Molecular phylogeography and reproductive biology of the freshwater snail Tarebia granifera in Thailand and Timor (Cerithioidea, Thiaridae): morphological disparity versus genetic diversity

The freshwater thiarid gastropod Tarebia granifera (Lamarck, 1816), including taxa considered either congeneric or conspecific by earlier authors, is widespread and abundant in various lentic and lotic water bodies in mainland and insular Southeast Asia, with its range extending onto islands in the Indo-West-Pacific. This snail is, as one of the most frequent and major first intermediate host, an important vector for digenic trematodes causing several human diseases. As a typical thiarid T. granifera is viviparous and parthenogenetic, with various embryonic stages up to larger shelled juveniles developing within the female’s subhemocoelic (i.e non-uterine) brood pouch. Despite the known conchological disparity in other thiarids as well as this taxon, in Thailand Tarebia has been reported with the occurrence of one species only. In light of the polytypic variations found in shell morphology of freshwater snails in general and this taxon in particular, the lack of a modern taxonomic-systematic revision, using molecular genetics, has hampered more detailed insights to date, for example, into the locally varying trematode infection rates found in populations of Tarebia from across its range in Thailand as well as neighboring countries and areas. Here, we integrate evidence from phylogeographical analyses based on phenotypic variation (shell morphology, using biometry and geometric morphometrics) with highly informative and heterogeneous mtDNA sequence data (from the gene fragments cytochrome c oxidase subunit 1 and 16 S rRNA). We evaluate both the morphological and molecular genetic variation (using several phylogenetic analyses, including haplotype networks and a dated molecular tree), in correlation with differences in the reproductive biology among populations of Tarebia from various water bodies in the north, northwest, central, and south of Thailand, supplementing our respective analyses of parasite infections of this thiarid by cercaria of 15 trematode species, reported in a parallel study. Based on the comparison of topotypical material from the island of Timor, with specimens from 12 locations as reference, we found significant, albeit not congruent variation of both phenotype and genotype in Tarebia granifera, based on 1,154 specimens from 95 Thai samples, representing a geographically wide-ranging, river-based cross-section of this country. Our analyses indicate the existence of two genetically distinct clades and hint at possible species differentiation within what has been traditionally considered as T. granifera. These two lineages started to split about 5 mya, possibly related to marine transgressions forming what became known as biogeographical barrier north of the Isthmus of Kra. Grounded on the site-by-site analysis of individual Tarebia populations, our country-wide chorological approach focussing on the conchologically distinct and genetically diverse lineages of Tarebia allows to discuss questions of this either reflecting subspecific forms versus being distinct species within a narrowly delimited species complex. Our results, therefore, provide the ground for new perspectives on the phylogeography, evolution and parasitology of Thai freshwater gastropods, exemplified here by these highly important thiarids

Freshwater pulmonate snails and their potential role as trematode intermediate host in a cercarial dermatitis outbreak in Southern Thailand

This study aimed to investigate the pulmonate snail species in the vicinity of the cercarial dermatitis outbreak area in southern Thailand. In 2020, an outbreak of cercarial dermatitis was reported in Chana district, Songkhla Province, caused by the ruminant schistosome Schistosoma indicum and its snail intermediate host Indoplanorbis exustus. In the present study, 1,175 pulmonate snails were collected between October 2021 and October 2022 from five provinces covering 34 locations in southern Thailand. Seven pulmonate snail species were identified based on shell morphology, including Amerianna carinata, Gyraulus bakeri, G. convexiusculus, G. hubendicki, Physella acuta, Indoplanorbis exustus, and Radix rubiginosa. Among these snails, eight species, and five types of cercariae were identified, viz. type (i) Echinostome cercariae consisted of Echinoparyphium recurvatum, Echinostoma spiniferum, and E. revolutum, type (ii) Brevifurcate-apharyngeate cercariae consisted of Schistosoma indicum and S. spindale, type (iii) Brevifurcate-pharyngeate-clinostomatoid-cercariae was represented by Clinostomum giganticum, type (iv) Longifurcate–pharyngeate cercariae (strigea cercaria) was Diplostomum baeri eucaliae, and type (v) Ophthalmoxiphidiocercaria. Among the seven pulmonated snail species, three were found to be infected, viz. G. convexiusculus, I. exustus, and R. rubiginosa, with infection rates of 1.14% (2/176), 0.25% (2/802), and 4.02% (7/174), respectively

