Paranannizziopsis spp. Infection in Wild Vipers, Europe.

We describe the detection of Paranannizziopsis sp. fungus in a wild population of vipers in Europe. Fungal infections were severe, and 1 animal likely died from infection. Surveillance efforts are needed to better understand the threat of this pathogen to snake conservation.

Effect of Myxoma Virus Species Jump on Iberian Hare Populations.

The myxoma virus species jump from European rabbits (Oryctolagus cuniculus) to Iberian hares (Lepus granatensis) has raised concerns. We assess the decline suffered by Iberian hare populations on the Iberian Peninsula and discuss the association between the effect of myxomatosis and the average abundance index, which we estimated by using hunting bags.

Spatial genetic structure in European marbled newts revealed with target enrichment by sequence capture.

European marbled newts come in two species that have abutting ranges. The northern species, Triturus marmoratus, is found in France and the northern part of the Iberian Peninsula, whereas the southern species, T. pygmaeus, is found in the southwestern corner of the Iberian Peninsula. We study the intraspecific genetic differentiation of the group because morphological data show geographical variation and because the Iberian Peninsula is a recognized center of speciation and intraspecific genetic diversity for all kinds of organisms, amphibians included. We use target enrichment by sequence capture to generate c. 7 k nuclear DNA markers. We observe limited genetic exchange between the species, which confirms their distinctiveness. Both species show substantial genetic structuring that is only in part mirrored by morphological variation. Genetically differentiated groups are found in the south (T. marmoratus) and west (T. pygmaeus) of the species ranges. Our observations highlight the position of the Iberian Peninsula as a hotspot for genetic differentiation.

The symbolic role of the underground world among Middle Paleolithic Neanderthals.

Cueva de Ardales in Málaga, Spain, is one of the richest and best-preserved Paleolithic painted caves of southwestern Europe, containing over a thousand graphic representations. Here, we study the red pigment in panel II.A.3 of "Sala de las Estrellas," dated by U-Th to the Middle Paleolithic, to determine its composition, verify its anthropogenic nature, infer the associated behaviors, and discuss their implications. Using optical microscopy, scanning electron microscopy coupled with energy dispersive X-ray spectroscopy, micro-Raman spectroscopy, and X-ray diffraction, we analyzed a set of samples from the panel and compared them to natural coloring materials collected from the floor and walls of the cave. The conspicuously different texture and composition of the geological samples indicates that the pigments used in the paintings do not come from the outcrops of colorant material known in the cave. We confirm that the paintings are not the result of natural processes and show that the composition of the paint is consistent with the artistic activity being recurrent. Our results strengthen the hypothesis that Neanderthals symbolically used these paintings and the large stalagmitic dome harboring them over an extended time span.

Tracking movements of meagre (Argyrosomus regius) during the spawning season: Preliminary indications of off estuary spawning.

Understanding the spawning behavior of meagre (Argyrosomus regius) is crucial for fisheries management and conservation. Meagre forms large spawning aggregations in estuaries, yet details of its spawning grounds remain elusive. We tagged 41 individuals and monitored their movements throughout several spawning seasons. Detections inside estuaries were limited. Fish detected inside the Tejo estuary during the spawning season performed regular movements toward coastal areas, suggesting potential spawning outside estuaries. Our findings underscore the significance of understanding meagre's migratory patterns for effective fisheries management and conservation strategies, supporting the establishment of spatial and seasonal closures during the spawning season.

Stable isotope analyses of amino acids reveal the importance of aquatic resources to Mediterranean coastal hunter-gatherers.

Determining the degree to which humans relied on coastal resources in the past is key for understanding long-term social and economic development, as well as for assessing human health and anthropogenic impacts on the environment. Prehistoric hunter-gatherers are often assumed to have heavily exploited aquatic resources, especially those living in regions of high marine productivity. For the Mediterranean, this view has been challenged, partly by the application of stable isotope analysis of skeletal remains which has shown more varied coastal hunter-gatherer diets than in other regions, perhaps due to its lower productivity. By undertaking a more specific analysis of amino acids from bone collagen of 11 individuals from one of the oldest and best-known Mesolithic cemeteries in the Mediterranean, at El Collado, Valencia, we show that high levels of aquatic protein consumption were achieved. By measuring both carbon and nitrogen in amino acids, we conclude that some of the El Collado humans relied heavily on local lagoonal fish and possibly shellfish, rather than open marine species. By contrast to previous suggestions, this study demonstrates that the north-western coast of the Mediterranean basin could support maritime-oriented economies during the Early Holocene.

