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The introduction of alien water frogs is perhaps one of the most underestimated herpetological conservation issues in Europe. The identification of distinct species is highly challenging at the phenotypic level, and artificial syntopy between various taxa and lineages may lead to diverse outcomes, including hybridisation and local extinction. In central Italy the native synklepton of Pelophylax bergeri (the parental taxon) and P. kl. (klepton) hispanicus (the hybridogenetic hybrid, which clonally transmits the genome of an extinct ridibundus-like taxon) is present. Until recently, data regarding the presence of alien water frogs in central Italy was scarce, and no alien taxa have been reported for Tuscany. In this study, four distinct non-native Pelophylax lineages have been identified via molecular analysis in the Cecina and Arno river basins and ascribed to the Marsh frog group (P. ridibundus sensu lato). Alien Pelophylax ridibundus, P. kurtmuelleri, and P. cf. bedriagae sensu stricto currently appear to be widespread in the Cecina basin. Furthermore, evidence of hybridisation with autochthonous taxa has been suggested by genetic analyses in four out of eight sampling localities. With a view to evaluate urgent conservation strategies, a greater sampling effort is required to assess the actual distribution and ecology of the alien lineages, and further research is necessary to measure their impact on the native hybridogenetic system of the central-southern Italian pool frogs.

The Socotra Archipelago is a unique hotspot of biodiversity, particularly for terrestrial reptiles (28 endemic out of 31 native species) whose ecology is known only in general terms. We describe the ecological preference and the daily activity of 21 species of terrestrial reptiles, geckos, lizards, skinks, snakes and chameleon, from the largest island of the archipelago. The database includes 6950 records collected by 260 standardized transects from 2007 to 2013. The sister species Haemodracon riebeckii–H. trachyrhinus with very distinct body sizes, overlapped in their distribution but segregated by habitat and substrate. Three pairs of sister species with great phylogenetic affinity and similar morphology (Pristurus insignis–P. insignoides; Pristurus guichardi–P. obsti; Hemidactylus dracaenacolus–H. granti) were only partially segregated by distribution, habitat, substrate and body size, in the order of decreasing frequency. On the contrary, the other, phylogenetically more distant species of geckos, widely distributed and less similar in morphology, overlapped completely in their habitat use and distributional range. Such patterns suggest that strictly related species reduced their competition by segregation in different habitats and temporal niches and by non-overlapping distribution within the relatively small island of Socotra. On the other hand, the activity rhythm was similar within each genus. The unique biodiversity of Socotra endured up to present, thanks to a human impact lower than for most other inhabited islands, but the pace of change has accelerated recently. Conservation measures are urgent, particularly for the species of endemic geckos with a distribution range as small as 10 km2., RV was supported by a contract from ‘Fundação para a Ciência e a Tecnologia’ (FCT), I.P. national funds from ‘Norma transitória’ (DL57/2016/CP1440/CT0002).

In northwestern Italy, traditional rice cultivation using permanent flooding has been replaced by cultivation that uses a reduced water depth and a shortened submersion period. Prey availability for the numerous herons and egrets using this intensive rice cultivation region changed as well, with a diminution of amphibians and an increase of crayfish (Procambarus clarkii) and oriental weatherfish (Misgurnus anguillicaudatus). The prey intake in 2014 was compared with data available for 1987 for the three most common species of waterbirds that forage in the paddies. Total biomass intake by Grey Herons (Ardea cinerea) feeding in the paddies decreased from 28 g dry mass/min in 1987 to 7 g in 2014, and by Black-crowned Night-Herons (Nycticorax nycticorax) from 8 g to 5 g, while the intake by Little Egrets (Egretta garzetta) remained similar (12 and 13 g in 1987 and 2014, respectively). The observed levels of prey intake allowed breeding Grey Herons to meet their energy needs by feeding exclusively in the rice paddies in 1987, but not in 2014. The other two study species could not meet their daily requirements for successful breeding by feeding in the paddies only, either in 1987 or 2014. Yet the three species bred successfully in the study area, and their reproductive output remained stable both in 1987 and 2014. Successful breeding was probable by the three species foraging in other habitats, such as rivers, small natural wetlands and irrigation canals, in the region.

