Genetic diversity and population structure of Physella acuta (Gastropoda: Physidae) in Thailand using mitochondrial gene markers: COI and 16S rDNA.

Physella acuta is a freshwater snail native to North America. Understanding the phylogeography and genetic structure of P. acuta will help elucidate its evolution. In this study, we used mitochondrial (COI and 16S rDNA) and nuclear (ITS1) markers to identify the species and examine its genetic diversity, population structure, and demographic history of P. acuta in Thailand. Phylogenetic and network analyses of P. acuta in Thailand pertained to clade A, which exhibits a global distribution. Analysis of the genetic structure of the population revealed that the majority of pairwise comparisons showed no genetic dissimilarity. An isolation-by-distance test indicates no significant correlation between genetic and geographical distances among P. acuta populations, suggesting that gene flow is not restricted by distance. Demographic history and haplotype network analyses suggest a population expansion of P. acuta, as evidenced by the star-like structure detected in the median-joining network. Based on these results, we concluded that P. acuta in Thailand showed gene flow and recent population expansion. Our findings provide fundamental insights into the genetic variation of P. acuta in Thailand.

Assessing the role of taxonomic impediment in the knowledge on geographic distribution of Physidae (Gastropoda: Hygrophila) in South America, using an intensive-data approach.

There is a marked disparity in the state of knowledge of Holartic x Neotropical species of the freshwater snail family Physidae; the incipiency of data on Neotropical physids reflecting the lower number of dedicated specialists. The gaps in the knowledge on Neotropical physids have led to historical uncertainty about species validity. Revisiting the species is essential to reduce taxonomic impediment and delineating their probable distribution is the first step to attain this purpose. We aimed at critically analyze occurrence records of South American physids, compiled through an intensive search in the literature, biodiversity and molecular databases. We present a provisional characterization of the distribution of this family in South America, considering the probable versus the poorly documented distribution of the species. The critical underrepresentation of South American physids in collections, molecular databases and literature reinforces the role of taxonomic impediment in delaying the advance of the knowledge on species diversity. Malacological collections represented the main source of records, evidencing the relevance of unpublished data associated to specimens to assess distributional information on neglected groups. As most of the species are represented by shells, the reassessment of species identity and distribution must be done, using molecular and anatomical criteria for species delimitation.

Identification of terrestrial gastropods families found in district Swat, Pakistan / Identificação de famílias de gastropódeos terrestres encontradas no distrito Swat, Paquistão

Very little is known about the malacofauna in district Swat of Khyber Pakhtunkhwa province, situated in the Northern part of Pakistan. The Swat valley possesses a variety of suitable environmental conditions, providing different types of habitats for both flora and fauna. Taken into consideration these points, the present study was conducted to explore the terrestrial snails diversity in agricultural and non-agricultural fields in the lower Swat valley. A total of 417 shells were collected from three sites: Kokarai, Kanju Township, and Deolai. Based on conchological features, three families: Bradybaenidae, Hygromiidae, and Zonitidae were identified using different terrestrial snail keys. In Kokarai and Deolai, shells of members of all the above-mentioned families were found from agricultural fields. In Kanju Township, shells of members of the family Bradybaenidae and Zonitidae were found. From these findings, it was concluded that district Swat possesses a significant malacofauna which was unexplored before and aided more to the existing literature on malacofauna. It also supports the idea that the agroecosystem is rich in species diversity. Further in-depth and extensive study is recommended to be carried out for a detailed taxonomy of the described terrestrial gastropod families.

