Galba Truncatula Research Articles

Multi‐Environment Quantification of Parasite and Intermediate Host DNA on Pasture for Fine‐Scale Disease Risk Assessment

ABSTRACTParasite transmission occurs in complex environments comprising multiple matrices. Trematode parasites of ruminant livestock such as the liver fluke, Fasciola hepatica and the rumen fluke, Calicophoron daubneyi, show affinity with freshwater environments shared with their amphibious snail intermediate host, Galba truncatula. Isolation of environmental DNA (eDNA) from these parasites and their snail hosts in water draining from grazing land provides opportunities for improved molecular diagnostic detection and can help identify infection risks at farm level. The detection and quantification of eDNA from other environmental matrices has received less attention but would improve the understanding of parasite dynamics on pasture. Our study has considerably extended eDNA sampling methods for the detection of parasitic trematodes of ruminant livestock and their snail intermediate host by including for the first time the analysis of soil and herbage environmental samples alongside water collections. A droplet digital PCR (ddPCR) workflow was developed to detect parasite and snail eDNA from soil, herbage, and water collected from livestock farms. For the first time, C. daubneyi eDNA was isolated from agricultural soil alongside water samples and G. truncatula eDNA was detected in water, soil, and herbage samples. No environmental samples were positive for F. hepatica eDNA. Assessing multiple environmental matrices increased the number of positive sites. Future implementation of eDNA detection methods alongside traditional parasite diagnostics can underpin more holistic evaluations of the environmental components of parasite epidemiology and facilitate adaptation to changing disease patterns.

The prevalence and infection rates of amphistome species in intermediate snail hosts: a systematic review and meta-analysis.

The systematic review and meta-analysis were conducted to determine the estimates of the prevalence and infection rates of natural and experimental infections of amphistome species in intermediate host snails (IHs) across different continents. A search of peer-reviewed literature on natural and experimental infections of freshwater snails with amphistome species was conducted from four electronic databases from 1984 to 2023. The estimates of the prevalence and/or infection rates were based on 36 eligible peer-reviewed articles, which met the inclusion criteria and reported on natural and experimental infections of amphistome species in freshwater snails. The results showed that a total of 1,67,081 snail species from the peer-reviewed articles were examined for natural infections and 7,659 snail species for experimental infections. The overall pooled prevalence of amphistome infections from naturally infected snails was 2% (95% CI: 0-4), while the overall pooled prevalence of amphistome infections from infections was 40% (95% CI: 18-64). The highest pooled prevalence of natural infection was 3%, which was recorded in Europe (95% CI: 1-7%). The highest overall prevalence of naturally infected amphistome was 6% (95% CI: 0-20%) for Paramphistomum epiclitum. The Americas had the highest pooled prevalence of experimental amphistome infection among freshwater snails (66%; 95% CI: 26-96%). The highest pooled infection rate of 65% (95% CI: 12-100%) was recorded for Paramphistomum cervi in experimental infections. Galba truncatula was the only snail that qualified for meta-analysis for natural infection with Calicophoron daubneyi, with a pooled prevalence of 3% (95% CI: 1-8%). Galba truncatula infected with C. daubneyi and P. cervi, and Bulinus tropicus infected with Calicophoron microbothrium in the experimental infection qualified for the meta-analysis, with an overall infection rate of 66% (95% CI: 34-92%) and 30% (95% CI: 0-74%), respectively. The pooled prevalence of amphistome species infection in the intermediate host (IH) snails based on detection techniques was higher with PCR compared to the dissection and shedding of cercariae. The results from the quality effects model revealed a high heterogeneity and publication bias between studies. This meta-analysis provided valuable insights into the prevalence and infection rates of amphistome species in snail IHs across different geographical regions.