Recent changes in rice crop management within Northern Italy rice district led to a reduction of seeding in flooding condition, which may have an impact on reservoir water management and on the animal and plant communities that depend on the flooded paddies. Therefore, monitoring and quantifying the spatial and temporal variability of water presence in paddy fields is becoming important. In this study we present a method to estimate dynamics of presence of standing water (i.e. fraction of flooded area) in rice fields using MODIS data. First, we produced high resolution water presence maps from Landsat by thresholding the Normalised Difference Flood Index (NDFI) made: we made it by comparing five Landsat 8 images with field-obtained information about rice field status and water presence. Using these data we developed an empirical model to estimate the flooding fraction of each MODIS cell. Finally we validated the MODIS-based flooding maps with both Landsat and ground information. Results showed a good predictability of water surface from Landsat (OA = 92%) and a robust usability of MODIS data to predict water fraction (R 2 = 0.73, EF = 0.57, RMSE = 0.13 at 1 × 1 km resolution). Analysis showed that the predictive ability of the model decreases with the greening up of rice, so we used NDVI to automatically discriminate estimations for inaccurate cells in order to provide the water maps with a reliability flag. Results demonstrate that it is possible to monitor water dynamics in rice paddies using moderate resolution multispectral satellite data. The achievement is a proof of concept for the analysis of MODIS archives to investigate irrigation dynamics in the last 15 years to retrieve information for ecological and hydrological studies.

At Anzali Lagoon, on the southwest coast of the Caspian Sea, Great Crested Grebes Podiceps cristatus were observed breeding both in monospecific colonies and in a mixed colony with Whiskered Terns Chlidonias hybridus. In the mixed colony, Great Crested Grebes bred later than in their monospecific colonies, while laying clutches of similar size, but significantly fewer of their nests disappeared during incubation and their chicks had higher survival rates up to at least 20 days old. Human disturbance and the resulting increases in predation likely caused greater losses at monospecific colonies of Great Crested Grebes where there is no common anti-predatory activity; while the strong anti-predatory behaviour of Whiskered Terns in the mixed colony might have offered enhanced protection to the chicks. Unfortunately, we could not separate the effects of the seasonal differences in human disturbance and of the protective presence of terns on the Great Crested Grebe's reproductive output.

The importance of targeting ecological and evolutionary processes in reserve design has been widely acknowledged in the literature but rarely implemented on islands. Using Socotran reptiles as models, we aim to relate richness of widespread and restricted-range species directly with landscape variables and to compare the impact of setting conservation targets for lineages versus species. Socotra Island is a UNESCO Natural World Heritage Site, containing high levels of endemism in relation to its area, especially of reptiles, the vertebrates with the most comprehensive available genetic data. We predicted the occurrences of reptile species using distribution models and used a novel approach to interpolate maps of spatial phylogenetic patterns. Patterns of intra and interspecifc diversity and differences between spatial outputs of lineage and species richeness were related to eco-geographic variables. We evaluated differences in target achievement for each conservation unit within protected areas (PAs) under the current Zoning Plan (ZP) using gap and reserve design analyses. Although intraspecific richness was strongly correlated with interspecific richness, differences in their spatial distribution reached ~30% in some areas. Differences were more pronounced for wide-ranging than restricted-range taxa. Gap analysis indicates that most conservation units are under-represented in sanctuaries and that intra and interspecific richness were significantly higher outside PAs. This work will guide local-scale conservation planning as the ZP is due to be re-evaluated. This is one of the few studies on islands using genetic data from an entire class of vertebrates to incorporate lineage diversity in reserve design. This study provides an alternative methodological framework for supporting the use of landscape and genetic tools in reserve design, circumventing the use of phylogenetic distances and deterministic spatial interpolation of lineage diversity that can be widely applied to other systems.