Muito pouco se sabe sobre a malacofauna no distrito de Swat, na província de Khyber Pakhtunkhwa, situada na parte norte do Paquistão. O vale do Swat possui uma variedade de condições ambientais adequadas, proporcionando diferentes tipos de habitats para a flora e a fauna. Levando em consideração esses pontos, o presente estudo foi conduzido para explorar a diversidade dos caracóis terrestres em campos agrícolas e não agrícolas no vale inferior de Swat. Um total de 417 conchas foram coletadas em três locais: Kokarai, Kanju Township e Deolai. Com base em características concológicas, três famílias: Bradybaenidae, Hygromiidae e Zonitidae foram identificadas usando diferentes chaves de caramujos terrestres. Em Kokarai e Deolai, conchas de membros de todas as famílias mencionadas acima foram encontradas em campos agrícolas. No município de Kanju, foram encontradas conchas de membros da família Bradybaenidae e Zonitidae. A partir dessas constatações, concluiu-se que o distrito de Swat possui uma malacofauna significativa que antes era inexplorada e ajudava mais a literatura existente sobre a malacofauna. Também apoia a ideia de que o agroecossistema é rico em diversidade de espécies. Recomenda-se a realização de um estudo mais aprofundado e extenso para uma taxonomia detalhada das famílias de gastrópodes terrestres descritas.

Distribución geográfica de la familia de caracoles terrestres Spiraxidae (Gastropoda: Pulmonata) en México / Geographic distribution of the land snail family Spiraxidae (Gastropoda: Pulmonata) in Mexico

Resumen Introducción: Spiraxidae es la familia de gasterópodos terrestres más diversa de México con una distribución principalmente neotropical. Sin embargo, la influencia de las condiciones ambientales de esta región en su distribución aún es poco conocida. Objetivo: Determinar las afinidades zoogeográficas y la influencia de factores ambientales en la distribución de Spiraxidae en México. Métodos: Compilamos registros de distribución de museos y usamos modelos para establecer áreas con composición de especies, concentración de especies, afinidades y relaciones con variables ambientales similares. Resultados: Encontramos 231 especies y subespecies, 96 en el estado de Veracruz y 74 en la Sierra Madre Oriental. La principal afinidad zoogeográfica fue neotropical. El bosque mesófilo de montaña tuvo la mayor cantidad de especies y subespecies (93). Tres de los grupos zonales tienen una composición particular de especies y el 67 % de las especies son especialistas de hábitat. La presencia de especies sigue un gradiente ambiental, con la cubierta de árboles de hoja perenne de hoja ancha y la precipitación media anual como variables determinantes. Las especies del norte tuvieron mayor tolerancia a la aridez y menor cobertura de hojas. Conclusiones: Spiraxidae tiene su mayor diversidad en los estados del Sur de México; sin embargo, la zona nororiental tiene más registros de especies. Estos caracoles se encuentran principalmente en bosques mesófilos de montaña y bosques tropicales siempre verdes. Las especies del norte de México tuvieron una mayor tolerancia a los sitios con poca lluvia y menos cobertura de árboles de hoja perenne de hoja ancha que las especies del sur.

Abstract Introduction: Spiraxidae is the most diverse family of terrestrial gastropods in Mexico with a mainly neotropical distribution. However, the influence of environmental conditions in this region on its distribution is still poorly known. Objective: To determine zoogeographic affinities and the influence of environmental factors on the distribution of Spiraxidae in Mexico. Methods: We compiled museum distribution records and used models to establish areas with similar species composition, species concentration, affinities and relationship with environmental variables. Results: We found 231 species and subspecies, 96 in Veracruz state and 74 in Sierra Madre Oriental. The main zoogeographic affinity was neotropical. Mountain mesophyll forest had the most species and subspecies (93). Three of zone groups have a particular species composition and 67 % of the species are habitat specialists. Species presence follows an environmental gradient, with broadleaf evergreen tree cover and average annual precipitation as determining variables. Northern species had greater tolerance to aridity and reduced leaf cover. Conclusions: Spiraxidae has its greatest diversity in the Southern states of Mexico; however, the Northeastern zone has more species records. These snails mostly occur in mesophyll mountain forest and tropical evergreen forest. Species from Northern Mexico had greater tolerance to sites with low rainfall and less broadleaf evergreen tree cover than Southern species.

