Potential intermediate hosts of Angiostrongylus cantonensis in the European Mediterranean region (Mallorca, Spain).

Angiostrongylus cantonensis is the main etiological agent of eosinophilic meningitis, a disease that often leads to severe neurological manifestations in mammals and birds. In recent years, the prevalence of this zoonotic nematode has dramatically increased as it expands into new territories beyond its native range in Southeast Asia and the Pacific Basin.Its arrival to Europe, the last continent to be invaded, has raised many questions concerning the parasite's life cycle, particularly in the Mediterranean region, where it is now endemic. This study aims to collect evidence about intermediate hosts (snail and slug species) involved in the transmission of the A. cantonensis in Mallorca. We have conducted a systematic surveillance of gastropods within 2 km radius areas, surrounding the specific locations where infected hedgehogs were found. We employed a sequence-based detection approach that included a species-specific PCR amplification followed by DNA sequencing of the internal transcribed spacer I (ITS-1). This conservative approach was essential to avoid cross reactions with the numerous metastrongylid species known to be circulating in Mallorca. Overall, we collected and identified 398 gastropods belonging to 17 species, of which 11% tested positive for A. cantonensis. These gastropods were collected from touristic settlements to agricultural lands. Five gastropod species: Theba pisana, Cornu aspersum, Deroceras reticulatum, Milax gagates and Otala lactea, had been previously incriminated as A. cantonensis intermediate hosts, while 6 species: Papillifera papillaris, Cochlicella acuta, Eobania vermiculata, Ganula lanuginosa, Milax nigricans and Rumina decollata, are newly recorded hosts. The findings of this study have important epidemiological implications, and further measures are discussed to prevent neuroangiostrongylosis cases.

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.

Evidence of autochthonous transmission of urinary schistosomiasis in Almeria (southeast Spain): An outbreak analysis.

BACKGROUND: Schistosomiasis is endemic in 78 countries belonging to tropical and subtropical areas. However, autochthonous transmission of urogenital schistosomiasis was reported in Corsica (France) in 2013. We present evidence of autochthonous transmission of urogenital schistosomiasis in Almería (Spain) in 2003. METHODS: Description of the outbreak in farmers and subsequent epidemiological studies aimed at searching for Bulinus snails and their genotypic characteristics. RESULTS: The outbreak affected 4 farmers out of a group of 5 people who repeatedly bathed that summer in an irrigation pool in the area. Two of them presented macroscopic hematuria with bilharziomas, showing the presence of Schistosoma eggs in bladder biopsies. Two others were asymptomatic but the serology for schistosomiasis was positive. In 2015, the presence of the vector Bulinus truncatus was demonstrated in Almería in water collections of appropriate characteristics. DNA sequencing proving that local B. truncatus species were base-to-base identical to B. truncatus from Senegal. CONCLUSIONS: We present a new outbreak of autochthonous transmission of urogenital schistosomiasis in Europe. Although no new cases of autochthonous transmission have been reported, some other cases may have occurred at that time or later on and be unnoticed as many cases of schistosomiasis are asymptomatic or present mild and unspecific symptoms.

Iberian Distribution of the Freshwater Snail Genus Bithynia Leach, 1818 (Mollusca: Truncatelloidea), Vector of Opisthorchiasis and Metorchiasis.

BACKGROUND: Opisthorchis felineus and Metorchis bilis are trematodes that cause opisthorchiasis and metorchiasis, respectively. The freshwater snails Bithynia hispanica and B. tentaculata are the respective intermediate hosts for these parasites in the Iberian Peninsula, where both parasites are present. METHODS: To study the distribution of these snail species, an exhaustive literature review and revision of museum collections was performed. RESULTS: A total of 370 localities were compiled and mapped. B. tentaculata were found throughout the Iberian Peninsula, both in Spain and Portugal, while B. hispanica was found only in the Mediterranean coast of Spain. CONCLUSION: Knowing the distribution of the Bithynia species found in the Iberian Peninsula is highlighted in terms of public health, as the presence of the snail is related to the presence of the disease and can be used as a tool to face future outbreaks.

Molecular phylogeny of the western Palaearctic Helicoidea (Gastropoda, Stylommatophora).

