Transversotrema hafniensis n. sp. infection in Poecilia reticulata by cercariae released from Melanoides tuberculata in Denmark.

BACKGROUND: Exotic and ornamental fish are highly popular companion animals resulting in a significant transcontinental trade of fish, invertebrates and aquatic plants. A major issue is the diseases associated with these organisms, as they have a major impact on health of the fish in both public and private household aquaria. A secondary issue is the trade with these products, which potentially may expand the distribution area and spread a range of diseases to new habitats. RESULTS: We here describe how Poecilia reticulata (guppy), produced in a private household aquarium, were invaded by cercariae of an exotic trematode released by imported Melanoides tuberculata snails. The fish presented with severe clinical signs (tremor, flashing, scraping of body against objects). A standard parasitological examination and morphometric identification showed scale pocket infections with a digenean trematode species within the genus Transversotrema. Molecular identification by PCR, sequencing and phylogenetic analyses of a 2646 bp sequence encoding ribosomal RNA (partial 18 S, ITS1, 5.8 S, ITS2, partial 28 S) was performed. The 1107 bp sequence of mitochondrial DNA (cox1) showed that the parasite differed from previously described Transversotrema species in M. tuberculata. Morphometrics of adult and larval specimens of this isolate also differed from previously described freshwater species within the genus. The new species was described and is named after Copenhagen, for its geographic origin. CONCLUSIONS: The genus Transversotrema comprises a range of species, adapted to a microhabitat in scalepockets of teleosts. A combination of morphological and molecular characterization techniques has been shown to provide a good differentiation between species. The fish were not purchased from a pet shop but produced in the home aquarium. This indicated that an infection pressure existed in the aquarium, where the source of infection was found to be exotic intermediate host snails M. tuberculata, which originally were imported and purchased from a pet shop. The potential spread of fish diseases associated with trade of fish and snails to new geographic regions, where climate conditions are favourable, is discussed.

Some trematodes including three new species from freshwater fishes of Venezuela.

The present paper comprises a systematic survey of trematodes found in 13 species of freshwater fishes in Venezuela collected in 1992, 1996 and 2001. The following 15 trematode species were recorded: Adults: Genarchella venezuelaensis sp. n., Thometrema dissimilis sp. n., Megacoelium spinicavum Thatcher et Varella, 1981, Doradamphistoma bacuense Thatcher, 1999, Crassicutis cichlasomae Manter, 1936, Parspina carapo Ostrowski de Núñez, Arredonto et Gil de Pertierra, 2011, Phyllodistomoides hoplerythrini sp. n. Larvae (metacercariae): Clinostomatopsis sorbens (Braun, 1899), Clinostomum marginatum (Rudolphi, 1819), C. detruncatum Braun, 1899, Ithyoclinostomum dimorphum (Diesing, 1850), Odhneriotrema microcephala (Travassos, 1922), Tylodelphys sp., Posthodiplostomum sp., Sphincterodiplostomum sp. All these parasites are reported from Venezuela for the first time and many of these findings represent new host records. The new species G. venezuelaensis sp. n., T. dissimilis sp. n. and P. hoplerythrini sp. n. were collected from the accessory respiratory organ of Loricariichthys brunneus (Hancock) (Loricariidae), from the stomach of Hoplerythrinus unitaeniatus (Spix et Agassiz) (Erythrinidae) and from the intestine of H. unitaeniatus, respectively. All parasites are briefly described and illustrated and problems concerning their morphology, taxonomy, hosts and geographical distribution are discussed. Megacoelium spinispecum Thatcher et Varella, 1981 is considered a junior synonym of M. spinicavum Thatcher et Varella, 1981, and Crassicutis opisthoseminis Bravo-Hollis et Arroyo, 1962 as a junior synonym of C. cichlasomae Manter, 1936.

Calicophoron daubneyi (Paramphistomidae) in deer of the Sumava National Park, Czech Republic - Consequence of prevalent rumen fluke infection in cattle.

