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Clinostomids are a group of digeneans in which substantial diversity has been recently discovered, with some metacercariae specific to their fish hosts. Genetic analysis has been instrumental in elucidating species diversity within this genus. Recently, seven COI lineages were reported in Argentina, along with three metacercarial morphotypes lacking molecular data. Two of these were found parasitising Rivulidae fishes. The discovery of Clinostomum metacercariae in Trigonectes aplocheiloides and Titanolebias monstrosus from temporary ponds in the western Chacoan region allowed us to redescribe the metacercariae previously identified by other authors and provide the first sequences of this lineage. The morphology of the metacercariae in both hosts matched previously reported descriptions. Genetic analysis clustered the new lineage with Clinostomum detruncatum , Clinostomum sp. 7, Clinostomum L1, and Clinostomum CRA. This discovery leaves only two morphological records of metacercariae to be characterised using DNA sequencing: one in another Rivulidae ( Neofundulus paraguayensis ) and one in a Loricaridae ( Hypostomus sp.). The present results represent the eighth clinostomid lineage sequenced from Argentina, highlighting the extensive diversity in South America and the many lineages yet to be described or identified, considering that only one of these lineages is formally described based on adult specimens found in the heron Ardea cocoi.

In this study, we describe a new species of Pseudoparacreptotrema (Allocreadiidae) from the mugilid Dajaus monticola collected in western Puerto Rico, where no allocreadiid has previously been reported, bringing the number of species in this genus to seven (five in D. monticola , two in Profundulus spp.). The new parasite species is distinguished from congeners by its overall size, oral-to-ventral sucker size ratio, pharynx size, cirrus sac, and oral lobe morphology, and by 0.64%-3.45% divergence in a 1019-bp alignment of 28S. We build on prior suggestions that the current concept of P. agonostomi likely includes multiple species and provide the first mitochondrial data (whole mitochondrial genome) as well as the complete nuclear rDNA array from Pseudoparacreptotrema to facilitate future phylogenetic work. Within the Allocreadiidae, phylogenetic analysis of mitochondrial genomes and 28S provides conflicting topologies for the placement of Pseudoparacreptotrema and Allocreadium. The 28S phylogeny of six species of Pseudoparacreptotrema resembles that of four lineages of D. monticola in that in both host and parasite, Pacific coastal lineages branch earliest, and a Caribbean lineage is more recently evolved.

Diplomonorchis micropogoni Nahhas & Cable, 1964 was considered a junior subjective synonym of Diplomonorchis leiostomi Hopkins, 1941 in 1969. Diplomonorchis leiostomi has since been widely reported from the coastal Western Atlantic between Delaware Bay and southern Brazil. Until now, taxonomically verifiable DNA sequence data for D. leiostomi has been available from an individual worm collected from the northern Gulf of Mexico. We generated a partial sequence of the 28S rRNA gene from D. leiostomi from the spot croaker, Leiostomus xanthurus Lacepède (type-host) from Beaufort, North Carolina, USA (type-locality) that differed at 31 of 1,246 bases from the available 28S sequence. This prompted a reevaluation of Diplomonorchis spp. identities from the northern Gulf of Mexico. We found D. leiostomi and D. micropogoni distinguishable by testes shape and size, and to a lesser degree by relative caecal length. Museum specimens of D. leiostomi, identified from the Gulf of Mexico represent a species complex containing D. leiostomi, D. cf. micropogoni and, a new species of Diplomonorchis. The sequences previously identified as D. leiostomi in GenBank (AY222137 & AY222252) are herein identified as D. cf. micropogoni. The new species is described from newly collected material herein. Phylogenetic analysis of 28S rRNA sequences from the species complex plus 46 species from the Monorchioidea Odhner, 1911 indicated all three Diplomonorchis spp. are closely related and form a clade with some species of Lasiotocus Looss, 1907. With the addition of the new species, and acceptance of D. micropogoni, there are currently 14 valid species in Diplomonorchis., Competing Interests: Declarations Conflict of interest All authors report no competing interest or conflict related to the research or writing. Ethical approval Auburn University institutional guidelines for care and use of animals were followed. Fish were collected under permits issued from the Mississippi Department of Marine Resources, Alabama Department of Conservation and Natural Resources Marine Division, and the Florida Fish and Wildlife Conservation Commission Division of Marine Fisheries Management, Special Activity License SAL-10-15-0942-SR., (© 2024. The Author(s).)