Interactions between Galba truncatula (Gastropoda, Lymnaeidae) and two land snail species on the acid soils of central France

ABSTRACT Interactions between Galba truncatula and two species of land snails (Cochlicopa lubrica, Succinea putris) that colonise lymnaeid habitats in June-July were studied by introducing G. truncatula into new sites (Experiment A) or removing it from its natural habitats (Experiment B). Snail counts were performed in late May or early June for the eight years following introduction or removal of G. truncatula. The results were compared to those observed at control sites where G. truncatula was absent. In sites where G. truncatula was introduced (Experiment A), the mean density of the lymnaeid peaked in the second year and gradually decreased in subsequent years. Conversely, the mean density of the land snails decreased sharply in the second year before increasing rapidly for S. putris and gradually for C. lubrica in subsequent years. Removal of G. truncatula from its natural habitats (Experiment B) had no significant effect on the mean density of C. lubrica. In contrast, that of S. putris peaked in the second year and decreased rapidly to lower values in subsequent years. Density interactions between G. truncatula and the two land snail species indicate that there is probably competition between these species for space in lymnaeid habitats in May and June.

Development of a qPCR Duplex Assay for simultaneous detection of Fascioloides magna and Galba truncatula in eDNA samples: Monitoring beyond boundaries

Parasites constitute a significant economic burden and highly impact environmental, public, and animal health. The emergence of many parasitic diseases is environmentally mediated and they share the same biogeography with humans and both domestic and wild animals. American liver fluke, Fascioloides magna – a trematode parasite of domestic and wild ungulates – is an example of the anthropogenic introduction of an “invasive alien species” in Italy and Europe. Multiple introductions to Europe have led to the biogeographical expansion of the parasite across the Danube region mainly provided by the presence of suitable habitats for all hosts involved in the parasite's life cycle, human-assisted transport, and drastic environmental events such as flooding. In Italy, it was introduced and established in La Mandria Regional Park (LMRP) near Turin in 1865 along with imported wapitis (Cervus elaphus canadensis) from North America (Bassi, 1875), but with no reported expansion to the surrounding areas. LMRP isolated F. magna focus, poses an important threat of possible expansion since the enclosed area is vulnerable to occasional bidirectional passage of roe deer. Additionally, tributary rivers to the Po river system, traversing the enclosed area, could further bolster the possibility of such spread. In this study, we developed a duplex qPCR assay for F. magna and its principal intermediate host Galba truncatula optimized for testing eDNA samples to meet the needs for surveillance of the parasite. Moreover, we validated the developed assay in natura by testing samples derived from filtered water and sediments collected inside and outside LMRP's fenced-off area. Our findings for the first time demonstrate the presence of F. magna's eDNA outside the park's internal fenced-off area.

Assessing the microbiota of the snail intermediate host of trematodes, Galba truncatula

BackgroundThe microbiome is known to play key roles in health and disease, including host susceptibility to parasite infections. The freshwater snail Galba truncatula is the intermediate host for many trematode species, including the liver and rumen flukes Fasciola hepatica and Calicophoron daubneyi, respectively. The snail-parasite system has previously been investigated. However, the specific interaction between the snail-associated microbiota and intra-snail developmental stages of trematodes has yet to be explored.MethodsGalba truncatula snails were collected from farms in Northern Ireland and trematode infection was diagnosed using PCR. High-throughput sequencing analysis of the bacterial 16S ribosomal DNA V3-V4 hypervariable regions was subsequently applied to characterise the microbiota of both uninfected and infected snails.Results We first showed that the snail harboured microbiota that was distinct for its environment. The microbiota of infected snails was found to differ significantly from that of uninfected snails. In particular, the bacterial genera Mycoplasma and Methylotenera were significantly more abundant in infected snails, while genera Sphingomonas and Nocardioides were predominantly associated with uninfected snails. ConclusionThese findings pave the way to future studies on the functional roles of bacteria in host-parasite relationships.Graphical

Destruction de Galba truncatula ou sauvegarde du mollusque : que faut-il faire sur les sols acides de la Haute-Vienne ?