The intensive rice cultivation area in northwestern Italy hosts the largest surface of rice paddies in Europe, and it is valued as a substantial habitat for aquatic biodiversity, with the paddies acting as a surrogate for the lost natural wetlands. The extent of submerged paddies strictly depends on crop management practices: in this framework, the recent diffusion of rice seeding in dry conditions has led to a reduction of flooded surfaces during spring and could have contributed to the observed decline of the populations of some waterbird species that exploit rice fields as foraging habitat. In order to test the existence and magnitude of a decreasing trend in the extent of submerged rice paddies during the rice-sowing period, MODIS remotely-sensed data were used to estimate the extent of the average flooded surface and the proportion of flooded rice fields in the years 2000–2016 during the nesting period of waterbirds. A general reduction of flooded rice fields during the rice-sowing season was observed, averaging − 0.86 ± 0.20 % per year (p-value < 0.01). Overall, the loss in submerged surface area during the sowing season reached 44 % of the original extent in 2016, with a peak of 78 % in the sub-districts to the east of the Ticino River. Results highlight the usefulness of remote sensing data and techniques to map and monitor water dynamics within rice cropping systems. These techniques could be of key importance to analyze the effects at the regional scale of the recent increase of dry-seeded rice cultivations on watershed recharge and water runoff and to interpret the decline of breeding waterbirds via a loss of foraging habitat.

In countries with emerging and developing economies the need to promote development and the lack of information on the status of the Near Threatened Eurasian otter Lutra lutra have given rise for concern about the conservation status of the species. In Albania information about the distribution of this otter dates from 1985. In 2013 we resurveyed 31 sites previously surveyed in 1985, and a further 42 sites throughout the country. At each site nine habitat variables of potential importance to otters were recorded and analysed. Overall the distribution pattern in 2013 did not differ from that recorded in 1985, although a reduction in marking intensity suggested a possible decline in otter numbers. Distribution of the otter has been influenced by land use and human density, suggesting man-induced habitat changes since the fall of communism may have affected the quality and fragmentation of otter habitats.

The condition-dependent sexual dimorphism model explains the evolution and maintenance of sexual dimorphism in traits targeted by sexual selection, and predicts that the magnitude of sexual dimorphism depends on the variability of individual condition, male traits being more variable than female corresponding traits. Most convincing examples concern insects, while studies among vertebrates are scanty because manipulating condition often is not possible, and the time to reach sexual maturity may be too long. Islands offer a unique opportunity to compare how the environment affects the expression of sexual dimorphism, since they represent ‘natural experimental sets’ in which different populations of the same species may experience alternative environmental constraints. We investigated the occurrence of context-dependent expression in sexual dimorphism of head shape in insular populations of the common wall lizards (Podarcis muralis) inhabiting the Tuscan Archipelago (Tyrrhenian Sea). Alternative models were formulated: H0 assumes that the sexual dimorphism is uninfluenced by islands, H1 assumes the only effect of phylogeny, H2A and H2B account for the biogeography of the archipelago (island size and distance from the mainland), while H3 assumes island-specific effects on sexual dimorphism. Models were compared using Akaike's information criterion adjusted for multivariate analyses. All hypotheses performed better than H0, but H3 largely outperformed all other alternative hypotheses, indicating that environmental features of islands play an additive effect to ontogenetic, biogeographic and genetic factors in defining variation in head shape sexual dimorphism. Our results support the hypothesis of a context-dependent sexual dimorphism in common wall lizards. © 2015 The Linnean Society of London, Biological Journal of the Linnean Society, 2015, 114, 552–565.

The concentrations of trace metals were assessed using feathers of cattle egrets (Bubulcus ibis), collected within two industrial areas of Pakistan, Lahore and Sialkot. We found, in order of descending concentration: Zinc (Zn), Iron (Fe), Nickel (Ni), Copper (Cu), Cadmium (Cd), and Manganese (Mn), Chromium (Cr), Arsenic (As), and Lithium (Li), without any significant difference (except Fe, Zn, and Ni) between the two areas. The concentrations of trace metals, we recorded were among the highest ever reported in the feathers of avian species worldwide. The concentrations of Cr, Pb, Cd were above the threshold that affects bird reproductive success. The high contamination by heavy metals in the two areas is due to anthropogenic activities as well to natural ones (for As and Fe). The bioaccumulation ratios in eggs and feathers of the cattle egret, their prey, and the sediments from their foraging habitats, confirmed that avian feathers are a convenient and non-destructive sampling tool for the metal contamination. The results of this study will contribute to the environmental management of the Lahore and Sialkot industrial areas.