Three new species of Gyrodactylu s were identified from the body surface of the Triplophysa species from the Qinghai-Tibet Plateau, Gyrodactylus triplorienchili n. sp. on Triplophysa orientalis in northern Tibet, G. yellochili n. sp. on T. sellaefer and T. scleroptera and G. triplsellachili n. sp. on T. sellaefer and T. robusta in Lanzhou Reach of the Yellow River. The three newly identified species share the nemachili group species' characteristic of having inturning hamulus roots. Gyrodactylus triplorienchili n. sp. shared a quadrate sickle heel and a thin marginal hook sickle, two morphological traits that set them apart from G. yellochili n. sp. However, they may be identified by the distinct shapes of the sickle base and marginal hook sickle point. Gyrodactylus triplsellachili n. sp. had much larger opisthaptoral hard part size than the other two species. The three new species show relatively low interspecific differences of 2.9-5.3% p-distance for ITS1-5.85-ITS2 rDNA sequences. Phylogenetic analysis indicated that the three new species formed a well-supported monophyletic group (bp = 99) with the other nemachili group species.

We described Creptotrema cambeve n. sp. from the intestine of the Brazilian catfish Cambeva davisi (Haseman, 1911) (= Trichomycterus davisi ) based on integrative analyses using morphological and molecular approaches. Creptotrema cambeve n. sp. closely resembles morphologically Creptotrema paraense Vicente, Santos & Souza, 1978, which was described from a siluriform fish in Northern Brazil; however, the two species differ mainly in the oral and ventral sucker sizes and the distribution of the vitellaria. The phylogenetic analyses of the 28S rDNA placed the sequences of the new species in a monophyletic clade together with all other Creptotrema spp. and as sister taxon of Creptotrema cruste Alcantara, Ebert, Ferreira-Silva, Forti, Morais, Pérez-Ponce de León & Silva, 2024, a parasite from a Brazilian anuran. Genetic divergences between the new species and other Creptotrema spp. varied from 0.2% to 4.3%, further corroborating the distinction of the new taxon. Creptotrema cambeve n. sp. is the 18th nominal Creptotrema species known from South America and the 22nd erected in the genus. This is the first study reporting a trematode in Cambeva spp. hosts and the second parasitological survey carried out for Ca. davisi , a poorly known small endemic fish.

Phylogenetic studies of aberrant species are of considerable scientific interest because their taxonomic rank in traditional systems based on morphological characters is not infrequently overestimated. Apopharynx bolodes (Braun, ) is one of the few psilostomid digeneans devoid of the pharynx. This is considered a sufficient basis for assigning it and similar species to the subfamily Apopharynginae. We found A. bolodes in Fulica atra Linnaeus, 1758 from Belarus, described it morphologically, and genotyped it by the 28S rRNA gene and the ITS2 region. It is the first molecular data on A. bolodes and the first record of this digenean species in Belarus. The phylogenetic analysis based on partial sequences of the 28S rRNA gene showed that A. bolodes is closely related to the Sphaeridiotrema spp. (Sphaeridiotrematinae). However, this phylogenetic inference has not received yet support with data on the ITS2 region.

Despite a great effort made for almost 90 years, the diversity of freshwater fish trematodes in Mexico is still far from being fully known. The addition of molecular data to the description of trematode diversity in the last two decades added the potential to establish more robust species limits and a more accurate biodiversity estimation, but also led in some instances to the recognition of cryptic species complexes. Here, we used sequences of the large subunit of the nuclear ribosomal gene (28S rRNA) as barcodes, and morphological data, to assess the diversity of freshwater fish trematodes from a lake within a tropical rainforest. Eighty freshwater fish specimens of eight species were studied, and 120 trematode specimens were collected. Morphologically, specimens were allocated into nine genera; molecular phylogenetic analyses along with sequence divergence data provided evidence for recognising 11 trematode taxa, six adults and five metacercariae; six of them were identified to species level. Geographical distribution and host association patterns are briefly discussed for each trematode taxa.

The present study aims to describe a new genus and species of microphalloid digenean parasite of two species of bats of the genus Peropteryx from the Mexican Neotropics (in the states of Chiapas and Yucatán). Morphological and molecular data (28s rDNA ribosomal gene sequences) were used to study Digeneans. Sagittatrema zutzi gen. nov. sp., nov., is diagnosed morphologically by having a sagittiform body, a genital pore in the midline of the body, posterior to the ventral sucker, and a cirrus sac running through much of the diameter of the ventral sucker. The nine sequences generated from the 28S rDNA gene were used to examine the phylogenetic affinities of this new taxon within the superfamily Microphalloidea Ward, 1901, using Maximum Likelihood and Bayesian Inference analyses. Both analyses resulted in trees with similar topologies and formed a well-supported clade (Bt = 100; pp = 1) with the Sagittatrema sequences. Because of the new genus's phylogenetic position and that some sister families to the proposed taxa, like Pleurogenidae and Prosthogonimidae, are polyphyletic, we prefer to consider Sagittatrema as a genus incertae sedis within Microphalloidea. A full systematic review of microphalloids is needed to confirm their phylogenetic position.