Faut-il éliminer la Limnée tronquée dans les prairies marécageuses de la Haute-Vienne comme le demandent les éleveurs de bovins ou d'ovins, ou encore faut-il sauvegarder le mollusque comme le réclament certains défenseurs de la biodiversité animale ? Pour résoudre ce dilemme, les auteurs ont rappelé dans un premier temps les tentatives qui ont été effectuées pour contrôler le mollusque depuis les années 1970 dans le département et se sont ensuite interrogés sur la situation actuelle en suggérant une troisième voie possible. En raison de la localisation de nombreux habitats de la limnée à la périphérie du réseau d'évacuation de l'eau pluviale dans les prairies, de leur nombre qui diminue avec le réchauffement climatique et de leur faible superficie, l'isolement de tous les sites colonisés par le mollusque semble être la meilleure solution. Cette méthode peut s'appliquer aux sols acides comme ceux du Limousin car 99,8 % des habitats de la Limnée tronquée ne sont colonisés que par cette espèce. Par contre, dans les régions sédimentaires, cette technique est plus difficile à utiliser car les habitats plus étendus sont souvent colonisés par d'autres espèces de pulmonés dulçaquicoles en plus de Galba truncatula.

Isolation and molecular identification of liver fluke cercariae in freshwater snails of Chaharmahal and Bakhtiari province, Iran

ABSTRACT Liver flukes are parasitic flatworms that can infect humans and other mammals. In this study, freshwater snails from Chaharmahal and Bakhtiari province in Iran were investigated for the presence of liver fluke larvae. The snails were collected from various rivers and analysed using standard morphological and molecular techniques. Additionally, molecular identification of the collected snails was conducted through polymerase chain reaction (PCR) and sequencing of the COI gene. The freshwater snails were identified as Physella acuta (Draparnaud, 1805), Galba truncatula (O.F. Müller, 1774), Radix euphratica (Mousson, 1874), Radix auricularia (Linnaeus, 1758) and Planorbis planorbis (Linnaeus, 1758). The presence of larval stages of Fasciola spp. in individual snails was examined using stereomicroscopy and a crushing method. Morphological and molecular identification based on the ITS-1 marker for identifying Fasciola spp. revealed that 10.9% (59 out of 549) of R. auricularia and 5.8% (37 out of 630) of P. acuta were infected. A blastn analysis of the collected samples revealed that all isolates belonged to F. hepatica, indicating a potential risk of liver fluke infection in the study area. These findings emphasise the importance of regular surveillance and control measures to prevent the spread of these parasites.

Infection Rates of Fasciola Intermediate Host Snail Species and Their Distribution in Africa: A Systematic Review and Meta-Analysis.

This systematic review and meta-analysis aimed to collate the infection rates of Fasciola spp. in intermediate host snails and their distribution in Africa. The overall infectivity prevalences of Galba truncatula, Radix natalensis, and Pseudosuccinea columella are 52%, 8%, and 3%, respectively. The intermediate host snails native to Africa (R. natalensis and G. truncatula) have been examined more than the invasive P. columella. The studies included in the review ranged from 1999 to 2022. North Africa has the highest prevalence of G. truncatula, with an infection rate of 52%. The review reveals that naturally infected intermediate host snails (G. truncatula, R. natalensis, and P. columella) are found in various regions of Africa. G. truncatula accounts for 22% (from three countries) of the studies included in the review and it was only found in the North African region with the highest overall infection rate of 52%. More studies on infection rate and distribution are needed to effectively control and prevent future transmissions.

A comprehensive study on the infectivity of $Galba truncatula$ (O.F. Müller, 1774), an intermediate host of Fasciola sp. reported in Diyarbakır hospitals