The population bottleneck of the Iberian wolf impacted genetic diversity but not admixture with domestic dogs: A temporal genomic approach.

After decades of intense persecution, the Iberian wolf subspecies faced a severe bottleneck in the 1970s that considerably reduced its range and population size, nearly leading to its extinction in central and southern Iberian Peninsula. Such population decline could have impacted the genetic diversity of Iberian wolves through different processes, namely genetic drift and dynamics of hybridization with domestic dogs. By contrasting the genomes of 68 contemporary with 54 historical samples spanning the periods before and immediately after the 1970s bottleneck, we found evidence of its impact on genetic diversity and dynamics of wolf-dog hybridization. Our genome-wide assessment revealed that wolves and dogs form two well-differentiated genetic groups in Iberia and that hybridization rates did not increase during the bottleneck. However, an increased number of hybrid individuals was found over time during the population re-expansion, particularly at the edge of the wolf range. We estimated a low percentage of dog ancestry (~1.4%) in historical samples, suggesting that dog introgression was not a key driver for wolf extinction in central and southern Iberia. Our findings also unveil a significant decline in genetic diversity in contemporary samples, with the highest proportion of homozygous segments in the genome being recently inherited. Overall, our study provides unprecedented insight into the impact of a sharp decline on the Iberian wolf genome and refines our understanding of the ecological and evolutionary drivers of wolf-dog hybridization in the wild.

Genomic phylogeography illuminates deep cyto-nuclear discordances in midwife toads (Alytes).

The advent of genomic methods allows us to revisit the evolutionary history of organismal groups for which robust phylogenies are still lacking, particularly in species complexes that frequently hybridize. In this study, we conduct RAD-sequencing (RAD-seq) analyses of midwife toads (genus Alytes), an iconic group of western Mediterranean amphibians famous for their parental care behavior, but equally infamous for the difficulties to reconstruct their evolutionary history. Through admixture and phylogenetic analyses of thousands of loci, we provide a comprehensive phylogeographic framework for the A. obstetricans complex, as well as a fully resolved phylogeny for the entire genus. As part of this effort, we carefully explore the influence of different sampling schemes and data filtering thresholds on tree reconstruction, showing that several, slightly different, yet robust topologies may be retrieved with small datasets obtained by stringent SNP calling parameters, especially when admixed individuals are included. In contrast, analyses of incomplete but larger datasets converged on the same phylogeny, irrespective of the reconstruction method used or the proportion of missing data. The Alytes tree features three Miocene-diverged clades corresponding to the proposed subgenera Ammoryctis (A. cisternasii), Baleaphryne (A. maurus, A. dickhilleni and A. muletensis), and Alytes (A. obstetricans complex). The latter consists of six evolutionary lineages, grouped into three clades of Pliocene origin, and currently delimited as two species: (1) A. almogavarii almogavarii and A. a. inigoi; (2) A. obstetricans obstetricans and A. o. pertinax; (3) A. o. boscai and an undescribed taxon (A. o. cf. boscai). These results contradict the mitochondrial tree, due to past mitochondrial captures in A. a. almogavarii (central Pyrenees) and A. o. boscai (central Iberia) by A. obstetricans ancestors during the Pleistocene. Patterns of admixture between subspecies appear far more extensive than previously assumed from microsatellites, causing nomenclatural uncertainties, and even underlying the reticulate evolution of one taxon (A. o. pertinax). All Ammoryctis and Baleaphryne species form shallow clades, so their taxonomy should remain stable. Amid the prevalence of cyto-nuclear discordance among terrestrial vertebrates and the usual lack of resolution of conventional nuclear markers, our study advocates for phylogeography based on next-generation sequencing, but also encourages properly exploring parameter space and sampling schemes when building and analyzing genomic datasets.

Prevalence of Oestrus ovis in small ruminants from the eastern Iberian Peninsula. A long-term study.