A substantial parallel increase in prevalence and geographical spread of the rumen fluke, Calicophoron daubneyi, in livestock in western and central Europe has been recognized in the recent past. In the course of the examination of rectum feces of 471 red deer (Cervus elaphus) and one sika deer (Cervus nippon) from the Fascioloides magna endemic Sumava National Park in the years 2021 and 2022, rumen fluke eggs were detected in four red deer (0.8%) and the sika deer and identified as eggs of C. daubneyi by molecular analysis. Subsequent examination of rectal fecal samples of 247 beef cattle from 22 herds of 14 farms located in or nearby the national park revealed rumen fluke eggs in 53 samples (21.5%) originating from 16 herds of 11 farms, molecularly identified as C. daubneyi eggs as well. One C. daubneyi egg positive red deer and three C. daubneyi egg positive cattle samples also contained fasciolid eggs, respectively, which were detected in 9.5% or 3.6% of the total samples from red deer or cattle, respectively. Results of this investigation reveal the first finding of C. daubneyi in sika deer worldwide and in red deer in mainland Europe and add to the growing number of reports on C. daubneyi in livestock in Europe. Considering that the ratio of cattle excreting rumen fluke eggs exceeded that of deer substantially, it can reasonably be assumed that the C. daubneyi infections in deer are a consequence of the prevalent infection in cattle, illustrating a pathogen spillover event from livestock into wildlife.

Morphological and molecular data on Pseudozoogonoides ugui Shimazu, 1974 (Digenea: Microphalloidea: Zoogonidae) ex Pseudaspius hakonensis (Günther, 1877) and taxonomic problems in Zoogoninae genera.

New morphological and molecular data were generated for trematodes recovered from the intestines of the fish Pseudaspius hakonensis from two locations in the south of the Russian Far East. Morphologically, these trematodes are identical to Pseudozoogonoides ugui (Microphalloidea: Zoogonidae) from Japan. According to results of phylogenetic analysis based on 28S rDNA sequence data, P. ugui was closely related to Zoogonoides viviparus, and P. subaequiporus appears as a sister taxon to these two species. Genetic distance values, calculated based on both 28S rDNA and ITS2 rDNA, between P. ugui and Z. viviparus represents an interspecific differentiation level. Our results have an ambiguous explanation, indicating that the implication of the presence of one or two compact vitellarial aggregations for the differentiation of Zoogonoides and Pseudozoogonoides should be reconsidered or that our results open up the question of the taxonomical status of trematodes previously denoted as Z. viviparus and P. subaequiporus.

Expanding the swimmer's itch pool of the Benelux: a first record of the neurotropic Trichobilharzia regenti and potential link to human infection.

BACKGROUND: Swimmer's itch, an allergic contact dermatitis caused by avian and mammalian blood flukes, is a parasitic infection affecting people worldwide. In particular, avian blood flukes of the genus Trichobilharzia are infamous for their role in swimmer's itch cases. These parasites infect waterfowl as a final host, but incidental infections by cercariae in humans are frequently reported. Upon accidental infections of humans, parasite larvae will be recognized by the immune system and destroyed, leading to painful itchy skin lesions. However, one species, Trichobilharzia regenti, can escape this response in experimental animals and reach the spinal cord, causing neuroinflammation. In the last few decades, there has been an increase in case reports across Europe, making it an emerging zoonosis. METHODS: Following a reported case of swimmer's itch in Kampenhout in 2022 (Belgium), the transmission site consisting of a private pond and an adjacent creek was investigated through a malacological and parasitological survey. RESULTS: Six snail species were collected, including the widespread Ampullaceana balthica, a well-known intermediate host for Trichobilharzia parasites. Shedding experiments followed by DNA barcoding revealed a single snail specimen to be infected with T. regenti, a new species record for Belgium and by extension the Benelux. Moreover, it is the most compelling case to date of the link between this neurotropic parasite and cercarial dermatitis. Additionally, an Echinostomatidae sp. and Notocotylus sp. were isolated from two other specimens of A. balthica. However, the lack of reference DNA sequences for these groups in the online repositories prevented genus- and species-level identification, respectively. CONCLUSIONS: The presence of T. regenti in Belgium might have severe clinical implications and its finding highlights the need for increased vigilance and diagnostic awareness among medical professionals. The lack of species-level identification of the other two parasite species showcases the barcoding void for trematodes. Overall, these findings demonstrate the need for a Belgian framework to rapidly detect and monitor zoonotic outbreaks of trematode parasites within the One Health context.

Integrative taxonomy of Anaporrhutum mundae sp. nov. (Trematoda: Gorgoderidae), a parasite of the Munda round ray Urotrygon munda (Urotrygonidae) in Costa Rica.

