Augusto Gentilli

Male-male combats in a polymorphic lizard: residency and size, but not color, affect fighting rules and contest outcome.

Theoretical models predict that the outcome of dyadic agonistic encounters between males is influenced by resource-holding potential, resource value, and intrinsic aggressiveness of contestants. Moreover, in territorial disputes residents enjoy a further obvious competitive advantage from the residency itself, owing to the intimate familiarity with their territory. Costs of physical combats are, however, dramatically high in many instances. Thus, signals reliably reflecting fighting ability of the opponents could easily evolve in order to reduce these costs. For example, variation in color morph in polymorphic species has been associated with dominance in several case studies. In this study, we staged asymmetric resident-intruder encounters in males of the common wall lizard Podarcis muralis, a species showing three discrete morphs (white, yellow, and red) to investigate the effects of asymmetries in color morph, residency, and size between contestants on the outcome of territorial contests. We collected aggression data by presenting each resident male with three intruders of different color morph, in three consecutive tests conducted in different days, and videotaping their interactions. The results showed that simple rules such as residency and body size differences could determine the outcome of agonistic interactions: residents were more aggressive than intruders, and larger males were competitively superior to smaller males. However, we did not find any effect of color on male aggression or fighting success, suggesting that color polymorphism in this species is not a signal of status or fighting ability in intermale conflicts.

Photographic identification in reptiles: a matter of scales

Photographic identification is a promising marking technique alternative to the toe-clipping, since it is completely harmless, cheap, and it allows long time identification of individuals. Its application to ecological studies is mainly limited by the time consuming to compare pictures within large datasets and the huge variation of ornamentation patterns among different species, which prevent the possibility that a single algorithm can effectively work for more than few species. Scales of Reptiles offer an effective alternative to ornamentations for computer aided identification procedures, since both shape and size of scales are unique to each individual, thus acting as a fingerprint like ornamentation patterns do. We used the Interactive Individual Identification System (I 3 S) software to assess whether different individuals of two species of European lizards ( Podarcis muralis and Lacerta bilineata ) can be reliably photographically identified using the pattern of the intersections among pectoral scales as fingerprints. We found that I 3 S was able to identify different individuals among two samples of 21 individuals for each species independently from the error associated to the ability of the operators in collecting pictures and in digitizing the pattern of intersections among pectoral scales. In a database of 1043 images of P. muralis collected between 2007 and 2008, the software recognized 98% of recaptures within each year, and 99% of the recaptures between years. In addition, 99% and 96% of matches were ranked among the top five, and no more than 5 minutes were needed for digitizing and processing each image. The lepidosis of reptiles is a reliable alternative to ornamentation patterns in photographic identification of reptiles, which can be effectively analysed using the I 3 S software. This result represents a significant improvement in photographic identification of reptiles since (i) this procedure can be easily extended to most other species of reptiles, (ii) all kind of individuals within a species can be marked (i.e., young, subadults and adults) despite the differences in ornamentation patterns, and (iii) it is the only technique for species, like the western green lizard, which lack a clear ornamentation pattern.

Molecular phylogeography of the asp viper Vipera aspis (Linnaeus, 1758) in Italy: evidence for introgressive hybridization and mitochondrial DNA capture

Owing to its temperature dependence and low vagility, the asp viper (Vipera aspis) is an interesting model species to study the effects of Pleistocene climatic fluctuations on vertebrate genomes. We genotyped 102 specimens from the whole Italian distribution range at three mitochondrial DNA regions (2278 characters, total) and six microsatellite DNA loci (Short Tandem Repeats, STR). The molecular phylogeny was constructed according to Bayesian, Neighbour Joining, Maximum Parsimony and Maximum Likelihood procedures. All methods grouped individuals of the three morphological subspecies (V. a. aspis, V. a. francisciredi, V. a. hugyi) into five different haploclades. Specimens assigned to hugyi clustered in two highly differentiated clades, one being sister group to the complex comprising the second clade of hugyi (i.e., a paraphyletic status), plus two clades of francisciredi. The Bayesian clustering of the STR variability disclosed only two groups, the first including aspis and francisciredi, the second all hugyi. Introgressive hybridization and capture of francisciredi-like lineages in the hugyi mitochondrial genome were suggested to explain the discordance between mitochondrial and nuclear data. The phylogeographic pattern was compatible with population contractions in three glacial refuges. Plausibility of derived hypothesis was checked using coalescence simulations as post hoc tests. Long-term drift and serial founder effects, rather than selection, appeared the main factors affecting the genetic make-up of the Italian asp viper.

