Different facets of the same niche: Integrating citizen science and scientific survey data to predict biological invasion risk under multiple global change drivers.

Citizen science initiatives have been increasingly used by researchers as a source of occurrence data to model the distribution of alien species. Since citizen science presence-only data suffer from some fundamental issues, efforts have been made to combine these data with those provided by scientifically structured surveys. Surprisingly, only a few studies proposing data integration evaluated the contribution of this process to the effective sampling of species' environmental niches and, consequently, its effect on model predictions on new time intervals. We relied on niche overlap analyses, machine learning classification algorithms and ecological niche models to compare the ability of data from citizen science and scientific surveys, along with their integration, in capturing the realized niche of 13 invasive alien species in Italy. Moreover, we assessed differences in current and future invasion risk predicted by each data set under multiple global change scenarios. We showed that data from citizen science and scientific surveys captured similar species niches though highlighting exclusive portions associated with clearly identifiable environmental conditions. In terrestrial species, citizen science data granted the highest gain in environmental space to the pooled niches, determining an increased future biological invasion risk. A few aquatic species modelled at the regional scale reported a net loss in the pooled niches compared to their scientific survey niches, suggesting that citizen science data may also lead to contraction in pooled niches. For these species, models predicted a lower future biological invasion risk. These findings indicate that citizen science data may represent a valuable contribution to predicting future spread of invasive alien species, especially within national-scale programmes. At the same time, citizen science data collected on species poorly known to citizen scientists, or in strictly local contexts, may strongly affect the niche quantification of these taxa and the prediction of their future biological invasion risk.

Five decades of breeding populations census for 12 species of colonial waterbirds in northwestern Italy.

Colonial waterbirds, a major biodiversity element occurring in the core of ultra-anthropized Europe, are ideal indicators of the wellness of inland wetlands. Nonetheless, there is a critical knowledge gap in their trend and population status. We present an uninterrupted 47 years-long dataset of the breeding populations of 12 species of colonial waterbirds (Ardeidae, Phalacrocoracidae, Plataleidae, Threskiornitidae) throughout a 58,000 km2 agricultural region in the higher Po basin (NW Italy). A trained team of collaborators censused with standardized field techniques the number of nests of each species at 419 colonies in the 1972-2018 period, summing up a total of 236,316 records. Data cleaning and standardization were performed for each census year, ensuring robust and consistent data. This dataset is among the largest ever collected for a guild of European vertebrates. It has already been used to describe the factors influencing population trends, and still offers opportunities to explore a wide range of key ecological processes such as biological invasions, global change consequences and biodiversity impact of agricultural practices.

The spreading of the invasive sacred ibis in Italy.

The spreading of invasive species in new continents can vary from slow and limited diffusion to fast colonisations over vast new areas. We studied the sacred ibis Threskiornis aethiopicus along a 31-year period, from 1989 to 2019, with particular attention to the first area of release in NW Italy. We collected data on species distribution through observations by citizen science projects, population density by transects with distance method, breeding censuses at colonies, and post breeding censuses at roosts. The birds counted at winter roosts in NW Italy increased from a few tens up to 10,880 individuals in 2019. Sacred ibises started breeding in 1989, with a single nest in north-western Italy. The number of breeders remained very low until 2006, when both overwintering and breeding sacred ibises started to increase exponentially and expand their range throughout northern Italy with isolated breeding cases in central Italy. In 2019, the number of nests had increased to 1249 nests in 31 colonies. In NW Italy, the density of foraging birds averaged 3.9 ind./km2 in winter and 1.5 ind./km2 in the breeding period, with a mean size of the foraging groups of 8.9 and 2.1 birds respectively. Direct field observations and species distribution models (SDM) showed that foraging habitats were mainly rice fields and wetlands. A SDM applied to the whole Italian peninsula plus Sardinia and Sicily showed that the variables best related to the SDM were land class (rice fields and wetlands), altitude, and the temperature seasonality. The areas favourable for species expansion encompass all the plains of Northern Italy, and several areas of Tuscany, Latium, Sardinia, and Apulia.

Combining molecular and landscape tools for targeting evolutionary processes in reserve design: An approach for islands.

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 richness 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.

Fishes in the desert: mitochondrial variation and phylogeography of Danakilia (Actinopterygii: Cichlidae) and Aphanius (Actinopterygii: Cyprinodontidae) in the Danakil Depression of northeastern Africa.