A new species of trematode of anaporrhutine gorgoderid, from the gill chambers of the Munda round ray Urotrygon munda in Costa Rica is described, based on an integrative taxonomic approach that includes the use of light and scanning electron microscopy, ITS2 and 28S rDNA sequencing, and phylogenetic analysis. Anaporrhutum mundae sp. nov. can be distinguished from congeneric species by a combination of morphological traits and particularly by having the genital pore opening at the level of the intestinal bifurcation. The new species also can be distinguished from all other species of Anaporrhutum, except A. euzeti Curran, Blend & Overstreet, 2003, by having fewer testicular follicles per testis. Anaporrhutum mundae sp. nov. also differs from A. euzeti in its forebody shape and by having different morphology and location of the vitellaria. The study of the tegumental surface of A. mundae sp. nov., as revealed by scanning electron microscopy, allowed detection of new morphological characters for a member of Anaporrhutinae that may be of taxonomic value. These are: a stylet cavity dorsal to the oral sucker with a large penetration gland opening on each side of the cavity and small penetration gland openings located ventral to the stylet cavity, arranged in a circle around the mouth. This represents the first record of an Anaporrhutum species from Costa Rica. Further, A. mundae sp. nov. represents the first parasite described or reported in this host.

Small intestinal flukes of the genus Metagonimus (Digenea: Heterophyidae) in Europe and the Middle East: A review of parasites with zoonotic potential.

The heterophyid trematode Metagonimus romanicus (Ciurea, 1915) (Digenea) is redescribed on the basis of type material from domestic dogs (Canis familiaris) in Romania, vouchers from experimentally infected cats (Felis catus) and adults recovered from golden hamsters (Mesocricetus auratus) infected with metacercariae from scales of chub (Squalius cephalus) and common nase (Chondrostoma nasus) (Cypriniformes: Leuciscidae) in Hungary. This trematode, endemic to Europe and neighbouring regions (northwestern Türkiye), was previously misidentified as M. yokogawai (Katsurada, 1912), a zoonotic parasite of humans in East Asia. However, the two species differ considerably both genetically and morphologically, e.g., in the position of the ventral sucker, the presence of the prepharynx, the anterior extent of the vitelline follicles and the posterior extent of the uterus. Metagonimus ciureanus (Witenberg, 1929) (syn. Dexiogonimus ciureanus Witenberg, 1929), described from domestic cats and dogs in Israel, is a valid species distributed in the Middle East and Transcaucasia, which is also confirmed by molecular data. It differs from all Metagonimus species, including M. romanicus, in having symmetrical testes instead of the oblique testes of the other congeners. The zoonotic significance of M. romanicus and M. ciureanus is unclear, but appears to be low in Europe, mainly because raw or undercooked, whole fish with scales are generally not consumed. Accidental infection of fishermen by metacercariae in the scales when cleaning fish is more likely, but has never been reported. Remains of cyprinoids with scales infected with metacercariae of Metagonimus spp. can be an important natural source of infection for dogs, cats, and other carnivores, which can serve as a reservoir for these parasites.

Title: Petits trématodes intestinaux du genre Metagonimus (Digenea : Heterophyidae) en Europe et au Moyen-Orient : revue de parasites à potentiel zoonotique. Abstract: Le trématode Heterophyidae Metagonimus romanicus (Ciurea, 1915) (Digenea) est redécrit sur la base de matériel type provenant de chiens domestiques (Canis familiaris) en Roumanie, de vouchers issus de chats (Felis catus) infectés expérimentalement et d'adultes collectés chez des hamsters dorés (Mesocricetus auratus) infectés par des métacercaires provenant d'écailles de chevesne commun (Squalius cephalus) et de nase commun (Chondrostoma nasus) (Cypriniformes : Leuciscidae) de Hongrie. Ce trématode, endémique d'Europe et des régions voisines (nord-ouest de la Turquie), avait été précédemment identifié à tort comme étant M. yokogawai (Katsurada, 1912), un parasite zoonotique des humains en Asie de l'Est. Cependant, les deux espèces diffèrent considérablement sur le plan génétique et morphologique, par exemple par la position de la ventouse ventrale, la présence du prépharynx, l'étendue antérieure des follicules vitellins et l'étendue postérieure de l'utérus. Metagonimus ciureanus (Witenberg, 1929) (syn. Dexiogonimus ciureanus Witenberg, 1929), décrite chez des chats et des chiens domestiques en Israël, est une espèce valide répartie au Moyen-Orient et en Transcaucasie, ce qui est également confirmé par des données moléculaires. Cette espèce diffère de toutes les espèces de Metagonimus, y compris M. romanicus, par ses testicules symétriques au lieu des testicules obliques des autres congénères. L'importance zoonotique de M. romanicus et M. ciureanus n'est pas claire, mais semble faible en Europe, principalement parce que les poissons n'y sont généralement pas consommés crus ou insuffisamment cuits et entiers avec les écailles. L'infection accidentelle des pêcheurs par des métacercaires présents dans les écailles lors du nettoyage du poisson est plus probable mais n'a jamais été signalée. Les restes de poissons cyprinoïdes avec les écailles, infectés par des métacercaires de Metagonimus spp. peuvent être une source naturelle importante d'infection pour les chiens, les chats et autres carnivores, qui peuvent servir de réservoir à ces parasites.