ummary Local recruitment and immigration play an important part in the dynamics and growth of animal populations. However, their estimation and incorporation into open populations models is, in most cases, problematic. We studied factors affecting the growth of a recently established colony of Eurasian spoonbill (Platalea leucorodia) and assessed the contribution of local recruits, i.e. birds born in the colony, and immigrants, i.e. birds of unknown origin, to colony growth. We applied an integrated population model that accounts for uncertainty in breeding state assignment and merges population surveys, local fecundity and individual longitudinal data of breeding and non-breeding birds, to estimate demographic rates and the relative role of recruitment and immigration in driving the local dynamics. We also used this analytical framework to assess the degree of support for the ʻperformance-basedʼ and ʻconspecific attractionʼ hypotheses as possible mechanisms of colony growth. Among the demographic rates, only immigration was positively and significantly correlated with population growth rate. In addition, the number of immigrants settling in the colony was positively correlated with colony size in the previous and current year, but was not correlated with fecundity of the previous year. Our results suggest that the variation in immigration affected colony dynamics and that conspecific attraction likely triggered the relevant role of immigration in the growth of a recently formed waterbird colony, supporting the need of including immigration in population analysis. This article is protected by copyright. All rights reserved.

The breeding herons and egrets in northwestern Italy derive a large portion of their food resources from rice fields. In order to investigate the possible effects of changes in rice cultivation techniques on the trends of the breeding population of these birds, we studied their diet and the prey availability in rice fields, and we compared the results for 2013 with the situation in past decades, by taking advantage of similar investigations conducted by our research team since 1978. The diet of five species of herons and egrets changed sharply after 1995, with the bulk of the diet now made up of completely new prey species, in particular the allochthonous Louisiana crayfish and Oriental weatherfish, that only recently colonized our study area, and with a lower frequency of the amphibians that were a staple prey before 2000, but whose abundance in rice fields has declined. Since 2000, the decrease in the populations of some breeders, particularly the most abundant Grey Heron and Little Egret, has c...

Complex and ritualized displays require an assemblage of structural, neuronal, and muscular adaptations, and habitat structure may affect the effectiveness of signals to convey information. Therefore, ornament exaggeration through sexual selection may be severely constrained by the costs imposed by natural selection. We investigated this compromise in the courtship behavior of Mediterranean Tortoises by assessing the variability of the acoustic and behavioral signals used among species, and the relationship between courtship and species body size. Marginated, Greek, and Hermann's Tortoises base their courtship on the same kinds of displays, and differences among species rely mainly on the relative importance of aggressive displays with respect to acoustic signals. We found that the divergence of courtship patterns among species was also related to body size. Greek Tortoises, intermediate in size between the other two species, showed an intermediate courtship aggression. Body size in Mediterranean...

This study provided the first hand data of trace elements into fish muscles (N = 65) collected from river Chenab in Pakistan during 2013, using inductively coupled plasma mass spectrometry (ICP-MS). We monitored the health risk associated with consumption of contaminated fish of river Chenab, by the local population. The mean concentrations (μg/g, wet weight), in descending order were: Zn (35.5–54.4), Cu (1.38–4.57), Mn (2.43–4.5), As (0.23–1.21), Cr (0.21–0.67), Ni (0.14–0.34), Pb (0.14–0.31), Co (0.09–0.12), Cd (0.07–0.12) with higher concentration to be observed in the herbivore fish species (i.e., Cirrhinus reba and Catla catla). The levels of trace elements in different fish species found in this study were compared with similar data worldwide, and with the international standards for consumption. The concentration (μg/g) of arsenic in many cases (>65%) exceeded the FAO/WHO expert committee on food additives permissible limits. From the human health point of view, this study highlights that the local inhabitants, (i.e., fisher folk communities and population frequently consuming fish at about 100 g/day) along the river Chenab are exposed chronically to arsenic pollution with carcinogenic (10−4 to 10−6) and non-carcinogenic (THQ>1) risks, especially from the intake of Cirrhinus reba.

