THE COCCIDIA SPECIES PFEIFFERINELLA ELLIPSOIDES EXHIBITS SPATIAL AND SPECIES-SPECIFIC VARIATION IN PREVALENCE AMONG FRESHWATER SNAILS.

Freshwater snails are commonly studied within the context of their role as intermediate hosts for digenetic trematodes. However, there are fundamental data deficiencies related to our understanding of directly transmitted parasites, such as coccidia, for freshwater snails. Because variation in coccidia pathogenicity and transmission among snail species likely has major impacts on snail community structure, we aimed to investigate the spatial distribution and prevalence of coccidia in several freshwater snail species throughout the Ozark and Ouachita Mountains ecoregions in Arkansas. We opportunistically collected 220 freshwater snails from 24 Ozark sites in summer 2022 and scanned fecal slides for the presence of coccidia. In summer 2023, we surveyed an additional 146 snails from 19 Ouachita sites. To test for apparent interactions among coccidia and trematodes, we scanned feces from a subset of snails (Physa and Planorbella in the Ozarks) that did not have concurrent trematode infections and from those that did. We observed oocysts that morphologically conformed to Pfeifferinella ellipsoides in 2 of the 9 snail taxa from 7 of the 43 sites. Planorbella trivolvis was infected at 2 of 6 sites in the Ozarks and 0 of 5 sites in the Ouachitas. Physa species were infected at 6 of 14 sites in the Ozarks and 0 of 12 sites in the Ouachitas. In the Ozarks, Pl. trivolvis had an overall prevalence of 0.13 (6 of 47), whereas individuals in the genus Physa had an overall prevalence of 0.08 (8 of 97). Our chi-square and Fisher exact tests revealed no significant evidence for trematode-coccidia competition or synergism within the two snail species. There were no other species infected, and we did not observe any coccidia in the snails from the Ouachitas. Our survey of 366 snails among 9 taxa and 43 sites represents the largest survey for freshwater snail coccidia to date and indicates that both Pl. trivolvis and Physa spp. may be primary hosts and/or reservoir hosts for Pf. ellipsoides in freshwater snail communities. The highly aggregated distribution of Pf. ellipsoides in northwestern Arkansas requires further investigation. Our results led to proposal of several hypotheses for additional research, including questions regarding the variation of coccidia host specificity and virulence.

An update on snail and trematode communities in the Sanyati Basin of Lake Kariba: New snail and trematode species but no human schistosomes.

BACKGROUND: The construction of Lake Kariba brought about a rise in the incidence of schistosomiasis in its surrounding towns of Kariba (Zimbabwe) and Siavonga (Zambia). After extensive control programs in Kariba, schistosomiasis prevalence dropped significantly. The objective of this study was to revisit the same localities sampled by Chimbari et al. (2003), and provide an update on the snail community and prevalence of trematodes in the Northern shore of Lake Kariba while focusing on planorbid species. METHODS: Monthly sampling of snails at 16 sites along the Northern shoreline of Lake Kariba, near Kariba town, was undertaken for one year. Minimum one specimen per morphotype was identified using molecular barcoding (sequencing a fragment of cytochrome c oxidase I subunit (COI)). The infection status of snails was assessed by Rapid Diagnostic PCRs (RD-PCR), and trematode infections were genotyped by sequencing COI and 18S rDNA markers. RESULTS: We collected and identified seven snail species: Bulinus truncatus, Bulinus forskalii, Gyraulus sp., Physella acuta, Bellamya sp., Radix affinis plicatula and Pseudosuccinea columella. Physella acuta was the most abundant snail species (comprising 56.95% of the total snail count) and present at all sites. The B. truncatus population was found to be infected with the stomach fluke Carmyerius cruciformis, a Petasiger sp. and a trematode species belonging to the family Notocotylidae. No Schistosoma sp. infections were detected in our collected snail specimens. CONCLUSIONS: We report B. truncatus as an intermediate snail host for Carmyerius cruciformis, and the presence of three non-schistosome trematode species that have not been reported in Lake Kariba before. Furthermore, we detect a possible shift in the snail community when compared to the report by Chimbari et al. (2003): this is the first record of Gyraulus sp. in Lake Kariba, and we did not observe the previously reported B. pfeifferi, B. globosus and Radix natalensis. Although this shift in snail communities might have contributed to the absence of Schistosoma spp. detection in this study, further monitoring of final and intermediate hosts across the Kariba basin is essential to prove a decrease of schistosomiasis in the area.