This study aims to comprehensively investigate the trematode infection in $Galba truncatula$(O. F. Müller, 1774) a widely distributed intermediate host in the Dicle River, following the previously reporting of Fascioliasis cases in Diyarbakır hospitals. While previous literature has documented the presence of $F. hepatica$ and $F. gigantica$ in individuals treated in Diyarbakır hospitals, the origin of the trematodes remains unspecified. In this study, G. truncatula samples were collected from three densely populated areas along the Dicle River, specifically Hevsel Gardens, and subjected to histopathological examination to determine the parasite's infectivity. The analysis revealed that the snails were not infected with trematodes; however, the presence of the host and its wide dispersion in the Dicle River pose a significant risk of disease in the future. Hevsel Gardens, a location where sewage and wastewater from the northern suburbs of Diyarbakır converge with the river, is extensively used for activities that involve direct contact with G. $truncatula$. These activities include vegetable cultivation, fishing, farming, husbandry, and swimming, and pose an escalating risk of potential contagion. The study acknowledges the undetermined source of the parasite but highlights its preliminary nature, emphasizing the urgency of proactive measures. The findings will enhance our understanding of the health risks linked to G. $truncatula$ and underscore the significance of implementing effective control measures in a timely manner.

A proposed ectochory of Galba truncatula snails between wallow sites enhances transmission of Fascioloides magna at gemenc, in Hungary.

Each year temporary puddles are formed on compacted earth roads as a consequence of heavy rains and subsequent flooding in the large floodplain forests of the Danube River, Hungary, Central Europe, Hungary. After the receding of floodwaters, the muddy puddles persist from spring to mid-fall, where they are densely populated by Galba truncatula snails on an annual basis. These snails are the sole intermediate host of Fascioloides magna liver fluke of deer in the forest. According to field observations, G. truncatula is a very rare species on banks of river branches and lakes within the forest but always appears in large aggregations in these semi-natural potholes. Red deer (Cervus elaphus) leave their droppings in puddles while they wallow in the mud, therefore the snails encounter the miracidia of F. magna frequently. Snails are not able to dig themselves into the tamp soil of roads therefore they are destroyed by the wheels of vehicles which drive down the roads from autumn to the end of winter. Therefore, snails colonize these puddles regularly every spring. Since there is no connection between the distant puddles, it is supposed that deer and wild boars repeatedly introduce the snails into the puddles each year carrying snails with the help of mud, which is stuck on their fur while they wallow. This method of transport is supported by the fact that shell remnants of snails can be found on the bark of rubbing trunks situated nearby wallows. It seems that the sequential creation of wallow sites and the repeated introduction of G. truncatula enhance the likelihood for the trematode to infect both hosts.

Molecular detection of Fasciola, Schistosoma and Paramphistomum species from freshwater snails occurring in Gauteng and Free State provinces, South Africa

Trematodiases are diseases caused by snail-borne trematode parasites that infect both animals and humans. Fascioliasis, schistosomiasis and paramphistomosis are some of these diseases and they affect millions of livestock, leading to significant economic losses. The aim of the study was to document freshwater snails occurring in selected study sites in the Free State and Gauteng provinces as well as identify and detect larval trematodes that they harbour. Samples were collected from a total of five study sites within two provinces of South Africa. Morphological features were used to identify snail species and were further confirmed genetically by polymerase chain reaction (PCR), sequencing and phylogenetic analysis. The larval trematodes were also detected by PCR, PCR-Restriction Length Fragment Polymorphism (PCR-RLFP), sequencing and phylogenetic analysis. A total of 887 freshwater snails were collected from Free State (n = 343) and Gauteng (n = 544). Five different genera of snails as well as species in the Succineidae family were documented. The snails in descending order of abundance were identified as: Physa (P.) spp. (51%), Succineidae spp. (20%), Galba (G.) truncatula (12%), Pseudosuccinea (Ps.) columella (10%), Planorbella (Pl.) duryi (6%) and Bulinus (B.) truncatus (1%). Approximately 272 DNA pools were created for genetic identification of snails and detection of trematode parasites. Schistosoma species were not detected from any of the snail species. A total prevalence of 46% was obtained for Fasciola hepatica in the identified snail species across all study sites. Overall, the highest prevalence of F. hepatica was obtained in Physa species (24%), whilst the lowest was observed in B. truncatus snails (1%). Forty three percent (43%) of the snail samples were PCR positive for Paramphistomum DNA. This is the first report of P. mexicana in South Africa. Fasciola hepatica was confirmed from all obtained snail species per study site. This is the first reported detection of F. hepatica in Pl. duryi and P. mexicana snails as well as the first confirmation of natural infection from P. acuta in South Africa.