The Danakil Depression in northeastern Africa represents one of the harshest arid environments on Earth, yet two genera of fishes, Danakilia (Cichlidae) and Aphanius (Cyprinodontidae), share its sparse aquatic habitats. The evolutionary history of these fishes is investigated here in the context of genetic, geological and paleoenvironmental information. We collected samples from seven sites and assessed phylogeographic relationships using concatenated COI and cytb mtDNA genes. Danakilia morphospecies show low differentiation at mitochondrial markers, but variation is partitioned between a northern cluster containing D. dinicolai plus three undescribed riverine populations, and a southern cluster including two creek populations of D. franchettii separated by the hypersaline waters of Lake Afrera. Aphanius displayed four genetically distinct clades (A. stiassnyae in Lake Afrera; one distributed across the entire area; one in Lake Abaeded; and one in the Shukoray River), but without clear large-scale geographic structure. However, Danakil Aphanius are clearly differentiated from A. dispar sensu stricto from the Sinai Peninsula. Geological evidence suggests that after the Late Pleistocene closure of the Danakil-Red Sea connection, increased post-glacial groundwater availability caused the formation of a brackish paleo-lake flooding the entire region below the -50 m contour. Fish populations previously isolated in coastal oases during glaciation were able to mix in the paleo-lake. Subsequently, in a more arid phase starting ∼7300 BP, paleo-lake regression isolated fishes in separate drainages, triggering their still ongoing diversification.

Conspecific and not performance-based attraction on immigrants drives colony growth in a waterbird.

Local recruitment and immigration play an important part in the dynamics and growth of animal populations. However, their estimation and incorporation into open population 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 'performance-based' and 'conspecific attraction' 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.

Human exposure to trace metals and arsenic via consumption of fish from river Chenab, Pakistan and associated health risks.

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.

Mercury contamination in deposited dust and its bioaccumulation patterns throughout Pakistan.

Mercury (Hg) contamination of environment is a major threat to human health in developing countries like Pakistan. Human populations, particularly children, are continuously exposed to Hg contamination via dust particles due to the arid and semi-arid climate. However, a country wide Hg contamination data for dust particles is lacking for Pakistan and hence, human populations potentially at risk is largely unknown. We provide the first baseline data for total mercury (THg) contamination into dust particles and its bioaccumulation trends, using scalp human hair samples as biomarker, at 22 sites across five altitudinal zones of Pakistan. The human health risk of THg exposure via dust particles as well as the proportion of human population that are potentially at risk from Hg contamination were calculated. Our results indicated higher concentration of THg in dust particles and its bioaccumulation in the lower Indus-plain agricultural and industrial areas than the other areas of Pakistan. The highest THg contamination of dust particles (3000ppb) and its bioaccumulation (2480ppb) were observed for the Lahore district, while the highest proportion (>40%) of human population was identified to be potentially at risk from Hg contamination from these areas. In general, children were at higher risk of Hg exposure via dust particles than adults. Regression analysis identified the anthropogenic activities, such as industrial and hospital discharges, as the major source of Hg contamination of dust particles. Our results inform environmental management for Hg control and remediation as well as the disease mitigation on potential hotspots.

Testing the island effect on phenotypic diversification: insights from the Hemidactylus geckos of the Socotra Archipelago.

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.

Unexpectedly High Levels of Cryptic Diversity Uncovered by a Complete DNA Barcoding of Reptiles of the Socotra Archipelago.

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.

Spatial distribution of dust-bound trace elements in Pakistan and their implications for human exposure.

This study aims to assess the spatial patterns of selected dust-borne trace elements alongside the river Indus Pakistan, their relation with anthropogenic and natural sources, and the potential risk posed to human health. The studied elements were found in descending concentrations: Mn, Zn, Pb, Cu, Ni, Cr, Co, and Cd. The Index of Geo-accumulation indicated that pollution of trace metals were higher in lower Indus plains than on mountain areas. In general, the toxic elements Cr, Mn, Co and Ni exhibited altitudinal trends (P < 0.05). The few exceptions to this trend were the higher values for all studied elements from the northern wet mountainous zone (low lying Himalaya). Spatial PCA/FA highlighted that the sources of different trace elements were zone specific, thus pointing to both geological influences and anthropogenic activities. The Hazard Index for Co and for Mn in children exceeded the value of 1 only in the riverine delta zone and in the southern low lying zone, whereas the Hazard Index for Pb was above the bench mark for both children and adults (with few exceptions) in all regions, thus indicating potential non-carcinogenic health risks. These results will contribute towards the environmental management of trace metal(s) with potential risk for human health throughout Pakistan.

Human exposure to toxic metals via contaminated dust: Bio-accumulation trends and their potential risk estimation.