Island colonization is often assumed to trigger extreme levels of phenotypic diversification. Yet, empirical evidence suggests that it does not always so. In this study we test this hypothesis using a completely sampled mainland-island system, the arid clade of Hemidactylus, a group of geckos mainly distributed across Africa, Arabia and the Socotra Archipelago. To such purpose, we generated a new molecular phylogeny of the group on which we mapped body size and head proportions. We then explored whether island and continental taxa shared the same morphospace and differed in their disparities and tempos of evolution. Insular species produced the most extreme sizes of the radiation, involving accelerated rates of evolution and higher disparities compared with most (but not all) of the continental groups. In contrast, head proportions exhibited constant evolutionary rates across the radiation and similar disparities in islands compared with the continent. These results, although generally consistent with the notion that islands promote high morphological disparity, reveal at the same time a complex scenario in which different traits may experience different evolutionary patterns in the same mainland-island system and continental groups do not always present low levels of morphological diversification compared to insular groups., This work was funded by CGL2012-36970 from the Ministerio de Economía y Competitividad, Spain (co-funded by FEDER). The JGP was also supported by the JAE-predoc grant of the CSIC and JS was supported by the Ministry of Culture of the Czech Republic under grant DKRVO 2016/15, National Museum, 00023272.

Few DNA barcoding studies of squamate reptiles have been conducted. Due to the significance of the Socotra Archipelago (a UNESCO Natural World Heritage site and a biodiversity hotspot) and the conservation interest of its reptile fauna (94% endemics), we performed the most comprehensive DNA barcoding study on an island group to date to test its applicability to specimen identification and species discovery. Reptiles constitute Socotra’s most important vertebrate fauna, yet their taxonomy remains under-studied. We successfully DNA-barcoded 380 individuals of all 31 presently recognized species. The specimen identification success rate is moderate to high, and almost all species presented local barcoding gaps. The unexpected high levels of intra-specific variability found within some species suggest cryptic diversity. Species richness may be under-estimated by 13.8–54.4%. This has implications in the species’ ranges and conservation status that should be considered for conservation planning. Other phylogenetic studies using mitochondrial and nuclear markers are congruent with our results. We conclude that, despite its reduced length (663 base pairs), cytochrome c oxidase 1, COI, is very useful for specimen identification and for detecting intra-specific diversity, and has a good phylogenetic signal. We recommend DNA barcoding to be applied to other biodiversity hotspots for quickly and cost-efficiently flagging species discovery, preferentially incorporated into an integrative taxonomic framework., This article is an output of project CGL2012-36970 led by SC from the Ministerio de Economía y Competitividad (www.mineco.gob.es), Spain (co-funded by FEDER) with the support of Secretaria d’Universitats i Recerca del Departament d’Economia i Coneixement de la Generalitat de Catalunya (2014-SGR-1532; http://universitatsirecerca.gencat.cat/ca). MSR is funded by a FPI grant from the Ministerio de Economía y Competitividad, Spain (BES-2013-064248). RV and XS are supported by Postdoctoral grants from FCT, Fundação para a Ciência e Tecnologia (SFRH/BPD/79913/2011 and SFRH⁄BPD⁄73176/2010, respectively) financed by The European Social Fund and the Human Potential Operational Programme, POPH/FSE (www.fct.pt).

In seasonally fluctuating environments, timing of reproduction is a crucial determinant of fitness. Studies of birds show that late breeding attempts generally result in offspring of lower reproductive value, with lower recruitment and long-term survival prospects. Several proximate mechanisms, including a seasonal decline of immune system functioning, may lead to a seasonal decline of offspring fitness. We investigated seasonal variation in offspring quality by subjecting first- and second-brood chicks of a sexually size dimorphic species, the European starling Sturnus vulgaris, to an immune challenge with a bacterial endotoxin (LPS), and evaluated their growth and physiological response in terms of total plasma antioxidant capacity (TAC), concentration of reactive oxygen metabolites and hematocrit. LPS challenge did not affect chick growth or oxidative status. However, hematocrit of second-brood chicks was higher in LPS chicks compared to controls. Body mass halfway through the rearing period (days 8–9 post-hatching), TAC and hematocrit were lower among second- vs. first-brood chicks. Interestingly, sexual dimorphism in body mass at days 8–9 post-hatching markedly differed between broods, first-brood males being 4.7% and second-brood males 22.7% heavier than their sisters, respectively. Pre-fledging mortality occurred among second-brood chicks only and was strongly female-biased. Our findings suggest that starling chicks, even if in poor conditions, are little affected by a bacterial challenge, at least in the short-term. Moreover, our study indicates that sex differences in body size, possibly mediated by sex-specific maternal investment in egg size, may heavily impact on pre-fledging survival in a different way in the course of the breeding season, resulting in sex-specific seasonal decline of offspring fitness. Finally, we suggest that levels of circulating antioxidants should be regarded among the proximate causes of the association between timing of fledging and long-term survival in avian species.

