Search

Piaractus brachypomus (Pacú) is the main native fish species cultivated in Peru and holds great potential for growth in aquaculture from the Peruvian Amazon. Between October 2021 and January 2022 in two fish producing farms in the Amazon region of San Martín in Peru, P. brachypomus individuals were examined for parasite evaluation. A total of 6366 monogeneans were isolated from the gills of 30 fish, revealing a prevalence of 100%, with an abundance and mean intensity of 212 parasites per fish. Monogeneans were morphologically identified as Mymarothecium viatorum and Anacanthorus penilabiatus. The genetic divergence in the 28S rDNA gene found among A. penilabiatus sequences was 0.1% and among Anacanthorus spp. it ranged from 0.9% to 7.5%. The genetic divergence found among the M. viatorum sequences was 0.3%. These finding represents the first molecular data of M. viatorum and A. penilabiatus in Peru using the 28S rDNA gene of these monogeneans. The new sequences obtained will contribute to future studies on the phylogenetic relationships among dactylogyrids. However, further research with a broader range of host-parasite samples and additional genetic markers is needed to clarify these relationships and provide stronger support for the phylogenetic positions., Competing Interests: Declaration of competing interest The authors of the manuscript “Molecular Identification of Mymarothecium viatorum and Anacanthorus penilabiatus in Extensive Native Fish Farming Systems of the Peruvian Amazon” declare that this information has not been previously published, has not been submitted to another journal for publication, and the publication of this manuscript is approved by all authors and by the institutions linked in this document. If the manuscript is accepted for publication it will not be considered for republication in any modified version., (Copyright © 2024 The Authors. Published by Elsevier B.V. All rights reserved.)

The Manila clam Ruditapes philippinarum on the west coast of Korea harbors several digenetic trematodes. However, most studies in this region have been restricted to a few sampling sites and the current species designation of some trematodes is largely based on morphology, leaving the molecular phylogenetic position among the Digenea unsolved. Thus, we first provide both morphology and molecular phylogeny of some components in the trematodes community in the Manila clam based on a large-scale survey of 26 sites on the west coast, where well-developed tidal flats serve as large commercial clam culture beds. Our study revealed that the trematodes community in the clams consisted of at least 5 species that belong to 3 families (Himasthlidae, Gymnophallidae, Baccigeridae) and 1 superfamily (Monorchioidea). The life mode of the 5 different species included the metacercaria and sporocyst, with one species (Parvatrema duboisi) utilizing the clams as both the first and/or second intermediate host. Trematode infection prevalences were not evenly distributed among the study sites, although the reasons behind this are yet to be determined. Morphological identification was confirmed with the molecular analyses based on ITS and 28S rDNA; phylogenetic analysis also revealed that Cercaria pectinata infecting the clam gonad should be referred to as Bacciger bacciger hereafter. The present preliminary study provides a crucial baseline that could be further developed in a future study on the digenean trematodes community in the Manila clam., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Inc. All rights reserved.)

Objectives: An integrative taxonomic description of Aponurus laguncula (Lecithasteridae), a digenean parasitic species of Chaetodipterus faber (Acanthuriformes) from Brazilian Southeast, is provided. Morphological techniques, as whole mounted slides, histology and scanning electron microscopy, and molecular analyses supported that integrative description., Methods: Fifteen digenean specimens were stained in hydrochloric carmine and mounted on permanent slides. Two specimens were stained in hematoxylin and eosin following histological routine processing. Four parasites were dehydrated through a graded ethanol series, critical point dried with carbon dioxide and coated with gold to scanning electron microscopy analysis. Sequence of the large ribosomal subunit (28S rDNA) gene was generated and used to construct a phylogeny based on maximum likelihood and Bayesian inference analyses., Results: Morphological description and morphometric data obtained in present study were in accordance with previous studies of the species. Use of another morphological techniques, as scanning electron microscopy and histology, corroborated the observed features of whole mounted slides. Also, they provided a better observation of previous reported characteristics and new features reporting, such as an elongated hermaphroditic duct, a smooth tegument and cells that compose the prostatic gland. The molecular sequence obtained in the present study formed a robust clade with available sequences of species of Aponurus., Conclusions: The integrative taxonomic approach successfully combined morphological observations, including both previously reported features and new descriptions from histological and electron microscopy analyses, with molecular data to identify these specimens as A. laguncula. Moreover, the detailed characterization of structures, such as the gonads in A. laguncula, that would be challenging to analyze using a single technique, was possible. Further molecular studies with less conserved genetic markers should be conducted to understand phylogenetic relationships between Aponurus species., (© 2024. The Author(s), under exclusive licence to Springer Nature Switzerland AG.)

Rastrelliger brachysoma (Bleeker, 1851), the short mackerel, is a dietary staple and of significant economic demand in Southeast Asia and Thailand. However, the demand for short mackerel has precipitated an overfishing crisis, leading to a depletion of fish stocks. Overfishing, coupled with parasitism, may result in a decline in the population of R. brachysoma. Digenetic trematode infection is prevalent in marine fish and has a considerable impact on the overall health of the fish. Here, to identify digenetic trematodes infecting R. brachysoma, we aim to determine the identity, prevalence, and intensity of digenean infections in R. brachysoma from the Gulf of Thailand. A total of 194 short mackerel were obtained from Chon Buri Province, where digeneans were isolated and identified. The molecular identity of the digeneans was confirmed using the nuclear 28S rRNA gene. Of the 194 short mackerel, 100% were found to be infected with digeneans, comprising of Lecithocladium, Prodistomum, Opechona, and Aphanurus. Lecithocladium was the most prevalent (98%) and had the highest intensity of infection (37 mean intensity), followed by Prodistomum (75% prevalence and 17 mean intensity). Our study thus presents the first evidence of digeneans infecting the economically important short mackerel from the Gulf of Thailand. The high infection rate of digenetic trematodes may have implications on the health of R. brachysoma, further driving their population decline. These data underscore the importance of safeguarding fisheries resources in the Gulf of Thailand, and downstream conservation efforts are crucial for evidence-based management decisions to safeguard the long-term sustainability of fish resources., (© 2024. The Author(s).)

Three new species of Gyrodactylus are described from three species of bitterling in Donghu Lake, China: Gyrodactylus ocellorhodei n. sp. from Rhodeus ocellatus; G. sinenorhodei n. sp. from Rhodeus sinensis; and G. acheilorhodei n. sp. from Acheilognathus macropterus. All the three new species showed similar opisthaptor morphology, especially the marginal hooks: all had a slender and perpendicular sickle shaft, and flat sickle base with distinct heel and inner arch which was different from the G. rhodei-group species parasitic on bitterling. Multivariate analyses based on hamulus and marginal hooks suggested that these three new species cannot be completely distinguished, despite some morphology divergence observed in certain less reliable morphometric features, such as hamulus root length, ventral bar total length and process shape. These three new species shared an identical 18S ribosomal RNA gene sequence, while the variation in the Internal Transcribed Spacers (ITS1-ITS2) sequence among them (8.4-11.2%, K2P) far exceeded the 1% ITS sequence difference that had been suggested as a threshold for species delimitation of Gyrodactylus. Phylogenetic analysis based on ITS1-ITS2 showed that all these sequenced Gyrodactylus spp. parasitic on the subfamily Acheilognathinae host formed a monophyletic group. However, a clear differentiation (18.9-20.9%, K2P of ITS1-ITS2) could be found between the subgroup from China (G. ocellorhodei n. sp., G. sinenorhodei n. sp. and G. acheilorhodei n. sp.) and that from Europe (G. rhodei)., Competing Interests: Declaration of competing interest The authors declare no conflict of interest., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Haliotrema Pteroisi: Paperna, 1972 (Monogenoidea: Dactylogyridae) was found parasitizing the gill lamellae of devil firefish, Pterois miles (Bennet) (Perciformes: Scorpaenidae), in the Red Sea off Safaga (26°44'N, 33°56'E), Egypt. The parasite species was described based on morphological features of available specimens and transferred to PlatycephalotremaKritsky and Nitta, 2019 (Dactylogyridae) as Platycephalotrema pteroisi (Paperna, 1972) n. comb. The occurrence of Pl. pteroisi off Safaga, Egypt, represented a range extension for the helminth of about 160 km to the southwest of the southern end of the Gulf of Aqaba. The transfer of the species to Platycephalotrema based on an evaluation of morphological features was supported by an analysis of molecular sequences of the 28S rDNA gene of Pl. pteroisi and 49 other dactylogyrid species. Maximum-likelihood, Bayesian inference, and maximum parsimony analyses of this dactylogyrid sequence data revealed H. pteroisi to nest with significant support within the clade of Platycephalotrema spp. During the literature review of dactylogyrid species infecting scorpionfishes, it was determined that Ancyrocephalus sp. of Dyer et al. from luna lion fish Pterois lunulata Temminck and Schlegel collected off Okinawa-jima, Japan represented an undescribed species of Platycephalotrema., (© American Society of Parasitologists 2024.)