Lateglacial to Mid-Holocene history of Vasby Mose, eastern Sjælland, Denmark

We retrieved a sediment core from Vasby Mose, a calcareous fen on eastern Sjælland, Denmark. The record spans the period from c. 14 700 to 6800 cal. years BP. During the Lateglacial, Vasby Mose was a lake where minerogenic sandy and clayey sediments accumulated. In the early Lateglacial, from c. 14 700 to 13 200 cal. years BP, a tundralike open, treeless vegetation with Betula nana and Dryas octopetala was found in the region. During the Younger Dryas, a rich flora of aquatic plants was found in the lake. In the Early to Mid-Holocene, Vasby Mose was a spring-fed calcareous fen, with deposition of peat and tufa. The flora included the sedges Carex rostrata and Carex paniculata, the aquatic plant Menyathes trifoliata, the calciphilous reed plant Cladium mariscus and the today nationally extinct bryophyte Meesia triquetra. The fauna included aquatic mollusc taxa such as Pisidium sp., Valvata spp. And Bithynia tentaculate and terrestrial or semi-terrestrial species such as Galba truncatula, Euconolus cf. alderi, Succinea/Oxyloma, Zonitoides nitidus and Vallonia pulchella. The Preboreal oscillation and other Early Holocene climate events are seen as short-lived, wet intervals.

Fasciolosis in the Mediterranean island of Corsica (France): Insights from epidemiological and malacological investigations

Fasciolosis is a re-emergent parasitic disease of worldwide significance with a major global impact on livestock health and production. In the French Mediterranean island of Corsica, fasciolosis has been recognized for a long time but little is known about its dynamic as the main investigations are outdated. Three compartments - definitive domestic hosts, intermediate hosts and environment - involved in fasciolosis transmission were studied by applying an integrative and extensive approach: (1) farm and abattoir surveys, (2) snail sampling, identification and infection prospection, and (3) snail habitat analysis; and (4) a questionnaire-based survey to inquire about husbandry practices and environmental risks. Our results indicate a significant circulation of the liver flukes in Corsican livestock, with 90% (252/279) of the sampled farms testing positive for anti-F. hepatica antibodies. At the abattoir, 46% (67/149) of cattle were positive for F. hepatica antibodies and eggs were present in the bile of 19% (26/139) bovines. In addition, high prevalence of Dicrocoelium dendriticum (69%) was observed in slaughtered cattle. Malacological surveys registered the occurrence of several lymnaeid species in a variety of habitats throughout the island. In particular, we report for the first time the presence of the invasive lymnaeid snail Pseudosuccinea columella in Corsica, a potential intermediate host for F. hepatica. We also found that the presence of Galba truncatula and, to a lesser extent, that of Peregriana peregra, is associated with altitude. Fasciola hepatica DNA was detected in the latter species occurring at two different sites. Finally, a questionnaire-based study revealed risky management practices among Corsican farmers, low perception of transmission and a suboptimal use of flukicide treatments as main control strategy. Our results show that animal fasciolosis in Corsica is characterised by a significant circulation and a favourable epidemiological scenario for transmission to occur.

Schistosomatidae from the trematode fauna of aquatic and semi-aquatic birds in Uzbekistan