Males and females of Hermann’s tortoise (Testudo hermanni) exhibit a conspicuous yellow patch on both their cheeks, whose origin and potential function are totally unknown. In this study, we measured the extent and the colour intensity of these patches in 29 male and 19 female tortoises in order to test for sexual difference in these features. In addition, we analysed the relationships between patch features and body condition to investigate the possible function of these ornaments as status signals. We detected symmetric yellow patches in all sampled females, while five males did not show at all the ornament, and two had a yellow patch only on the left cheek. Although head and scale size, as well as RGB values, did not differ between sexes, female patches were significantly larger than those of males. In addition, the extent of cheek patches was correlated to female body condition, suggesting that these ornaments may have evolved as honest signals of quality through sexual selection processes driven by female–female contests over rank or by male mate choice.

Many animals use chemical and visual cues to obtain reliable information about potential food resources. Most reptiles have evolved highly specialized chemosensory and visual capacities, in particular colour vision, to accomplish this task. By presenting animals with flowers and coloured cardboard discs in two-choice experiments, we examined whether both male and female Hermann's tortoises (Testudo hermanni) were able to distinguish between different colours, and show some preference for certain colours. We found that both sexes responded and discriminated equally between different colours, with some differences between flowers and discs. In particular, the red was preferred in the disc experiment, mainly by males, but was disregarded by both sexes in the flower experiment, where red was represented by poppy, Papaver rhoeas. This suggests that Hermann's tortoise probably relies on a mixed system of olfactory and visual abilities to detect food in order to optimize the intake of essential nutrient...

Parental decisions can determine offspring experience of environmental conditions. Such 'maternal' effects act both before and after hatching via, e. g., egg quality or the social milieu predisposed by parents. Resource availability may constrain the expression of adaptive maternal effects, and the specific pattern of allocation of these effects among offspring depending on their sex or birth order can result in different fitness payoffs to parents. Declining egg mass with laying order observed in several bird species may constitute an adaptive strategy of parental favouritism towards early hatching offspring with larger reproductive value but may also result from nutritional constraints on laying effort. A previous study has shown that the small size of the third, last laid (c-)egg in yellow-legged gull (Larus michahellis) clutches depends on food availability and that food-supplemented mothers increase the size of their female but not male c-eggs. Here, we show that increased mass of c-eggs laid by females supplemented with food after clutch initiation depends on increased albumen mass, which, in turn, enhances the size of daughters at hatching. Because asynchronous hatching results in a competitive disadvantage of c-chicks, present results suggest that mothers relieved from nutritional constraints enhance the size of daughters to compensate for their larger susceptibility to hatching last. The study also confirms the role of egg albumen content in determining hatchling size, previously experimentally detected only in one species in the wild. The effect of increased egg mass on offspring size persisted at least until day 8 after hatching, when, however, it did not vary with sex, suggesting intense negative selection on small female c-chicks in control broods. Hence, maternal effects mediated by egg albumen content had persistent effects on offspring size. © 2010 Springer-Verlag.

Maternal effects through albumen quality are largely unexplored, despite the fundamental role that albumen exerts as source of proteins and water, as well as for antimicrobial defence of the embryo. We analysed the variation of two major albumen antimicrobials, avidin and lysozyme, by extracting samples from freshly laid eggs of the yellow-legged gull (Larus michahellis) and by correlating their levels to egg features. Lysozyme concentration increased along the laying sequence, while avidin concentration decreased. Both antimicrobials declined during the season. In addition, avidin concentration declined from first- to last-laid male eggs, whereas the opposite was true among the female eggs. We also analysed chick body mass and size and immune response, in relation to albumen antimicrobial levels in their original egg while controlling for potential covariation between egg quality and rearing conditions by cross-fostering eggs between nests. Tarsus length decreased with avidin concentration, particularly early in the season. Avidin concentration negatively predicted tarsus length of chicks and the phytohaemagglutinin response of females, but not males. However, chick phenotype did not covary with lysozyme albumen concentration. This is the first study where maternal effects mediated by albumen antimicrobials are investigated in relation to both sex and egg features in any wild bird species. Whether the observed patterns of variation in antimicrobial concentration are the by-product of maternal physiological constraints, or reflect adaptive allocation strategies, cannot be ascertained. The covariation between chick cell-mediated immunity and albumen avidin concentration might be causal, according to the documented effects of albumen proteins on immunity in other species.