The genome sequence of the Whirlpool Ramshorn snail, Anisus vortex (Linnaeus, 1758).

We present a genome assembly from an individual Anisus vortex (the Whirlpool Ramshorn snail; Mollusca; Gastropoda; Hygrophila; Lymnaeoidea; Planorbidae). The genome sequence is 869.5 megabases in span. Most of the assembly is scaffolded into 18 chromosomal pseudomolecules. The mitochondrial genome has also been assembled and is 13.57 kilobases in length.

Experimental infection with Schistosoma mansoni from Belém, Pará, Brazil: Strains newly isolated vs. laboratory maintained.

BACKGROUND: Schistosomiasis is a neglected tropical disease that occurs in locations with inadequate sanitation conditions. The geographic distribution of Schistosoma mansoni trematode depends directly on the presence of its intermediate host, Biomphalaria mollusks. Studies involving recently isolated and laboratory strains are not common due to the difficulty in cycle maintenance. This study evaluated the susceptibility and infectivity responses in intermediate and definitive hosts with strains of S. mansoni, one isolated and kept in laboratory environment for 34 years (BE) and the other recently collected (BE-I) METHODS: For experimental infection, a total of 400 B. glabrata mollusks were divided in four infection groups. Thirty mice were divided in two groups for infection with the two strains. RESULTS: It was possible to notice differences about S. mansoni infection in both strains. The laboratory strain was more harmful to freshly collected mollusks. Differences in the patterns of infection in mice could be observed. CONCLUSION: Particularities occurred in each group of infection by S. mansoni strains, despite having the same geographic origin. Effects from the parasite-host interaction are visible in terms of infection in definitive and intermediate hosts.

First report of Biomphalaria tenagophila (d'Orbigny, 1835) (Gastropoda/Planorbidae) in Pará State, Amazon region of Brazil.

INTRODUCTION: Mollusks belonging to Biomphalaria genus are intermediate hosts of Schistosoma mansoni. In the Pará State, Northern Region of Brazil, there are reports of B. glabrata, B. straminea, B. schrammi, B. occidentalis, and B. kuhniana occurrence. Here, we report for the first time the presence of B. tenagophila in Belém, capital of Pará state. METHODS: A total of 79 mollusks were collected and examined to search for possible S. mansoni infection. The specific identification was made by morphological and molecular assays. RESULTS: No specimens parasitized by trematode larvae were detected. For the first time the presence of B. tenagophila in Belém, capital of Pará state, was reported. CONCLUSION: The result increases the knowledge about Biomphalaria mollusks occurrence in the Amazon Region and specifically alerts on the possible role of B. tenagophila in schistosomiasis transmission in Belém.

The complete mitochondrial genome of the file ramshorn snail Planorbella pilsbryi (Mollusca: Gastropoda: Hygrophila: Planorbidae).

The file ramshorn snail Planorbella pilsbryi Baker, 1926 (Gastropoda: Hygrophila: Planorbidae) is a widespread herbivorous North American freshwater snail found in diverse habitats, including standing and moving water bodies. Genome skimming by Illumina sequencing allowed the assembly of a complete nuclear rRNA repeat sequence and a complete circular mitogenome of 13,720 bp from P. pilsbryi consisting of 75.3% AT nucleotides, 22 tRNAs, 13 protein-coding genes, 2 rRNAs and a control region in the typical order found in panpulmonate snails. Planorbella pilsbryi COXI features a rare TTG start codon while COXII, CYTB, ND2, ND3, and ND5 exhibit incomplete stop codons completed by the addition of 3' A residues to the mRNA. Phylogenetic reconstruction of mitochondrial protein-coding gene and rRNA sequences places P. pilsbryi as sister taxon to Planorbella duryi (Planorbidae) within family Planorbidae, which is consistent with previous phylogenetic hypotheses.

Miocene geologic dynamics of the Australian Sahul Shelf determined the biogeographic patterns of freshwater planorbid snails (Miratestinae) in the Indo-Australian Archipelago.