Species-level delineation of digenetic trematodes is complex and can be best achieved by integrative taxonomy using both genetic characterisation and morphological analysis. Two new Bucephalidae species of the genus Rhipidocotyle Diesing, 1858 are described here based on specimens collected from the intestine of Sphyraena putnamae Jordan & Seale following this approach. Adults of R. siphonyaka n. sp. and R. nolwe n. sp. possess tentacles and a tegument with scales. They are distinguished from their congeners by the arrangement of the digestive structures, the extent of the uterus relative to vitelline fields, and the arrangement of the reproductive structures. Rhipidocotyle siphonyaka n. sp. differs from R. nolwe n. sp. in having the pharynx and mouth positioned in the pre-uterine field, tandem testes, longer body length, and shorter pre-vitelline and post-testicular distance. Rhipidocotyle siphonyaka n. sp. differs from its congeners in having a tube-like intestinal caecum, pharynx and mouth opening positioned in the pre-vitelline field. Rhipidocotyle nolwe n. sp. appears to be similar, morphologically and morphometrically, to Rhipidocotyle khalili (Nagaty, 1937). Despite their similarities, R. nolwe n. sp. has a shorter body length and egg size. Moreover, the molecular analysis of 28S and ITS rDNA fragments indicate that R. siphonyaka n. sp. and R. nolwe n. sp. are closely related phylogenetically but distinct from one another and other Bucephalidae for which molecular data are available.

Members of the genus Maritrema Nicoll, 1907 include endoparasites of aquatic birds and mammals, distributed worldwide. Adult specimens were collected from the intestines of three bird species (the great black hawks, Buteogallus urubitinga Gmelin; laughing gull, Leucophaeus atricilla Linnaeus; and the willet, Tringa semipalmata Gmelin) in three localities along the Gulf of Mexico. Photogenophores were sequenced for the large subunit (LSU) from nuclear rDNA, and the new sequences were aligned with other microphalloid sequences available in GenBank. The maximum likelihood and Bayesian inference analyses revealed three independent lineages, one corresponding to a previously described species, Maritrema corai Hernández-Orts, Pinacho-Pinacho, García-Varela & Kostadinova, 2016, and two representing two undescribed species. These two new species are described in the current study. Maritrema itzamnai n. sp. can be morphologically differentiated from its four congeneric species occurring in coastal areas of Mexico by having smaller oral (20-29 × 20-38 μm) and ventral (20-39 × 19-33 μm) suckers. In addition, Maritrema itzamnai n. sp. possess annular vitellarium instead of a horseshoe-shaped vitellarium. Maritrema kukulkanni n. sp. can be distinguished from its congeneric species reported from Mexico by possessing a larger oesophagus (44-117μm) and by having a vitellarium distributed in both the hindbody and forebody. Maritrema corai is the third species recorded in this study parasitizing the great black hawk ( Buteogallus urubitinga ); this represents a new host and locality record, expanding its distribution range from the Mexican Pacific Ocean to the Gulf of Mexico.

This study focuses on the opecoelid genus Podocotyle, particularly the species in the northern European seas (White, Barents and Pechora), with additional data from few isolates from the Far East. The research combines molecular analyses (28S, 5.8S+ITS2 rDNA, and cox1 mtDNA) and morphological examinations. Molecular analyses suggested that there are most likely four species of Podocotyle in the northern European seas and three more in the Far East. Morphological redescriptions are provided for three northern European species: P. atomon, P. reflexa, and P. odhneri, with the latter recognized as a valid species. The study also verified the life cycle of P. atomon and identified Lacuna vincta (Littorinidae) as the first intermediate host for P. reflexa. In the Sea of Okhotsk, L. turrita is the first intermediate host for two apparently undescribed Podocotyle species. The results also highlight the morphological variability of P. atomon, emphasizing the need for an integrative approach in the taxonomy and biodiversity studies., (© 2024. The Author(s), under exclusive licence to Springer Nature B.V.)

Two new species of the genus Allocreadium were isolated from the intestines of the Lake minnow Rhynchocypris percnura caught in the backwater of the Komissarovka River in the South of the Russian Far East. The morphology of A. anastasii n. sp. corresponds to that of Allocreadium sp. from Lake Khar (Mongolia) and Allocreadium sp. Belous, 1952 from the Primorsky region of Russia except for the preacetabular anterior border of the vitelline follicles in A. anastasii n. sp. from the Komissarovka River vs. at anterior half of ventral sucker in Allocreadium sp. Genetic analysis revealed the identity of A. anastasii n. sp. to Allocreadium sp. 1 from the Nezhinka River and Lake Khar. Allocreadium macrolecithum n. sp. was differentiated from Palaearctic Allocreadium spp. by having the following features: respectively large vitelline follicles extending from posterior extremity to anterior margin of the ventral sucker; relatively short caeca reaching the border of middle and posterior thirds of hindbody; and small testes in the middle of hindbody. Interspecific genetic p-distances between Allocreadium spp. were 0.16-7.23% in 28S gene and 18.62-31.54% in Cox1 mtDNA gene. In the phylogenetic tree reconstructed with Maximum parsimony and Bayesian Inference methods, A. anastasii n. sp. and A. macrolecithum n. sp. were nested into different species groups of the genus Allocreadium - sister to A. khankaiense and A. bursense , respectively. Modified dichotomous keys were prepared for 31 Palaearctic species of Allocreadium including A. crassum , A. dogieli , A. papilligerum , A. bursense , A. anastasii n. sp., and A. macrolecithum n. sp.

