated in pairs, we generated a phylogenetic tree, fully consistent with the phylogenetic relationships of Podarcis as determined by parallel analyses based on partial mitochondrial (mt) DNA sequences. Although the exact mechanisms triggering lymphocyte responses in lizard two-way xenogeneic MLR are not fully understood, our results show the potential use of cell-mediated immune responses as an additional approach to mtDNA analysis, for species delimitation within specific lizard taxa.

Sex determination by three-dimensional geometric morphometrics of the vault and midsagittal curve of the neurocranium in a modern Greek population sample.

The aim of this study is to assess sexual dimorphism of adult crania in the vault and midsagittal curve of the vault using three-dimensional geometric morphometric methods. The study sample consisted of 176 crania of known sex (94 males, 82 females) belonging to individuals who lived during the 20th century in Greece. The three-dimensional co-ordinates of 31 ecto-cranial landmarks and 30 semi-landmarks were digitized using a MicroScribe 3DX contact digitizer. Generalized Procrustes analysis (GPA) was used to obtain size and shape variables for statistical analysis. Shape, size and form analyses were carried out by logistic regression and three discriminant function analyses. Results indicate that there are shape differences between sexes. Females in the region of the parietal bones are narrower and the axis forming the frontal and occipital bones is more elongated; the frontal bone is more vertical. Sex-specific shape differences give better classification results in the vault (79%) compared with the midsagittal curve of the neurocranium (68.8%). Size alone yielded better results for cranial vault (82%), while for the midsagittal curve of the vault the result is poorer (68.1%). As anticipated, the classification accuracy improves when both size and shape are combined (89.2% for vault, and 79.4% for midsagittal curve of the vault). These latter findings imply that, in contrast to the midsagittal curve of the neurocranium, the shape of the cranial vault can be used as an indicator of sex in the modern Greek population.

Sex determination by three-dimensional geometric morphometrics of craniofacial form.

SUMMARY: The purpose of the present study is to define which regions of the cranium, the upper-face, the orbits and the nasal are the most sexually dimorphic, by using three-dimensional geometric morphometric methods, and investigate the effectiveness of this method in determining sex from the shape of these regions. The study sample consisted of 176 crania of known sex (94 males, 82 females) belonging to individuals who lived in Greece during the 20(th) century. The three-dimensional co-ordinates of 31 ecto-cranial landmarks were digitized using a MicroScribe 3DX contact digitizer. Goodall's F-test was performed in order to compare statistical differences in shape between males and females. Generalized Procrustes Analysis (GPA) was used to obtain size and shape variables for statistical analysis. Shape, Size and Form analyses were carried out by logistic regression and discriminant function analysis. The results indicate that there are shape differences between the sexes in the upper-face and the orbits. The highest shape classification rate was obtained from the upper-face region. The centroid size of the caraniofacial and the orbital regions was smaller in females than males. Moreover, it was found that size is significant for sexual dimorphism in the upper-face region. As anticipated, the classification accuracy improves when both size and shape are combined. The findings presented here constitute a firm basis upon which further research can be conducted.