Coping with drought? Effects of extended drought conditions on soil invertebrate prey and diet selection by a fossorial amphisbaenian reptile.

Arid climates are characterized by a summer drought period to which animals seem adapted. However, in some years, the drought can extend for unusually longer periods. Examining the effects of these current extreme weather events on biodiversity can help to understand the effects of climate change, as models predict an increase in drought severity. Here, we examined the effects of "unusual" extended drought on soil invertebrate prey availability and on diet composition (based on fecal contents) and diet selection of a fossorial amphisbaenian, the checkerboard worm lizard Trogonophis wiegmanni. Weather data show interannual variations in summer drought duration. The abundance and diversity of soil invertebrates in spring were high, and similar to those found in a "normal" early autumn, after some rain had ended with the summer drought. In contrast, in years with "unusual" extended drought, abundance, and diversity of soil invertebrates in early autumn were very low. Also, there were seasonal changes in amphisbaenians' diet; in autumn with drought, prey diversity, and niche breadth decreased with respect to spring and autumns after some rain had fallen. Amphisbaenians did not eat prey at random in any season, but made some changes in prey selection that may result from drought-related restrictions in prey availability. Finally, in spite that amphisbaenians showed some feeding flexibility, their body condition was lower in autumn than in spring, and much lower in autumn with drought. If extended drought became the norm in the future, amphisbaenians might suffer important negative effects for their health state.

DNA metabarcoding the diet of Podarcis lizards endemic to the Balearic Islands.

Dietary studies are essential to unravel the functioning of ecosystems and ultimately to understand biodiversity. This task, which at first may seem simple, becomes especially complex in those cases of omnivorous species with highly variable diets. In this regard, the emergence of next-generation DNA sequencing methodologies represents a powerful tool to address the problem. Here we implement a high-throughput metabarcoding strategy based on the analysis of four molecular markers aimed at sequencing both mitochondrial (animal prey) and chloroplast (diet plants) genome fragments from fecal samples of two lizard species endemic to the Balearic Archipelago (Podarcis lilfordi and P. pityusensis) obtained through non-invasive methods. The results allowed for the characterization of their diets with a high degree of taxonomic detail and have contributed a large number of new trophic records. The reported diets are based mainly on the consumption of arthropods, mollusks and plants from a diversity of taxonomic orders, as well as carrion and marine subsidies. Our analyses also reveal inter- and intra-specific differences both in terms of seasonality and geographical distribution of the sampled lizard populations. These molecular findings provide new insights into the trophic interactions of these threatened endemic lizards in their unique and isolated ecosystems.

Effects of Anthropogenic Disturbance of Natural Habitats on the Feeding Ecology of Moorish Geckos.

Urbanization and anthropic influences can drastically modify a natural habitat and transform it into an easily recognizable "urban habitat". Human activities can also induce less severe modifications of what apparently might still look like natural habitats. Therefore, these subtle alterations may be hidden but can still cause important negative effects on plant and animals. In contrast, some species seem able to take advantage of these anthropic alterations. Here, we examined the possible effects of the anthropogenic disturbance of an apparent natural habitat on the feeding ecology and body condition of Moorish geckos, Tarentola mauritanica. For this, we compared microhabitat structure, invertebrate availability, the diet composition (estimated from fecal contents), diet selection patterns and body condition of the two populations of geckos inhabiting two contiguous small islands. These islands have similar environmental characteristics, but highly contrasting differences in urbanization and anthropogenic influence. We found that, although the abundance of potential invertebrate prey was similar on both habitats, the diversity of invertebrate prey was lower in the altered habitat. As a consequence, although composition of the diet of geckos was similar on both islands, the diversity of prey and food niche breadth were lower in the altered habitat, and patterns of diet selection changed. However, these inter-habitat differences did not seem to affect the body size and body condition of geckos. We discuss how flexibility in feeding ecology may allow some species to cope with small anthropic disturbances of the habitat.

Long-Term Seed Dispersal within an Asymmetric Lizard-Plant Interaction.

During the last 24 years, the mutualistic interaction between the dead horse arum, Helicodiceros muscivorus, and the Balearic lizard, Podarcis lilfordi, was studied on Aire Island (Balearic Islands, Spain). From a small population of a hundred plants, the dead horse arum expanded extraordinarily throughout the island, reaching the highest known densities of the species and occupying areas of the island where it was not previously present. The current abundance of plants is a direct effect of the frugivorous activity of the Balearic lizard, which is the main, if not the only, effective seed disperser of the plant on Aire Island. However, abiotic factors predominated over biotic factors in driving abundance of plants. Over the years, plant densities varied significantly depending on the aridity of the island, with higher densities recorded in drier years. Lizards' frugivorous activity and dispersal intensity was inversely correlated with annual rainfall. We found higher dispersal intensity in years with lower rainfall. We propose that the years of lower rainfall are those in which there is a lower prey availability. In such years, lizards compensate the shortage of other trophic resources with a more intense consumption of dead horse arum fruits. The mutualistic interaction is therefore asymmetric, since there is a greater influence of the frugivorous activity of the lizards on the plants than of the plants on lizards. It is, in short, a system chronically out of balance.