Morph-specific immunity in male Podarcis muralis

Permanent colour polymorphism in lizards is maintained by complex interactions between environmental pressures and physiological traits (such as immune responsiveness) that differ among morphs. In this study we investigated whether T-cell mediated immune response vary among male colour morphs in the trimorphic (white, yellow and red) common wall lizard, Podarcis muralis . We found that yellow males showed a lower immune response compared to both red and white males, whose responses were similar. Thus, immune responsiveness is morph-specific in male common wall lizards, suggesting that this physiological trait could play an important role in maintaining colour polymorphism in this species. Moreover, immune responsiveness significantly increased with increasing male size, irrespective of colour morph, indicating that it could be regarded as a condition-dependent trait.

Sex-, morph- and size-specific susceptibility to stress measured by haematological variables in captive common wall lizard Podarcis muralis.

In polymorphic species of animals, colour morphs may show alternative physiological properties, and hence evolve or be maintained as an indirect response to selection exerted on these physiological attributes. In this study, we investigated if different colour morphs (white, red and yellow) of the polymorphic common wall lizard differed in their physiological responses to a long-term stress by determining variation between capture and release in leukocytes profiles, haemoparasite loads and body condition of male and females maintained in captivity throughout the breeding season. We found that most blood parameters of lizards varied significantly following captivity, and this variation was sex-, morph- and size-dependent. In particular, the heterophil:lymphocyte ratio (H:L), a sensitive measure of immunodepression and long-term stress, varied significantly among yellow females, larger individuals significantly increasing and smaller individuals decreasing their H:L ratio after captivity. This trend was reversed in red females, where smaller individuals presented raised H:L index at release. Our study indicated that response to long-term stressful conditions, such as those induced by captivity, differed among common wall lizard colour morphs, implying a sex-, size-(i.e. age) and morph-specific sensitivity to stress, and hence a different physiological profile of colour morphs, which may contribute to the maintenance of colour polymorphism in this species.

Colour Polymorphism and Alternative Breeding Strategies: Effects of Parent’s Colour Morph on Fitness Traits in the Common Wall Lizard

Colour polymorphism (CP) is widespread in animals, but mechanisms underlying morph evolution and maintenance are not completely resolved. In reptiles, CP is often genetically based and associated with alternative behavioural strategies, mainly in males for most cases. However, female colour morphs also display alternative reproductive strategies associated with behavioural and physiological traits, which may contribute to maintain CP in the population. Both sexes of the common wall lizard (Podarcis muralis) show three pure colour morphs, white, yellow and red. Here, we looked for the effects of male and female colour morphs on fitness traits of captive-breeding pairs. All yellow-throated females laid clutches of many small eggs and produced many light offspring, behaving as r-strategists, whereas white-throated females laid clutches of few large eggs and produced few heavy offspring, behaving as K-strategists. Red-throated females adopted a conditional Kr-strategy depending on their size/age. These basic female strategies were modulated in relation to mate morph: white females had the best fitness gain in terms of viable offspring when mated to red males; mating between yellow morphs yielded a greater breeding success than all other morph crosses, but also lighter offspring; finally, red females produced heavy progeny when paired with red or white males, and light offspring in pair with yellow males. Thus, correlation between CP and traits relevant to fitness combined with non-random mating, either assortative or disassortative, could increase the potential for CP to contribute to divergent evolution in the common wall lizard.