We examined gelatinous zooplankton from off eastern Australia for lepocreadiid trematode metacercariae. From 221 specimens of 17 species of cnidarian medusae and 218 specimens of four species of ctenophores, infections were found in seven cnidarian and two ctenophore species. Metacercariae were distinguished using cox1 mtDNA, ITS2 rDNA and morphology. We identified three species of Prodistomum Linton, 1910 [P. keyam Bray & Cribb, 1996, P. orientale (Layman, 1930), and Prodistomum Type 3], two species of Opechona Looss, 1907 [O. kahawai Bray & Cribb, 2003 and O. cf. olssoni], and Cephalolepidapedon saba Yamaguti, 1970. Two species were found in cnidarians and ctenophores, three only in cnidarians, and one only in a ctenophore. Three Australian fishes were identified as definitive hosts; four species were collected from Scomber australasicus and one each from Arripis trutta and Monodactylus argenteus. Transmission of trematodes to these fishes by ingestion of gelatinous zooplankton is plausible given their mid-water feeding habits, although such predation is rarely reported. Combined morphological and molecular analyses of adult trematodes identified two cox1 types for C. saba, three cox1 types and species of Opechona, and six cox1 types and five species of Prodistomum of which only two are identified to species. All three genera are widely distributed geographically and have unresolved taxonomic issues. Levels of distinction between the recognised species varied dramatically for morphology, the three molecular markers, and host distribution. Phylogenetic analysis of 28S rDNA data extends previous findings that species of Opechona and Prodistomum do not form monophyletic clades., Competing Interests: Declaration of competing interest The authors declare that they have no conflict of interest., (Copyright © 2024 The Authors. Published by Elsevier B.V. All rights reserved.)

Background: Biological invasions pose risks to the normal functioning of ecosystems by altering the structure and composition of several communities. Molluscs stand out as an extensively studied group given their long history of introduction by either natural or anthropogenic dispersal events. An alien population of the lymnaeid species Orientogalba viridis was first sighted in 2009 in southern Spain. In its native range (Australasian), this species is one of the main intermediate hosts of Fasciola hepatica, a major worldwide trematode parasite largely affecting humans, domestic animals and wildlife., Methods: We collected field populations of O. viridis from its native (Malaysia) and invaded (Spain) ranges. We performed detailed morphoanatomical drawings of the species and screened for natural infection of parasites. Individuals were molecularly characterized using ITS2 for comparison with existing sequences in a fine phylogeography study. We founded experimental populations at two different conditions (tropical, 26 °C and temperate, 21 °C) to study the life-history traits of exposed and non-exposed individuals to different F. hepatica isolates., Results: We found a 9% natural prevalence of trematode infection (98% similarity with a sequence of Hypoderaeum conoideum [Echinostomatidae]) in the Spanish field population. The haplotypes of O. viridis found in our study from Spain clustered with Australian haplotypes. Experimental infection with F. hepatica was successful in both experimental conditions but higher in tropical (87% prevalence) than in temperate (73%). Overall lifespan, however, was higher in temperate conditions (mean 32.5 ± 7.4 weeks versus 23.3 ± 6.5) and survivorship remained above 70% during the first 20 weeks. In parasite-exposed populations, life expectancy dropped from an overall 37.75 weeks to 11.35 weeks but still doubled the time for initial cercariae shedding. Cercariae shedding started at day 23 post-exposure and peaked between days 53 and 67 with an average of 106 metacercariae per snail., Conclusions: Whether O. viridis will succeed in Europe is unknown, but the odds are for a scenario in which a major snail host of F. hepatica occupy all available habitats of potential transmission foci, ravelling the epidemiology of fasciolosis. This research provides a comprehensive understanding of O. viridis biology, interactions with parasites and potential implications for disease transmission dynamics, offering valuable insights for further research and surveillance., (© 2024. The Author(s).)

Troglocephalinae n. subfam. is proposed for Spinuris Doran, 1953, Neoheterocotyle Hargis, 1955, Anoplocotyloides Young, 1967, Troglocephalus rhinobatidis Young, 1967 (previously incertae sedis), Nonacotyle pristis Ogawa, 1991, Mehracotyle insolita Neifar, Euzet & Ben Hassine, 2002, Scuticotyle cairae n. gen. et sp., and Brancheocotyle imbricata n. gen. et sp. All members of the proposed new subfamily are gill parasites of shovelnose rays of the order Rhinopristiformes. The subfamilies Heterocotylinae Chisholm, Wheeler & Beverley-Burton, 1995, and Dasybatotreminae Bychowsky, 1957, are amended to exclude Spinuris, Nonacotyle, Neoheterocotyle, and Anoplocotyloides and Mehracotyle, respectively. Heterocotylinae includes gill parasites of members of the orders Myliobatiformes and Torpediniformes. Dasybatotreminae includes parasites of the gills and pharyngeal cavity of members of the orders Myliobatiformes and Rajiformes. A revised phylogeny of the Monocotylidae Taschenberg, 1879 is presented and discussed, based on 28S rDNA sequences, including new sequences for Myliocotyle pteromylaei Neifer, Euzet & Ben Hassine, 1999, Heterocotyle tokoloshei Vaughan & Chisholm, 2010, Neoheterocotyle robii Vaughan & Chisholm, 2010, and the two newly proposed species and genera. Additional locality records are also provided for Monocotylidae from off South Africa., (© 2024. The Author(s).)

Glypthelmins quieta is a frog trematode native to North and Central America. This trematode was recently detected in Japan in the American bullfrog Lithobates catesbeianus, which was introduced from North America to Japan. As the first intermediate host of G. quieta, typically a snail, has not yet been identified in Japan, we conducted a snail survey in eastern Japan to screen for an intermediate host using DNA barcoding based on the nuclear 28S ribosomal RNA and mitochondrial cytochrome c oxidase subunit 1. We sampled 3 different snail species, Orientogalba ollula, Physella acuta, and Sinotaia quadrata histrica (157 individuals in total), and only the freshwater snail Physella acuta, which is also believed to have been introduced from North America to Japan, had sporocysts of G. quieta in its hepatopancreas. The introduction of the intermediate and definitive hosts from North America may have facilitated the invasion of G. quieta into Japan.

Monogenean trematodes, particularly those belonging to the Diplectanidae family, are significant metazoan parasites with substantial implications for aquaculture expansion. This study, investigatied the occurrence, prevalence, and pathological impact of Diplectanum spp. in European seabass (Dicentrarchus labrax) across three distinct Egyptian fish farms. During 2021-2022, we sampled 1800 European seabass (Dicentrarchus labrax) from three Egyptian fish farms (600 fish per farm). Farms 1 and 2 used semi-intensive earthen pond systems, while Farm 3 utilized an intensive floating cage system. Employing Clinical, post-mortem, parasitological, and molecular examination technique. Pathological lesions were identified, including skin and gill discoloration, emaciation, and internal organ abnormalities. Seasonal prevalence exhibited significant variations between farms, with highest rates observed in spring and Farm 3 reached an overall peak prevalence of 84.67 %. Parasitological examination distinguished two Diplectanum species morphologically, while molecular techniques exhibited limited specificity. Histopathology unveiled damage to gill, liver, spleen, kidney, and intestine, attributed to Diplectanum haptors including inflammation and internal bleeding, potentially leading to secondary infections. Molecular identification via PCR targeting ITS and 28SrDNA genes, revealing similar band sizes for the two Diplectanum species, indicating limited intraspecific genetic diversity. The study emphasizes investigating parasitic infections' prevalence and impact in aquaculture, necessitating robust molecular techniques for species differentiation. This study underscores the importance of investigating the prevalence and impact of parasitic infections in aquaculture. It highlights the need for robust molecular techniques to differentiate species. By focusing on Diplectanum spp. infections in D. labrax, the study offers valuable insights into managing parasites in aquaculture effectively., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper, (Copyright © 2024 Elsevier B.V. All rights reserved.)