Male and female offspring can differ in their susceptibility to pre-natal (e.g. egg quality) and post-natal (e.g. sib–sib competition) conditions, and parents can therefore increase their individual fitness by adjusting these maternal effects according to offspring sex. In birds, egg mass and laying/hatching order are the main determinants of offspring viability, but these effects can act differently on each sex. In a previous study, relatively large last-laid (c-)eggs of yellow-legged gulls ( Larus michahellis ) were more likely to carry a female embryo. This suggests compensatory allocation of maternal resources to daughters from c-eggs, which suffer reduced viability. In the present study, we supplemented yellow-legged gulls with food during the laying period to experimentally test whether their nutritional conditions were responsible for the observed covariation between c-egg sex and mass. As predicted, food supplementation enhanced female c-eggs' mass more than that of male c-eggs. Thus, this experiment indicates that mothers strategically allocated their resources to c-eggs, possibly in order to compensate for the larger susceptibility of daughters to hatching (and laying) order. The results also suggested that mothers decided on resource allocation depending on the sex of already ovulated c-eggs, rather than ovulating ova of either sex depending on food availability.

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.

The occurrence of variation in body size and reproductive traits of Vipera aspis was assessed by analysing 74 reproductive females of different populations, collected throughout a large part of the distribution range of the taxon, from central-western France to central Italy. Six populations were analysed, two of plain habitats, in France and Italy, characterized by a Continental climate, whereas the other four derived from two coastal and two inland, hilly Italian habitats, respectively, showing a Mediterranean climate. Females of the French area showed the smallest mean body size, whereas the pre- and post-partum body masses of females from the coolest, central Italic area were significantly higher. Litter size varied among habitats and was significantly correlated with maternal body size. If the snout-vent length feature varied largely among offspring of different habitats (with the longest size occurring in representatives of the coastal, central Italian area), the average of both body mass and total mass of offsprings did not exhibit any clear pattern among populations colonizing different habitats. Nevertheless, relative litter mass was higher in the French continental populations, and lower in the coolest, Mediterranean, Italian ones. It is worth emphasizing the positive correlation between the snout-vent length feature of females and the total litter mass to environmental factors, such as hottest month temperature and total rainfall. The available literature records that female body size affects offspring size and fecundity. If the present study bolsters this correlation, it also rules out any other effect of female body size on the offspring characteristics analysed. Finally, evidence is provided for the role of climatic factors on life-history traits (e.g. pre-partum body mass) of asp vipers, although confounding effects pursued, for example, by food availability may occur.

Sperm competition models predict that males should adjust their sperm expenditure according to the risk and intensity of sperm competition, increasing it when competing with at least one other ejaculate and decreasing it when the number of competing ejaculates rises above two. However, when the raffle to fertilize eggs is strongly loaded due to sperm removal by subsequent rivals, it could be hypothesized that first-mating males progressively decrease their sperm expenditure and/or defer copulation with an increasing number of nearby competitors. Here, by varying experimentally the number of competitors around a mating pair, we analysed mating behaviour of both sexes and sperm expenditure in relation to variation in the immediate risk of sperm competition in the freshwater crayfish Austropotamobius italicus , a species where males remove all or part and feed on sperm laid by previous mating males. We found that male mating behaviour varied significantly with increasing number of rivals, because interactions between focal males and competitors, as well as male refusals to copulate, increased with number of the latter. As a consequence, the probability to reach an effective copulation decreased with increasing number of competitors. However, males released similar amounts of sperm independently of the number of surrounding competitors. Thus, the observed lack of variation in sperm expenditure in relation to the number of surrounding rivals suggests that freshwater crayfish males are unable to adjust the size of their ejaculates to short-term variation in the immediate risk of sperm competition.