A tribute to Hubert Saint Girons: niche separation between Vipera aspis and V. berus on the basis of distribution models

Interspecific competition in contact areas is a major topic in ecological studies. A number of studies were carried out on European vipers, focusing on contact areas between two or three species characterized as appropriate by ecological needs more or less similar. The aim of this study is to extend the analysis of this topic to the case of Vipera aspis and V. berus in an alpine area of northern Italy, by comparing suitability models to verify which ecological factors affect their occurrence and to assess a possible niche separation. Potential distribution was modelled using the maximum entropy method, using six non-correlated ecogeographical variables as predictors. The models fitted well for both species (mean AUC = 0.926; 87.4% of testing data correctly classified). The most informative variables were: habitat, altitude and solar radiation for the asp viper; altitude and habitat for the adder. Deciduous woods, meadows and urban areas had a positive effect on V. aspis distribution as wetlands, meadows and rocks vegetation did on V. berus . However, the variable best separating the species was the elevation, the adder occurring more frequently at higher altitude than the asp viper. Our data showed that the two vipers were mutually exclusive, as already observed by Saint Girons in 1975. Vipera aspis is more thermophilic and lives at low altitude, while Vipera berus lives under cool and humid areas typical of alpine pastures. A similar pattern were found in the contact areas between European vipers belonging to the V. aspis and Pelias group respectively.

The karyology of Vipera aspis , V. atra , V. hugyi , and Cerastes vipera

In the current paper we show the results obtained using standard and banding staining methods (Ag-NOR-, CMA3-, C-banding and sequential colorations (or Alu I digestions) + CMA3 + DAPI) in specimens of Cerastes vipera, Vipera aspis, V. atra, and V. hugyi. Cerastes vipera presented chromosomal characters, primitive in snakes, as a karyotype of 2n = 36 chromosomes, with 16 biarmed macrochromosomes and 20 microchromosomes, NORs on one microchromosome pair and absence of cytologically evident sex chromosomes, at least with the methods used. The three taxa of Vipera studied showed chromosomal characters either derived, or primitive or at an initial stage of differentiation. All three species showed a karyotype (derived) of 2n = 42 chromosomes with 22 macro- and 20 micro-chromosomes; they all showed NORs on one micro-chromosome pair and presented Z and W chromosomes at an initial stage of differentiation. Sexchromosomes Z and W, were in fact homomorphic, but the former was near all euchromatic, while the W chromosome was almost completely heterochromatic. All the three taxa of Vipera resulted, however, karyologically diversified, mainly due to the number of macro-chromosomes pairs with a centromeric, CMA3 positive heterochromatin: almost all the pairs in V. aspis, two pairs in V. atra and absent in V. hugyi.

Rana dalmatina and R. latastei: Habitat selection, fluctuation in egg clutch deposition and response to exceptional floods in northern Italy

Abstract A ten year census was carried out on Rana dalmatina and R. latastei egg masses deposited in 36 reproductive sites in a 10‐hectare area, subject to periodic flooding, on the banks of the Ticino River. The vegetation type surrounding reproductive sites was noted. There were no signs of decline in the populations of either frog species. The number of egg clutches fluctuated widely from year to year but no correlation between clutch number and either flooding or climate parameters were detected. R. dalmatina preferred partly open areas for reproductive sites, whereas R. latastei selected areas characterized by hygrophilous vegetation and a high tree canopy.

Seasonal variations of plasma testosterone among colour-morph common wall lizards (Podarcis muralis)

Sexual steroids influence reproductive behaviours and promote secondary sexual traits. In male lizards, increasing levels of testosterone (T) bolster conspicuous colouration, stimulate territoriality, and trigger antagonistic interactions among rivals. Moreover, in colour polymorphic species, reproductive strategy, aggressiveness and T levels can differ between morphs. Therefore, T level is considered as an important mechanism that regulates the expression of colour polymorphism and sexual behaviours of males. But in the polymorphic territorial wall lizard (Podarcis muralis), a lack of relationship between morphs and aggressiveness challenges the notion that T plays such a role. To examine this issue, we compared adult T levels among three colour morphs (white, yellow and red) through repeated sampling during the mating season. High T levels were observed at the onset of the mating season followed by a significant decrease, a pattern documented in other lizard species. Mean T levels did not differ among morphs. However, yellow males maintained significantly higher T levels over time and displayed a stronger subsequent decline. Overall, in this species, seasonal T patterns differ among morphs, not mean values. Previous studies revealed that T suppresses the immune response; suggesting that a strong initial investment promoted by high T levels may trade-off against immunity (maintenance). Further experimental investigations are required to clarify the relationship between T and reproductive effort in polymorphic species that exhibit complex temporal pattern of T levels.