Reproducción y limitaciones de pesca recomendadas para el caracol marino Hexaplex princeps (Neogastropoda: Muricidae) en Guerrero, México / Reproduction and recommended fishing limitations for the marine snail Hexaplex princeps (Neogastropoda: Muricidae) in Guerrero, Mexico

Abstract Introduction: The snail Hexaplex princeps is the most important gastropod in the fishery of Guerrero, but capture is not legally regulated for lack of enough studies about its reproduction, growth and other fishing aspects. Objective: To assess fishery data for H. princeps in Guerrero. Methods: From October 2016 to December 2017, we analyzed histological samples of the gonads from 6 to 13 individuals per month. We also collected data on pre-reproductive organisms captured in Costa Chica (2013 and 2014), Costa Grande (2017) and Acapulco (2019 and 2020). Results: Both sexes have five stages of gonadal development; most reproductive activity was from December to July, related to sea surface temperatures of 26 to 28 °C. The LC M in females was 94.1 mm (males: 86.1 mm). There was a high percentage of juvenile organisms in the catches (88.6 %). Conclusions: To let these snails reproduce at least once, we recommend a fishing ban from December to July, and that the minimum size of commercial capture be set at 94.1 mm.

Resumen Introducción: El caracol Hexaplex princeps es el gasterópodo más importante en la pesquería de Guerrero, pero su captura no está regulada legalmente por falta de suficientes estudios sobre su reproducción, crecimiento y otros aspectos de la pesca. Objetivo: Evaluar los datos de pesca de H. princeps en Guerrero. Métodos: De octubre 2016 a diciembre 2017, analizamos muestras histológicas de las gónadas de 6 a 13 individuos por mes. También recolectamos datos sobre organismos pre-reproductivos capturados en Costa Chica (2013 y 2014), Costa Grande (2017) y Acapulco (2019 y 2020). Resultados: Ambos sexos tienen cinco etapas de desarrollo gonadal; la mayor actividad reproductiva fue de diciembre a julio, relacionada con las temperaturas superficiales del mar, que se encuentran entre 26 y 28 °C. El LC M en hembras fue de 94.1 mm (machos: 86.1 mm). Hubo un alto porcentaje de organismos juveniles en las capturas (88.6 %). Conclusiones: Para que estos caracoles se reproduzcan al menos una vez, se recomienda una veda de diciembre a julio y que la talla mínima de captura comercial se fije en 94.1 mm.

Liguus Landscapes: Amateur Liggers, Professional Malacology, and the Social Lives of Snail Sciences.

Malacologists took notice of tree snails in the genus Liguus during the last decades of the nineteenth century. Since then, Liguus have undergone repeated shifts in identity as members of species, states, shell collections, backyard gardens, and engineered wildernesses. To understand what Liguus are, this paper examines snail enthusiasts, collectors, researchers, and conservationists-collectively self-identified as Liggers-in their varied landscapes. I argue that Liguus, both in the scientific imaginary and in the material landscape, mediated knowledge-making processes that circulated among amateur and professional malacologists across the United States and Cuba during the twentieth century. Beginning with an examination of early Liggers' work in Florida and Cuba, this paper demonstrates how notions of taxonomy and biogeography informed later efforts to understand Liguus hybridization and conservation. A heterogeneous community of Liggers has had varied and at times contradictory commitments informed by shifting physical, social, and scientific landscapes. Genealogizing those commitments illuminates the factors underpinning a decision to undertake the until now little-chronicled large-scale and sustained transplantation of every living Floridian form of Liguus fasciatus into Everglades National Park. The social history of Liggers and Liguus fundamentally blurs distinctions between professional scientists and amateur naturalists. The experiences of a diverse cast of Liggers and their Liguus snails historicize the complex character of human-animal relations and speak to the increasing endangerment of many similarly range-restricted invertebrates.

Persistence of Schistosoma haematobium transmission among school children and its implication for the control of urogenital schistosomiasis in Lindi, Tanzania.