Exploring the evolutionary potential of parasites: Larval stages of pathogen digenic trematodes in their thiarid snail host Tarebia granifera in Thailand

Minute intestinal flukes from several distinct families of endoparasitic platyhelminths are a medically important group of foodborne trematodes prevalent throughout Southeast Asia and Australasia. Their lifecycle is complex, with freshwater snails as primary intermediate hosts, with infecting multiple species of arthropods and fish as second intermediate hosts, and with birds and mammals including humans as definitive hosts. In Southeast Asian countries, the diversity of snail species of the Thiaridae which are frequently parasitized by trematode species is extremely high. Here, the thiarid Tarebia granifera in Thailand was studied for variation of trematode infections, by collecting the snails every two months for one year from each locality during the years 2004–2009, and during 2014–2016 when snails from the same localities were collected and new localities found. From ninety locations a total of 15,076 T. granifera were collected and examined for trematode infections. With 1,577 infected snails the infection rate was found to be 10.46 %. The cercariae were categorized into fifteen species from eight morphologically distinguishable types representing several distinct families, viz. (i) virgulate xiphidiocercariae (Loxogenoides bicolor, Loxogenes liberum and Acanthatrium histaense), (ii) armatae xiphidiocercariae cercariae (Maritreminoides caridinae and M. obstipus); (iii) parapleurophocercous cercariae (Haplorchis pumilio, H. taichui and Stictodora tridactyla); (iv) pleurophocercous cercariae (Centrocestus formosanus); (v) megarulous cercariae (Philophthalmus gralli); (vi) furcocercous cercariae (Cardicola alseae, Alaria mustelae and Transversotrema laruei); as well as (vii) echinostome-type cercariae, and (viii) gymnocephalous-type cercariae. In addition, a phylogenetic marker (internal transcribed spacers 2, ITS2) was employed in generic and infrageneric level classifications of these trematodes, using sequences obtained from shed cercariae isolated from T. granifera specimens of the second study period collected in various regions in Thailand. We obtained ITS2 sequences of cercariae from nine species (of seven types): Loxogenoides bicolor, Loxogenes liberum, Maritreminoides obstipus, Haplorchis taichui, Stictodora tridactyla, Centrocestus formosanus, Philophthalmus gralli, as well as from one species each of echinostome cercariae and gymnocephalous cercariae. Thus, this analysis combines the parasites’ data on morphology and geographical occurrence with molecular phylogeny, aiming to provide the groundwork for future studies looking into more details of the parasite-snail evolutionary relationships

Survey of Stenomelania Fisher, 1885 (Cerithioidea, Thiaridae): The potential of trematode infections in a newly-recorded snail genus at the coast of Andaman Sea, South Thailand

Stenomelania snails (Fisher 1885) have been reported from the coastal regions of the Indian Ocean and the Pacific Ocean, spanning India to Australia. Here, the species diversity and distribution of these snails in the south of Thailand are recorded. The snails were also examined for trematode infections in 13 locations in three Provinces, viz. Krabi, Trang and Satun, along the coast of the Andaman Sea. A total of 1,551 snails were in five morphs tentatively identified as Stenomelania aspirans, S. crenulata, S. punctata, S. torulosa and the closely-related Neoradina prasongi. With 10 infected snails, the trematode infection rate was 0.64%. The cercariae were categorised into three species from two morphologically-distinguishable types, viz. parapleurolophocercous cercariae (Haplorchis taichui and Procerovum cheni) and xiphidiocercariae (Loxogenoides bicolor), through the morphological characterisation of the larval stage. These trematodes were also analysed using the internal transcribed spacer subunit II region to confirm the species identity at generic and infrageneric levels