Avian clutch size has been reported to show a poleward increase, a pattern that is well supported in landbirds but less so in waterbirds. We analyzed geographic variation in clutch and brood size among seven species of Nearctic herons and egrets (Ardeidae) over a latitudinal range spanning from 25° to 52°N. We included in the analysis data from 71 published references that reported information on both clutch and brood size. We found a strong and coherent latitudinal increase in clutch size in all species, but no unequivocal latitudinal variation in brood size. This resulted in a decreased index of productivity (brood size:clutch size ratio, excluding nest failures) with increasing latitude. In addition, we detected habitat-related variation in clutch size, which was, on average, smaller in salt-water than in freshwater habitats. The latitudinal decrease of the productivity index, which was not associated with latitudinal variation in nest-failure rates at either the clutch or brood stage, contras...

Summary 1Egg size is an important maternal trait that can have major consequences on offspring phenotype. However, the effects of the variation of different components of cleidoic eggs have been little investigated. 2Here, we addressed whether a reduction of the relative egg albumen content within the natural range of variation affects viability, time to hatching, early post-natal begging displays, morphology and immune response of yellow-legged gull Larus michahellis chicks. 3Egg mass strongly positively predicted chick size and mass at all ages, while time to hatching positively predicted tarsus length and immune response, irrespective of albumen removal. Variation in time to hatching may thus affect immune system maturation. 4Albumen removal resulted in a lower embryonic viability and increased time to hatching of individual eggs. The probability that an egg originated a chick surviving until 8 days of age increased with original egg mass among controls, but not among chicks hatching from eggs with reduced albumen content (‘albumen chicks’). 5Begging rate increased with laying order among albumen chicks while it decreased among controls. Concomitantly, begging rate decreased with egg mass among controls while it did not vary among albumen chicks. Surprisingly, albumen removal did not affect body mass or tarsus length except at 8 days of age, when control chicks were lighter than albumen chicks. 6In conclusion, our study indicates that a reduction of the relative egg albumen content can have complex effects on offspring development, behaviour and viability of a semiprecocial bird, suggesting that the relative albumen content of the eggs represents an important mechanism of maternal effects.

The assessment of the biological effects on aquatic vertebrate species is frequently employed to monitor water pollution, as it provides significant information on bioavailability and actual concentration levels. In anamniote vertebrates (fish and amphibians), significant correlations have been observed between exposure to contaminants - both natural and experimental - and blood modification. We investigated the changes in some circulating blood cell parameters of green frog (Rana snk esculenta) tadpoles and adults collected at two sample rice fields, one heavily polluted and the other relatively unpolluted. The frequency of eosinophilic leucocytes, mitotic, anucleated and micronucleated erythrocytes was evaluated also regarding the haemopoietic/haemocatheretic and NOS expression of the liver. Haematological indicators in polluted samples were found to be significantly different from controls as regards both larval and adult exposure, and provided information on long-term background pollution of the habitats under investigation. The population of the polluted area showed evident effects of chronic exposure to contaminants, to a degree which could lead to sub-lethal alterations of their health status. The general nature of responses to this kind of stress emphasizes the role of amphibian peripheral blood as a sensitive indicator regarding contamination in aquatic environments.

Maternal hormones in vertebrate eggs can mediate important forms of maternal effects. However, the function of hormone transfer to the eggs is still debated, especially because long-term fitness consequences have been little studied. We investigated the effect of prenatal exposure to physiologically elevated yolk testosterone (T) levels on reproduction of female pheasants ( Phasianus colchicus ) in captivity. We found that females hatching from T-injected eggs (T-females) had a lower egg-laying rate than controls, and their eggs were more frequently infertile than those laid by control females. There were no effects of prenatal maternal treatment on egg size and yolk T concentration, but eggs carrying a female embryo laid by T-females had smaller yolks than eggs with a male embryo, while there was no sex difference in yolk size among the eggs laid by control females. Progeny sex ratio was unaffected by maternal treatment. These findings suggest that the transfer of high androgen levels to the eggs by the mother is constrained by complex trade-offs between direct effects on her daughters’ reproduction and by trans-generational differential consequences on male and female descendants.