Diet of the Insular Lizard, Podarcis lilfordi (Günther, 1874): Complementary Morphological and Molecular Approaches.

The diets of insular lizards are extremely varied, depending on the different environmental characteristics of each island population. This is particularly evident in the case of the populations of small coastal islets of the Balearic Islands, where the Balearic lizard, Podarcis lilfordi, is found. The study of trophic ecology carried out by means of traditional tools, such as morphological analysis of feces, has made it possible to detect numerous prey and nutritional elements. However, these methods are clearly insufficient, as some rare groups are not detected. It is also difficult to identify remains of marine subsidies or of foods contributed to these small islands by other predators, such as seabirds. The current study demonstrates the advantages of combining morphological diet analysis with the molecular study of individual feces samples obtained from the same populations. We obtained a greater diversity of prey groups using the combined methodologies, with each method identifying prey items that were not detected using the other method. Particularly, the study of diets at the molecular level identified plant species consumed by lizards that were, occasionally, not identified in morphological analyses. Conversely, the traditional morphological study of an equivalent number of fecal samples allowed for the identification of several prey groups that had not been detected in the molecular study. From this viewpoint, the advantages and disadvantages of each methodology are discussed.

Faecal Microbiota Divergence in Allopatric Populations of Podarcis lilfordi and P. pityusensis, Two Lizard Species Endemic to the Balearic Islands.

Gut microbial communities provide essential functions to their hosts and are known to influence both their ecology and evolution. However, our knowledge of these complex associations is still very limited in reptiles. Here we report the 16S rRNA gene faecal microbiota profiles of two lizard species endemic to the Balearic archipelago (Podarcis lilfordi and P. pityusensis), encompassing their allopatric range of distribution through a noninvasive sampling, as an alternative to previous studies that implied killing specimens of these IUCN endangered and near-threatened species, respectively. Both lizard species showed a faecal microbiome composition consistent with their omnivorous trophic ecology, with a high representation of cellulolytic bacteria taxa. We also identified species-specific core microbiota signatures and retrieved lizard species, islet ascription, and seasonality as the main factors in explaining bacterial community composition. The different Balearic Podarcis populations are characterised by harbouring a high proportion of unique bacterial taxa, thus reinforcing their view as unique and divergent evolutionary entities.

Scat piling and strong frugivory of the Balearic lizard, Podarcis lilfordi (Günther, 1874).

BACKGROUND: In lacertid lizards from Mediterranean islands, frugivory is common, particularly under prey scarcity, a characteristic of small islands. In several populations, the diet of the Balearic lizard, Podarcis lilfordi, is extremely variable and includes fleshy fruits. However, frugivory is sporadic and there are very few examples of dominant fruit consumption. RESULTS: We describe the existence of an extraordinary fruit consumption of a single plant species, the juniper, Juniperus phoenicea, by the Balearic lizard, P. lilfordi. In addition, for the first time in Lacertidae, we describe the existence of scat piling in the population of these lizards inhabiting Cabrera Island (Balearic Islands, Spain). Scat piling was detected in an isolated location with hundreds of scats deposited by several individuals at a particular place. CONCLUSIONS: The high population density of lizards at the island of Cabrera and the great versatility of foraging behavior of this species allows us to hypothesize that scat piles could act as an attractor for numerous individuals, that is, as inadvertent social information. If that hypothesis is correct, it would result in the concentration of several individuals foraging on a single or a few ripening plants. We cannot, however, rule out that individuals concentrated due to the scarcity of ripening plants in other areas, without any influence of the presence of several lizards, as attractors to the site. Our findings modify previous descriptions of the diet of the Balearic lizard in Cabrera made with smaller samples. In some places and periods of the year, frugivory on a single plant species can be extremely intense and only large sample sizes of scats allow to find these particular trends in the foraging ecology of insular lizards.

Genomic signatures of drift and selection driven by predation and human pressure in an insular lizard.

Genomic divergence was studied in 10 small insular populations of the endangered Balearic Islands lizard (Podarcis lilfordi) using double digest restriction-site associated DNA sequencing. The objectives were to establish levels of divergence among populations, investigate the impact of population size on genetic variability and to evaluate the role of different environmental factors on local adaptation. Analyses of 72,846 SNPs supported a highly differentiated genetic structure, being the populations with the lowest population size (Porros, Foradada and Esclatasang islets) the most divergent, indicative of greater genetic drift. Outlier tests identified ~ 2% of loci as candidates for selection. Genomic divergence-Enviroment Association analyses were performed using redundancy analyses based on SNPs putatively under selection, detecting predation and human pressure as the environmental variables with the greatest explanatory power. Geographical distributions of populations and environmental factors appear to be fundamental drivers of divergence. These results support the combined role of genetic drift and divergent selection in shaping the genetic structure of these endemic island lizard populations.