Monogeneans are parasitic flatworms that represent a significant threat to the aquaculture industry. Species like Neobenedenia melleni (Capsalidae) and Rhabdosynochus viridisi (Diplectanidae) have been identified as causing diseases in farmed fish. In the past years, molecular research on monogeneans of the subclass Monopisthocotylea has focused on the generation of genomic and transcriptomic information and the identification in silico of some protein families of veterinary interest. Proteomic analysis has been suggested as a powerful tool to investigate proteins in parasites and identify potential targets for vaccine development and diagnosis. To date, the proteomic dataset for monogeneans has been restricted to a species of the subclass Polyopisthocotylea, while in monopisthocotyleans there is no proteomic data. In this study, we present the first proteomic data on two monopisthocotylean species, Neobenedenia sp. and R. viridisi, obtained from three distinct sample types: tissue, excretory-secretory products (ESPs), and eggs. A total of 1691 and 1846 expressed proteins were identified in Neobenedenia sp. and R. viridisi, respectively. The actin family was the largest protein family, followed by the tubulin family and the heat shock protein 70 (HSP70) family. We focused mainly on ESPs because they are important to modulate the host immune system. We identified proteins of the actin, tubulin, HSP70 and HSP90 families in both tissue and ESPs, which have been recognized for their antigenic activities in parasitic flatworms. Furthermore, our study uncovered the presence of proteins within ESPs, such as annexin, calcium-binding protein, fructose bisphosphate aldolase, glutamate dehydrogenase, myoferlin, and paramyosin, that are targets for immunodiagnostic and vaccine development and hold paramount relevance in veterinary medicine. This study expands our knowledge of monogeneans and identified proteins that, in other platyhelminths are potential targets for vaccines and drug discovery., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

A new species of hemiurid trematode found on the gills and in the aorta of the pelagic thresher Alopias pelagicus from the eastern Pacific, off Costa Rica, is described based on an integrative taxonomic approach that includes the use of light and scanning electron microscopy, and 28S rDNA sequencing. Phylogenetic analysis was also performed to explore, for the first time, the relationships of a member of the subfamily Otiotrematinae within the suborder Hemiurata. Paronatrema davidbowiei n. sp. can be distinguished from the congeners by having tegumental spines on the dorsal surface of the forebody, papillae on the oral sucker, and different morphology or number of testicular follicles. BLAST analysis revealed that sequences of Paronatrema davidbowiei n. sp. had the highest degree of similarity with Hirudinella spp. (Hirudinellidae). Results from Maximum Likelihood and Bayesian phylogenetic analyses, returning trees with the exact same topology and strong branch support, distinguished between the two superfamilies included in the suborder Hemiurata: Azygioidea and Hemiuroidea. Our analysis placed the new species in a clade with Copiatestes filiferus, the only existing sequence of the family Syncoeliidae., Competing Interests: Declaration of competing interest The authors declare that there is no conflict of interest., (Copyright © 2024. Published by Elsevier B.V.)

Echinochasmids are a group of globally distributed digeneans, and the adults are found in the intestines of birds, mammals and reptiles. In the Neotropical region of Mexico, adult specimens were obtained from seven fish-eating bird species in six localities, whereas specimens of Stephanoprora aylacostoma were obtained experimentally in Argentina. Morphologically, the new specimens from the Neotropical region of Mexico were identified as Stephanoprora uruguayense and an undescribed species of Echinochasmus. Sequences for two nuclear (large subunit (28S) and internal transcribed spacer from DNA ribosomal (ITS1-5.8S-ITS2)) molecular markers were generated and analysed together with other sequences downloaded from GenBank. The phylogenies obtained with each molecular marker indicated that Echinochasmus is paraphyletic and agreed with previous phylogenetic studies. The first cluster included the type species (E. coaxatus, which has 24 head-collar spines) plus three congeneric species. The second cluster contained species of Echinochasmus plus Stephanoprora, including the species analysed herein, S. uruguayense, S. aylacostoma (with 22 head-collar spines) and Echinochasmus sp. (with 20 head-collar spines), which formed three independent subclades, allowing us to recognize a lineage that was described morphologically as a new species. Echinochasmus ostrowskiae n. sp. can be distinguished from its congeners by having a head collar with 20 spines in a single row, seven spines on each edge and three angle spines, and a pharynx with an irregular edge and by the body, egg and collar spine sizes. Additionally, new host and locality records for S. uruguayense are presented, expanding its geographical distribution range in the Americas., Competing Interests: Declaration of competing interest The authors declare there are no conflicts of interest., (Copyright © 2024 The Authors. Published by Elsevier B.V. All rights reserved.)

In Japan, trematodes of the family Ochetosomatidae are not naturally distributed. However, the introduced ochetosomatid Ochetosoma kansense (Crow, 1913) has been reported from the oral cavity of native snakes in western Japan since 2010s. In this study, trematodes were isolated from the oral cavities of the native Japanese snakes, Elaphe quadrivirgata (Boie, 1826), E. climacophora (Boie, 1826), and Rhabdophis tigrinus (Boie, 1826), in the central Kanto region of eastern Japan. Morphological and molecular analyses of the isolated trematodes revealed that all trematodes were identifiable to a newly introduced ochetosomatid species to Japan, O. elongatum (Pratt, 1903), which originated from North America; Lechriorchis tygarti Talbot, 1933 was synonymized with O. elongatum based on identical molecular data and morphological similarity. To identify first intermediate hosts of O. elongatum, seven freshwater snail species were examined in eastern Japan. Molecular analysis was used to identify O. elongatum sporocysts in the freshwater snail Physella acuta (Draparnaud, 1805), which also originated from North America. The other six species did not host O. elongatum, suggesting that Ph. acuta is the only first intermediate host of O. elongatum in Japan. Although O. elongatum has been detected in Japan, its invasion route and period of introduction are unclear. Frequent imports of freshwater snails and wild snakes from North America, after the 1990s and 2005, respectively, presumably introduced O. elongatum in Japan., Competing Interests: Declaration of competing interest None., (Copyright © 2024 Elsevier B.V. All rights reserved.)

While biogeographic patterns of free-living organisms are well documented, the biogeography of parasitic fauna remains largely unclear. Due to morphological similarities, parasites are often difficult to identify without the aid of molecular genetics, further complicating the interpretation of their biogeographic patterns. We investigated trematode parasites infecting the East Asian freshwater snail Semisulcospira libertina to understand their biogeography and to evaluate how molecular approaches influence the interpretation of biogeographic patterns of the trematode fauna. We identified 46 genetically delimited species from 19 morphologically distinguishable trematodes infecting S. libertina and found that their species richness was negatively correlated to latitude. We also found that potential definitive host (fishes) richness and host body size were positively correlated with trematode species richness, suggesting that host attributes are essential factors shaping the biogeographic pattern in trematodes. These trends were observed irrespective of species identification methods, demonstrating that classical morphological identification can also effectively identify the latitudinal gradient pattern in trematodes. We further detected the distance decay of similarity in trematode communities, although this trend was only detectable in the biogeographic dataset based on molecular identification. Our study showed that morphological identification sufficiently reflects the latitudinal richness gradient while molecular identification is essential to estimate accurate local species richness and increase the resolution of the large-scale pattern of population similarities in the trematode communities., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024. Published by Elsevier B.V.)

Background: Gastropods of the genus Biomphalaria (Family Planorbidae) are exploited as vectors by Schistosoma mansoni, the most common causative agent of human intestinal schistosomiasis. Using improved genomic resources, overviews of how Biomphalaria responds to S. mansoni and other metazoan parasites can provide unique insights into the reproductive, immune, and other systems of invertebrate hosts, and their responses to parasite challenges., Results: Using Illumina-based RNA-Seq, we compared the responses of iM line B. glabrata at 2, 8, and 40 days post-infection (dpi) to single infections with S. mansoni, Echinostoma paraensei (both digenetic trematodes) or Daubaylia potomaca (a nematode parasite of planorbid snails). Responses were compared to unexposed time-matched control snails. We observed: (1) each parasite provoked a distinctive response with a predominance of down-regulated snail genes at all time points following exposure to either trematode, and of up-regulated genes at 8 and especially 40dpi following nematode exposure; (2) At 2 and 8dpi with either trematode, several snail genes associated with gametogenesis (particularly spermatogenesis) were down-regulated. Regarding the phenomenon of trematode-mediated parasitic castration in molluscs, we define for the first time a complement of host genes that are targeted, as early as 2dpi when trematode larvae are still small; (3) Differential gene expression of snails with trematode infection at 40dpi, when snails were shedding cercariae, was unexpectedly modest and revealed down-regulation of genes involved in the production of egg mass proteins and peptide processing; and (4) surprisingly, D. potomaca provoked up-regulation at 40dpi of many of the reproduction-related snail genes noted to be down-regulated at 2 and 8dpi following trematode infection. Happening at a time when B. glabrata began to succumb to D. potomaca, we hypothesize this response represents an unexpected form of fecundity compensation. We also document expression patterns for other Biomphalaria gene families, including fibrinogen domain-containing proteins (FReDs), C-type lectins, G-protein coupled receptors, biomphalysins, and protease and protease inhibitors., Conclusions: Our study is relevant in identifying several genes involved in reproduction that are targeted by parasites in the vector snail B. glabrata and that might be amenable to manipulation to minimize their ability to serve as vectors of schistosomes., (© 2024. The Author(s).)