BACKGROUND: Despite twelve rounds of school-based preventive chemotherapy for schistosomiasis in endemic areas of Tanzania such as Mtama district, Lindi: the burden of Schistosoma haematobium infection has remained highly conceivable due to re-infections. The factors associated with continuity of S.haematobium transmission in Mtama district, Lindi have not been fully established. This study investigated the burden and factors contributing to the ongoing transmission of S.haematobium infection in the endemic district of Mtama, Lindi. METHODS: A quantitative cross-sectional survey was carried out among 649 school-age children in the Mtama district to determine the burden and factors associated with continuity of S.haematobium infection transmission. A single urine specimen was obtained from each pupil and tested for macro- and microhaematuria, presence of S.haematobium ova, as well intensity of infection; this was complemented with a survey of Bulinus spp snail intermediate hosts and their infectivity. A structured questionnaire was employed to gather information on individual and environmental risk factors for S.haematobium transmission. Summary statistics were computed for individual variables; while a univariate and multivariate logistic regression analysis was performed to assess the association between risk factors with S.haematobium infection. RESULTS: Prevalence of S.haematobium infection by macro- and microhaematuria was 13.1% and 46.2% respectively. The prevalence of S.haematobium ova was 52.7%; intensity of infection was light in 53.1%, and heavy in 46.9%. Snail intermediate hosts were Bulinus globosus and B.nasutus, whose infectivity was 2.2% and 1.3%, respectively. Among the assessed risk factors, long residency (10-13 years) in the area was a significant risk factor for the continuity of S.haematobium transmission (AOR: 21.79, 95% CI: 1.37-346.4). CONCLUSIONS: The observed 52.7% prevalence of S.haematobium infection represents unacceptably high prevalence after 12 rounds of preventive chemotherapy. Therefore, an urgent need for the implementation of integrated multiple control interventions in the Mtama district; is considered to be imperative.

Comparing the genetic diversity and population structure of sister marine snails having contrasting habitat specificity.

BACKGROUND: To grasp the processes of spatial genetic structuring in open and connectable marine environments is the principal study goal in molecular biological studies. Comparative seascape genetics using multiple species are a powerful approach to understand the physical geographic and oceanographic effects on genetic variation. Besides, species-specific ecological traits such as dispersal abilities and habitat specificity are important factors for spatial genetic structuring. METHODS AND RESULTS: We focused on the sister marine snail species Tegula kusairo and T. xanthostigma around the Japanese mainland, which have contrasting habitat specificities for wave strength. Tegula kusairo only inhabits sheltered coastal environments, while T. xanthostigma is found mainly on wave-exposed rocky shores facing the open sea. We estimated their genetic diversity indices and levels of population differentiation based on mtDNA. We found that the genetic diversity of T. kusairo was lower than that of T. xanthostigma, while their level of population genetic differentiation was higher than that of T. xanthostigma. Namely, the species specific to weak wave environments had a higher level of population genetic differentiation than the species specific to strong wave action. CONCLUSION: Ecological traits linked not only to dispersal abilities but also to habitat specificity can influence genetic variation in a pair of closely related sister species distributed in the same seascape.

Divergence together with microbes: A comparative study of the associated microbiomes in the closely related Littorina species.

Any multicellular organism during its life is involved in relatively stable interactions with microorganisms. The organism and its microbiome make up a holobiont, possessing a unique set of characteristics and evolving as a whole system. This study aimed to evaluate the degree of the conservativeness of microbiomes associated with intertidal gastropods. We studied the composition and the geographic and phylogenetic variability of the gut and body surface microbiomes of five closely related sympatric Littorina (Neritrema) spp. and a more distant species, L. littorea, from the sister subgenus Littorina (Littorina). Although snail-associated microbiomes included many lineages (207-603), they were dominated by a small number of OTUs of the genera Psychromonas, Vibrio, and Psychrilyobacter. The geographic variability was greater than the interspecific differences at the same collection site. While the microbiomes of the six Littorina spp. did not differ at the high taxonomic level, the OTU composition differed between groups of cryptic species and subgenera. A few species-specific OTUs were detected within the collection sites; notably, such OTUs never dominated microbiomes. We conclude that the composition of the high-rank taxa of the associated microbiome ("scaffolding enterotype") is more evolutionarily conserved than the composition of the low-rank individual OTUs, which may be site- and / or species-specific.