DULCITRANSVERSOTREMA N. GEN. (DIGENEA: TRANSVERSOTREMATIDAE) AND FIRST MORPHOLOGICAL AND NUCLEOTIDE-BASED CONFIRMATION OF THE LIFE CYCLE OF A TRANSVERSOTREMATID IN NORTH AMERICA.

We propose Dulcitransversotrema n. gen. for the freshwater transversotrematids Dulcitransversotrema patialense (Soparkar, 1924) n. comb. and Dulcitransversotrema chauhani (Agrawal and Singh, 1960) n. comb. Dulcitransversotrema is unique by the combination of lacking an oral sucker and an extensively coiled uterus as well as having testes that are not deeply lobed and that abut or nearly abut the inner posterolateral margins of the digestive tract (vs. deeply lobed testes that abut the cyclocoel region of the digestive tract only or that do not abut the ceca), a pre-oral genital pore (vs. post-oral genital pore), an ovary that is anterior to and abuts or nearly abuts the sinistral testis (vs. an ovary that is separated from the sinistral testis by some distance or having an ovary that is median and immediately pre-testicular), non-embryonated eggs (vs. embryonated eggs), typically having none or sparse vitelline follicles between the testes and ceca (vs. having a wholly inter-cecal vitellarium comprising a pair of highly compacted clusters of vitelline follicles anterolateral to the testes or having numerous vitelline follicles between the testes and ceca), an oblong, median, and primarily inter-testicular vitelline reservoir (vs. an extensively elongate, pre-testicular vitelline reservoir that arches anteriorly around the sinistral testis), and a subterminal excretory pore opening on the dorsal body surface between the level of the cyclocoel and posterior body end (vs. a terminal excretory pore). We describe the redia and cercaria of Dulcitransversotrema cf. patialense from the red-rimmed melania snail, Melanoides tuberculata (Müller, 1774) (Cerithioidea: Thiaridae), and their corresponding adult from beneath the scales of the zebrafish, Danio rerio (Hamilton, 1822) (Cypriniformes: Danionidae). All hosts were sampled from a spring-fed earthen pond at a private aquaculture facility near Ruskin, Florida. No transversotrematid life cycle was known from North America previously. The large subunit ribosomal DNA and ribosomal internal transcribed spacer 2 sequences of D. cf. patialense from Florida were most similar to those from D. patialense infecting red-rimmed melania from Mayagüez, Puerto Rico and differed by 134 and 69 nucleotides, respectively. Both phylogenetic analyses recovered Dulcitransversotrema as monophyletic and sister to a clade comprising Transversotrema spp. plus Crusziella formosa Cribb, Bray, and Barker, 1992.

Description of Saccocoelioides miguelmontesi n. sp. (Digenea: Haploporidae) from characid fishes in the Iguazu River Basin based on morphological and molecular evidence.

Here we describe a new species of the genus Saccocoelioides found parasitizing Astyanax dissimilis Garavello & Sampaio, Psalidodon bifasciatus (Garavello and Sampaio) and Bryconamericus ikaa Casciotta, Almirón & Azpelicueta from the Iguazu National Park, Misiones province, Argentina. Saccocoelioides miguelmontesi n. sp. was studied based on morphological and molecular (28S rDNA and COI mtDNA sequences) data. The COI mtDNA tree indicated that the specimens collected from the three fish hosts are conspecific, with an intragroup p-distance of 0%. The new species shows an intermediate morphological configuration between the diminutive and robust forms described for Saccocoelioides by Curran (2018). Although, in the 28S rDNA tree, it is placed in a well-supported clade with the two robust species analysed (S. elongatus and S. magnus; p-distance of 1 and 2%, respectively), it differs from the robust group by the range of body size, mature egg size, oral and ventral sucker size, sucker ratio, oral sucker to pharynx ratio, and post-cecal or post-testis/body length percentage. Our results led us to redefine the robust group as having eggs shorter or equal in length to the pharynx. Saccocoelioides miguelmontesi n. sp. the 10th species reported from Argentina and the 7th species within the robust group.