The complex geological and climatic processes that have shaped the Indo-Australian Archipelago since the Cenozoic likely also gave rise to its species-rich biota. Strictly freshwater organisms might be particularly suitable for understanding the influence of these abiotic factors on their biogeography in such a insular setting as their distribution may reflect past abiotic events at large and small geographical scales. We here investigate the historical biogeography of the Miratestinae, a subfamily of Planorbidae. These freshwater gastropods are widely distributed in the eastern IAA from Australia, New Guinea, the Moluccas, and Sulawesi to the Philippines. The first comprehensive molecular phylogeny of the Miratestinae was inferred based on two mitochondrial and two nuclear genetic markers using maximum likelihood and Bayesian inference. Four species delimitation methods were applied to identify molecular operational taxonomic units (MOTUs). Divergence times were inferred using an uncorrelated lognormal relaxed-clock model by applying a taxon- and marker-specific substitution rate. Ancestral geographic ranges were estimated based on the dated phylogeny using BioGeoBEARS. The species delimitation revealed a total of 23 MOTUs, 16 of which might represent species new to science. The BioGeoBEARS analyses suggest an Australian origin for the Miratestinae at c. 22 Ma and identified jump dispersal to be the main process of colonization. The first colonization events from Australia to the IAA occurred in the Middle-Late Miocene (12-13 Ma), whereas intra-island diversification took mainly place since the Late Miocene-Pliocene. Colonization and diversification events remarkably coincide with major geologic events that shaped the geography of the region. The increasing availability of landmasses along the Sahul Shelf likely promoted stepping-stone dispersal to New Guinea, Sulawesi and the Philippines as early as the islands emerged. Major geological and climatic events such as the amalgamation of the island Sulawesi, the regional aridification in Australia or the uplift of massive mountain ranges in New Guinea likely played a considerable role for intra-island diversification.

A preliminary study on the distribution of breeding sites of BIOMPHALARIA GLABRATA in the municipality of peri mirim, a low endemicity area for schistosomiasis in northeast Brazil

Information on areas colonized by snails that transmit Schistosoma mansoni is essential for planning schistosomiasis control measures. Therefore, the purpose of this study was to map the natural breeding sites of Biomphalaria spp. in the municipality of Peri Mirim, Maranhão, Brazil. The snails were manually collected and the breeding sites were geo-referenced (seven in the urban area and five in flooded fields), from November 2017 to March 2018. In the laboratory, the snails were examined for the presence of larval stages of S. mansoni identified by morphology and internal anatomy analysis. While no snails were found in the urban area, sixteen B. glabrata were collected in the flooded fields. No S. mansoni or other trematode infections were detected. Wild rodents, most likely Holochilus sciureus that could act as S. mansoni reservoirs, were also found in the flooded fields. These data indicate possible risk areas for further outbreaks of schistosomiasis transmission in the municipality of Peri Mirim.

Size and survival of two freshwater snail species in relation to shedding of cercariae of castrating Echinostoma spp.

Trematode-induced castration of snails is widespread and can lead to other life history changes of snails such as changes in trajectories of size and growth or survival. The changes produced likely depend on whether the parasite or host controls allocation of host resources remaining after partial or complete cessation of host current reproduction by castrating trematodes. Documenting host life history changes, like changes in host size in response to castration, is a first step in assessing whether these changes are beneficial to the parasite (increasing transmission success) or to the host (outliving the infection) or to neither. Herein, we test for differences in size and survival among individuals of two snail species in relation to infection by Echinostoma spp. trematodes. Active shedding of Echinostoma spp. was associated with castration of all Stagnicola elodes snails from a site in Eastern Ontario. Snails actively shedding cercariae were not different in size from non-shedding, egg-laying snails but had a higher mortality than egg-laying snails. Active shedding of Echinostoma spp. cercariae was also associated with castration of nearly all Helisoma trivolvis monitored, from a site in Southwestern Ontario. Actively shedding, non-laying H. trivolvis hosts were smaller on average than non-shedding egg-laying hosts, but both non-laying and egg-laying snails survived equally well. We discuss these results in light of what is known about effects of castration on snail hosts in terms of growth and survival for these and other trematode species and speculate on whether changes in size or survival benefits parasite or host.

