×

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.

×

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 previ- ous 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 representa- tion 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.

×

In this article an introduction is given on the geological history leading to the separation of Podarcis lilfordi (GÜNTHER, 1874) and Podarcis pityusensis (BOSCÁ, 1883) as separate species, as well as a Holocene sea level rise model which combined with bathymetric data leads to an estimation of recent divergence time in populations of the Balearic lizards.

×

New data on estimated divergence times of the populations of lacertid lizards in the Balearic Islands are provided in this second update of the October 2011 article: Estimating recent divergence time in populations of Podarcis lilfordi (GÜNTHER, 1874) and Podarcis pityusensis (BOSCÁ, 1883) (VAN DEN BERG 2011), which received its first update May 2012. In most cases better estimations of divergence times were available by using the NAVIONICS SonarCharts™webapp.

×

This is our fifth report in a series on our whereabouts while collecting data for a future revision of the present subspecific order of the endemic Balearic sisterspecies Podarcis lilfordi (GÜNTHER, 1874) and Podarcis pityusensis (BOSCÁ, 1883), which data are stored in our free accessible database at www.pityusensis.nl (VAN DEN BERG & ZAWADZKI 2011 ; VAN DEN BERG et al. 2013 ; VAN DEN BERG et al. 2014a ; VAN DEN BERG et al. 2014b). During this trip from the 9th of May until the 23rd of May 2015, we were able to collect data on a few mainland Ibiza locations, as well as the following adjacent islands: Punta Galera, Es Canaret, Illa de la Xanga (Sal Rossa), Pouet de Ses Illetes, Formentera, Purroig, Es Vedrà, Rates, Malví Pla (North), Malví Rodó (South), S’Espardell, Calders, Penjats and Ses Margalides. We continue with the ventral coloration as a possible determining key as introduced in our 2014 trip report (VAN DEN BERG et al. 2014b). Images of the anal shields are presented of each lizard for purposes of illustration.

×

Phylogenetic relationships and timings of major cladogenesis events are investigated in the Balearic Island lizards Podarcis lilfordi and Podarcis pityusensis using 2675 bp of mitochondrial and nuclear DNA sequences. Partitioned Bayesian and Maximum Parsimony analyses provided a well-resolved phylogeny with high node-support values. Bayesian MCMC estimation of node dates was investigated by comparing means of posterior distributions from different subsets of the sequence against the most robust analysis which used multiple partitions and allowed for rate heterogeneity among branches under a rate-drift model. Evolutionary rates were systematically underestimated and thus divergence times overestimated when sequences containing lower numbers of variable sites were used (based on ingroup node constraints). The following analyses allowed the best recovery of node times under the constant-rate (i.e., perfect clock) model: (i) all cytochrome b sequence (partitioned by codon position), (ii) cytochrome b (codon position 3 alone), (iii) NADH dehydrogenase (subunits 1 and 2; partitioned by codon position), (iv) cytochrome b and NADH dehydrogenase sequence together (six gene–codon partitions), (v) all unpartitioned sequence, (vi) a full multipartition analysis (nine partitions). Of these, only (iv) and (vi) performed well under the rate-drift model. These findings have significant implications for dating of recent divergence times in other taxa. The earliest P. lilfordi cladogenesis event (divergence of Menorcan populations), occurred before the end of the Pliocene, some 2.6 Ma. Subsequent events led to a West Mallorcan lineage (2.0 Ma ago), followed 1.2 Ma ago by divergence of populations from the southern part of the Cabrera archipelago from a widely-distributed group from north Cabrera, northern and southern Mallorcan islets. Divergence within P. pityusensis is more recent with the main Ibiza and Formentera clades sharing a common ancestor at about 1.0 Ma ago. Climatic and sea level changes are likely to have initiated cladogenesis, with lineages making secondary contact during periodic landbridge formation. This oscillating cross-archipelago pattern in which ancient divergence is followed by repeated contact resembles that seen between East-West refugia populations from mainland Europe.