Two new species of Monopisthocotyla (Dactylogyridea: Dactylogyridae) parasitizing the nasal cavities of Bryconops melanurus (Characiformes: Iguanodectidae) from coastal drainages of the Eastern Amazon, Brazil.

The present study integrates molecular and morphological data to support the proposal of new species of Telethecium Kritsky, Van Every & Boeger, 1996 and Diaphorocleidus Jogunoori, Kritsky & Venkatanarasaiah, 2004 from the nasal cavities of Bryconops melanurus (Bloch) of the coastal drainages of the Eastern Amazon. Telethecium tiquira sp. n. is characterized by possessing a male copulatory organ (MCO) with two circular sclerotized brims on the base, a coiled tubular shaft having 1 counterclockwise rings, an accessory piece with enlarged base, pincer-shaped at the distal portion; a sclerotized calyx-shaped vaginal vestibule, and hooks with proximal shank dilatation comprising 3/4 of the shank length. Also, Telethecium tiquira sp. n. can be easily distinguished from other species of the genus by the absence of a protruding bag located at the level of the copulatory complex. Diaphorocleidus forficata sp. n. is characterized by having a MCO with two counterclockwise rings, circular sclerotized tandem brim associated with the base of the MCO; accessory piece non-articulated with the MCO, bifurcate, pincer-shaped; vaginal pore sinistral-ventral with opening marginal, vaginal canal sclerotized, elongated, comprising one loop in the proximal portion before entering to the seminal receptacle; ventral anchor with shaft elongated and evenly curved on the axis; point short and slightly curved, and hooks similar in shape and size, hooks with proximal dilatation comprising approximately of the shank length. Furthermore, D. forficata sp. n. is supported by phylogenetic analysis based on sequences of the partial 28S rDNA gene, which placed D. forficata sp. n. in a well-supported clade of Diaphorocleidus spp. of characiform fishes. Thus, the two new species described here expand our knowledge about the diversity of monopisthocotylan parasites from the nasal cavities of Neotropical fishes. The findings of this study provide valuable insights into the biodiversity of the region and highlight the importance of further research in this area.

An integrative taxonomy study reveals a rare new species of the genus Creptotrema (Trematoda: Allocreadiidae) in an endangered frog in South America.

During an ecological study with a near-endangered anuran in Brazil, the Schmidt's Spinythumb frog, Crossodactylus schmidti Gallardo, 1961, we were given a chance to analyze the gastrointestinal tract of a few individuals for parasites. In this paper, we describe a new species of an allocreadiid trematode of the genus Creptotrema Travassos, Artigas & Pereira, 1928, which possesses a unique trait among allocreadiids (i.e., a bivalve shell-like muscular structure at the opening of the ventral sucker); the new species represents the fourth species of allocreadiid trematode parasitizing amphibians. Besides, the new species is distinguished from other congeners by the combination of characters such as the body size, ventral sucker size, cirrus-sac size, and by having small eggs. DNA sequences through the 28S rDNA and COI mtDNA further corroborated the distinction of the new species. Phylogenetic analyses placed the newly generated sequences in a monophyletic clade together with all other sequenced species of Creptotrema. Genetic divergences between the new species and other Creptotrema spp. varied from 2.0 to 4.2% for 28S rDNA, and 15.1 to 16.8% for COI mtDNA, providing robust validation for the recognition of the new species. Even though allocreadiids are mainly parasites of freshwater fishes, our results confirm anurans as hosts of trematodes of this family. Additionally, we propose the reallocation of Auriculostoma ocloya Liquin, Gilardoni, Cremonte, Saravia, Cristóbal & Davies, 2022 to the genus Creptotrema. This study increases the known diversity of allocreadiids and contributes to our understanding of their evolutionary relationships, host-parasite relationships, and biogeographic history.

Strobilurin fungicide increases the susceptibility of amphibian larvae to trematode infections.