There are several species of gymnophallid digeneans in the genus Parvatrema that are unique in developing metacercariae that reproduce by parthenogenesis in the second intermediate host. Transmission of these digeneans takes place in coastal ecosystems of the North Pacific and North Atlantic seas. The first intermediate hosts are bivalves, the second ones are gastropods, and the definitive hosts are migratory birds. We integrated data accumulated over 25 years of research and differentiated a complex of five closely related species. They differ in the molluscan second intermediate hosts, distribution ranges, and life cycles patterns. The type I life cycle includes two generations of parthenogenetic metacercariae, followed by development of metacercariae which are invasive for the definitive host. In the type II life cycle, the number of generations of parthenogenetic metacercariae is unlimited, and they can also produce cercariae. These cercariae emerge into the environment and can infect new individuals of the second intermediate host. We conclude that the type I life cycle is a derived option that has evolved as a better fit to transmission in the unstable conditions in the intertidal zone. Another evolutionary trend in Parvatrema is transition from inhabiting the extrapallial space of the gastropod second intermediate host to endoparasitism in its mantle and internal organs. rDNA sequence analysis highlighted that Parvatrema spp. with parthenogenetic metacercariae form a monophyletic clade and suggested the Pacific origin of the group, with two transfers to the North Atlantic and colonisation of new second intermediate host species. Apparently the group formed in the late Pliocene-Pleistocene and diversified as a result of recurrent isolation in inshore refugia during glacial periods. We argue that parthenogenetic metacercariae in Parvatrema may serve as a model for early digenean evolution, demonstrating the first steps of adopting the molluscan first intermediate host and becoming tissue parasites., (Copyright © 2024 Australian Society for Parasitology. Published by Elsevier Ltd. All rights reserved.)

Freshwater snails are integral to local ecosystems as a primary food source for various vertebrate species, thereby contributing significantly to ecological food webs. However, their role as intermediate hosts also makes them pivotal in the transmission of parasites. In Australia, research on freshwater snails has predominantly focused on their role as intermediate hosts for livestock parasites, while there has been limited exploration of the impact of these parasites on snail health and population dynamics. The aim of this study was to determine parasitic infection in freshwater snails. This study was conducted in the south-eastern region of Australia, in 2022. A total of 163 freshwater snails from four different species were collected and examined in the Murrumbidgee catchment area in the southeastern part of Australia during the Southern Hemisphere summer and autumn months (February to May). The species included Isidorella hainesii, Glyptophysa novaehollandica, Bullastra lessoni (endemic species), and Physella acuta (an introduced species). Through the analysis of sequence data from the various regions of the nuclear ribosomal DNA, we determined that the Digenea species in this study belonged to three distinct species, including Choanocotyle hobbsi, Petasiger sp. and an unidentified species belonging to Plagiorchioidea. Additionally, analysis of the sequences from Nematoda found in this study, revealed they could be categorized into two separate taxa, including Krefftascaris sp. and an unidentified nematode closely associated with plant and soil nematodes. This research holds significant implications for the future understanding and conservation of Australian freshwater ecosystems. Most parasites found in the present study complete their life cycle in snails and turtles. As many of freshwater snail and turtle species in Australia are endemic and face population threats, exploring the potential adverse impacts of parasitic infections on snail and turtle health, is crucial for advancing our understanding of these ecosystems and also paving the way for future research and conservation efforts. While none of the native snail species in the present study have been listed as endangered or threatened, this may simply be attributed to the absence of regular population surveys., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Author(s). Published by Elsevier Inc. All rights reserved.)

Trematodes of the genus Leucochloridium exhibit an unusual transmission strategy among mollusks (intermediate host). The fully developed sporocyst, housing encysted metacercariae, displays vivid coloration and rhythmic activity in the snail's tentacle, mimicking insect larvae. These strategies attract insectivorous birds, their final hosts, thereby increasing the chances of completing their life cycle. In South America, the reports of adults and larval stages of Leucochloridium are scarce. Brown-banded broodsac of Leucochloridium sp. were obtained from Omalonyx unguis collected in a shallow lake from Corrientes Province, Argentina. Here, we morphologically characterized the larval stages (broodsac and metacercaria), identified the parasite through DNA sequences from nuclear 28S-rRNA (28S) and the mitochondrial cytochrome c oxidase I (COI) genes, and explored its evolutionary affinities with the Leucochloridium species available in GenBank. The present broodsac displays brown bands, with a yellowish background in the first two-thirds and yellowish-white in the last third. Based on morphological comparisons, the broodsac and metacercaria described in this study could not be conclusively categorized under any known South American species of Leucochloridium. In relation to the phylogenetic reconstructions, Leucochloridium sp. consistently clustered with L. perturbatum, and species delimitation analyses resulted in recognized Leucochloridium sp. from Argentina as a distinct species. The DNA sequences obtained in this study constitute the first genetic data generated for sporocyst broodsacs in South America. Future studies, incorporating morphology, genetic, and biological data, will be essential for both species identification and the elucidation of leucochloridiid diversity in the region., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024. Published by Elsevier Inc.)

We herein provide a supplemental description of Nomasanguinicola dentata (Paperna, 1964) Warren and Bullard, 2023 (Digenea: Sanguinicolidae) and provide a revised 28S phylogeny to test relationships among freshwater fish blood flukes. We examined the heart of three African sharptooth catfish, Clarias gariepinus (Burchell, 1822) Teugles, 1982 from the Kavango River (northeastern Namibia) that was infected with adults of N. dentata. This blood fluke differs from N. canthoensis by having a body 5.3-6.7 longer than wide (vs. 3.5-4.6), an anterior esophageal swelling 7-8% (vs. 14-24%) of total esophageal length, a posterior esophageal swelling 3-5% (vs. 8-10%) of total esophageal length, a pre-cecal (vs. wholly post-cecal) testis, and an ovary that does not extend laterally beyond the nerve cords. The 28S sequence for N. dentata differed from that of N. canthoensis by 144 bp (9% difference). The phylogenetic analysis recovered these species as sister taxa and Sanguinicolidae as monophyletic. This is the first report of a fish blood fluke from sub-Saharan Africa, and the first report of a species of Nomasanguinicola from Africa in ∼40 yrs., Competing Interests: Declaration of competing interest No author has a competing interest., (Copyright © 2023. Published by Elsevier B.V.)

The Cyathocotylidae Mühling, 1898 is a family of primitive diplostomoid trematodes important for understanding the evolution of the superfamily Diplostomoidea. However, cyathocotylids remain poorly studied with the use of molecular techniques. In this study we sequenced the 5.8S + ITS2 region, 28S rRNA , and cox1 genes of two cyathocotylid species and obtained new morphological data on them. We propose Georduboisia nom. nov. instead of the preoccupied name Duboisia Szidat, 1936 (junior homonym of Duboisia Stremme, 1911). Adults of Georduboisia cf. teganuma (Ishii, 1935) and Paracoenogonimus ovatus Katsurada, 1914 were collected from fish-eating birds in the south of the European part of Russia. Georduboisia cf. teganuma was very similar to G. teganuma but differed from it in the shape of the testes. The 28S rRNA gene dataset provided the best-resolved phylogeny of the Cyathocotylidae to date. In the phylogram based on partial sequences of this gene, P. ovatus was close to members of Holostephanoides Dubois, 1983, Neogogatea Chandler & Rausch, 1947 and Gogatea Szidat, 1936. Georduboisia cf. teganuma clustered with members of Cyathocotyle Mühling, 1896 and Holostephanus Szidat, 1936. Phylogenetic analysis based on the 5.8S + ITS2 dataset showed that adults of P. ovatus examined in our study were conspecific with the metacercariae from the musculature of fish collected in Hungary and Italy. It also revealed probable misidentifications of larvae and adults of cyathocotylids whose sequences are deposited in GenBank NCBI.

Ochoterenatrema Caballero, 1943 is a genus of lecithodendriid digeneans that prior to this study included 8 species parasitic in bats in the Western Hemisphere. Species of Ochoterenatrema possess a unique morphological feature in form of the pseudogonotyl on the sinistral side of the ventral sucker. In this study, we describe 2 new species of Ochoterenatrema from bats in Ecuador. The new species are readily differentiated from their congeners by a combination of morphological characters, including the distribution of vitelline follicles, length of oesophagus, sucker ratio and the body shape, among other features. We have generated partial nuclear 28S rDNA and mitochondrial cox1 gene DNA sequences from both new species. The newly obtained sequences were used to differentiate among species and study the phylogenetic interrelationships among Ochoterenatrema spp. The internal topology of the clade was weakly supported, although the cox1 tree was much better resolved than the 28S tree. Comparison of sequences revealed 0-1.2% interspecific divergence in 28S and 3.3-20.5% interspecific divergence in cox1 among Ochoterenatrema spp. The new findings demonstrate that bats in South America likely harbor multiple additional undescribed species of Ochoterenatrema. More extensive sampling from broader geographic and host ranges, especially in North America, should allow for a better understanding of the evolution of host associations and morphological traits of this lineage of lecithodendriid digeneans., (© 2024. The Author(s).)