Molecular species delimitation refines the taxonomy of native and nonnative physinine snails in North America.

Being able to associate an organism with a scientific name is fundamental to our understanding of its conservation status, ecology, and evolutionary history. Gastropods in the subfamily Physinae have been especially troublesome to identify because morphological variation can be unrelated to interspecific differences and there have been widespread introductions of an unknown number of species, which has led to a speculative taxonomy. To resolve uncertainty about species diversity in North America, we targeted an array of single-locus species delimitation methods at publically available specimens and new specimens collected from the Snake River basin, USA to generate species hypotheses, corroborated using nuclear analyses of the newly collected specimens. A total-evidence approach delineated 18 candidate species, revealing cryptic diversity within recognized taxa and a lack of support for other named taxa. Hypotheses regarding certain local endemics were confirmed, as were widespread introductions, including of an undescribed taxon likely belonging to a separate genus in southeastern Idaho for which the closest relatives are in southeast Asia. Overall, single-locus species delimitation was an effective first step toward understanding the diversity and distribution of species in Physinae and to guiding future investigation sampling and analyses of species hypotheses.

Origin and significance of two pairs of head tentacles in the radiation of euthyneuran sea slugs and land snails.

The gastropod infraclass Euthyneura comprises at least 30,000 species of snails and slugs, including nudibranch sea slugs, sea hares and garden snails, that flourish in various environments on earth. A unique morphological feature of Euthyneura is the presence of two pairs of sensory head tentacles with different shapes and functions: the anterior labial tentacles and the posterior rhinophores or eyestalks. Here we combine molecular phylogenetic and microanatomical evidence that suggests the two pairs of head tentacles have originated by splitting of the original single tentacle pair (with two parallel nerve cords in each tentacle) as seen in many other gastropods. Minute deep-sea snails of Tjaernoeia and Parvaplustrum, which in our phylogeny belonged to the euthyneurans' sister group (new infraclass Mesoneura), have tentacles that are split along much of their lengths but associated nerves and epidermal sense organs are not as specialized as in Euthyneura. We suggest that further elaboration of cephalic sense organs in Euthyneura closely coincided with their ecological radiation and drastic modification of body plans. The monotypic family Parvaplustridae nov., superfamily Tjaernoeioidea nov. (Tjaernoeiidae + Parvaplustridae), and new major clade Tetratentaculata nov. (Mesoneura nov. + Euthyneura) are also proposed based on their phylogenetic relationships and shared morphological traits.

MALDI-TOF mass spectrometry for the identification of freshwater snails from Senegal, including intermediate hosts of schistosomes.

Freshwater snails of the genera Biomphalaria, Bulinus, and Oncomelania are intermediate hosts of schistosomes that cause human schistosomiasis, one of the most significant infectious neglected diseases in the world. Identification of freshwater snails is usually based on morphology and potentially DNA-based methods, but these have many drawbacks that hamper their use. MALDI-TOF MS has revolutionised clinical microbiology and has emerged in the medical entomology field. This study aims to evaluate MALDI-TOF MS profiling for the identification of both frozen and ethanol-stored snail species using protein extracts from different body parts. A total of 530 field specimens belonging to nine species (Biomphalaria pfeifferi, Bulinus forskalii, Bulinus senegalensis, Bulinus truncatus, Bulinus globosus, Bellamya unicolor, Cleopatra bulimoides, Lymnaea natalensis, Melanoides tuberculata) and 89 laboratory-reared specimens, including three species (Bi. pfeifferi, Bu. forskalii, Bu. truncatus) were used for this study. For frozen snails, the feet of 127 field and 74 laboratory-reared specimens were used to validate the optimised MALDI-TOF MS protocol. The spectral analysis yielded intra-species reproducibility and inter-species specificity which resulted in the correct identification of all the specimens in blind queries, with log-score values greater than 1.7. In a second step, we demonstrated that MALDI-TOF MS could also be used to identify ethanol-stored snails using proteins extracted from the foot using a specific database including a large number of ethanol preserved specimens. This study shows for the first time that MALDI-TOF MS is a reliable tool for the rapid identification of frozen and ethanol-stored freshwater snails without any malacological expertise.