The article discusses the infection of wetland birds in Uzbekistan with the trematodes Schistosomatidae Stiles et Hassall, 1898. The research covered 282 individuals from the main groups of birds represented by the orders Pelecaniformes, Ciconiformes, Anseriformes, Gruiformes, Podicipediformes and Charadriiformes. The Schistosomatidae fauna of Uzbekistan includes 13 species: Bilharziella polonica (Kowalewsky, 1895), Trichobilharzia ocellata (La Valette, 1854), T. filiformis (Szidat, 1938), T. kowalewskii (Ejsmont, 1929), T. tatianae (Spasskaja, 1953), Macrobilharzia macrobilharzia Trawassos, 1923, Ornithobilharzia canaliculata (Rudolphi, 1819), O. baeri Fain, 1955, Dendritobilharzia pulverulenta (Braun, 1901), D. loossi Skrjabin, 1924, D. anatinarum Cheatum, 1941, Gigantobilharzia acotylea Odhner, 1910 and Gigantobilharziella monocotylea (Szidat, 1930). Predominant are representatives of the genera Trichobilharzia and Dendritobilharzia. The species composition of Schistosomatidae is most diverse in birds from the lower reaches of the Amu Darya, where a high rate of infection with larvae of some of these trematodes was also recorded in aquatic molluscs. 11 species of molluscs were registered in the studied regions: Galba truncatula (Muller, 1774), Stagnicola corvus (Gmelin, 1791), Radix auricularia (Linnaeus, 1758), Lymnaea stagnalis (Linnaeus, 1758), Physa fontinalis (Linnaeus, 1758), Physella acuta (Draparnaud, 1805), Planorbis planorbis (Linnaeus, 1758), P. tangitarensis Germain, 1918, Anisus spirorbis (Linnaeus, 1758), Gyraulus albus (Muller, 1774), Melanoides kainarensis Starobogatov et Izzatullaev, 1980, from the families Lymnaeidae (4 species), Planorbidae (4 species), Physidae (2 species) and Thiaridae (1 species). They are identified as intermediate hosts of Schistosomatidae and were infected with 7 species of flukes. The total rate of infection with larval stages of Schistosomatidae in molluscs was about 2.0%. The highest infection rate was observed in Melanoides kainarensis – 5.3%. Morpho-biological indicators for mature forms of B. polonica from different bird species (Anas platyrhynchos and Oxyura leucocephala) were confirmed by molecular genetic studies. The study identified foci of birds’ infection with Schistosomatidae and the occurrence of human cercarial dermatitis.

Changes in the Populations of Two Lymnaeidae and Their Infection by Fasciola hepatica and/or Calicophoron daubneyi over the Past 30 Years in Central France.

Field investigations were carried out during three periods (from 1976 to 1997, in 2013-2014, and in 2020-2021) on 39 cattle-raising farms on acidic soils to track changes in the populations of two Lymnaeidae (Galba truncatula and Omphiscola glabra) and their infection with Fasciola hepatica and/or Calicophoron daubneyi. Compared to the survey between 1976 and 1997 on these farms, there was a significant decrease in the number of the two lymnaeid populations and the size of the G. truncatula populations in both 2013-2014 and 2020-2021. This decline was significantly faster in the last nine years than it was before 2013. The area of habitats colonized by G. truncatula showed no significant variation over the years, while that of habitats with O. glabra significantly decreased in the period covered by the three surveys. The prevalence of F. hepatica infection in snails significantly decreased over the years, while C. daubneyi infection increased over time in both lymnaeid species. These changes are due to the use of triclabendazole to treat fasciolosis in ruminants since the 1990s, and are probably a consequence of the successive heatwaves that have occurred since 2018 in the region.

Potential Hybridization of Fasciola hepatica and F. gigantica in Africa-A Scoping Review.