Clinostomum is a cosmopolitan genus of trematodes that infect piscivorous birds, freshwater molluscs, freshwater fish and amphibians. Herein, a novel species of Clinostomum is described based on morphological and molecular data from an adult in the oral cavity of the great blue heron Ardea herodias and metacercariae collected from the gills and skin of American bullfrog tadpoles Rana catesbeiana. The novel species shares similar qualitative and quantitative morphological features with a congener, Clinostomum marginatum, which has overlap in host and geographic distribution. The most notable morphological difference when compared to C. marginatum is the greater posterior testis length of the novel species. Molecular data resolved similarities with morphological comparisons to nominal species and supports the establishment of a novel species. Molecular data include partial small ribosomal subunit (18S rRNA gene), ribosomal internal transcribed spacer regions (ITS1, 5.8S rRNA gene, and ITS2), partial large ribosomal subunit (28S rRNA gene), cytochrome c oxidase subunit 1 gene (cox1), and nicotinamide adenine dinucleotide dehydrogenase subunit 1 gene (nad1) sequences. Phylogenetic analyses place the novel species in a sister clade to C. marginatum. Morphological and molecular data, combined with phylogenetic analyses support the establishment of Clinostomum dolichorchum n. sp., (© 2024. The Author(s), under exclusive licence to Springer Nature B.V.)

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., (© 2024. The Author(s).)

Dicrocoeliid trematodes were detected from Iwasaki's snail-eating snake Pareas iwasakii in Iriomote Island, Okinawa Prefecture, Japan, and described as a new species Paradistomum dextra n. sp. in the present study. This new species can be distinguished from the type series of the other members of the genus based on size of eggs and morphological characteristics of body, oral and ventral suckers, and reproductive organs. However, the new species was hard to distinguish from Paradistomum megareceptaculum infecting snakes in Japan, including Iriomote Island where is the type locality of the new species, because it is closely similar to some part of the broad range of morphological variations in P. megareceptaculum. On the other hand, a partial sequence of 28S ribosomal DNA clearly distinguished these two species. Moreover, the new species' host snake Pareas iwasakii is reported to exclusively feed on land snails while host snakes of P. megareceptaculum feed on small vertebrates, indicating that the new species is also ecologically different from P. megareceptaculum. We also redescribed P. megareceptaculum based on adults sampled in this study and past studies to record the morphological variations of this species., (© 2024. The Author(s), under exclusive licence to Springer Nature B.V.)

The deepest recorded depth for trematodes currently stands at approximately 6200 m. This depth record was achieved solely through sequence datasets of Lepidapedon sp. obtained from a gastropod. Given that trematodes of this genus typically use fish as definitive hosts, the origin of the trematode sequence was thought to be larval stages. However, the specific species remained unclear owing to the absence of reported adult-stage sequences. In the present study, we definitively identified the deepest trematode as Lepidapedon oregonense by comparing 28S ribosomal DNA sequences from adult worms from the macrourid fish Coelorinchus gilberti with data from the gastropod in the previous study.

The genus Ancyrocephalus sensu lato is a large assemblage of species of dactylogyrid monopisthocotyleans without clear taxonomic boundaries. Despite an urgent need for revision, only three representatives of this taxon have been molecularly characterised so far. We found specimens of Ancyrocephalus curtus , a previously non-genotyped species, in gills of Perccottus glenii caught in the River Syumnyur, Amur Basin, Russia. The aim of this study was to assess the phylogenetic position of this parasite using partial sequences of 28S rRNA gene. In the phylogenetic tree, A. curtus appeared as a sister taxon to the dactylogyrine genus Gobioecetes. The new molecular evidence supports the hypothesis about the non-monophyletic status of Ancyrocephalus sensu lato.

Presenting new molecular and scanning electron microscope (SEM) features, this study gives additional data to the better knowledge of Thaparocleidus vistulensis (Siwak, 1932) (Monopisthocotyla, Ancylodiscoididae), a parasite of the European catfish Silurus glanis Linnaeus, 1758 (Siluriformes, Siluridae) cultured in a commercial fish farm in Hungary. In addition, notes on the early development of sclerotized anchors are also provided. The main morphological difference of T. vistulensis compared to other congeneric species is associated with the male copulatory organ, which exhibits 5-7 loops in the middle of the penis length and a long open V-shaped sclerotized accessory piece, dividing terminally into two parts, securing the terminal part of the penis tube. The present study provides for the first time molecular characterization data based on the 2694 bp long nucleotide sequence of rDNA (ITS1, 5.8S, ITS2, and flanked with partial 18S and partial 28S) submitted in GenBank with the accession number OR916383. A phylogenetic tree based on ITS1 sequences supports a well-defined clade including T. vistulensis, forming a sister group with T. siluri, a species-specific monopisthocotylan parasite to S. glanis. The morphological characterization of T. vistulensis, especially for the male copulatory organ, together with the molecular data in the present study, extends knowledge about this monopisthocotylan species and provides new information for future phylogeny studies., (© 2024. The Author(s).)

Two new species of Urocleidoides are described from the gills of Pseudanos trimaculatus (Characiformes: Anostomidae) from the coastal drainages of the Eastern Amazon, Brazil. Urocleidoides itabocaensis n. sp. is characterized by having a sclerotized, tubular, sigmoid male copulatory organ (MCO), a circular sclerotized tandem brim associated with the base of the MCO; an accessory piece articulated with the MCO, a V-shaped, divided into two subunits, distal subunit spoon-shaped; and a vaginal pore dextroventral with opening marginal. Urocleidoides omphalocleithrum n. sp. is characterized by presenting a C-shaped or sigmoid MCO; an accessory piece articulated with the MCO, L-shaped, divided into two subunits, distal subunit gutter-shaped; a vaginal pore dextroventral with opening marginal; and a ventral bar broadly V-shaped, with anteromedial projection. Molecular phylogenetic analysis based on partial 28S rDNA and COI mtDNA genes indicate that U. itabocaensis n. sp. and U. omphalocleithrum n. sp. are closely related and appear as a sister group to other Urocleidoides species (U. paradoxus, U. digitabulum and U. sinus) parasitizing anostomid fishes. This study represents the first record of monogenoids from the gills of P. trimaculatus for the Eastern Amazon., (© 2024. The Author(s), under exclusive licence to Springer Nature B.V.)

The humpback whale (Megaptera novaeangliae) is a cosmopolitan migratory, seasonal mysticete that frequents the Brazilian coast. Strands of specimens may occur during the migratory stay in the country. In 2021 and 2022, three live humpback whales stranded on the coast of Rio Grande do Sul and Santa Catarina states in southern Brazil. After euthanasia, specimens were necropsied, and organs were thoroughly examined for lesions. Grossly, in all three cases, the liver exhibited multifocal, irregular, firm, white areas on the hepatic capsule, which extended into the parenchyma. On the cut surface, the livers were yellow to pale brown with orangish to greenish areas, the bile ducts were prominent, thickened, and severely dilated, and leaf-shaped flukes were found inside of them. Additionally, one case showed moderate atrophy of the right hepatic lobe. The histological findings included dilation of bile ducts, hyperplasia of the bile duct epithelium, marked inflammatory infiltration of lymphocytes, plasma cells, and eosinophils, and portal fibrosis. The parasite Brachycladium goliath was both morphologically and molecularly identified based on diagnostic key for trematodes and the original description of the species, and the amplification and sequencing of the ITS-2 region, respectively. Even though hepatic injury was not the primary cause of stranding, it may have contributed to the debilitation of the whales. To the authors' knowledge, this is the first study that reports M. novaeangliae as a definitive host of B. goliath and that describes the lesions caused by the parasite in cetaceans., (© 2024. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

Pleustonic organisms form an important part of pelagic ecosystems by contributing to pelagic trophic chains and supporting connectivity between oceanic habitats. This study systematically analysed the trematode community harboured by pleustonic molluscs and cnidarians from offshore Queensland, Australia. Four mollusc and three cnidarian species were collected from beaches of North Stradbroke Island, Queensland. Two mollusc species and all three cnidarians harboured large numbers of hemiuroid metacercariae (Trematoda: Hemiuroidea). Eight taxa from four hemiuroid families (Accacoeliidae, Didymozoidae, Hemiuridae and Sclerodistomidae) were distinguished via molecular sequencing. Four of those taxa were identified to species. All trematode taxa except one didymozoid were shared by two or more host species; five species occurred in both gastropods and cnidarians. It is hypothesised that the life-cycles of these hemiuroids are highly plastic, involving multiple opportunistic pathways of metacercarial transmission to the definitive hosts. Transmission and the use of pleuston by hemiuroids likely varies with sea surface use and ontogenetic trophic shifts of apex predators. The small number of trematode species found in pleuston is consistent with significant ecological specificity, and the inference that other pelagic trematodes use alternative pathways of transmission that do not involve pleustonic organisms. Such pathways may involve i) pelagic hosts exclusively; ii) benthic or demersal hosts exclusively, consumed by apex predators during their dives; or iii) both benthic and pelagic hosts in transmission chains dependent on vertical migrations of prey. The influence of the connectivity of open-ocean ecosystems on parasite transmission is identified as an area in critical need of research., (Copyright © 2023 The Authors. Published by Elsevier Ltd.. All rights reserved.)