In light of the morphological and molecular data for cyclocoelids observed from the air sacs of Mareca strepera (Linnaeus) (Anatidae, Anseriformes) caught in the southern region of the Russian Far East, we suggest new insights into the systematics of the family Cyclocoelidae. A comparative morphological and phylogenetic analyzes revealed that new cyclocoelids represented the new genus and species Paracyclocoelum lobatum. Based on the 28S rRNA gene fragment we showed the significant genetic divergence of P. lobatum from the type species of the type genus for the family, Cyclocoelum mutabile (Zeder, 1800) Brandes, 1892 and along with the confusing morphological similarity by the prepharyngeal position of the genital pore it most likely indicate homologous development of the reproductive system of Paracyclocoelum and Cyclocoelum. Here, we provide a new dichotomous key for five cyclocoelid genera from the subfamily Cyclocoelinae including Paracyclocoelum n. g. The new genus Paracyclocoelum had sister relationship to the cyclocoelin genus Circumvitellatrema. Based on the polyphyletic interrelationships of Cyclocoelum and Circumvitellatrema the Cyclocoelinae were assigned with the status sensu lato., (© 2024. The Author(s), under exclusive licence to Springer Nature B.V.)

The trematode family Cyathocotylidae infects various hosts worldwide, including birds, mammals, reptiles, and fish. However, the lack of molecular data from adult worms hinders phylogenetic, epidemiological, and host association studies. This study aims to identify the common cyathocotylid trematodes infecting African catfish in Egypt using morphological and molecular evidence. Out of 142 Clarias gariepinus, 123 fish (86.6%) harbored cyathocotylid metacercariae, with a mean metacercarial intensity of 201 ± 38.5/g. Cyathocotylid metacercariae prevalence gradually rose as host size increased. Although there was no significant difference between groups, larger fish had a higher mean metacercarial intensity. The prevalence and intensity were unrelated to the fish gender. Histopathological examination of metacercariae-infected catfish revealed varying degrees of degenerative changes, including intermuscular edema leading to muscle fiber dispersion and atrophy, involving 11% to over 81% of muscle sections. We identified three cyathocotylid metacercariae and eight cyathocotylid adult species from experimental infection using morphometric and molecular data, including internal transcribed spacer (ITS) and/or mitochondrial cytochrome c oxidase subunit 1 (cox1) sequences. We determined the phylogenetic position of these cyathocotylid samples. The ITS sequence analysis linked the isolated Cyathocotylidae sp. 1 and 2 metacercariae to Prohemistomum vivax adults. Mesostephanus appendiculatoides and Paracoenogonimus ovatus were reported for the first time in Egypt. These findings may provide valuable genetic data for future molecular epidemiological and phylogenetic studies of cyathocotylid trematodes., (© 2024. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

Encotyllabe percussa n. sp. is proposed based on morphology and DNA sequences analysis of ribosomal (18S, 28S) and mitochondrial (COI) gene fragments. Encotyllabe percussa n. sp. was found infecting the spangled emperor Lethrinus nebulosus (n = 75) with higher prevalence from Dibba, Musandam (Gulf of Oman) than in Dhofar Salalah (Indian Ocean), Oman (p = 0.03). The general morphology of E. percussa n. sp. resembles E. caballeroi, E. chironemi and E. spari, which exhibit pre-equatorial testes. However, E. percussa n. sp. shows unique morphological characteristics distinguishing from congeneric species: the large hamuli bear notch allocated externally between the first half proximal of the root, and the small hamuli exhibit semicircular shape with undivided roots. Phylogenetic relationships within the Encotyllabe genus remain unresolved. However, the tree topology with the 28S showed overall consistency with the principal component analysis arrangement (PCA) derived from the morphological analysis. Which showed that the large and small hamuli, marginal hooks, ovary, testes (length and width) and peduncle are currently the most important morphological traits within the genus. Cytochrome c oxidase subunit I (COI) gene fragment showed high interspecific genetic divergence adding unambiguous resolution to discriminate/designate species identity. Interrelations within the genus support the identity of Encotyllabe percussa as a new species. This is the first species characterized with three gene fragments, the second congeneric species described in L. nebulosus and the first recorded in Oman., (© 2024. The Author(s), under exclusive licence to Springer Nature B.V.)