It sounds like food: Phonotaxis of a diurnal lizard.

Foraging diurnal lizards are well known for their use of visual and chemical cues to detect prey. We already showed that the Balearic lizard is able to detect prey using visual and chemical cues, even from airborne odors. In this study we carried out a field experiment to test if lizards can detect prey using acoustic cues. Our results show that Podarcis lilfordi is able to detect flies trapped inside opaque cups, only using acoustic cues. To our knowledge, this is the first known case of phonotaxis of a diurnal lizard. Thus, P. lilfordi can detect, from far away, current pollinators trapped inside floral chambers of the dead horse arum, Helicodiceros muscivorus. This is another behavioral trait displayed by the Balearic lizard during its complex interaction with the dead horse arum.

Spontaneous magnetic alignment behaviour in free-living lizards.

Several species of vertebrates exhibit spontaneous longitudinal body axis alignment relative to the Earth's magnetic field (i.e., magnetic alignment) while they are performing different behavioural tasks. Since magnetoreception is still not fully understood, studying magnetic alignment provides evidence for magnetoreception and broadens current knowledge of magnetic sense in animals. Furthermore, magnetic alignment widens the roles of magnetic sensitivity in animals and may contribute to shed new light on magnetoreception. In this context, spontaneous alignment in two species of lacertid lizards (Podarcis muralis and Podarcis lilfordi) during basking periods was monitored. Alignments in 255 P. muralis and 456 P. lilfordi were measured over a 5-year period. The possible influence of the sun's position (i.e., altitude and azimuth) and geomagnetic field values corresponding to the moment in which a particular lizard was observed on lizards' body axis orientation was evaluated. Both species exhibited a highly significant bimodal orientation along the north-northeast and south-southwest magnetic axis. The evidence from this study suggests that free-living lacertid lizards exhibit magnetic alignment behaviour, since their body alignments cannot be explained by an effect of the sun's position. On the contrary, lizard orientations were significantly correlated with geomagnetic field values at the time of each observation. We suggest that this behaviour might provide lizards with a constant directional reference while they are sun basking. This directional reference might improve their mental map of space to accomplish efficient escape behaviour. This study is the first to provide spontaneous magnetic alignment behaviour in free-living reptiles.

Omnivory of an Insular Lizard: Sources of Variation in the Diet of Podarcis lilfordi (Squamata, Lacertidae).

Through 17 years and from a sample of 7,790 faecal pellets and 26,346 prey items, we studied the diet of the Balearic lizard Podarcis lilfordi in Aire Island (Menorca, Balearic Islands, Spain). We analysed the diet in terms of prey frequencies, as well as by their volume and biomass contributions. The diet of the Balearic lizard was extremely variable through the years, months and areas under study. The dominance of small clumped prey, particularly ants, was confirmed. However, the main contribution by volume corresponded to beetles, with a relevant role for Diplopoda and terrestrial Isopoda during some months and at particular areas of the island. Several prey items were probably captured at the base of shrubs, under stones or inside rock crevices. Therefore, our estimations of electivity would only be reliable for epigeal and flying prey. The capacity of the Balearic lizard to include marine subsidies in its diet, such as coastal crustaceans, is noteworthy. Also, its consumption of carrion from carcasses of gulls and rabbits and leftovers from human visitors is remarkable. Juvenile conspecifics can also be a sporadic food resource, especially during the second half of summer, whereas the consumption of vegetal matter is constant for each whole year. The shifts of vegetal exploitation among areas of the island and months take place according to availability of different plant species at each area or during a given period. Thus, lizards are able to conduct a thorough monitoring of plant phenology, exploiting a large variety of plant species. Omnivory does not imply the indiscriminate inclusion of any edible food in its diet. Rather, the inclusion of several food items means the adoption of a wide range of foraging behaviours adapted to the exploitation of each food resource.

Local enhancement and social foraging in a non-social insular lizard.

Even in solitary foragers, conspecifics can provide reliable information about food location. The insular lizard Podarcis lilfordi is a solitary species with high population densities that sometimes aggregate around rich food patches. Its diet includes novel and unpredictable resources, such as carcasses or plants, whose exploitation quickly became widespread among the population. We tested the use of social information by lizards through some field experiments in which they had to choose one of the two pieces of fruit. Probably due to local enhancement, lizards preferred to feed on the piece of fruit where conspecifics or lizard-shaped models were already present. Conspecifics' behaviour, but also their mere presence, seems to be a valuable source of information to decide where to feed. Lizards also showed a strong attraction to conspecifics, even in the absence of food. Maybe the presence of a group is interpreted as an indirect cue for the presence of food. The group size was not important to females, but males had a significantly higher attraction towards groups with three conspecifics. We discuss some characteristics of P. lilfordi at Aire Island that can explain the development of the observed social foraging, as well as their possible consequences.