The global rise in fungal pathogens has driven the increased usage of fungicides, yet our understanding of their ecotoxicity remains largely limited to acute toxicity. While such data is critical for projecting the risk of fungicide exposure to individual species, the contamination of natural systems with fungicides also has the potential to alter species interactions within communities including host-parasite relationships. We examined the effects of the fungicide pyraclostrobin on the susceptibility of larval American bullfrogs (Rana catesbeiana) to trematode (echinostome) infections using a controlled laboratory experiment. Following a 2-wk exposure to 0, 1.0, 5.2, or 8.4 µg/L of pyraclostrobin, tadpoles were then exposed to parasites either in the 1) presence (continued/simultaneous exposure) or 2) absence (fungicide-free water) of pyraclostrobin. We found that when exposed to pyraclostrobin during parasite exposure, meta cercariae counts increased 4 to 8 times compared to control tadpoles. Additionally, parasite loads were approximately 2 times higher in tadpoles with continued fungicide exposures compared to tadpoles that were moved to fresh water following fungicide exposure. This research demonstrates that fungicides at environmentally relevant concentrations can indirectly alter host-parasite interactions, which could elevate disease risk. It also underscores the need for studies that expand beyond traditional toxicity experiments to assess the potential community and ecosystem-level implications of environmental contaminants.

Swimmer's itch control: Timely waterfowl brood relocation significantly reduces an avian schistosome population and human cases on recreational lakes.

Swimmer's itch (SI) is a dermatitis in humans caused by cercariae of avian and mammalian schistosomes which emerge from infected snails on a daily basis. Mitigation methods for SI have long been sought with little success. Copper sulfate application to the water to kill the snail hosts is the historically employed method, but is localized, temporary, and harmful to many aquatic species. Here, we test an alternative method to control Trichobilharzia stagnicolae, a species well-known to cause SI in northern Michigan and elsewhere in North America. Summer relocation of broods of the only known vertebrate host, common merganser (Mergus merganser), greatly reduced snail infection prevalence the following year on two large, geographically separated lakes in northern Michigan. Subsequent years of host relocation achieved and maintained snail infection prevalence at ~0.05%, more than an order of magnitude lower than pre-intervention. A Before-After-Control-Intervention (BACI) study design using multiple-year snail infection data from two intervention lakes and three control lakes demonstrates that dramatic lake-wide reduction of an avian schistosome can be achieved and is not due to natural fluctuations in the parasite populations. The relevance of reducing snail infection prevalence is demonstrated by a large seven-year data set of SI incidence in swimmers at a high-use beach, which showed a substantial reduction in SI cases in two successive years after relocation began. In addition, data from another Michigan lake where vertebrate-host based intervention occurred in the 1980's are analyzed statistically and show a remarkably similar pattern of reduction in snail infection prevalence. Together, these results demonstrate a highly effective SI mitigation strategy that avoids the use of environmentally suspect chemicals and removes incentive for lethal host removal. Biologically, the results strongly suggest that T. stagnicolae is reliant on the yearly hatch of ducklings to maintain populations at high levels on a lake and that the role of migratory hosts in the spring and fall is much less significant.

In-herd prevalence of Fasciola hepatica and Calicophoron / Paramphistomum spp. infections in German dairy cows with comparison of two coproscopical methods and establishment of real-time pyrosequencing for rumen fluke species differentiation.

Infections with liver and rumen flukes are among the most frequent parasitic diseases in cattle worldwide. In Europe, the predominant liver fluke species is Fasciola hepatica, and the recently rapidly spreading rumen flukes are mostly Calicophoron daubneyi and occasionally Paramphistomum leydeni. In this study, 1638 faecal samples from individual dairy cows from 24 northern and 18 southern German farms as well as one central German farm, all preselected for potential F. hepatica infection, were examined to determine in-herd prevalences of liver and rumen fluke infections. Furthermore, individual faecal egg counts (FECs) were determined in the northern and central German cows. On farms with patent F. hepatica infections, the mean in-herd prevalence was 15.8% in northern Germany, 41.6% in southern Germany and 14.0% in the central German farm. Rumen fluke infections resulted in high in-herd prevalences in all regions with a mean prevalence of 46.0% in northern, 48.4% in southern and 40.0% in central Germany. Individual FECs varied between 0.1 and 4.1 (mean 0.4) eggs per gram faeces (EPG) for F. hepatica and between 0.1 and 292.4 (mean 16.9) EPG for rumen flukes. Mean in-herd prevalence and mean FECs did not differ significantly between mono- and coinfected farms for either fluke species. Comparison of the classical sedimentation technique and the Flukefinder® method on a subset of 500 faecal samples revealed a similar number of positive samples, however, Flukefinder® mean FECs were three to four times higher for liver and rumen fluke eggs, respectively, with an increasing gap between EPG levels with rising egg counts. Fluke egg size measurement confirmed P. leydeni eggs on average to be larger in length and width (161.0 µm x 87.1 µm) than those of C. daubneyi (141.8 µm x 72.9 µm). However, due to overlap of measurements, morphological species identification based on egg size proved unreliable. For accurate identification, a real-time pyrosequencing approach was established, offering the advantage over classical Sanger sequencing of unambiguously identifying rumen fluke mixed species infections. Real-time pyrosequencing confirmed C. daubneyi (78.1% [50/64]) as the predominant rumen fluke species in Germany, while P. leydeni was detected in 12.5% (8/64) of sampled cows. A total of 9.4% (6/64) cows were infected with both C. daubneyi and P. leydeni, representing the first finding of a mixed infection in domestic ruminants in Europe to date.