A new species of Gyrodactylus is described from the gills of the near-threatened Clanwilliam sawfin, Cheilobarbus serra (Smiliogastrinae) collected from the Matjies River, Cape Fold Ecoregion, Western Cape Province, South Africa. Morphometry and morphology of the haptoral hard parts (hamuli, bars and marginal hooks) of Gyrodactylus serrai n. sp. differ from the other known species of the genus in the smaller size of hamuli and the shape and size of marginal hooks. Furthermore, ITS rDNA for the new species is unique among available Gyrodactylus spp. data in GenBank. Based on the uncorrected p-distances, G. serrai n. sp. is genetically most closely related to Gyrodactylus moroccensis Rahmouni, 2023 and Gyrodactylus pseudomoroccensis Rahmouni, 2023 from two species of Luciobarbus (Barbinae) from northern Africa, with interspecific divergence of 8.7% and 8.8%, respectively. The presence of a median ridge in the terminal part of the ventral bar membrane at G. serrai n. sp. most probably represents a morphological link to the North African Gyrodactylus spp. that suggests a morphogenetic association across the African continent as a result of ancient waterways that facilitated the dispersion of cyprinids and their parasite fauna or an independent evolution event retaining similarities from a common ancestor. The description of Gyrodactylus serrai n. sp. represents only the second species of Gyrodactylus described from an endemic South African cyprinid host, underscoring the need for focused research on this group of fishes to provide a sound understanding of the parasitic communities of these highly threatened and poorly studied hosts., (© 2024. The Author(s).)

This study aimed to investigate the occurrence of larval trematode infections in bithyniid snails across five provinces in southern Thailand. A total of 1,413 Bithynia siamensis snails were collected between October 2021 and October 2022 using handpicking and scooping methods. Among these, 844 were identified as B. s. siamensis and 569 as B. s. goniomphalos. The snail samples were examined for parasitic infections in the laboratory using shedding and crushing methods. Among the 27 sampling sites, snails from 6 sites were infected with various trematode species (infection rate: 4.95%, 70/1,413). Evaluation of the morphological and internal organ characteristics of the cercariae allowed for the categorization of the trematodes into five species of trematodes (belonging to four families). The identified species included Echinochasmus pelecani (family: Echinostomatidae), Echinostoma revolutum (family: Echinostomatidae), Haematoloechus similis (family: Haematoloechidae), Loxogenoides bicolor (family Lecithodendriidae), and Stictodora tridactyla (family: Heterophyidae), and the infection rates for these species were 3.26% (46/1,413), 0.35% (5/1,413), 0.42% (6/1,413), 0.78% (11/1,413), and 0.14% (2/1,413), respectively. The cercariae from the identified trematode species were studied for DNA analysis. The phylogenetic lineage reveals relationships among the species, confirming the morphological distinctions., (© 2024. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

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.)

Rhopalocercous Cercaria duplicata von Baer, 1827 develops in an intermediate host, the unionid bivalve Anodonta anatina (L.), but its adult form has been unknown. We examined eight fish species occurring in the presence of a highly infested population of A. anatina in the Zesławice reservoir (S Poland). Gravid Phyllodistomum specimens were obtained from the ureters of ide, Leuciscus idus (L.) and common rudd, Scardinius erythrophthalmus (L.). One of the rudd specimens was doubly infected, a trematode was also found in the urinary bladder. In addition, a gravid Phyllodistomum specimen was found in the ureter of a tench Tinca tinca (L.), caught in Lake Ilmėdas (Lithuania). In order to clarify the phylogenetic position of larval and adult gorgoderids and to establish their life cycle, ITS2 and 28S rDNA sequences were analysed. The analysis showed that adult Phyllodistomum specimens located in the ureters are conspecific with C. duplicata. The trematode found in the urinary bladder of S. erythrophthalmus was P. folium (Olfers, 1816). It is suggested that adult stages of C. duplicata should be referred to as Phyllodistomum duplicatum n. comb. The intercaecal position of the uterus and the deeply-lobed ovary are the main features distinguishing it from other Phyllodistomum species. Host specificity and ecology are discussed., (© 2024. The Author(s).)

The type species of the genus Halipegus Looss, 1899, Halipegus ovocaudatus (Vulpian, 1859) is a generalist parasite of different species of amphibians, mainly anurans. To date, the phylogenetic position of this species has not been established. Specimens of H. ovocaudatus, were collected from three species of water frogs, Pelophylax ridibundus (Pallas), P. esculentus (Linnaeus), and P. lessonae (Camerano) inhabiting the Middle Volga region (European Russia). In our study, we provide detailed morphological redescription of these digeneans and combine it with molecular phylogenetic analyses based on partial sequences of the 28S rDNA and coI mtDNA genes. The phylogenetic position of H. ovocaudatus among the derogenids has been established as a sister clade to other representatives of the Halipeginae. We compared pairwise distances based on partial sequences of coI mtDNA genes of H. ovocaudatus and other representatives of Derogenidae. It was shown, that adults of H. ovocaudatus from different hosts are representatives of the same species. Specimens parasitizing three frog hosts from several localities differ slightly in body shape and size, which probably reflects host-induced intraspecific variability., (© 2024. The Author(s), under exclusive licence to Springer Nature B.V.)