The occurrence of Fasciola gigantica and F. hepatica in Africa is well documented; however, unlike in Asia, there is a paucity of information on the existence of hybrids or parthenogenetic species on the continent. Nonetheless, these hybrid species may have beneficial characteristics, such as increased host range and pathogenicity. This study provides evidence of the potential existence of Fasciola hybrids in Africa. A literature search of articles published between 1980 and 2022 was conducted in PubMed, Google Scholar, and Science Direct using a combination of search terms and Boolean operators. Fasciola species were documented in 26 African countries with F. hepatica being restricted to 12 countries, whilst F. gigantica occurred in 24 countries, identified based on morphological features of adult Fasciola specimens or eggs and molecular techniques. The co-occurrence of both species was reported in 11 countries. However, the occurrence of potential Fasciola hybrids was only confirmed in Egypt and Chad but is suspected in South Africa and Zimbabwe. These were identified based on liver fluke morphometrics, assessment of the sperms in the seminal vesicle, and molecular techniques. The occurrence of intermediate host snails Galba truncatula and Radix natalensis was reported in Ethiopia, Egypt, South Africa, Tanzania, and Uganda, where F. hepatica and F. gigantica co-occurrences were reported. The invasive Pseudosuccinea columella snails naturally infected with F. gigantica were documented in South Africa and Egypt. In Zimbabwe, P. columella was infected with a presumed parthenogenetic Fasciola. This suggests that the invasive species might also be contributing to the overlapping distributions of the two Fasciola species since it can transmit both species. Notwithstanding the limited studies in Africa, the potential existence of Fasciola hybrids in Africa is real and might mimic scenarios in Asia, where parthenogenetic Fasciola exist in most Asian countries. In South Africa, aspermic F. hepatica and Fasciola sp. have been reported already, and Fasciola hybrids have been reported? in Chad and Egypt. Thus, the authors recommend future surveys using molecular markers recommended to identify Fasciola spp. and their snail intermediate hosts to demarcate areas of overlapping distribution where Fasciola hybrids and/or parthenogenetic Fasciola may occur. Further studies should also be conducted to determine the presence and role of P. columella in the transmission of Fasciola spp. in these geographical overlaps to help prevent parasite spillbacks.

Identification of factors associated with Fasciola hepatica infection risk areas on pastures via an environmental DNA survey of Galba truncatula distribution using droplet digital and quantitative real‐time PCR assays

AbstractEnvironmental DNA (eDNA) is a powerful tool for identifying the spatial and temporal presence and density of species in a range of aquatic habitats. The analysis of eDNA has a wide range of application, one of which may be to inform of Fasciola hepatica infection risk on pastures based on the detection of its eDNA as well as that of its intermediate snail host, Galba truncatula eDNA. Here, droplet digital PCR (ddPCR) and quantitative real‐time PCR (qPCR) assays were developed to detect the eDNA of F. hepatica, and its intermediate snail host, G. truncatula in water samples collected from pastures grazed by cattle and/or sheep. Environmental factors associated with species presence, as detected via an eDNA survey, were identified using zero‐inflated linear mixed models. Sixty‐four habitats were sampled across six farms in Ceredigion, Wales, UK, with ddPCR and qPCR identifying 42 and 33 habitats to be positive for G. truncatula eDNA, respectively. G. truncatula eDNA was significantly less likely to be detected in habitats fully shaded by trees, those that contained black or dark brown soils and habitats that contained deep water pools (p < 0.05). Significantly higher G. truncatula eDNA concentrations were observed in habitats that tend to dry up during Summer (i.e., temporary habitats) (p < 0.05). ddPCR also identified five habitats to be positive for F. hepatica eDNA; however, questions remain regarding the utility of F. hepatica eDNA detection due to a lack of specificity toward infective F. hepatica larval stages. The results of this study inform of factors which influences G. truncatula distribution and ecology on pastures and also provided practical information for farmers to aid F. hepatica control in their flocks and herds.

Occurrence of Fasciola (Digenea: Fasciolidae) Species in Livestock, Wildlife and Humans, and the Geographical Distribution of Their Intermediate Hosts in South Africa-A Scoping Review.