×

EL ESTUDIO ABORDADO EN LA TESIS VERSA SOBRE LA CARACTERIZACION TAXONOMICA DE LA LAGARTIJA DE LAS PITIUSAS PODARCIS PITVUSENSIS. SE ANALIZAN LAS DISTINTAS POBLACIONES DESDE TRES ASPECTOS DISTINTOS: EL ANALISISBIOMETRICO EL ANALISIS ELECTROFORETICO DE DISTINTAS PROTEINAS Y EL ANALISIS COLORIMETRICO. LOS ANALISIS ESTADISTICOS APLICADOS SOBRE LAS VARIANTES BIOMETRICAS DEMUESTRAN LA EXISTENCIA DE DIVERSOS GRUPOS DE POBLACIONES MUY SEMEJANTES ENTRE SI. LA VARIABILIDAD DE LA ESPECIE NO SOLO ES FENOTIPICA SINO QUE TAMBIEN ES GENETICA DETECTANDOSE UNA DIVERSIDAD EN ESTOS CARACTERES SUPERIOR A LA ESPERADA EN REPTILES. SE OBSERVA UNA ALTA HETEROSIS QUE ES CARACTERISTICA DE LA ESPECIE LO QUE PARECE DEMOSTRAR QUE ESTA SE ENCUENTRA EN LAS PRIMERAS FASES DE COLONIZACIONY ADAPTACION A LOS DIFERENTES HABITATS OFRECEN LAS ISLAS QUE OCUPA. SE CONSTATA LA ACCION DEL EFECTO FUNDADOR Y LA DERIVA GENETICA EN LOS TRES ASPECTOS CONSIDERADOS EN LA TESIS ASI COMO UNA TENDENCIA EVOLUTIVA HACIA EL AUMENTO DE TAMAÑO SIEMPRE QUE NO EXISTA UNA PRESION SELECTIVA CONTRARIA. SUCEDE LO MISMO CON EL MELANISMO. CONSIDERANDO EL ESTADO EVOLUTIVO ACTUAL DE LA ESPECIE REFLEJADO EN LA PLASTICIDAD DE LAS DISTINTAS POBLACIONES DE ESTALAARTIJA Y EN LA ADAPTACION QUE MANIFIESTAN EN CADA NICHO CONCRETO ASI COMO EL CONCEPTO RESTRICTIVO ACTUAL DE SUBESPECIE LA AUTORA CONSIDERA QUE SOLO PUEDEN CONSIDERARSE SEIS TAXONES SUBESPECIFICOS O SUBESPECIES DE LA LAGARTIJA DE LAS PITIUSAS.

×

In this paper are presented the results obtained with 45 populations of Podarcis pityusensis. Several multivariant technics are performed: discriminant analysis, cluster analysis and canonic analysis of populations. These populations present gradual change in all their biometric characteristics, and also a great interpopulation variability, that invalidate th statistical methods to discriminate between all of them. In different analysis very related population groups are found, that suggests they are constitute the same subspecies, in the actual taxonomic sense. These groups always inhabits islands with the same geological age, it is therefore necessary to complete the study with othr biological reflections, all of which are getting ready.

×

The dorsal, side and belly colours of 118 specimens of Podarcis pityusensis have been analysed with physical methods. Twenty samples from 16 islands have been chosen, 3 of then from Eivissa Island (Ibiza), 2 from Formentera Island and 2 from Espardell Island. The results reveal great colouration differences between the same island samples and sometimes similar colouration between different islands (different subspecies). specimens. This feature seems to show the colour criteria is not useful in taxonomic subjects for this species.

×

The variation in morphological and colouring features shown by the insular lacertid populations of Podarcis pityusensis is discussed from the point of view of their adaptive advantages to specific insular ecosystems. Insularity factors, i.e. area and island-age, have been found to be related to average body size, and the average luminosity of each population. Populations tend to show a size increase, a greater morphological homogeneity and darker dorsal colouring on smaller and older islands. Genetic drift seems to play a secondary role, whereas a positive selection in favour of melanism and giantism is observed. Both features are not linked as cause and effect, but seem to share a common cause: isolation and time enough to allow selection to take place. Predation, though slight in degree, does exist, and seems to be one of the selective pressure favouring melanism, together with the parallel trend towards an increase in body size and the need to an effective thermoregulation during the early hours of the day.