Low genetic diversity and the phylogeny of Achatina fulica, an intermediate host of Angiostrongylus cantonensis in Thailand, inferred from 16S mitochondrial sequences.

The giant African land snail, Achatina fulica, is a pulmonate land snail of the Achatinidae family. The snail is native of East Africa and has spread to tropical and subtropical areas in different parts of the world. Achatina fulica is an important agricultural pests and the intermediate host for Angiostrongylus spp., especially Angiostrongylus cantonensis, which causes eosinophilic meningoencephalitis in humans. However, information on the genetic diversity and phylogenetic relationships among A. fulica populations in Thailand is limited. The goal of this study was to evaluate the genetic diversity and analyze haplotype network relationships between A. fulica populations in Thailand and other areas of the world. We collected A. fulica from 22 provinces in 6 regions of Thailand and analyzed a phylogenetic tree and haplotype network based on a fragment of 239 base pairs of the 16S rRNA sequence. The maximum likelihood, neighbor joining, and Bayesian inference trees for the 133 A. fulica 16S rRNA sequences revealed only one group, closely related only to A. fulica haplotype C, in all regions of Thailand. The haplotype network analysis showed that haplotype C is associated with F, P, E, Q, H and D. The genetic distance between the haplotype and A. fulica isolates in Thailand varies from 0 and 0.024, with a total of 21 variable sites. Haplotype C is predominant in Thailand and associated with other haplotypes from several areas. As inferred from 16S rDNA sequences, this study demonstrated low genetic diversity in A. fulica in Thailand. Low genetic diversity in the population can increase susceptibility to A. cantonensis infection, which may be associated with the current distribution of A. cantonensis.

Deep genetic structure at a small spatial scale in the endangered land snail Xerocrassa montserratensis.

Species with small geographic ranges do not tend to have a high genetic structure, but some land snail species seem to be an exception. Xerocrassa montserratensis, an endangered land snail endemic to Catalonia (northeastern Iberian Peninsula), is an excellent model to study the processes affecting the phylogeography of specialized species of conservation concern. This species is restricted to xerophilous stony slopes and occurs within a small and fragmented area of ca. 500 km2. We sequenced the COI barcode region of 152 individuals from eight sites covering the entire range of the species. We found four genetic groups mostly coincident with their geographic distribution: a central ancestral group containing shared haplotypes among five localities and three groups restricted to a single locality each. Two of these derived groups were geographically and genetically isolated, while the third and most differentiated group was not geographically isolated. Geomorphologic and paleoclimatic processes during the Pleistocene can explain the divergence found between populations of this low dispersal species with historical fragmentation and secondary contacts. Nonetheless, recent passive large dispersal through streams was also detected in the central group. Overall, our study uncovered four evolutionary units, partially matching morphologically described subspecies, which should be considered in future conservation actions.

Land snail biogeography and endemism in south-eastern Africa: Implications for the Maputaland-Pondoland-Albany biodiversity hotspot.

Invertebrates in general have long been underrepresented in studies on biodiversity, biogeography and conservation. Boundaries of biodiversity hotspots are often delimited intuitively based on floristic endemism and have seldom been empirically tested using actual species distributions, and especially invertebrates. Here we analyse the zoogeography of terrestrial malacofauna from south-eastern Africa (SEA), proposing the first mollusc-based numerical regionalisation for the area. We also discuss patterns and centres of land snail endemism, thence assessing the importance and the delimitation of the Maputaland-Pondoland-Albany (MPA) biodiversity hotspot for their conservation. An incidence matrix compiled for relatively well-collected lineages of land snails and slugs (73 taxa in twelve genera) in 40 a priori operational geographic units was subjected to (a) phenetic agglomerative hierarchical clustering using unweighted pair-group method with arithmetic means (UPGMA), (b) parsimony analysis of endemicity (PAE) and biotic element analysis (BEA). Fulfilling the primary objective of our study, the UPGMA dendrogram provided a hierarchical regionalisation and identified five centres of molluscan endemism for SEA, while the PAE confirmed six areas of endemism, also supported by the BEA. The regionalisation recovers a zoogeographic province similar to the MPA hotspot, but with a conspicuous westward extension into Knysna (towards the Cape). The MPA province, centres and areas of endemism, biotic elements as well as the spatial patterns of species richness and endemism, support the MPA hotspot, but suggest further extensions resulting in a greater MPA region of land snail endemism (also with a northward extension into sky islands-Soutpansberg and Wolkberg), similar to that noted for vertebrates. The greater MPA region provides a more robustly defined region of conservation concern, with centres of endemism serving as local conservation priorities.