Oestrus ovis is an obligate parasite that causes myiasis in domestic ruminants, being commonly found in the Mediterranean area. From 2009 to 2019 a total of 3476 heads of culling sheep and goats from the Mediterranean coast of Spain were examined for the presence of O. ovis. The total prevalence was 56.3%, significantly higher in sheep than in goats (61.2% and 43%, respectively). Differences were found in the mean annual prevalence, with the highest value being registered in 2018 (61.7%) and the lowest in 2012 (50.3%). Autumn, for sheep, and winter, for goats, were the seasons with the highest number of infested specimens. Temperature, but not rainfall, was found to be associated with prevalence (p < 0.05). Most L1 were found in the anatomic region I (septum, meatus, and ventral conchae), while L2 and L3 were mainly located in regions II (nasopharynx, ethmoid labyrinth, and dorsal conchae), and III (sinuses). The overall intensity was 12.8 larvae per head, significantly higher in sheep (13.3) than in goats (3.5). Our results confirm the high prevalence of O. ovis in sheep and goats in this geographic area over the last decade, with the trend increasing in recent years in association with higher mean temperatures.

First helminthological data on the Iberian adder, Vipera seoanei.

The helminth fauna present in the gut contents of Iberian adders, Vipera seoanei (Squamata: Viperidae), were characterised and analysed in respect to biological and eco-geographic factors that may affect the occurrence and diversity of helminths in this species. A total of 317 samples of preserved stomachs and intestines, covering the distributional range of V. seoanei, were examined. Similar to other Vipera species from the Iberian Peninsula, the helminth fauna was also impoverished in V. seoanei, but unlike other Vipera species from Central and East Europe, helminths were mostly found in adult vipers, and occurred in vipers located at the periphery of the species range, characterised by low elevation, high temperature and precipitation levels, and abundant pastures.

Molecular phylogeny of the genus Chondrina (Gastropoda, Panpulmonata, Chondrinidae) in the Iberian Peninsula.

Chondrina Reichenbach, 1828 is a highly diverse genus of terrestrial molluscs currently including 44 species with about 28 subspecific taxa. It is distributed through North Africa, central and southern Europe, from Portugal in the West to the Caucasus and Asia Minor in the East. Approximately 70% of the species are endemic to the Iberian Peninsula constituting its main center of speciation with 34 species. This genus includes many microendemic taxa, some of them not yet described, confined to limestone habitats (being strictly rock-dwelling species). They are distributed on rocky outcrops up to 2000 m.a.s.l. It is a genus of conical-fusiform snails that differ mainly in shell characters and in the number and position of teeth in their aperture. So far, molecular studies on Chondrina have been based exclusively on the mitochondrial Cytochrome Oxidase subunit I region (COI). These studies gave a first view of the phylogeny of the genus but many inner nodes were not statistically supported. The main objective of the study is to obtain a better understanding of the phylogeny and systematics of the genus Chondrina on the Iberian Peninsula, using multilocus molecular analysis. Partial sequences of the COI and 16S rRNA genes, as well as of the nuclear Internal Transcribed Spacer 1 (ITS1-5.8S) and Internal Transcribed Spacer 2 (5.8S-ITS2-28S) were obtained from individuals of all the extant Chondrina species known from the Iberian Peninsula. In addition to this, the newly obtained COI sequences were combined with those previously published in the GenBank. Phylogenetic relationships were inferred using maximum likelihood and Bayesian methods. The reconstructed phylogenies showed high values of support for more recent branches and basal nodes. Moreover, molecular species delimitation allowed to better definethe studied species and check the presence of new taxa.

Genetic traces of hybrid zone movement across a fragmented habitat.