We assessed the levels of potentially toxic trace metals, Zinc (Zn), Lead (Pb), Manganese (Mn), Copper (Cu), Nickel (Ni), Chromium (Cr), Cobalt (Co), and Cadmium (Cd), in dust, hair, nail and serum, sampled in rural, urban and industrial areas of Punjab, Pakistan. Trace metals occurrence in all samples, in descending order, was: Zn, Pb, Mn, Cu, Cr, Ni, Co, Cd. The samples from the urban areas showed significantly higher concentration of toxic trace metals (Zn, Ni, Cr, Co, Mn, and Cd) than those from industrial (which conversely had higher levels of Pb and Cu), and than samples from rural areas. Bioaccumulation patterns showed that dust exposure is one of the major routes into human body for Cd, Pb, Co, Mn and Cr, while the burden of Zn, Cu, and Ni can be more linked to dietary sources. The concentrations of trace metals in the samples from Punjab were comparable and/or higher than those reported worldwide. In many cases, the levels of Zn, Cr, Pb, Ni and Cd in hair and nail were beyond the ATSDR threshold guideline values that may cause some serious health effects. Hazard Index (HI) calculated for trace metal concentrations in the human population of Punjab points particularly to health risks from Cd (for children in urban and industrial areas) and from Pb (for all sub-groups).

Avian feathers as a non-destructive bio-monitoring tool of trace metals signatures: a case study from severely contaminated areas.

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.

Cryptic diversity within the Anatololacerta species complex (Squamata: Lacertidae) in the Anatolian Peninsula: evidence from a multi-locus approach.

The rapid development of innovative molecular tools for characterizing biodiversity is leading to an extensive and sometimes unexpected renovation of taxonomic classifications. Particularly, for species having allopatric or parapatric distributions or resulting from recent speciation processes, the absence of clear phenotypic differentiation may hinder the recognition of closely related taxa, while intraspecific polymorphism may be confused with the presence of more than one single species. In the present work, we apply different phylogenetic methods in order to infer relationships within the genus Anatololacerta, and to assess the taxonomy of this morphologically diversified group of lizards endemic to western and southern Anatolia and some neighboring Aegean islands. According to morphology, three species have been recognized (Anatololacerta anatolica, A. oertzeni and A. danfordi) as well as several subspecies, but small variation at immunological markers led some authors to join all the populations into one single taxon, A. danfordi. By selecting both mitochondrial and nuclear informative markers, we tested the effectiveness of classical "gene tree" (i.e. Bayesian Inference) vs. innovative (i.e. coalescent-based) "species tree" methods in resolving the Anatololacerta taxonomic enigma, as a case in point for similar studies on species complexes resulting from non-obvious and cryptic diversification patterns. According to our results, the gene tree method failed in resolving phylogenetic relationships among clades, whereas the multi-locus species tree approach, coupled with species delimitation methods, allowed the identification of four well distinct species. These species probably diversified in different allopatric refugia located in southern and western Anatolia, where isolated populations may have persisted during Pleistocene glacial cycles.

Cattle egrets as a biosentinels of persistent organic pollutants exposure.

We investigated selected chlorinated pollutants (ß-HCH, γ-HCH, DDDs, DDEs, o,p'-DDT, p,p'-DDT, heptachlor, aldrin, dieldrin, and endrin) in the Lahore and the Sialkot districts of Pakistan, using eggs of cattle egret (Bubulcus ibis) collected during May and June 2007. The pollutant with highest level and frequency was ΣDDT, followed by ß-HCH, γ-HCH, heptachlor, aldrin, dieldrin, and endrin in descending order. The concentration(s) were significantly higher in Sialkot heronry for all the pollutants (except p,p'-DDT) than in Lahore. The values for DDTs, ß-HCH, γ-HCH, and heptachlor were significantly higher (p < 0.05) in the egg(s) than in sediment(s) and in the chicks' diet, due to biomagnification. Among DDTs analogues, p,p'-DDD was the major contaminant with >60 % of total DDT burden, reflecting the widespread aged as well as recent use of DDT as well as anaerobic degradation (DDD/DDE > 1 in many cases) in the nearby paddy soils. In few samples, p,p'-DDT/(DDD + DDE) > 0.5 suggested the recent emission patterns from surrounding contaminated areas of demolished DDT units and obsolete pesticide stores. The higher levels of HCHs (i.e., ß-HCH) in the samples collected from Sialkot indicate exposure from long-term agricultural use. Overall, concentrations of all studied POPs were less than the threshold levels known to affect reproduction. Nevertheless, total DDTs and/or HCHs burdens in some eggs contained concentrations of greater than what would educe adverse effects on birds. This is among few studies on OCPs exposure to avian species, which provide the evidence of Pakistan's contribution toward the Global POPs emission.