One, two or three? Integrative species delimitation of short-range endemic Hemicycla species (Gastropoda: Helicidae) from the Canary Islands based on morphology, barcoding, AFLP and ddRADseq data.

Hemicycla mascaensis and H. diegoi are short-range endemics that occur allopatrically in small areas in the Teno Mountains in the western part of Tenerife (Canary Islands). Both taxa have been recognised as distinct species based on differences in shell morphology and genital anatomy. Preliminary molecular analyses using mitochondrial markers suggested a potential paraphyly of H. diegoi with regard to H. mascaensis. We here use multilocus AFLP data and ddRADseq data as well as distribution data, data on shell morphology and genital anatomy to assess the status of these taxa using phylogenetic analyses, species tree reconstruction and molecular species delimitation based on the multispecies coalescent as implemented in BFD* and BPP in an integrative approach. Our analyses show that, based on the analysis of multilocus data, the two taxa are reciprocally monophyletic. Species delimitation methods, however, tend to recognise all investigated populations as distinct species, albeit neither lending unambiguous support to any of the species hypotheses. The comparison of the anatomy of distal genital organs further suggests differentiation within H. mascaensis. This highlights the need for a balanced weighting of arguments from different lines of evidence to determine species status and calls for cautious interpretations of the results of molecular species delimitation analyses, especially in organisms with low active dispersal capacities and expected distinct population structuring such as land snails. Taking all available evidence into account, we favour to recognise H. mascaensis and H. diegoi as distinct species, acknowledging, though, that the recognition of both taxa as subspecies (with possibly a third yet undescribed) would also be an option as morphological differentiation is within the limits of other land snail species that are traditionally subdivided into subspecies.

Insights on foodborne zoonotic trematodes in freshwater snails in North and Central Vietnam.

Foodborne zoonotic trematode (FZT) infections are common neglected tropical diseases in Southeast Asia. Their complicated life cycles involve freshwater snails as intermediate hosts. A cross-sectional study was conducted in Yen Bai and Thanh Hoa provinces in North and Central Vietnam, to investigate the diversity of cercariae of potential FZT and to construct the phylogenetic relationship of trematode cercariae based on the Internal Transcribed Spacer 2 (ITS2) region. Among 17 snail species collected from various habitats, 13 were infected by 10 cercarial groups among which parapleurolophocercous, pleurolophocercous, and echinostome cercariae were of zoonotic importance. The monophyletic tree separated cercarial sequences into different groups following the description of the cercariae families in which Haplorchidae, Opisthorchiidae, Echinochasmidae, and Echinostomatidae are important families of FZT. The overall prevalence was different among snail species and habitats and showed a seasonal trend. Parapleurolophocercous and echinostome cercariae emerged as the most common cercariae in snails in Yen Bai, while monostome, echinostome, and megalura cercariae were most common in Thanh Hoa. Using a molecular approach, we identified Parafossarulus striatulus as the first intermediate snail host of Clonorchis sinensis in Thac Ba Lake. Melanoides tuberculata and Bithynia fuchsiana were we identified preferred intermediate snail hosts of a diverse range of trematode species including intestinal flukes (i.e., Haplorchis pumilio and Echinochasmus japonicus) in Yen Bai and Thanh Hoa, respectively.

Evolution of reproductive strategies in the species-rich land snail subfamily Phaedusinae (Stylommatophora: Clausiliidae).