Environmental influence on abundance and infection patterns of snail intermediate hosts of liver and intestinal flukes in North and Central Vietnam.

Liver and intestinal flukes (LIF) are important groups of foodborne zoonotic trematodes (FZTs) in Southeast Asia, including Vietnam. Their complex life cycles require specific freshwater snail species as the obligatory first intermediate hosts. In 2019, we conducted a longitudinal study in Yen Bai and Thanh Hoa provinces in North and Central Vietnam, respectively, to investigate the diversity of LIF and their infection prevalence in relation to snail host abundance and environmental factors. Using a combination of morphological and molecular identification techniques, we identified 10 LIF species infecting 11 snail host species. We observed significant seasonal variation in the mean abundance of several snail host species, with the majority of snails collected during the spring. We also detected seasonal changes in LIF species composition, with the highest species richness reported in the spring. Clonorchis sinensis and Fasciola gigantica, two medically important human liver flukes in Asia, were found only in the spring in Yen Bai. Our study revealed that not all snail host species have the same probability of becoming infected, and we recorded seasonal variations in the prevalence of LIF infection in different snail species in relation to water parameters.

Idiopyrgus souleyetianus (Gastropoda: Tomichiidae) as an intermediate host of the liver fluke Amphimerus sp. (Trematoda: Opisthorchiidae) in Brazil.

Despite the importance of fish-borne trematodes of the family Opisthorchiidae as causative agents of human liver fluke disease, studies on these parasites outside Asia are relativally scarce. In South America, human focus of amphimerosis is known in Ecuador since the mid-20th century, and Amphimerus spp. have also been reported in wild and domestic mammals. Nevertheless, the knowledge on the snails that act as the first intermediate host of these potentially zoonotic parasites are scarce. Herein, a new cercaria of the pleurolophocercous morphotype found in the freshwater snail Idiopyrgus souleyetianus from Brazil was subjected to morphological and molecular studies. Multigene phylogenetic analyses based on 28S, 5.8S-ITS-2 and Cox-1 sequences enabled the identification of Amphimerus sp., a species distinct from that reported in humans from Ecuador. This cercariae was morphologically compared with other opisthorchiid cercariae known. The possible occurrence of human amphimerosis in Brazil is discussed.

Effectiveness of health education in improving knowledge, attitude and practice related to foodborne zoonotic trematodes in Vietnam, with a particular focus on Clonorchis sinensis.

OBJECTIVE: Foodborne zoonotic trematodes (FZT), which infect the liver, lungs and intestines of humans, are an emerging public health concern in tropical countries including Vietnam. In northern Vietnam, Clonorchis sinensis is recognised as the most important species of the FZT. Up to now, small-scale studies conducted in the country have indicated that health education could improve participants' knowledge and practices related to clonorchiasis, however strong evidence is still lacking. We conducted an intervention trial between 2020 and 2021 in four communes in Yen Bai and Thanh Hoa province, aiming to evaluate the impact of an extended educational package on knowledge, attitude and practices related to clonorchiasis, as well as on knowledge on the large liver fluke and minute intestinal flukes. METHODS: To this end, baseline and post-intervention questionnaires were conducted. Generalised estimating equations models were run to analyse the impact of the intervention on knowledge, attitudes and practices over time in the intervention versus control group. Finally, 172 people in the intervention group and 162 in the control group were included for analysis. RESULTS: Results demonstrated that the intervention significantly improved clonorchiasis knowledge, attitudes and practices, with higher odds for a correct knowledge response (odds ratio (OR) = 2.80, 95% confidence interval (CI) = 1.84-4.27, p-value < 0.001), increased average mean attitude score (mean = 0.363, 95%CI = 0.182-0.544, p-value < 0.001), and a reduced odds of consuming raw fish (OR = 0.15, 95%CI = 0.06-0.40, p = 0.002) in the intervention group compared to the control group post-intervention versus baseline. Additionally, participants in the intervention group exhibited enhanced basic knowledge of the large liver fluke and minute intestinal flukes post-intervention. CONCLUSIONS: These findings demonstrate the potential of health education programs in preventing clonorchiasis in endemic areas, emphasising the importance of continued health education as a critical component of integrated control programs for clonorchiasis.