Freshwater mollusks from three reservoirs of Piauí, northeastern Brazil / Moluscos límnicos de três reservatórios do Piauí, nordeste do Brasil

Abstract: The pronounced dry season determines the characteristics of the semiarid region of Brazil. Numerous small reservoirs are built to overcome this condition, accumulating multiple uses and causing important changes in the landscape and the local biota. Considering the limited amount of information about the malacofauna of reservoirs, mainly in the Northeast region of the country, this inventory provides a list of mollusks from three important reservoirs located in the city of São Julião, state of Piauí. The collections were carried out monthly between May/2017 and April/2018, sampling a total of 11,149 mollusks, corresponding to the species Melanoides tuberculata (Müller, 1774) (n = 9,724), Biomphalaria straminea (Dunker, 1848) (n = 1,361) and Pomacea lineata (Spix, 1827) (n = 64). The richness of mollusks from the three reservoirs was low, similar to those described in studies conducted in other regions of Northeast Brazil. There was a significant difference in the abundance of M. tuberculata and B. straminea among the studied reservoirs. These environments are not connected and are installed in urban and rural areas, presenting different ecological conditions. The relationship between the human population and the reservoirs, associated with the lack of sanitation, increases the risks of spreading waterborne diseases besides the development of environmental imbalance by the introduction of exotic species.

Resumo: O período de seca pronunciado marca a região semiárida brasileira. Para superação dessa condição inúmeros reservatórios de pequeno porte são construídos, acumulando múltiplos usos e causando alterações importantes na paisagem e na biota local. Considerando as poucas informações sobre a malacofauna de reservatórios, principalmente na região nordeste do país, este trabalho apresenta o inventário das espécies de moluscos provenientes de três importantes reservatórios localizados no município de São Julião, estado do Piauí. As coletas foram realizadas mensalmente entre os meses de maio/2017 e abril/2018. Foi amostrado um total de 11.149 moluscos, correspondentes as espécies Melanoides tuberculata (Müller, 1774) (n=9,724), Biomphalaria straminea (Dunker, 1848) (n=1,361) e Pomacea lineata (Spix, 1827) (n=64). A riqueza de moluscos amostrados nos três reservatórios foi baixa, não diferindo daquelas descritas em estudos realizados em outras regiões do Nordeste do Brasil. Houve diferença significativa na abundância de M. tuberculata e B. straminea entre os reservatórios estudados os quais não são conectados e encontram-se instalados em áreas urbana e rurais, com condições ambientais distintas. A relação entre a população humana da região e os reservatórios, juntamente com a falta de saneamento representam riscos à propagação de doenças transmitidas pela água e desequilíbrio ambiental pela introdução de espécies exóticas.

Musculature of the penial complex: A new criterion in unravelling the phylogeny of Hygrophila (Gastropoda: Pulmonata).

The taxonomy of freshwater pulmonates (Hygrophila) has been in a fluid state warranting the search for new morphological criteria that may show congruence with molecular phylogenetic data. We examined the muscle arrangement in the penial complex (penis and penis sheath) of most major groups of freshwater pulmonates to explore to which extent the copulatory musculature can serve as a source of phylogenetic information for Hygrophila. The penises of Acroloxus lacustris (Acroloxidae), Radix auricularia (Lymnaeidae), and Physella acuta (Physidae) posses inner and outer layers of circular muscles and an intermediate layer of longitudinal muscles. The inner and outer muscle layers in the penis of Biomphalaria glabrata consist of circular muscles, but this species has two intermediate longitudinal layers separated by a lacunar space, which is crossed by radial and transverse fibers. The muscular wall of the penis of Planorbella duryi is composed of transverse and longitudinal fibers, with circular muscles as the outer layer. In Planorbidae, the penial musculature consists of inner and outer layers of longitudinal muscles and an intermediate layer of radial muscles. The penis sheath shows more variation in muscle patterns: its muscular wall has two layers in A. lacustris, P. acuta, and P. duryi, three layers in R. auricularia and Planorbinae and four layers in B. glabrata. To trace the evolution of the penial musculature, we mapped the muscle characters on a molecular phylogeny constructed from the concatenated 18S and mtCOI data set. The most convincing synapomorphies were found for Planorbinae (inner and outer penis layers of longitudinal muscles, three-layered wall of the penis sheath). A larger clade coinciding with Planorbidae is defined by the presence of radial muscles and two longitudinal layers in the penis. The comparative analysis of the penial musculature appears to be a promising tool in unraveling the phylogeny of Hygrophila.