×

An electrophoretic comparison of ten colonies of Podarcis pityusensis Bosca, 1882 (Lacertidae) from Ibiza, Formentera and neighbouring islets (Balearic Is., Spain). 71 animals from ten colonies (vide list in text) of the Ibiza wall lizard Podarcis pi- tyusensis Bosca, 1882 were analysed by electrophoresis, on 12 % starch gel, and 16 gene- tic loci were studied, by checking against samples of Podarcis muralis muralis. The genetic frequency resulting in NEI`s identity index (I), reveals no clear differen- ces between the various colonies. However, itcan be seen on the resultant dendogram that P. p. carlkochi (affinis group) is distinct from the pityusensis sensu stricto group colonies. By other means we found characteristic allels for Podarcis pityusensis in four loci: GOT-1, IDM-1, G-6-PD-2 and 6-PGD, which differed from Podarcis muralis allels.

×

We examined the diets of several species of Podarcis from different western Mediterranean islands. We tested the prediction of more generalized diets in lizards from poor environments. We also examined the extert and mode of herbivory in relation to arthropod diversity in the diet. Lizards from the Tuscan Archipelago, P. sicula from Menorca, and P. tiliguerta from Corsica all show a rather generalized diet. In contrast, P. lilfordi and P. pilyusensis from the Balearic Islands and, to a lesser extent, P. hispanica from Benidorm Island, appear much more stenophagous, including a high proportion of clumped prey in their diets. Plant matter is a common food item in most of the population studied, specially in Balearic lizards. A marked seasonal shift in the degree of herbivory was observed in these species where high-energy plant parts are frequently consumed during summer. In one of the Balearic Islands (Nitge), the bulk of the summer diet was formed by pollen and nectar of one particular plant species. Herbivory, consumption of clumped prey and other dietary peculiarities of insular populations of Mediterranean lizards indicate a very flexible feeding behaviour compared to mainland congenerics. This could result from a lack of predation pressure allowing high population densities to be reached in islets with poor trophic resources. Differences found in dietary characteristics among island populations can be interpreted as a consequence of different current ecological conditions, as well as historical and biogeographical differences among the islands.

×

While the use of faecal pellets is widely accepted as a primary methodological source of data for dietary studies, a recent paper advocated for the use of gut contents. This was due to the fact that faecal samples would give biased results of the diet of arthropod predators, due to a lower representation of soft-bodied prey in faecal pellets. To test this assumption, we compared the spring diet of several populations of two insular lizards from the Balearic Islands (Spain), Podarcis lilfordi and Podarcis pityusensis, using both faecal pellets and gut contents. Our results do not support the supposed bias of dietary analyses based on faecal pellets. Indeed, soft-bodied prey and particularly insect larvae are often equally represented in faecal pellets and gut contents. Alternatively, soft bodied prey are represented in different proportions in gut contents and faecal pellets, but in some cases with higher proportions being observed in the gut contents, and in other cases with higher proportions in faecal samples. We conclude that faecal pellets can be a reliable source of information for dietary studies.

×

The parasitic female of the nematode Strongyloides ophiusensis sp. n.. fund in the intestine of lizard Podarcis pityusensis (Bosca, 1883) from Balearic Islands (West Mediterranean), is described. This species differs from other similar Strongyloides species in the body size, the morphology of the stoma, the structure of the ovaries and in the stage of development of the eggs, and also in some ecological and chorological characters.

×

The knowledge of the helminth communities of reptiles and their ecological relationships with their hosts are until the present not well known. Some general researches have been made only on American herps (AHO 1990). Recently, ROCA & HORNERO (1991a, 1991b) attempted similar kind of researches from Mediter- ranean insularlizards.