The Helicoidea is one of the most diverse superfamilies of terrestrial land snails. In this study we present a molecular phylogeny of the western Palaearctic Helicoidea obtained by means of neighbor joining, maximum likelihood and Bayesian analysis of the mitochondrial 16S rRNA gene fragment and the nuclear rRNA gene cluster including the 3' end of the 5.8S gene, the complete ITS2 region and 5' end of the large subunit 28S. Most of the morphologically-defined families were confirmed. We propose a revised phylogenetic classification so that families, subfamilies and tribes are monophyletic. The family Hygromiidae sensu Hausdorf and Bouchet (2005) is divided into three clades which are here given familial rank: Canariellidae and Geomitridae, which are recognized for the first time at familial rank, and Hygromiidae s.str. (including Ciliella and Trochulus) that is here restricted. The subfamilies Ciliellinae, Geomitrinae, Hygromiinae, Monachainae and Trochulinae recognized in current classifications were not recovered as monophyletic groups. The family Cochlicellidae is here given tribe rank (Cochlicellini) belonging to the Geomitridae. We describe a new tribe, Plentuisini. Three subfamilies are recognized within Helicidae: Ariantinae, Helicinae (including Theba) and Murellinae. New classification indicates that free right ommatophore retractor muscle arose only once within Geomitridae. The anatomy of the auxiliary copulatory organs of the reproductive system of families, subfamilies and tribes is highlighted. We estimate the origin of the Helicoidea at the end of the Early Cretaceous and its families as Late-Cretaceous to Paleogene. Western Palaearctic Helicoidea belongs to two different lineages that diverged around 86Ma ago, both starting their diversification at the end of the Cretaceous (around 73-76Ma). Radiation of some western Helicoidean families started during the Eocene.

Land snails as a diet diversification proxy during the early upper palaeolithic in Europe.

Despite the ubiquity of terrestrial gastropods in the Late Pleistocene and Holocene archaeological record, it is still unknown when and how this type of invertebrate resource was incorporated into human diets. In this paper, we report the oldest evidence of land snail exploitation as a food resource in Europe dated to 31.3-26.9 ka yr cal BP from the recently discovered site of Cova de la Barriada (eastern Iberian Peninsula). Mono-specific accumulations of large Iberus alonensis land snails (Ferussac 1821) were found in three different archaeological levels in association with combustion structures, along with lithic and faunal assemblages. Using a new analytical protocol based on taphonomic, microX-Ray Diffractometer (DXR) and biometric analyses, we investigated the patterns of selection, consumption and accumulation of land snails at the site. The results display a strong mono-specific gathering of adult individuals, most of them older than 55 weeks, which were roasted in ambers of pine and juniper under 375°C. This case study uncovers new patterns of invertebrate exploitation during the Gravettian in southwestern Europe without known precedents in the Middle Palaeolithic nor the Aurignacian. In the Mediterranean context, such an early occurrence contrasts with the neighbouring areas of Morocco, France, Italy and the Balkans, where the systematic nutritional use of land snails appears approximately 10,000 years later during the Iberomaurisian and the Late Epigravettian. The appearance of this new subsistence activity in the eastern and southern regions of Spain was coeval to other demographically driven transformations in the archaeological record, suggesting different chronological patterns of resource intensification and diet broadening along the Upper Palaeolithic in the Mediterranean basin.

Temporal speciation pattern in the western Mediterranean genus Tudorella P. Fischer, 1885 (Gastropoda, Pomatiidae) supports the Tyrrhenian vicariance hypothesis.

The land snail genus Tudorella shows a peculiar disjunct distribution around the western Mediterranean coasts. Despite high phenotypic plasticity, only two species with a disputed number of subspecific taxa are currently recognised. We delimited the species with mitochondrial (COI & 16S) and nuclear (ITS-1) markers based on the unified species concept and suggested that there are eight species in the genus, two of them currently undescribed. Applying Bayesian phylogenetic model selection, we tested four different biogeographic hypotheses that could be causal for the current distribution pattern of extant Tudorella species. A scenario involving vicariance events resulting from the repeated splits of the Tyrrhenian plate with subsequent dispersal events over land bridges during the Pliocene received greatest support in the data.