Theoretical and empirical studies suggest that the structure and position of hybrid zones can change over time. Evidence for moving hybrid zones has been directly inferred by repeated sampling over time, or indirectly through the detection of genetic footprints left by the receding species and the resulting asymmetric patterns of introgression across markers. We here investigate a hybrid zone formed by two subspecies of the Iberian golden-striped salamander, Chioglossa lusitanica, using a panel of 35 nuclear loci (31 SNPs and 4 allozymes) and one mitochondrial locus in a transect in central Portugal. We found concordant and coincident clines for most of the nuclear loci (n = 22, 63%), defining a narrow hybrid zone of ca. 6 km wide, with the centre positioned ca. 15 km south of the Mondego River. Asymmetric introgression was observed at another 14 loci. Their clines are displaced towards the north, with positions located either close to the Mondego River (n = 6) or further northwards (n = 8). We interpret these profiles as genetic traces of the southward displacement of C. lusitanica lusitanica by C. l. longipes over the wider Mondego River valley. We noted the absence of significant linkage disequilibrium, and we inferred low levels of effective selection per locus against hybrids, suggesting that introgression in the area of species replacement occurred under a neutral diffusion process. A species distribution model suggests that the C. lusitanica hybrid zone coincides with a narrow corridor of fragmented habitat. From the position of the displaced clines, we infer that patches of locally suitable habitat trapped some genetic variants that became disassociated from the southward moving hybrid zone. This study highlights the influence of habitat availability on hybrid zone movement.

Metal accumulation by Alyssum serpyllifolium subsp. malacitanum Rivas Goday (Brassicaceae) across different petrographic entities in South-Iberian ultramafic massifs: plant-soil relationships and prospects for phytomining.

NOVELTY STATEMENT: To date, studies of hyperaccumulation in plant tissues on ultramafic rocks have not considered the great diversity of petrographic entities in the world's outcrops. One of them is the one that we studied in Spain with more than eight petrographic entities and different soils. Our hypothesis is that the different chemical compositions of the soils in ultramafic rocks significantly affect the hyperaccumulation of metals by specialized plants, which may have consequences for phytomining. For this purpose, individuals, populations, and different soils have been tested and the results have been subjected to the corresponding statistical tests. The obtained knowledge reflects the different behavior of the studied plant not only for the Ni: the obtained results for Sr and for Ba revealed interesting results for the hyperaccumulation in Alyssum of both metals.

Mapping the spatial variability of rainfall from a physiographic-based multilinear regression: model development and application to the Southwestern Iberian Peninsula.

A physiographic-based multilinear regression model supported by GIS was developed to estimate spatial rainfall variability in the Southwest Iberian Peninsula. The area study includes a wide diversity of landscape features and comprises four Portuguese regions and one Spanish province (totalizing 28,860 km2). The region suffers a very strong Mediterranean influence, with a major cleavage between winter and summer seasons. Thus, the analysis was carried out separately for the wet (October to March) and dry (April to September) semesters. From an initial set of 10 explanatory physiographic variables, five were selected to be used in the multilinear regression, as they allowed generating models by map algebra that fitted well with the last 40 years of monthly rainfall data records. These records were obtained from 163 weather stations, filtered from an initial set of 230 (142 stations in Portugal and 88 in Spain). The correlation between the physiographic-based multilinear regression model and a model obtained by interpolation from rainfall historical data showed to be good or very good in approximately 75% of the area under study. Results show that physiographic-based models can be effectively used to estimate rainfall where there is a lack of rain gauges, or to densify spatial resolution of rainfall between rain gauges.

The genetic scenario of Mercheros: an under-represented group within the Iberian Peninsula.

BACKGROUND: The general picture of human genetic variation has been vastly depicted in the last years, yet many populations remain broadly understudied. In this work, we analyze for the first time the Merchero population, a Spanish minority ethnic group that has been scarcely studied and historically persecuted. Mercheros have been roughly characterised by an itinerant history, common traditional occupations, and the usage of their own language. RESULTS: Here, we examine the demographic history and genetic scenario of Mercheros, by using genome-wide array data, whole mitochondrial sequences, and Y chromosome STR markers from 25 individuals. These samples have been complemented with a wide-range of present-day populations from Western Eurasia and North Africa. Our results show that the genetic diversity of Mercheros is explained within the context of the Iberian Peninsula, evidencing a modest signal of Roma admixture. In addition, Mercheros present low genetic isolation and intrapopulation heterogeneity. CONCLUSIONS: This study represents the first genetic characterisation of the Merchero population, depicting their fine-scale ancestry components and genetic scenario within the Iberian Peninsula. Since ethnicity is not only influenced by genetic ancestry but also cultural factors, other studies from multiple disciplines are needed to further explore the Merchero population. As with Mercheros, there is a considerable gap of underrepresented populations and ethnic groups in publicly available genetic data. Thus, we encourage the consideration of more ethnically diverse population panels in human genetic studies, as an attempt to improve the representation of human populations and better reconstruct their fine-scale history.