Aggregation is a fundamental feature of parasite distribution in the host population, but the biological implications of the aggregation indices used to describe the relationships between the populations of parasites and hosts are not evident. It is speculated that the form of distribution in each case is predicated on the host's varying resistance to the infection, which is hard to control, making it difficult to adequately interpret the index values. This paper examines several cases from trout farms in Russian Karelia to explore the monogenean Gyrodactylus spp. infection in rainbow trout of varying ages. The genetic homogeneity of cage-reared fish and the direct life cycle of the helminths make the relationship between the species more lucid than in natural host-parasite systems. The results give no ground to speak of any specific patterns: as well as in the natural systems, the infection rates in trout vary widely, i.e., the helminth distribution has not become more uniform; the observed distributions in all cases are adequately approximated by the negative binomial model; the positive abundance-occupancy relationships (AORs) and abundance-variance relationships (AVRs) common for parasitic systems apply to the basic infection parameters. The form of the negative binomial distribution is shaped by two parameters-k and θ, the former being a metric of the infection variability, which depends on the host's individual resistance, and the latter representing the parasites' reproduction and establishment success rates. A rise in the parameter k indicates increased aggregation and a higher parameter θ points to a more uniform frequency distribution. These parameters can be used as a representative tool for monitoring the parasite communities in salmonid fishes, including in aquaculture., (© 2024. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

We examined several American alligators, Alligator mississippiensis (Daudin, 1802) (Crocodilia: Alligatoridae) from Louisiana, Alabama, and South Carolina in August 2022. The intestine of one alligator from Alabama was infected by Dracovermis occidentalis Brooks and Overstreet, 1978 (Platyhelminthes: Digenea: Liolopidae Odhner, 1912), a seldom collected and incompletely described trematode that lacks a representative nucleotide sequence. Liolopidae comprises 5 genera and 15 species: Liolope spp. infect giant salamanders; Helicotrema spp. infect turtles and lizards; Harmotrema spp. infect snakes; Paraharmotrema spp. infect turtles; and Dracovermis spp. infect crocodilians. Based on our study of the newly collected specimens and the holotype of D. occidentalis, we redescribe D. occidentalis, correct errors in its original description, and provide an updated phylogeny for Liolopidae that, for the first time, includes Dracovermis Brooks and Overstreet, 1978. Our specimens were identified as D. occidentalis by having testes in the posterior 1/3 of the body, a pretesticular cirrus sac, a spined and eversible cirrus, a bipartite seminal vesicle, and a post-acetabular vitellarium. A phylogenetic analysis of the D1-D3 domains of the nuclear large subunit ribosomal DNA (28S) recovered Liolopidae as monophyletic; however, low taxon sampling in the group precludes hypothesis-testing about liolopid-vertebrate cophyly. This is the first collection for morphological study of the type species for Dracovermis since the genus was proposed 46 years ago, the first record of a liolopid from Alabama, and the first phylogenetic analysis that includes Dracovermis., (© 2024. The Author(s).)

Parasites belonging to the trematode family Clinostomidae have a global distribution. Freshwater fish in Bangladesh frequently serve as hosts for different clinostomid species. During the metacercaria stage, the digenetic trematode Euclinostomum heterostomum exhibits a specific preference for Channidae species, where it undergoes encystment within the liver, kidneys, and muscles of these hosts. The study aimed to identify E. heterostomum in Channa punctata through combined morphological and molecular analysis. Light and scanning electron microscopy were conducted to investigate the morphological characteristics of the species. The histological examination of the infected liver tissue revealed the presence of encysted flukes, inflammatory leukocyte infiltrates, and degeneration and loosening of hepatic tissue. In molecular analysis, the generated internal transcribed spacer 1 (ITS1), 5.8S ribosomal DNA and ITS2 regions sequence (GenBank accession no: OR591452) of 830 bp showed 100% identity with Euclinostomum heterostomum identified in India (MT785786). The phylogenetic reconstruction provided substantial evidence of genetic similarities (0-1% genetic distance) among different isolates of the genus Euclinostomum, suggesting that there is a shared heritage., (© 2024. The Author(s), under exclusive licence to Springer Nature B.V.)