This review was conducted to provide an update on the status of the occurrence of Fasciola species in livestock, wildlife and humans, and the geographical distribution of snail intermediate host (IH) species in South Africa. The literature search was conducted on four electronic databases using the Boolean operators in combination with predetermined search terms for thematic analysis. Results showed that Fasciola species have been reported in six out of nine provinces of South Africa in the last six decades (1960–2021), with both F. hepatica and F. gigantica infecting vertebrate hosts and F. hepatica and Fasciola spp infecting humans. Results also showed that most studies relied on morphological identification of eggs and flukes without molecular confirmation, which might have led to the misidentification of specimens, especially when immature. Fasciola hepatica has been documented in Limpopo, Mpumalanga, and KwaZulu-Natal provinces. The occurrences of Galba truncatula as the probable snail IH for F. hepatica in the three provinces has been documented while Pseudosuccinea columella has only been documented in Mpumalanga and KwaZulu-Natal provinces. The occurrence of F. gigantica to date has been reported in Mpumalanga and KwaZulu-Natal provinces, with overlapping distribution with F. hepatica. Radix natalensis, the main IH of F. gigantica has been documented in all the three provinces, while the two alien Radix species (R. auricularia and R. rubiginosa) were documented in KwaZulu-Natal province and have been implicated elsewhere with the transmission of F. gigantica. The presence of Fasciola spp eggs and antibodies in humans were documented in the Eastern Cape and the Western Cape provinces, where both P. columella and G. truncatula are known to be present. The prevalence of Fasciola spp infection in livestock ranged from 9.1 to 37.67 %, with an estimated annual financial loss ranging from R44930.26-129901 in cattle production in the Eastern Cape province of South Africa. This review reaffirms the scarcity of information on the occurrence and burden of fasciolosis in South Africa, and further highlights the importance of future research covering all provinces of the country and assessing the public health significance of the disease in resource-poor livestock communities in the areas where the parasite is endemic.

Growth, Laying, and Survival Rates of the Galba truncatula Snails Infected with Fasciola hepatica

Fasciolosis is one of the most important parasitic diseases in ruminants in Algeria, of which the intermediate host is Galba truncatula (G. truncatula) snail. The current study aimed to investigate the prevalence of Fasciola sp. in naturally infected G. truncatula snails using multiplex PCR. Secondly, it was targeted toward examining the rate of growth, survival, and laying of the snails in experimental conditions during 6 weeks of rearing in three tanks. This study was conducted in two different regions of Algeria, namely El Tarf and Constantine. The investigated tanks 1, 2, and 3 consisted of 12 (size 3-4 mm), 30 (size 5-6 mm), and 30 (size 7-8 mm) snails, respectively. Fasciola sp. DNA was detected in 33.33% of G. truncatula snails (25% in Constantine and 42.85% in El Tarf). The total survival rates in the first, second, and third tanks were 50%, 43.3%, and 40%, respectively. The obtained results indicated that the growth rate of the snail depended on its initial size (the smaller the initial size, the higher the weekly growth rate). The total growth rates were 3, 1.7, and 1.1 mm in tanks 1, 2, and 3, respectively. The use of multiplex PCR indicated a relatively high level of infestation of the snails by Fasciola sp. Snails larger than 7 mm had the highest lay rate. Further studies are needed to investigate other snails that may be infested with Fasciola sp.

Plant molluscicide based on Smolevka white (Silene Latifolia) as prevention of pastoral helminthiasis of animals

Soft-bodied is the intermediate host of helminthiases, in the body of which several development stages of larval forms of helminths occur. There is the highest population density of mollusks in the areas of ruminant grazing, which leads to mass infection of animals with trematodes. To destroy the intermediate host of helminths in agricultural production, molluscicidal remedies of synthetic and plant origin are used. The work aimed to determine the molluscicidal effectiveness of a plant remedy based on Silene Latifolia in conditions of natural pastures. The material for work was the green mass of the plant S. Latifolia obtained in the warm season from roots, leaves, stems, flowers, and seeds. By grinding this plant, a powder with a particle size of 1-3 mm was obtained. Then, the powder was extracted with ethyl alcohol. The obtained product (concentrate) was an amorphous gel-like mass of dark green color with a specific smell and well soluble in water. Fieldwork in natural pastures was carried out on 5 biotopes with an area of 4-25 m2. Three species of gastropods were recorded from freshwater mollusks in the biotopes: Planorbis planorbis, Planorbarius corneus, Physa fontinalis, Lymnaea truncatula, and L. palustris. The results of experiments conducted in the conditions of pastures indicate a high molluscicidal activity of the studied plant agent on pond fish, intermediate hosts of trematodes pathogens. The effectiveness of the developed molluscicide on gastropods, when treated with a working solution (10.0 g/l) is from 98.1 to 100%.