Most of the present knowledge on animal reproductive mode evolution, and possible factors driving transitions between oviparity and viviparity is based on studies on vertebrates. The species rich door snail (Clausiliidae) subfamily Phaedusinae represents a suitable and unique model for further examining parity evolution, as three different strategies, oviparity, viviparity, and the intermediate mode of embryo-retention, occur in this group. The present study reconstructs the evolution of reproductive strategies in Phaedusinae based on time-calibrated molecular phylogenetics, reproductive mode examinations and ancestral state reconstruction. Our phylogenetic analysis employing multiple mitochondrial and nuclear markers identified a well-supported clade (including the tribes Phaedusini and Serrulinini) that contains species exhibiting various reproductive strategies. This clade evolved from an oviparous most recent common ancestor according to our reconstruction. All non-oviparous taxa are confined to a highly supported subclade, coinciding with the tribe Phaedusini. Both oviparity and viviparity occur frequently in different lineages of this subclade that are not closely related. During Phaedusini diversification, multiple transitions in reproductive strategy must have taken place, which could have been promoted by a high fitness of embryo-retaining species. The evolutionary success of this group might result from the maintenance of various strategies.

Cercarial trematodes in freshwater snails from Bangkok, Thailand: prevalence, morphological and molecular studies and human parasite perspective.

We investigated the prevalence, morphological characters and molecular classifications of trematode cercariae in freshwater snails randomly collected from 59 sampling localities in Bangkok from May 2018 to March 2019. We used a crushing technique to observe the cercarial stage inside each snail body and amplified the internal transcribed spacer 2 regions of cercarial DNA using polymerase chain reaction methodology. The associated phylogenetic tree was reconstructed using Bayesian inference analyses. A total of 517 of 15 621 examined snails were infected with trematode cercariae, and the infected snails were classified into 11 species of seven families with a 3.31% overall prevalence of the infection. The Bithynia siamensis siamensis snail displayed the highest prevalence of infection (16.16%), whereas the Physella acuta snail exhibited the lowest prevalence (0.08%) of infection. Eight morphological types of cercariae were observed. The highest prevalence of infection was observed in mutabile cercaria (1.86%). Based on molecular investigations, the phylogram revealed eight cercaria types assigned to at least nine digenean trematode families, of which five belong to groups of human intestinal flukes. Although, with the exception of schistosome cercaria, trematode cercariae are not known to directly damage humans, understanding the general biology of trematode cercariae (including diversity, distribution, infection rates and host range) is important and necessary for the prevention and control of parasitic transmission that impacts aquatic cultivations, livestock farming and human health.

Systematics and geographical distribution of Galba species, a group of cryptic and worldwide freshwater snails.

Cryptic species can present a significant challenge to the application of systematic and biogeographic principles, especially if they are invasive or transmit parasites or pathogens. Detecting cryptic species requires a pluralistic approach in which molecular markers facilitate the detection of coherent taxonomic units that can then be analyzed using various traits (e.g., internal morphology) and crosses. In asexual or self-fertilizing species, the latter criteria are of limited use. We studied a group of cryptic freshwater snails (genus Galba) from the family Lymnaeidae that have invaded almost all continents, reproducing mainly by self-fertilization and transmitting liver flukes to humans and livestock. We aim to clarify the systematics, distribution, and phylogeny of these species with an integrative approach that includes morphology, molecular markers, wide-scale sampling across America, and data retrieved from GenBank (to include Old World samples). Our phylogenetic analysis suggests that the genus Galba originated ca. 22 Myr ago and today comprises six species or species complexes. Four of them show an elongated-shell cryptic phenotype and exhibit wide variation in their genetic diversity, geographic distribution, and invasiveness. The remaining two species have more geographically restricted distributions and exhibit a globose-shell cryptic phenotype, most likely phylogenetically derived from the elongated one. We emphasize that no Galba species should be identified without molecular markers. We also discuss several hypotheses that can explain the origin of cryptic species in Galba, such as convergence and morphological stasis.