Trematodes and their snail hosts have developed intimate parasite-host associations, with snails supporting a diverse and often species-specific trematode fauna. In the faucet snail, Bithynia tentaculata (Caenogastropoda, Littorinimorpha), a unique trematode fauna has been recorded recently. However, knowledge of the exact species identity, phylogenetic relationships, and geographical distribution remains limited as many of the species belong to groups with unclear or controversial taxonomical assignment. To contribute to our knowledge of the trematodes, we investigated the trematode fauna of B. tentaculata by examining a total of 556 snails from lakes in County Galway, Ireland. Using an integrative taxonomic approach including DNA sequence data analyses (28S rRNA gene, ITS1-5.8S-ITS2, ITS2, cox 1, nad 1) and morphological tools (taxonomical drawings and measurements), we identified nine trematode species of seven families, with seven species occurring as cercariae ( Cyathocotyle prussica , Lecithodendrium linstowi , Lecithodendrium sp., Asymphylodora progenetica , Sphaerostoma bramae , Metorchis xanthosomus , and Notocotylus sp.) and three species occurring as metacercariae ( A. progenetica , Parasymphylodora parasquamosa , and Sphaeridiotrema sp.). Except for S. bramae , all are new species records for Ireland and provide the most western distribution of these trematodes in Europe. The trematode species recorded are known to use a wide range of definitive hosts and have a wide geographical distribution; among them are species members of genera that are zoonotic ( Metorchis ) and pathogenic to wildlife ( Cyathocotyle , Sphaeridiotrema , and Notocotylus ). There remains an ongoing need for precise identification of the trematode species to ensure that wider ecological contexts are correctly understood and biodiversity and disease threats can be accurately evaluated.

Octomacrum spinum Dansby & Shoemaker, 1973, is reported for the first time from 23 of 146 (16%) highland stonerollers, Campostoma spadiceum collected from central and western Arkansas and southeastern Oklahoma, USA. Analysis of molecular data revealed that the SSU rRNA gene of specimens of O. spinum from Arkansas was identical to a number of Octomacrum spp. and the LSU rRNA gene was 97.46% similar to Octomacrum europaeum. We also provide a summary of the species of Octomacrum reported from North American and European fishes., (© 2024. The Author(s), under exclusive licence to Springer Nature B.V.)

Background: The Bucephalidae is a large family of digenean trematodes but most previous analyses of its phylogenetic position have relied on a single mitochondrial gene or morphological features. Mitochondrial genomes (mitogenomes) remain unavailable for the entire family. To address this, we sequenced the complete mitogenome of Dollfustrema vaneyi and analyzed the phylogenetic relationships with other trematodes., Results: The circular genome of Dollfustrema vaneyi spanned 14,959 bp and contained 12 protein-coding genes, 22 transfer RNA genes, 2 ribosomal RNA genes, and a major non-coding region. We used concatenated amino acid and nucleotide sequences of all 36 genes for phylogenetic analyses, conducted using MrBayes, IQ-TREE and PhyloBayes. We identified pronounced topological instability across different analyses. The addition of recently sequenced two mitogenomes for the Aspidogastrea subclass along with the use of a site-heterogeneous model stabilized the topology, particularly the positions of Azygiidae and Bucephalidae. The stabilized results indicated that Azygiidae was the closest lineage to Bucephalidae in the available dataset, and together, they clustered at the base of the Plagiorchiida., Conclusions: Our study provides the first comprehensive description and annotation of the mitochondrial genome for the Bucephalidae family. The results indicate a close phylogenetic relationship between Azygiidae and Bucephalidae, and reveal their basal placement within the order Plagiorchiida. Furthermore, the inclusion of Aspidogastrea mitogenomes and the site-heterogeneous model significantly improved the topological stability. These data will provide key molecular resources for future taxonomic and phylogenetic studies of the family Bucephalidae., (© 2024. The Author(s).)

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.)

Monogeneans of the genus Dactylogyrus Diesing, 1850, the largest genus in the family Dactylogyridae, mostly parasitize the gills of cyprinoid hosts; however, only 3 Dactylogyrus' mitochondrial genomes (mitogenomes) are studied so far. The aim of this research is to extend our understanding of the mitogenomes of Dactylogyrus. We sequenced the mitogenomes of D. crucifer and D. zandti isolated from Rutilus rutilus and Abramis brama orientalis in northwest China, and then we compared these mitogenomes with other monogeneans. We used Illumina NovaSeq to sequence the entire mitochondrial genomes of D. crucifer and D. zandti and characterized the mitogenomes to understand the gene structure, gene identity, the secondary structures of the 22 tRNA genes, and relative synonymous codon usage. We used the analytic Bayesian Information and Maximum Likelihood methods to determine their associated phylogenetic trees. The mitogenomes of D. crucifer and D. zandti were 14,403 and 18,584 bp, respectively. Organization and positioning of these genes were in accordance with Dactylogyrus lamellatus and Dactylogyrus tuba. The nucleotide composition of Dactylogyridae was different from other families of Monogenea, and the A+T count of genus Dactylogyrus (54 - 58.4 %) was lower than other genus species of the family Dactylogyridea (63.9 - 78.4 %) in protein-coding genes. Dactylogyrus members displayed a codon usage bias. The relative synonymous codon used by Dactylogyrus was not conserved and was lower than other monogeneans. The codon use patterns of closely-related species isolated from closely-related hosts were identical. Phylogenetic analyses using mitogenomic dataset produced Dactylogyrus isolated from host subfamily Leuciscinae formed a sister-group. Our results contributed significantly to an increased database of mitogenomes, more than 50 %, for Dactylogyrus that may help future studies of mitochondrial genes and codon uses for the analysis of monogenean phylogenetics., 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.)