×

Two monophyletic sister species of wall lizards inhabit the two main groups of Balearic Islands: Podarcis lilfordi from islets and small islands around Mallorca and Menorca and Podarcis pityusensis from Ibiza, Formentera and associated islets. Genetic diversity within the endangered P. lilfordi has been well characterized, but P. pityusensis has not been studied in depth. Here, 2430 bp of mtDNA and 15 microsatellite loci were analysed from P. pityusensis populations from across its natural range. Two main genetic groupings were identified, although geographical structuring differed slightly between the mtDNA and the nuclear loci. In general, individuals from islets/islands adjacent to the main island of Ibiza were genetically distinct from those from Formentera and the associated Freus islands for both mtDNA and the nuclear loci. However, most individuals from the island of Ibiza were grouped with neighbouring islets/islands for nuclear loci, but with Formentera and Freus islands for the mitochondrial locus. A time-calibrated Bayesian tree was constructed for the principal mitochondrial lineages within the Balearics, using the multispecies coalescent model, and provided statistical support for divergence of the two main P. pityusensis lineages 0.111–0.295 Ma. This suggests a mid-late Pleistocene intraspecific divergence, compared with an early Pleistocene divergence in P. lilfordi, and postdates some major increases in sea level between 0.4 and 0.6 Ma, which may have flooded Formentera. The program IMa2 provided a posterior divergence time of 0.089–0.221 Ma, which was similar to the multispecies coalescent tree estimate. More significantly, it indicated low but asymmetric effective gene copy migration rates, with higher migration from Formentera to Ibiza populations. Our findings suggest that much of the present-day diversity may have originated from a late Pleistocene colonization of one island group from the other, followed by allopatric divergence of these populations. Subsequent gene flow between these insular groups seems likely to be explained by recent human introductions. Two evolutionary significant units can be defined for P. pityusensis but these units would need to exclude the populations that have been the subjects of recent admixture.

×

Sequences of the cytochrome b gene were analysed for 47 samples of two species of Podarcis from the Balearic Islands (P. pityusensis from Pityuses and P. lilfordi, from Gymnesies archipelago). The average uncorrected distance between the two species studied was 9.7%. The sampled individuals from each species form reciprocally monophyletic units. Assuming an overall rate of change for cytochrome b of 2% per million years the nucleotide divergence of 9.7 ± 1.9% between P. lilfordi and P. pityusensis, could correspond to a divergence time of 4.95 ± 0.95 million years. The separation time between Gymnesies and Pityuses archipelagos according to geological hypotheses is around 5 million years, thus our results suggest the ancestors of the two present forms became isolated during this event. The most parsimonious networks suggest that currently accepted subspecies do not form monophyletic groups, and so should be reassessed. Despite our limited sampling the level of variability is much higher in P. lilfordi than P. pityusensis. Since they are sister taxa the relative age of each species is equal, therefore the differences might be due to historical population structure differences. However it seems that the forms on the islands are not genetically distinct units, implying that morphological differences are recent adaptations to their environments.

×

The alteration of a habitat such as that caused by the introduction of exotic animal species that displace the native ones, reducing their populations or leading them to extinction, can result in the disruption of species interactions which may have evolved during millions of years. The insular ecosystems are especially vulnerable to such disturbances, as they are usually less complex and have a lower number of species than the continental ones. In the present work I document the consequences of the disruption of plant-disperser interactions, caused by the introduction of carnivorous mammals in the Balearic Islands, for the populations of two shrub species: Cneorum tricoccon L., with a distribution restricted to the western Mediterranean, and Daphne rodriguezii Texidor, endemic from Menorca island. Whereas carnivores have replaced ⁄ the native dispersers (lizards) of the former, notably modifying its distribution -especially in Mallorca-, the plant-lizard mutualism disruption seems to have been dramatic for the second species, drastically reducing its populations except in an islet where lizards still persist.