Timing of molt of barn swallows is delayed in a rare Clock genotype.

Photoperiodic responses are major factors entraining circannual life-cycles, functioning to adaptively synchronize annual routines to seasonal fluctuations in ecological conditions. Photoperiodism in physiology and behaviour is enforced by genes, including the vertebrate Clock orthologues, which are associated, for example, with phenology of migration in fish and breeding in birds. However, the role of Clock in photoperiodic plumage molt processes is unknown. We analyzed variation in molt schedules in relation to Clock genotype, using the long-distance migratory barn swallow (Hirundo rustica) as a model and by identifying males and females using molecular sexing techniques. Consistently with previous studies, we found one very common (Q 7) and two rare (Q 6, Q 8) variants of a functionally significant Clock polyglutamine repeat. Molt schedules of primary wing feathers of swallows during their wintering period in Nigeria differed among Clock genotypes: rare (1.1%) Q 7/Q 8 heterozygotes had significantly delayed molt compared to the other genotypes. Molt schedules did not differ between males and females, and no differential association between molt and Clock in relation to sex emerged. The same rare Clock genotype that exhibited delayed breeding in Europe was here found to delay molt in Africa. Though based on a limited number of Q 7/Q 8 individuals from an otherwise very large sample, these novel results suggest that Clock is involved in the photoperiodic control of both molt and breeding, potentially also via reciprocal carry-over effects. If confirmed in species with higher Clock polymorphism, present results may have far-reaching consequences for the study of photoperiodic control of molt and expression of annual routines.

Sperm traits negatively covary with size and asymmetry of a secondary sexual trait in a freshwater crayfish.

In species where females mate promiscuously, the reproductive success of males depends both on their ability to acquire mates (pre-copulatory sexual selection) and ability of their ejaculates to outcompete those of other males (post-copulatory sexual selection). Sperm competition theory predicts a negative relationship between investment in body traits favouring mate acquisition (secondary sexual characters, SSCs) and investment in ejaculate size or quality, due to the inherent costs of sperm production. In contrast, the phenotype-linked fertility hypothesis posits that male fertilizing efficiency is reliably reflected by the phenotypic expression of male SSCs, allowing females to obtain direct benefits by selecting more ornamented males as copulation partners. In this study, we investigated the relationships between male SSCs and size and quality (viability and longevity) of ejaculates allocated to females in mating trials of the freshwater crayfish Austropotamobius italicus. We showed that the relative size of male weapons, the chelae, was negatively related to ejaculate size, and that chelae asymmetry, resulting from regeneration of lost chelipeds, negatively covaried with sperm longevity. Moreover, males allocated more viable sperm to mates from their own rather than different stream of origin. Our findings thus suggest that, according to sperm competition theory, pre-copulatory sexual selection for large weapons used in male fighting may counteract post-copulatory sperm competition in this crayfish species, and that investment in cheliped regeneration may impair ejaculate quality.

Origin and in situ diversification in Hemidactylus geckos of the Socotra Archipelago.

The Socotra Archipelago is an ancient continental fragment of Gondwanan origin and one of the most isolated landforms on Earth and a biodiversity hot spot. Yet, the biogeography and evolutionary history of its endemic fauna still remain largely overlooked. We investigate the origin, tempo and mode of diversification in the Hemidactylus geckos of the Socotra Archipelago. Concatenated and multilocus species coalescent analyses of Hemidactylus from Arabia and North Africa indicate that the Hemidactylus from Socotra do not form a monophyletic group and branch as three independent and well-supported clades instead. Both the chronogram inferred using the gene tree approach of BEAST and the age-calibrated multilocus species tree obtained using *BEAST suggest that the origin of Hemidactylus from Socotra may have involved a first vicariance event that occurred in the Early Miocene, followed by two independent transoceanic dispersal events that occurred more recently, during the Pliocene. Within Socotra, we analysed patterns of genetic diversity, the phylogeography and the demographic history in all seven nonintroduced species of Hemidactylus. Results based on two mitochondrial and two nuclear loci from 144 individuals revealed complex patterns of within-island diversification and high levels of intra-species genetic divergence. The interplay of both historical and ecological factors seems to have a role in the speciation process of this group of geckos. Interestingly, the case of H. forbesii and H. oxyrhinus, which inhabit the island of Abd al Kuri with an area of 133 km(2), may represent one of the most extreme cases of intra-island speciation in reptiles ever reported.

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.

Sex allocation in yellow-legged gulls (Larus michahellis) depends on nutritional constraints on production of large last eggs.

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.