AbstractMany parasite species use multiple host species to complete development; however, empirical tests of models that seek to understand factors impacting evolutionary changes or maintenance of host number in parasite life cycles are scarce. Specifically, one model incorporating parasite mating systems that posits that multihost life cycles are an adaptation to prevent inbreeding in hermaphroditic parasites and thus preclude inbreeding depression remains untested. The model assumes that loss of a host results in parasite inbreeding and predicts that host loss can evolve only if there is no parasite inbreeding depression. We provide the first empirical tests of this model using a novel approach we developed for assessing inbreeding depression from field-collected parasite samples. The method compares genetically based selfing rate estimates to a demographic-based selfing rate, which was derived from the closed mating system experienced by endoparasites. Results from the hermaphroditic trematode Alloglossidium renale , which has a derived two-host life cycle, supported both the assumption and the prediction of the mating system model, as this highly inbred species had no indication of inbreeding depression. Additionally, comparisons of genetic and demographic selfing rates revealed a mixed mating system that could be explained completely by the parasite's demography (i.e., its infection intensities).

The Cyathocotylidae Mühling, 1896 is a small but broadly distributed family of digeneans parasitic in a wide range of vertebrate definitive hosts, from fish to mammals. Despite existing taxonomic questions, only a few studies have generated DNA sequence data from cyathocotylids, and only a single publication included sequence data from a cyathocotylid parasitic in snakes. Four genera are known to infect snakes: GogateaLutz, 1935, Szidatia Dubois, 1938, MesostephanoidesDubois, 1951, and SerpentostephanusSudarikov, 1961. Members of these genera were known from only Asia and Africa. In the present study, we describe 2 new species of Gogatea from snakes in Australia and 1 from Vietnam. The new species from Vietnam described herein is the first member of the genus that lacks a ventral sucker. We used partial sequences of the nuclear large ribosomal subunit (28S) and mitochondrial cytochrome c oxidase subunit I (COI) genes to explore phylogenetic relationships among cyathocotylids and species differentiation. In addition, this is the first report of a cyathocotylid from snakes in Australia, thus posing interesting questions regarding the dispersal and historical biogeography of these parasites. Cyathocotylid genera from snakes have a long, convoluted taxonomic history. The genera Gogatea, Mesostephanoides, and Szidatia were distinguished from each other based on very small morphological differences. Therefore, the validity of Szidatia and Mesostephanoides was often questioned in the literature. Based on the detailed morphological analysis of our freshly collected high-quality specimens and comparison with published information, we synonymize Mesostephanoides and Szidatia with Gogatea., (© American Society of Parasitologists 2024.)

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.)

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.)

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.)

Members of the genus Scaphanocephalus mature in accipitrids, particularly osprey, Pandion haliaetus , with metacercaria causing Black Spot Syndrome in reef fishes. In most of the world, only the type species, Scaphanocephalus expansus (Creplin, 1842) has been reported. Recent molecular studies in the Western Atlantic, Mediterranean and Persian Gulf reveal multiple species of Scaphanocephalus , but have relied on 28S rDNA, mainly from metacercariae, which limits both morphological identification and resolution of closely related species. Here we combine nuclear rDNA with mitochondrial sequences from adult worms collected in osprey across North America and the Caribbean to describe species and elucidate life cycles in Scaphanocephalus . A new species described herein can be distinguished from S. expansus based on overall body shape and size. Phylogenetic analysis of the whole mitochondrial genome of Scaphanocephalus indicates a close relationship with Cryptocotyle . We conclude that at least 3 species of Scaphanocephalus are present in the Americas and 2 others are in the Old World. Specimens in the Americas have similar or identical 28S to those in the Mediterranean and Persian Gulf, but amphi-Atlantic species are unlikely in light of divergence in cytochrome c oxidase I and the lack of amphi-Atlantic avian and fish hosts. Our results provide insight into the geographic distribution and taxonomy of a little-studied trematode recently linked to an emerging pathology in ecologically important reef fishes.

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.)