Environmental DNA (eDNA) surveys promise to be a sensitive and powerful tool for the detection of trematodes. This can contribute to the limited studies on trematode ecology, specifically in aquatic ecosystems. Here, we developed species-specific primer and probe sets for Moliniella anceps, Opisthioglyphe ranae, and Plagiorchis multiglandularis cercariae and applied a novel eDNA qPCR assay to detect larval trematodes quantitatively. We evaluated the effectiveness of the assays using filtered lake water samples collected from different sites of Lake Fadikha and Kargat River Estuary in Lake Chany, Russia, showing high species specificity and sensitivity in all 3 assays. Further, all 3 assays had high efficiencies ranging from 94.9 to 105.8%. Moliniella anceps, O. ranae, and P. multiglandularis were detected in the environmental water samples through real-time PCR. Thus, we anticipate that our approach will be beneficial for biomonitoring, measuring, and managing ecological systems., (© American Society of Parasitologists 2024.)

Invasive snails are associated with ecological problems in freshwater bodies worldwide. However, their impact on the transmission of digenean infections remain underreported. In the present study, 1708 specimens representing four snail species were sampled from four impoundments in the Limpopo River system in South Africa. Gyraulus chinensis (Planorbidae), Physella acuta (Physidae) and Pseudosuccinea columella (Lymnaeidae), which are invasive, were found in all the sampling sites. In contrast, the native lymnaeid Radix natalensis occurred at only one study site. Digeneans were observed only from R. natalensis (prevalence = 49%) and Ps. columella (prevalence = 23%). Morphological and genetic analyses revealed four digeneans: Fasciola nyanzae, Orientocreadium sp., Petasiger sp. and Patagifer vioscai. Pseudosuccinea columella was infected by the four digeneans while R. natalensis harboured only Orientocreadium sp. and Petasiger sp. Partial sequences of Orientocreadium sp. from the current study differed from congeners whose DNA data are available on GenBank, by p-distances of at least 1.84 and 2.2% for 28S and the internal transcribed spacer (ITS) rDNA, respectively. Phylogenetic analyses demonstrated that the present species is sister to Orientocreadium batrachoides. Genetic and phylogenetic data based on 28S and ITS rDNA suggested that Petasiger sp. from the present study and isolates of three unidentified Petasiger spp. from Kenya, Hungary and Australia, were representatives of the same species. This is the first known report of Orientocreadium, Petasiger and Patagifer from Ps. columella. The occurrence of F. nyanzae in Ps. columella indicates spillback from R. natalensis. These findings echo the concerns raised in previous studies about the potential role of Ps. columella in the amplification of digenean diseases in its introduced range. Phylogenetic analyses of partial sequences of the cytochrome c oxidase subunit 1 mitochondrial gene (cox1) showed multiple lineages of Ps. columella in North and South America. Pseudosuccinea columella specimens from the present study belong to an invasive genotype that has spread globally and has been reported from Zimbabwe, Egypt, Portugal, Australia, Argentina, Colombia and New Mexico (USA). Physella acuta from the current study had a stronger genetic relationship with isolates from Canada and Iceland, than with isolates from other parts of Africa, suggesting several invasion routes into Africa. This is the first known DNA characterisation of G. chinensis from Africa. Phylogenetic reconstruction indicated multiple exit events of G. chinensis from Asia into Europe and Africa. South African isolates clustered in a recent branch containing isolates from the Czech Republic and Hong Kong, China. Considering the presence of invasive snails in all the sampling sites in the present study, it is necessary to investigate the factors that enhance their establishment and to monitor their effects on the native snail populations., (Copyright © 2024 The Author(s). Published by Elsevier Ltd.. All rights reserved.)

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.

As part of a parasitological survey, several specimens of two new monopisthocotylean species, Neotetraonchus celsomanueli sp. nov. and N. peruvianus sp. nov. (Dactylogyridea, Dactylogyridae), were collected from the gill filaments of the Peruvian sea catfish Galeichthys peruvianus (Siluriformes, Ariidae) off Puerto Pizarro, Tumbes region, Peru. Neotetraonchus celsomanueli sp. nov. is characterised by an MCO with a T-shaped distal end and an accessory piece that is ribbed and expanded proximally with a worm-shaped termination. Neotetraonchus peruvianus sp. nov. is typified by its MCO, which has a sledgehammer-shaped distal end and an accessory piece with a claw-shaped distal end. Additionally, N. peruvianus sp. nov. is characterised by its jellyfish-shaped onchium. A partial 28S rDNA sequence was obtained from N. celsomanueli sp. nov., and a phylogenetic analysis was conducted. This analysis revealed the phylogenetic position of Neotetraonchus celsomanueli sp. nov. within a clade comprising monopisthocotylean parasites of diadromous and marine ariid catfishes, including Hamatopeduncularia spp., Chauhanellus spp., Thysanotohaptor Kritsky, Shameem, Kumari & Krishnaveni, , and Neocalceostomoides spinivaginalis Lim, 1995. This finding brings the number of known Neotetraonchus species to seven and represents the first described Neotetraonchus species infecting marine catfishes from Peru.

Background: Prior research suggests that trematode rediae, a developmental stage of trematode parasites that reproduce clonally within a snail host, show evidence of division of labor (DOL). Single-species infections often have two morphologically distinct groups: small rediae, the 'soldiers', are active, aggressive, and do not appear to reproduce; large rediae, the 'reproductives', are larger, sluggish, and full of offspring. Most data supporting DOL come from trematodes infecting marine snails, while data from freshwater trematodes are more limited and generally do not supported DOL. The shorter lifespan typical of freshwater snails may partially explain this difference: defending a short-lived host at the expense of reproduction likely provides few advantages. Here, we present data from sixty-one colonies spanning twenty species of freshwater trematode exploring morphological and behavioral patterns commonly reported from marine trematodes believed to have DOL., Methods: Trematode rediae were obtained from sixty-one infected snails collected in central Vermont, USA. A portion of the COI gene was sequenced to make tentative species identifications ('COI species'). Samples of rediae were photographed, observed, and measured to look for DOL-associated patterns including a bimodal size distribution, absence of embryos in small rediae, and pronounced appendages and enlarged pharynges (mouthparts) in small rediae. Additional rediae were used to compare activity levels and likelihood to attack heterospecific trematodes in large vs . small rediae., Results: Many of the tests for DOL-associated patterns showed mixed results, even among colonies of the same COI species. However, we note a few consistent patterns. First, small rediae of most colonies appeared capable of reproduction, and we saw no indication (admittedly based on a small sample size and possibly insufficient attack trial methodology) that small rediae were more active or aggressive. This differs from patterns reported from most marine trematodes. Second, the small rediae of most colonies had larger pharynges relative to their body size than large rediae, consistent with marine trematodes. We also observed that colonies of three sampled COI species appear to produce a group of large rediae that have distinctly large pharynges., Conclusions: We conclude that these freshwater species likely do not have a group of specialized non-reproductive soldiers because small rediae of at least some colonies in almost every species do appear to produce embryos. We cannot rule out the possibility that small rediae act as a temporary soldier caste. We are intrigued by the presence of rediae with enlarged pharynges in some species and propose that they may serve an adaptive role, possibly similar to the defensive role of small 'soldier' rediae of marine trematodes. Large-pharynx rediae have been documented in other species previously, and we encourage future efforts to study these large-pharynx rediae., Competing Interests: The authors declare that they have no competing interests., (© 2024 Neal et al.)