Phylogeography of Bulinus truncatus (Audouin, 1827) (Gastropoda: Planorbidae) in Selected African Countries.

The transmission of some schistosome parasites is dependent on the planorbid snail hosts. Bulinus truncatus is important in urinary schistosomiasis epidemiology in Africa. Hence, there is a need to define the snails' phylogeography. This study assessed the population genetic structure of B. truncatus from Giza and Sharkia (Egypt), Barakat (Sudan) and Madziwa, Shamva District (Zimbabwe) using mitochondrial cytochrome oxidase subunit 1 gene (COI) and internal transcribed spacer 1 (ITS 1) markers. COI was sequenced from 94 B. truncatus samples including 38 (Egypt), 36 (Sudan) and 20 (Zimbabwe). However, only 51 ITS 1 sequences were identified from Egypt (28) and Sudan (23) (because of failure in either amplification or sequencing). The unique COI haplotypes of B. truncatus sequences observed were 6, 11, and 6 for Egypt, Sudan, and Zimbabwe, respectively. Also, 3 and 2 unique ITS 1 haplotypes were observed in sequences from Egypt and Sudan respectively. Mitochondrial DNA sequences from Sudan and Zimbabwe indicated high haplotype diversity with 0.768 and 0.784, respectively, while relatively low haplotype diversity was also observed for sequences from Egypt (0.334). The location of populations from Egypt and Sudan on the B. truncatus clade agrees with the location of both countries geographically. The clustering of the Zimbabwe sequences on different locations on the clade can be attributed to individuals with different genotypes within the population. No significant variation was observed within B. truncatus populations from Egypt and Sudan as indicated by the ITS 1 tree. This study investigated the genetic diversity of B. truncatus from Giza and Sharkia (Egypt), Barakat area (Sudan), and Madziwa (Zimbabwe), which is necessary for snail host surveillance in the study areas and also provided genomic data of this important snail species from the sampled countries.

Phylogeography of freshwater planorbid snails reveals diversification patterns in Eurasian continental islands.

BACKGROUND: Islands have traditionally been the centre of evolutionary biological research, but the dynamics of immigration and differentiation at continental islands have not been well studied. Therefore, we focused on the Japanese archipelago, the continental islands located at the eastern end of the Eurasian continent. While the Japanese archipelago is characterised by high biodiversity and rich freshwater habitats, the origin and formation mechanisms of its freshwater organisms are not clear. In order to clarify the history of the planorbid gastropod fauna, we conducted phylogenetic analysis, divergence time estimation, ancestral state reconstruction, and lineage diversity estimations. RESULTS: Our analyses revealed the formation process of the planorbid fauna in the Japanese archipelago. Most lineages in the Japanese archipelago have closely related lineages on the continent, and the divergence within the Japanese lineages presumably occurred after the late Pliocene. In addition, each lineage is characterised by different phylogeographical patterns, suggesting that immigration routes from the continent to the Japanese archipelago differ among lineages. Furthermore, a regional lineage diversity plot showed that the present diversity in the Japanese archipelago potentially reflects the differentiation of lineages within the islands after the development of the Japanese archipelago. CONCLUSIONS: Although additional taxon sampling and genetic analysis focused on each lineage are needed, our results suggest that immigration from multiple routes just prior to the development of the Japanese archipelago and subsequent diversification within the islands are major causes of the present-day diversity of the Japanese planorbid fauna.

Radula and mandible of Biomphalaria (Planorbidae) of São Paulo, Brazil

The radular apparatus of Biomphalaria occidentalis Paraense, 1981, Biomphalaria oligoza Paraense, 1971, Biomphalaria peregrina (d’Orbigny, 1835), Biomphalaria schrammi (Crosse, 1864) and Biomphalaria tenagophila (d’Orbigny, 1835) from the Metropolitan Region of São Paulo, Brazil were analyzed from Scanning Electron Microscope (SEM) micrographs. The data support the usage of characters of central, lateral and intermediate teeth in the species identification of Biomphalaria. The use of SEM provided additional quantitative and structural details to radula descriptions, which are extremely important for elucidating the taxonomy of Biomphalaria species...