Evolutionary morphology of haptoral anchors in monogenoids (Dactylogyridae) of marine catfish (Siluriformes: Ariidae) from the Atlantic coast of South America.

Exploring the phylogenetic signal of morphological traits using geometric morphometry represents a powerful approach to assess the relative weights of convergence and shared evolutionary history in shaping species' forms. We evaluated the phylogenetic signal in shape and size of ventral and dorsal haptoral anchors of 10 species of monogenoids (Hamatopeduncularia, Chauhanellus and Susanlimocotyle) occurring in marine catfish (Siluriformes: Ariidae) from the Atlantic coast of South America. The phylogenetic relationships among these species were mapped onto the morphospaces of shape and size of dorsal and ventral anchors. Two different tests (squared change-parsimony and Kmult) were applied to establish whether the spatial positions in the phylomorphospace were influenced by phylogenetic relationships. A significant phylogenetic signal was found between anchor form and parasite phylogeny. Allometric effects on anchor shape were non-significant. Phylogenetically distant species on the same host differed markedly in anchor morphology, suggesting little influence of host species on anchor form. A significantly higher level of shape variation among ventral anchors was also found, suggesting that the evolutionary forces shaping ventral anchor morphology may operate with differing intensities or exhibit distinct mechanisms compared to their dorsal counterparts. Our results suggest that phylogenetic relationships were a key driver of changes in shape (but not size) of anchors of monogenoids of South American ariids. However, it seems that the emergence of the digitiform haptor in Hamatopenducularia and in some species of Chauhanellus played an important role in the reduction in anchor size and may cause secondary losses of anchors in other groups of monogenoids.

"Our desire is to make this village intestinal worm free": Identifying determinants of high coverage of community-wide mass drug administration for soil transmitted helminths in Benin, India, and Malawi.

BACKGROUND: Soil-transmitted helminth infections (STH) are associated with substantial morbidity in low-and-middle-income countries, accounting for 2.7 million disability-adjusted life years annually. Current World Health Organization guidelines recommend controlling STH-associated morbidity through periodic deworming of at-risk populations, including children and women of reproductive age (15-49 years). However, there is increasing interest in community-wide mass drug administration (cMDA) which includes deworming adults who serve as infection reservoirs as a method to improve coverage and possibly to interrupt STH transmission. We investigated determinants of cMDA coverage by comparing high-coverage clusters (HCCs) and low-coverage clusters (LCCs) receiving STH cMDA in three countries. METHODS: A convergent mixed-methods design was used to analyze data from HCCs and LCCs in DeWorm3 trial sites in Benin, India, and Malawi following three rounds of cMDA. Qualitative data were collected via 48 community-level focus group discussions. Quantitative data were collected via routine activities nested within the DeWorm3 trial, including annual censuses and coverage surveys. The Consolidated Framework for Implementation Research (CFIR) guided coding, theme development and a rating process to determine the influence of each CFIR construct on cMDA coverage. RESULTS: Of 23 CFIR constructs evaluated, we identified 11 constructs that differentiated between HCCs and LCCs, indicating they are potential drivers of coverage. Determinants differentiating HCC and LCC include participant experiences with previous community-wide programs, communities' perceptions of directly observed therapy (DOT), perceptions about the treatment uptake behaviors of neighbors, and women's agency to make household-level treatment decisions. CONCLUSION: The convergent mixed-methods study identified barriers and facilitators that may be useful to NTD programs to improve cMDA implementation for STH, increase treatment coverage, and contribute to the successful control or elimination of STH. TRIAL REGISTRATION: The parent trial was registered at clinicaltrials.gov (NCT03014167).