Lymnaeid snails are some of the most widespread snails and are the first intermediate host of trematode parasites that affect human and livestock health. A full understanding of the genetic relationship of hosts and parasites is of paramount importance for effective parasite management. The present study assessed the prevalence of trematode larvae in lymnaeid snails and examined the genetic diversity of these snails collected across Thailand. We collected 672 lymnaeid snails from 39 locations in 22 provinces of six regions in Thailand. Subsequently, cercarial infection in the snails was observed by using the shedding method. Lymnaeid snails released 5 types of trematode cercariae, namely, xiphidiocercariae, echinostome cercariae I, echinostome cercariae II, furcocercous cercariae, and strigea cercariae. The phylogenetic analysis based on ITS2 and 28S rDNA sequences revealed 5 cercaria types assigned to four trematode families, of which two belong to the group of human intestinal flukes. Combination of shell morphology and sequence analysis of the mitochondrial COI and 16S rDNA genes, the lymnaeid snails were classified into two species, Radix rubiginosa and Orientogalba viridis. Moreover, the combined dataset of mtDNA genes (COI + 16S rDNA) from R. rubiginosa and O. viridis revealed 32 and 15 different haplotypes, respectively, of which only a few haplotypes were infected with cercariae. The genetic diversity and genetic structure revealed that R. rubiginosa and O. viridis experienced a bottleneck phenomenon, and showed limited gene flow between populations. Population demographic history analyses revealed that R. rubiginosa and O. viridis experienced population reductions followed by recent population expansion. These findings may improve our understanding of parasite-lymnaeid evolutionary relationships, as well as the underlying molecular genetic basis, which is information that can be used for further effective control of the spread of trematode disease., Competing Interests: Declaration of competing interest The authors declare there are no conflicts of interest., (Copyright © 2024 The Authors. Published by Elsevier B.V. All rights reserved.)

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 K mult ) 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.

Background: The construction of Lake Kariba brought about a rise in the incidence of schistosomiasis in its surrounding towns of Kariba (Zimbabwe) and Siavonga (Zambia). After extensive control programs in Kariba, schistosomiasis prevalence dropped significantly. The objective of this study was to revisit the same localities sampled by Chimbari et al. (2003), and provide an update on the snail community and prevalence of trematodes in the Northern shore of Lake Kariba while focusing on planorbid species., Methods: Monthly sampling of snails at 16 sites along the Northern shoreline of Lake Kariba, near Kariba town, was undertaken for one year. Minimum one specimen per morphotype was identified using molecular barcoding (sequencing a fragment of cytochrome c oxidase I subunit (COI)). The infection status of snails was assessed by Rapid Diagnostic PCRs (RD-PCR), and trematode infections were genotyped by sequencing COI and 18S rDNA markers., Results: We collected and identified seven snail species: Bulinus truncatus, Bulinus forskalii, Gyraulus sp., Physella acuta, Bellamya sp., Radix affinis plicatula and Pseudosuccinea columella. Physella acuta was the most abundant snail species (comprising 56.95% of the total snail count) and present at all sites. The B. truncatus population was found to be infected with the stomach fluke Carmyerius cruciformis, a Petasiger sp. and a trematode species belonging to the family Notocotylidae. No Schistosoma sp. infections were detected in our collected snail specimens., Conclusions: We report B. truncatus as an intermediate snail host for Carmyerius cruciformis, and the presence of three non-schistosome trematode species that have not been reported in Lake Kariba before. Furthermore, we detect a possible shift in the snail community when compared to the report by Chimbari et al. (2003): this is the first record of Gyraulus sp. in Lake Kariba, and we did not observe the previously reported B. pfeifferi, B. globosus and Radix natalensis. Although this shift in snail communities might have contributed to the absence of Schistosoma spp. detection in this study, further monitoring of final and intermediate hosts across the Kariba basin is essential to prove a decrease of schistosomiasis in the area., Competing Interests: Declaration of Competing Interest The authors declare that they have no competing interests., (Copyright © 2023. Published by Elsevier B.V.)

The genomic evolution of Polyopisthocotylea remains poorly understood in comparison to the remaining three classes of Neodermata: Monopisthocotylea, Cestoda, and Trematoda. Moreover, the evolutionary sequence of major events in the phylogeny of Neodermata remains unresolved. Herein we sequenced the mitogenome and transcriptome of the polyopisthocotylean Diplorchis sp., and conducted comparative evolutionary analyses using nuclear (nDNA) and mitochondrial (mtDNA) genomic datasets of Neodermata. We found strong mitonuclear discordance in the phylogeny of Neodermata. Polyopisthocotylea exhibited striking mitonuclear discordance in relative evolutionary rates: the fastest-evolving mtDNA in Neodermata and a comparatively slowly-evolving nDNA genome. This was largely attributable to its very long stem branch in mtDNA topologies, not exhibited by the nDNA data. We found indications that the fast evolution of mitochondrial genomes of Polyopisthocotylea may be driven both by relaxed purifying selection pressures and elevated levels of directional selection. We identified mitochondria-associated genes encoded in the nuclear genome: they exhibited unique evolutionary rates, but not correlated with the evolutionary rate of mtDNA, and there is no evidence for compensatory evolution (they evolved slower than the rest of the genome). Finally, there appears to exist an exceptionally large (≈6.3 kb) nuclear mitochondrial DNA segment (numt) in the nuclear genome of newly sequenced Diplorchis sp. A 3'-end segment of the 16S rRNA gene encoded by the numt was expressed, suggesting that this gene acquired novel, regulatory functions after the transposition to the nuclear genome. In conclusion, Polyopisthocotylea appears to be the lineage with the fastest-evolving mtDNA sequences among all of Bilateria, but most of the substitutions were accumulated deep in the evolutionary history of this lineage. As the nuclear genome does not exhibit a similar pattern, the circumstances underpinning this evolutionary phenomenon remain a mystery., (Copyright © 2024 Australian Society for Parasitology. Published by Elsevier Ltd. All rights reserved.)

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.

Strigea falconis is a common parasite of birds of prey and owls widely distributed in the Holarctic. We aimed to characterise S. falconis from Iceland via integrative taxonomic approach and to contribute to the understanding of its circulation in the Holarctic. We recovered adult S. falconis from two gyrfalcons (Falco rusticolus) collected in 2011 and 2012 in Iceland (Reykjanes Peninsula, Westfjords) and characterised them by morphological and molecular genetic (D2 of rDNA, cox1, ND1 of the mDNA) methods. We provide the first species record of S. falconis in Iceland which to the best of our knowledge is its northernmost distributional range. The presence of S. falconis in Iceland is surprising, as there are no suitable intermediate hosts allowing completion of its life cycle. Gyrfalcons are fully sedentary in Iceland; thus, the only plausible explanation is that they acquired their infection by preying upon migratory birds arriving from Europe. Our data indicate that the most likely candidates are Anseriformes and Charadriiformes. Also, we corroborate the wide geographical distribution of S. falconis, as we found a high degree of similarity between our haplotypes and sequences of mesocercariae from frogs in France and of a metacercaria from Turdus naumanni in Japan, and adults from Buteo buteo and Circus aeruginosus from the Czech Republic. The case of Strigea falconis shows the advantages of a complex life cycle and also depicts its pitfalls when a parasite is introduced to a new area with no suitable intermediate hosts. In Iceland, gyrfalcons are apparently dead-end hosts for S. falconis., (© 2024. The Author(s).)

Objectives: The present work aims to expand the knowledge of the digenean species Prosogonotrema bilabiatum (Sclerodistomidae), a parasite of Chaetodipterus faber (Acanthuriformes) from Brazil, with an integrative taxonomic approach, using light microscopy, scanning electron microscopy, histology, and molecular biology., Methods: Forty-one digenean specimens were stained with hydrochloric carmine for morphological studies. Eleven parasites were dehydrated through a graded ethanol series, critical point dried with carbon dioxide, and coated with gold for scanning electron microscopy analysis. Four specimens were processed following histological routine and stained with hematoxylin and eosin and Gomori trichrome. DNA extracted was amplified using 28S partial primer D1-D3. Maximum likelihood and Bayesian inference were performed for phylogenetic analysis., Results: Morphometric and morphological data of the specimens studied ranged in accordance as observed in previous descriptions of the species. Observations from scanning electron microscopy and histology corroborated with those observed in stained whole mounts. Molecular analysis showed that specimens of P. bilabiatum from Brazil clustered with another two sequences of this species from different hosts and localities, with a high node support value., Conclusions: The integrative taxonomic approach allowed to record and describe new characteristics of P. bilabiatum related to the tegument, the structure and the arrangement of its tissues. The use of molecular markers confirmed that specimens identified as P. bilabiatum from different hosts and localities are all conspecific. Further studies, mainly molecular with less conserved genetic markers, should be carried out to better understand the phylogenetic relationships of Prosogonotrema with Hemiuroidea., (© 2024. The Author(s), under exclusive licence to Springer Nature Switzerland AG.)