Small but diverse: larval trematode communities in the small freshwater planorbids Gyraulus albus and Segmentina nitida (Gastropoda: Pulmonata) from the Ruhr River, Germany.

In contrast to the well-studied trematode fauna of lymnaeid snails, only little is known about the role of small planorbid snails as first intermediate hosts for trematodes in temperate freshwater systems. This study aims at closing this gap by assessing the diversity and composition of larval trematode communities in Gyraulus albus and Segmentina nitida in a Central European reservoir system, and by providing an updated comprehensive review of the published trematode records of these snail hosts. A total of 3691 planorbid snails (3270 G. albus; 421 S. nitida) was collected in three consecutive years from four reservoirs of the River Ruhr catchment area in Germany. Gyraulus albus showed a higher overall trematode prevalence (11.7%) and more diverse trematode fauna (12 species) compared to S. nitida, which harboured three species and showed a lower trematode prevalence (1.7%). Altogether, 13 trematode species belonging to four families were identified in both hosts. Seven trematode species encountered in this study represent novel records for these hosts, and/or constitute first records of these larval stages from Germany. Trematode component communities in G. albus were stable across seasons and years, indicating excellent conditions for trematodes in this snail host and the continuous presence of the final hosts of the most dominant trematode species. Overall, this study reveals the importance of small planorbid snails, in particular G. albus, as first intermediate hosts for a species-rich trematode fauna in European freshwater systems, and highlights the parasites' contribution to the ecosystem's biodiversity.

The mitochondrial genome of the planorbid snail Planorbella duryi.

The complete mitochondrial genome of a freshwater planorbid snail, Planorbella duryi (Mollusca, Gastropoda) was recovered from de novo assembly of genomic sequences generated with the Illumina NextSeq500 platform. The P. duryi mitogenome (14,217 base pairs) is AT rich (72.69%) and comprises 13 protein-coding genes, two ribosomal subunit genes, and 22 transfer RNAs. The gene order is identical to that of Biomphalaria glabrata and other snail species in the family Planorbidae. This is the first full characterization of a mitochondrial genome of the genus Planorbella.

Putative new genera and species of avian schistosomes potentially involved in human cercarial dermatitis in the Americas, Europe and Africa.

New larval avian schistosomes found in planorbid snails from Brazil and USA were used for morphological and molecular studies. Eggs with a distinctive long polar filament were found in ducks infected experimentally with Brazilian cercariae. Similar eggs were reported previously in wild or experimentally infected anatids from Brazil, South Africa, and the Czech Republic. Molecular phylogenetic analyses showed that the North American and European schistosomes are sister taxa, which are both sister to the Brazilian species. However, these clades do not group with any named genus. Molecular data plus egg morphology suggest that these are new putative genera and species of avian schistosomes that can cause human cercarial dermatitis in the Americas, Africa and Europe.

Occurrence and effect of trematode metacercariae in two endangered killifishes from Greece.

We report digeneans (Diplostomidae, Crassiphialinae) in the endangered freshwater fishes Valencia letourneuxi and Valencia robertae, endemics of Western Greece. Digenean metacercariae occurred in two forms in the abdominal cavity, excysted and encysted, the latter attached to the gonads, liver and alimentary tract. Parasites were, using morphological and molecular techniques, identified as two representatives of Crassiphialinae, specifically part of the Posthodiplostomum-Ornithodiplostomum clade. The spatial, seasonal, and age class variation in parasite prevalence was examined. Autumn parasite prevalence varied between the six populations sampled (18.2 to 100%). Seasonal prevalence at the two sites sampled quadannually peaked in autumn and reached its lowest value in spring; prevalence increased with size to 100% in young adult fish. We did not find a correlation between prevalence and host sex. Overall parasites' weight averaged 0.64% of the host's, while parasite weight increased with host weight. A comparison of relative condition and hepatosomatic and gonadosomatic indices of infected and metacercariae-free specimens showed that infection did not have a significant effect on host body condition and reproduction. Regarding the parasite's life cycle, planorbid gastropods are proposed as potential first intermediate hosts in view of the host's diet and occurrence data of molluscs in the ecosystem. This is the first record of a diplostomid digenean in valenciid fishes and of representatives of the Posthodiplostomum-Ornithodiplostomum clade in a native Greek freshwater fish. Our findings are discussed in conjunction to fish conservation interventions, since parasites may contribute to the decline of endangered species.

