One Health monitoring reveals invasive freshwater snail species, new records, and undescribed parasite diversity in Zimbabwe.

BACKGROUND: Snail-borne trematodes afflict humans, livestock, and wildlife. Recognizing their zoonotic potential and possible hybridization, a One Health approach is essential for effective control. Given the dearth of knowledge on African trematodes, this study aimed to map snail and trematode diversity, focusing on (i) characterizing gastropod snail species and their trematode parasites, (ii) determining infection rates of snail species as intermediate hosts for medically, veterinary, and ecologically significant trematodes, and (iii) comparing their diversity across endemic regions. METHODS: A cross-sectional study conducted in 2021 in Chiredzi and Wedza districts in Zimbabwe, known for high human schistosomiasis prevalence, involved malacological surveys at 56 sites. Trematode infections in snails were detected through shedding experiments and multiplex rapid diagnostic polymerase chain reactions (RD-PCRs). Morphological and molecular analyses were employed to identify snail and trematode species. RESULTS: Among 3209 collected snail specimens, 11 species were identified, including schistosome and fasciolid competent snail species. We report for the first time the invasive exotic snail Tarebia granifera in Zimbabwe, which was highly abundant, mainly in Chiredzi, occurring at 29 out of 35 sites. Shedding experiments on 1303 snails revealed a 2.24% infection rate, with 15 trematode species identified through molecular genotyping. Five species were exclusive to Chiredzi: Bolbophorus sp., Schistosoma mansoni, Schistosoma mattheei, Calicophoron sp., and Uvulifer sp. Eight were exclusive to Wedza, including Trichobilharzia sp., Stephanoprora amurensis, Spirorchid sp., and Echinostoma sp. as well as an unidentified species of the Plagiorchioidea superfamily. One species, Tylodelphys mashonensis, was common to both regions. The RD-PCR screening of 976 non-shedding snails indicated a 35.7% trematode infection rate, including the presence of schistosomes (1.1%) Fasciola nyanzae (0.6%). In Chiredzi, Radix natalensis had the highest trematode infection prevalence (33.3%), while in Wedza, R. natalensis (55.4%) and Bulinus tropicus (53.2%) had the highest infection prevalence. CONCLUSIONS: Our xenomonitoring approach unveiled 15 trematode species, including nine new records in Zimbabwe. Schistosoma mansoni persists in the study region despite six mass deworming rounds. The high snail and parasite diversity, including the presence of exotic snail species that can impact endemic species and biomedically important trematodes, underscores the need for increased monitoring.

Ecosystem links: Anthropogenic activities, environmental variables, and macrophytes structure snail preferences in man-made waterbodies.

Freshwater snails act as obligate intermediate hosts for trematode parasites that cause trematodiases threatening public and veterinary health, and biodiversity conservation. While interest in snail control for trematodiases has re-emerged, their ecology remains poorly understood. We examined the relationship between ecosystem indicators - such as environmental variables, macroinvertebrates, macrophytes, and land use - and their correlation with snail abundance, diversity, and infection prevalence in 19 man-made ponds in eastern Zimbabwe. In total, 926 freshwater snails from 10 species were collected, with 547 individuals belonging to five schistosome-competent species: Bulinus tropicus, Bulinus truncatus, Bulinus globosus, Bulinus forskalii, and Biomphalaria pfeifferi. The remaining 379 snails comprised Radix natalensis, Gyraulus sp., and the exotic invasive species Melanoides tuberculata, Pseudosuccinea columella, and Physella acuta. Six cercarial types - mammalian schistosomes, avian schistosomes, longifurcate pharyngeates, echinostomes, amphistomes, and xiphidiocercariae - were isolated from 104 out of 926 snails (11.2 %). PCR revealed a significantly higher infection rate, with 70.2 % of snails testing positive for trematodes. Snail taxon diversity and infection rate significantly varied across land use types, with the lowest values observed in the commercial tobacco farm section, highlighting the potential adverse effects of agriculture on biodiversity. Ponds with extensive Lagarosiphon major (oxygen weed) coverage appeared to facilitate the presence and abundance of P. acuta and P. columella. Schistosome-competent snails such as B. truncatus and B. tropicus seemed to favor shallow water depths and more eutrophic sites characterized by high levels of nitrates, phytoplankton biomass, turbidity, and phycocyanin. These ponds were predominantly associated with the emergent macrophyte Cladium mariscus, revealing a potential association with important intermediate snail hosts. In conclusion, our study emphasizes the complex interplay among environmental factors, macrophyte composition, land use, and the abundance, diversity, and infection prevalence of freshwater snails, offering insights into potential strategies for targeted snail control and disease management in man-made waterbodies.

Exposing the Barcoding Void: An Integrative Approach to Study Snail-Borne Parasites in a One Health Context.

Trematodes are snail-borne parasites of major zoonotic importance that infect millions of people and animals worldwide and frequently hybridize with closely related species. Therefore, it is desirable to study trematodiases in a One Health framework, where human and animal trematodes are considered equally important. It is within this framework that we set out to study the snail and trematode communities in four artificial lakes and an abattoir in Zimbabwe. Trematode infections in snails were detected through multiplex PCR protocols. Subsequently, we identified snails by sequencing a partial mitochondrial cytochrome c oxidase subunit I (COI) fragment, and trematodes (adults from the abattoir and larval stages detected in snails) using COI and nuclear rDNA markers. Of the 1,674 collected snails, 699 were molecularly analyzed, in which we identified 12 snail and 19 trematode species. Additionally, three parasite species were sampled from the abattoir. Merely four trematode species were identified to species level through COI-based barcoding. Moreover, identification of members of the superfamilies Opisthorchioidea and Plagiorchioidea required a phylogenetic inference using the highly conserved 18S rDNA marker, as no related COI reference sequences were present in public databases. These barcoding challenges demonstrate a severe barcoding void in the available databases, which can be attributed to the neglected status of trematodiases. Adding to this, many available sequences cannot be used as different studies use different markers. To fill this gap, more studies on African trematodes, using a standardized COI barcoding region, are desperately needed.

A rapid diagnostic multiplex PCR approach for xenomonitoring of human and animal schistosomiasis in a 'One Health' context.

Studying the epidemiology of schistosomiasis-the most prevalent gastropod-borne human disease and an economic burden for the livestock industry-relies on adequate monitoring tools. Here we describe a molecular assay for detecting human and animal African schistosome species in their planorbid gastropod host (xenomonitoring) using a two-step approach. First, schistosome infections are detected and discriminated from other trematode infections using a multiplex polymerase chain reaction (PCR) that includes a trematode-specific marker (in 18S rDNA), a Schistosoma genus-specific marker (in internal transcribed spacer 2 [ITS2]) and a general gastropod marker (in 18S rDNA) as an internal control. Upon Schistosoma sp. detection, a second multiplex PCR is performed to discriminate among Schistosoma haematobium, Schistosoma mansoni, Schistosoma mattheei and Schistosoma bovis/Schistosoma curassoni/Schistosoma guineensis using markers of differential lengths in the cytochrome c oxidase subunit 1 (COX1) gene. The specificity of these assays was validated with adult worms, naturally infected gastropods and human urine and stool samples. Sensitivity was tested on experimentally infected snail specimens that were sacrificed 10 and 40 days post-infection in order to mimic a natural prepatent and mature infection, respectively. The assay provides a diagnostic tool to support the xenomonitoring of planorbid gastropods for trematode infections in a One Health context, with a focus on the transmission monitoring of schistosomiasis.