Trematodes of the family Allocreadiidae are primarily found in the intestines of freshwater fishes around the world. The family includes 15 genera and c. 130 species. The last 2 decades have witnessed an increase in the genetic library of its species. Molecular data have been crucial for species delimitation and species description within Allocreadiidae and for understanding their evolutionary and biogeographical history and classification. Here, the mitogenomes of 3 species of allocreadiids were obtained using high throughput sequencing methods. Mitogenomes were compared with other members of the order Plagiorchiida to determine their molecular composition, gene rearrangement and phylogenetic interrelationships. The complete circular mitogenomes of Allocreadium lobatum, Creptotrematina aguirrepequenoi and Wallinia mexicana were 14 424, 13 769 and 13 924 bp long respectively, comprising 12 protein-coding genes, 22 transfer RNA genes, 2 ribosomal RNA genes and 2 non-coding regions. Gene arrangements were identical to other Xiphidiatan trematodes. Phylogenetic analyses using the mitogenomes revealed Allocreadiidae as a monophyletic group closely related to other members of the suborder Xiphidiata; A. lobatum was yielded as the sister taxon of C. aguirrepequenoi + W. mexicana . Our study increases the complete mitochondrial genome library of trematodes and strengthens our understanding of the phylogenetic relationships and classification of this parasite group.

Purpose: Derogenes ruber Lühe, 1900, the type-species of the genus Derogenes Lühe, 1900, is a poorly known derogenid digenean. The original description of this species was not illustrated and aspects of the morphology of the parasite from the type-host remain scarce. Available records of this species were brief and/or lacked illustrations and were based on morphology alone. Additionally, molecular data for Derogenes spp. are warranted to untangle species complexes as they provide a better assessment of interspecific genetic divergence., Methods: Derogenes ruber is redescribed based on newly collected specimens from the gall bladder of its type-host Chelidonichthys lastoviza (Bonnaterre, 1788) collected in the Western Mediterranean off the Algerian coast during 2017-2019 and molecular data are provided using a partial fragment of the nuclear 28S ribosomal RNA gene (28S rRNA), the internal transcribed spacer 2 (ITS2) and a fragment of the mitochondrial cytochrome c oxidase subunit 1 (cox1) gene., Results: We herein provide a detailed illustrated redescription and morphometric data of D. ruber from its type-host C. lastoviza. We report a new geographical record (off Algeria) for it. Derogenes ruber is also genetically characterised for the first time. Species/lineages of Derogenes were recovered in five strongly supported reciprocally monophyletic clades: (i) D. ruber from C. lastoviza off Algeria; (ii) D. lacustris from Galaxias maculatus (Jenyns) off Argentina; (iii) Lineage "D. varicus DV1" (D. varicus sensu stricto) from fish hosts in the White and Barents seas and the North Sea; (iv) Lineage "D. varicus DV2" from mollusc hosts in the White Sea; and (v) Lineage "D. varicus DV3" from Eumicrotremus fedorovi Mandrytsa. in the Pacific Ocean. Hence, comparison of the newly generated sequences with other available data for Derogenes species supports the distinction of D. ruber confirming its taxonomic status and helping assess interspecific variation. Comparison of D. ruber with the closely related species Derogenes latus revealed overlaps in morphometric data and the validity of the latter species is questioned., Conclusion: The combination of morphological and molecular data provided for D. ruber provides a firm foundation for further investigations of Derogenes spp. Although we do describe herein material of D. ruber from the type-host, given that the occurrence of a single Derogenes species in various hosts has been challenged by molecular data, and both D. lacustris and D. varicus sensu stricto had been genetically proven to occur in various hosts, D. ruber and D. latus may be indeed synonymous. Additional sequencing effort on Derogenes spp. will strengthen systematic comparative studies and evolutionary relationships within the Derogenidae in general., (© 2023. The Author(s).)

Purpose: Species of the genus Philophthalmus are eye flukes with a complex taxonomy, which began to be improved with the help of molecular data only recently. However, most described species have never been placed into a phylogenetic context. In this study, eye flukes previously found on kelp gulls, Larus dominicanus, from Brazil and identified as Philophthalmus lacrymosus were subjected to molecular analysis., Methods: For the molecular analyses, we analyzed parasites found in six infected gulls (one worm per bird) collected from different municipalities of the state of Santa Catarina, Brazil. We carried out the amplification and sequencing of the partial region of the 28S and cox1 genes and the data obtained were compared with sequences available to philophthalmid species and subjected to phylogenetic analysis., Results: The isolates of P. lacrymosus from Brazil grouped in well-supported clades with five other species of Philophthalmus with sequences available for comparison. Interspecific divergences of 0.1-1.6% in 28S and 8.2-14.9% in cox1 were found in relation to other isolates of Philophthalmus spp. Two cox1 haplotypes differing in one nucleotide (0.1%) were found between the six eye flukes isolates in gulls from different localities. The Brazilian isolates grouped in a subclade with parasites identified as P. lacrymosus in Portugal; however, the molecular divergences found in cox1 (8.2-8.5%) strongly suggest that these isolates belong to different species. The phylogenetic trees obtained and the intergeneric divergences to species of the genera Cloacitrema and Parorchis did not support the validity of the genus Natterophthalmus, for which P. lacrymosus was proposed as the type species in the past., Conclusion: As P. lacrymosus was described from Brazil, we recommend that this name be applied to the South American isolates and that the Portuguese isolates be provisionally considered as Philophthalmus sp., a probable cryptic species. Moreover, data obtained supports the previous morphology-based synonymizing between Natterophthalmus and Philophthalmus. Considering our results and most of previous reports of P. lacrymosus in South America, we suggest this species presents a marine life cycle., (© 2023. The Author(s) under exclusive licence to Witold Stefański Institute of Parasitology, Polish Academy of Sciences.)

The Almaco jack (Seriola rivoliana) is a marine fish maintained in mariculture systems and frequently infested by monogenean parasites like Neobenedenia sp. Severe infestations can lead to high mortalities and economic losses for farmers. This study evaluated the effects of temperature on the immune response on Almaco jack infested with Neobenedenia sp. We exposed infested fishes at temperatures of 20 °C, 24 °C, and 30 °C for 20 days and took samples of different tissues at the beginning of the experiment, and after 3 and 20 days. The tissues considered were the skin, thymus, cephalic kidney, and spleen to evaluate the relative gene expression of different genes: Hsp70, IgM, IL-1β, IL-10, and MyD88. Our results showed an increase in IL-1β gene expression in the skin after 20 days of infestation but no significant effect of temperature on gene expression, despite increases in infestation rates with temperature. Therefore, relative genetic expression was controlled by the number of parasites and the days post-infestation. These results show that the parasite infestation induced a local response in the skin, but that temperature has an indirect effect on the immune system of Almaco jack., (© 2024. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

The freshwaters of Iraq harbour a high diversity of endemic and phylogenetically unique species. One of the most diversified fish groups in this region is cyprinoids, and although their distribution is relatively well known, their monogenean parasites have only rarely been investigated. Herein, we applied an integrative approach, combining morphology with molecular data, to assess the diversity and phylogeny of cyprinoid-associated monogenean parasites. A total of 33 monogenean species were collected and identified from 13 endemic cyprinoid species. The highest species diversity was recorded for Dactylogyrus (Dactylogyridae, 16 species) and Gyrodactylus (Gyrodactylidae, 12 species). Four species of Dactylogyrus and 12 species of Gyrodactylus were identified as new to science and described. Two other genera, Dogielius (Dactylogyridae) and Paradiplozoon (Diplozoidae), were represented only by 4 and 1 species, respectively. Phylogenetic analyses of the Dactylogyrus and Gyrodactylus species revealed that the local congeners do not form a monophyletic group and are phylogenetically closely related to species from other regions (i.e. Europe, North Africa and Eastern Asia). These findings support the assumption that the Middle East served as an important historical crossroads for the interchange of fauna between these 3 geographic regions.

Brachylaima spp. are trematodes that have a unique life cycle as they exclusively use land snails as the intermediate host. Although their intermediate host has been well studied, very little information is available about their definitive host, partly as isolation of its adult stage from wild animals is rare. We found three trematodes in the small intestine of a red fox (Vulpes vulpes schrencki) in Hokkaido, the northernmost island of Japan. The trematodes were identified as Brachylaima ezohelicis based on morphological features and genetic analysis, which is believed to have a definitive avian host. The morphological features of the isolated trematodes were consistent with B. ezohelicis samples grown in the definitive host except for body length. Our study suggests that B. ezohelicis uses mammals as definitive hosts as well as birds., (Copyright © 2023. Published by Elsevier B.V.)