Hybridization and cryptic speciation in the Iberian endemic plant genus Phalacrocarpum (Asteraceae-Anthemideae).

Understanding the role and impact of reticulation in phylogenetic inquiry has improved with extended use of high throughput sequencing data. Yet, due to the dynamism of genomes over evolutionary time, disentangling old hybridization events remains a serious challenge. Phalacrocarpum (DC.) Willk. is one of the 27 Iberian endemic plant genera, currently considered monotypic but including three subspecies. Its uncertain phylogenetic relationships within tribe Anthemideae (Asteraceae) point to an Early Miocene divergence from its sister group, and its persistent taxonomic instability has been proposed to be due to hybridization. We aim at understanding the evolutionary history of this genus using SNPs called from a genotyping-by-sequencing (GBS) analysis, Sanger sequences-from three plastid DNA regions (psbJ-petA, petB-petD, trnH-psbA) and the nuclear ribosomal ITS regions (cloned)-as well as leaf morphometric multivariate analysis. SNP data and Sanger sequences strongly support the unforeseen existence of a cryptic species in the eastern populations of P. oppositifolium subsp. anomalum. Broad molecular and morphometric patterns of variation found in conflictive populations from the Sanabria Valley region convincingly identify a recent previously undocumented hybrid zone. By contrast, evidence is less conclusive on relationships between subspecies hoffmannseggii, oppositifolium and a second conflictive group distributed along the Galician-Portuguese border (Orense massifs). Although genetic clustering analysis of SNP data suggests that the former subspecies was the maternal progenitor in hybridization events that gave rise to the other two groups, we found considerable uniqueness of ITS ribotypes and plastid haplotypes in them. This result, in the context of Pleistocene climatically-driven range shifts in NW Iberian Peninsula, can be due to periods of isolation, genetic bottlenecks and drift superimposed on old hybridization events. Our study confirms the idea that unravelling old hybridization events may be compromised by the suite of evolutionary processes accumulated subsequently, particularly in areas with a history of climatic instability.

Incorporating palaeogeography into ancestral area estimation can explain the disjunct distribution of land snails in Macaronesia and the Balearic Islands (Helicidae: Allognathini).

The systematics and biogeographical history of the Eastern Mediterranean and Macaronesian land snail tribe Allognathini (Helicidae: Helicinae) is investigated based on mitochondrial and nuclear DNA sequence data. Our molecular phylogenetic analyses indicate that the genus-group systematics of the tribe needs to be revised. We show for the first time that the narrow-range endemics Lampadia and Idiomela from the Madeira Archipelago belong to Allognathini and represent together the sister group of the diverse Canary Island Hemicycla radiation. We therefore suggest synonymising Lampadiini with Allognathini. Sister to these Macaronesian genera was the Balearic Island Allognathus radiation. Pseudotachea was not recovered as a monophyletic group and the two currently recognised species clustered in Iberus. Similarly, Adiverticula was not recovered as a monophyletic group and clustered in Hemicycla. We therefore suggest synonymising Pseudotachea with Iberus and Adiverticula with Hemicycla. The six genera in Allognathini, which we distinguish here (Cepaea, Iberus, Allognathus, Hemicycla, Idiomela and Lampadia), originated in Western to South-western Europe according to our ancestral area estimation and the fossil record. The disjunct distribution of the Balearic Islands and Macaronesian sister clades and the mainly Iberian Iberus clade that separated earlier can be explained by the separation of the Betic-Rif System from the Iberian Peninsula during the late Oligocene to early Miocene, along with independent Miocene dispersals to the Balearic Islands and Macaronesia from the Iberian Peninsula, where the ancestral lineage became extinct.

Stable isotope evidence of human diet in Mediterranean context during the Last Glacial Maximum.