The South-American distribution and southernmost record of Biomphalaria peregrina-a potential intermediate host of schistosomiasis.

Schistosomiasis remains a major parasitic disease, endemic in large parts of South America. Five neotropical species of Biomphalaria have been found to act as intermediate hosts of Schistosoma mansoni in natural populations, while others have been shown to be susceptible in experimental infections, although not found infected in the field. Among these potential intermediate hosts, Biomphalaria peregrina represents the most widespread species in South America, with confirmed occurrence records from Venezuela to northern Patagonia. In this study, we report the southernmost record for the species at the Pinturas River, in southern Patagonia, which finding implies a southward reassessment of the limit for the known species of this genus. The identities of the individuals from this population were confirmed through morphological examination, and by means of two mitochondrial genes, cytochrome oxidase subunit I (COI) and 16S-rRNA. With both markers, phylogenetic analyses were conducted in order to compare the genetic background of individuals from the Pinturas River with previously genetically characterized strains of B. peregrina from various South-American locations. In addition, we produced a potential distribution model of B. peregrina in South America and identified the environmental variables that best predict that distribution. The model was estimated through a maximum entropy algorithm and run with occurrence points obtained from several sources, including the scientific literature and international databases, along with climatic and hydrographic variables. Different phylogenetic analyses with either the COI or 16S-rRNA sequences did not conflict, but rather gave very similar topological organizations. Two major groups were identified, with sequences from the Pinturas River grouping together with haplotypes from subtropical and temperate regions. The model developed had a satisfactory performance for the study area. We observed that the areas with higher habitat suitability were found to be mainly linked to subtropical and temperate regions of South America between 15° and 45° south latitude, with different moderate- and low-suitability areas outside this range. We also identified the coldest temperatures as the main predictors of the potential distribution of this snail. Susceptibility surveys would be required to evaluate if southern populations of B. peregrina still retain their potential as intermediate hosts of S. mansoni.

Deep, multi-stage transcriptome of the schistosomiasis vector Biomphalaria glabrata provides platform for understanding molluscan disease-related pathways.

BACKGROUND: The gastropod mollusc Biomphalaria glabrata is well known as a vector for the tropical disease schistosomiasis, which affects nearly 200 million people worldwide. Despite intensive study, our understanding of the genetic basis of B. glabrata development, growth and disease resistance is constrained by limited genetic resources, constraints for which next-generation sequencing methods provide a ready solution. METHODS: Illumina sequencing and de novo assembly using the Trinity program was used to generate a high-quality transcriptomic dataset spanning the entirety of in ovo development in schistosomiasis-free B. glabrata. This was subjected to automated (KEGG, BLAST2GO) and manual annotation efforts, allowing insight into the gene complements of this species in a number of contexts. RESULTS: Excellent dataset recovery was observed, with 133,084 contigs produced of mean size 2219.48 bp. 80,952 (60.8 %) returned a BLASTx hit with an E value of less than 10-3, and 74,492 (55.97 %) were either mapped or assigned a GO identity using the BLAST2GO program. The CEGMA set of core eukaryotic genes was found to be 99.6 % present, indicating exceptional transcriptome completeness. We were able to identify a wealth of disease-pathway related genes within our dataset, including the Wnt, apoptosis and Notch pathways. This provides an invaluable reference point for further work into molluscan development and evolution, for studying the impact of schistosomiasis in this species, and perhaps providing targets for the treatment of this widespread disease. CONCLUSIONS: Here we present a deep transcriptome of an embryonic sample of schistosomiasis-free B. glabrata, presenting a comprehensive dataset for comparison to disease-affected specimens and from which conclusions can be drawn about the genetics of this widespread medical model. Furthermore, the dataset provided by this sequencing provides a useful reference point for comparison to other mollusc species, which can be used to better understand the evolution of this commercially, ecologically and medically important phylum.