The Final Gravettian in Europe overlapped with the cold and dry climatic event of Heinrich 2 (ca. 27-23.5 kyr cal BP), which caused the contraction of human distribution over refuge regions in the southern peninsulas of Europe. Here, we consider the human subsistence in the northeast Iberian Peninsula, where an extensive range of small to large prey was available. Four human remains from the Serinyà caves were investigated using the stable isotope ratios of carbon, nitrogen, and sulfur of bulk collagen (δ13Ccoll, δ15Ncoll, δ34Scoll) and of phenylalanine and glutamine amino acids (δ15NPhe, δ15NGlu). Direct AMS dating of the human and animal remains from the Final Gravettian levels of Mollet III, Reclau Viver, and Arbreda at Serinyà confirmed their chronological position from 27.5 to 22.6 kyr cal BP and the occurrence of four different human individuals. The δ13Ccoll and δ15Ncoll values showed a large contribution of terrestrial prey to the dietary protein of the individuals. The δ34Scoll values were consistent with a subsistence based on the local continental resources, without detectable contribution of marine resource. The δ15NPhe and δ15NGlu values confirm that freshwater resources were not a substantial component of the diet of the considered individuals. Contrast in the isotopic amounts in bulk collagen could be interpreted as the result of different proportions of terrestrial prey in human diet at Serinyà. Altogether, the isotopic investigation reveals the importance of terrestrial over aquatic resources in the subsistence of the studied Final Gravettian individuals from the Serinyà caves in northeastern Iberia during the Last Glacial Maximum. It would be consistent with a scenario of a productive enough terrestrial ecosystem to sustain hunter-gatherer subsistence in this refuge region.

Proctodeal extrusion as a defensive behavioral response in blister beetles (Coleoptera: Meloidae).

Defensive mechanisms in blister beetles (Coleoptera: Meloidae) include a wide variety of behavioral responses, chemical defense, and conspicuous external colorations. Although some of these mechanisms have been previously described, proctodeal extrusion, a defensive behavior involving the extrusion of inner abdominal membranes from the proctodeal region which appear intensely red or orange colored when the hemolymph is seen through them, has not been reported to date. Here, we tested the ability to display proctodeal extrusion in response to threat stimuli in wild populations of three blister beetle species inhabiting Central Spain: Berberomeloe majalis (Linnaeus, 1758), Berberomeloe comunero Sánchez-Vialas, García-París, Ruiz & Recuero, 2020, and Physomeloe corallifer (Germar, 1818). In addition, we observed and recorded various other defensive behaviors such as immobility, antennal threat display, autohemorrhage (reflex bleeding), defecation, and thanatosis (death feigning). The frequency at which proctodeal extrusion was observed differed among species, as did the stress intensity needed for extrusion and the probability of proctodeal extrusion in response to a particular threatening stimulus. Our findings indicate that, although proctodeal extrusion might be a widespread potential defensive mechanism in Meloidae, the ability to elicit it is not generalized across lineages. Physomeloe and Berberomeloe are endemic to the semi-arid Mediterranean region, and species adapted to such a climate would have developed strategies that limit hydric stress such as proctodeal extrusion, which mirrors the effect of autohemorrhage but without the fluid loss.

Phenological and intrinsic predictors of mite and haemacoccidian infection dynamics in a Mediterranean community of lizards.

Ectotherms are vulnerable to environmental changes and their parasites are biological health indicators. Thus, parasite load in ectotherms is expected to show a marked phenology. This study investigates temporal host­parasite dynamics in a lizard community in Eastern Spain during an entire annual activity period. The hosts investigated were Acanthodactylus erythrurus, Psammodromus algirus and Psammodromus edwardsianus, three lizard species coexisting in a mixed habitat of forests and dunes, providing a range of body sizes, ecological requirements and life history traits. Habitat and climate were considered as potential environmental predictors of parasite abundance, while size, body condition and sex as intrinsic predictors. Linear models based on robust estimates were fitted to analyse parasite abundance and prevalence. Ectoparasitic mites and blood parasites from two haemococcidian genera were found: Lankesterella spp. and Schellackia spp. Habitat type was the only predictor explaining the abundance of all parasites, being mostly higher in the forest than in the dunes. The results suggest that particularities in each host­parasite relationship should be accounted even when parasites infect close-related hosts under the same environmental pressures. They also support that lizard parasites can be biomarkers of environmental perturbation, but the relationships need to be carefully interpreted for each host­parasite assemblage.