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Digenean trematodes commonly use marine crustaceans as intermediate hosts, yet their distribution and host-specific dynamics remain underexplored in temperate European systems. In this study, metacercarial stages of Microphallus similis and M. primas were identified in the hepatopancreas of Carcinus maenas collected from two distinct locations in South Wales, UK across a 4-month period. M. similis exhibited consistently high prevalence (~ 76%) at both sites, while M. primas occurred only as a co-infection, with lower prevalence (2–9%) and infection intensity. At Oxwich Bay, crab size and coloration were significantly associated with trematode infection, whereas no such associations were observed at Mumbles Head. Phylogenetic analysis confirmed species-level divergence between M. similis and M. primas . Overall parasite burden was significantly higher at Mumbles Head, with larger crabs harbouring more metacercariae. Additionally, metacercariae from Mumbles Head were significantly larger than those from Oxwich Bay, suggesting site-specific variation in parasite development. The higher prevalence, intensity, and cyst size at Mumbles likely reflect habitat and host-community differences between sites, including greater availability of first-intermediate hosts and differences in crab size cohorts and microhabitats that may influence transmission.

Here, we present a comprehensive morphological and molecular phylogenetic analysis of Clinostomum sp. (Digenea: Clinostomidae) metacercariae parasitizing two freshwater fish species from Southeast Brazil: Serrasalmus spilopleura (piranha) and Callichthys callichthys (tambuatá). The morphological examination revealed distinct characteristics of metacercariae in each host. Using the cytochrome c oxidase I (COI) gene barcode region, we obtained DNA sequences that allowed for accurate phylogenetic placement. Phylogenetic analyses revealed that Clinostomum sp. HM41 (metacercariae), isolated from S. spilopleura, exhibited 86% similarity to Ithyoclinostomum yamagutii, while Clinostomum sp. HM125 (metacercariae), from C. callichthys, showed 98.7% similarity to Clinostomum sp. Cr_Ha1. The phylogenetic trees constructed through Bayesian Inference and Maximum Likelihood methods indicated high biodiversity within the Clinostomum genus and strong support for distinct lineages. These findings enhance our understanding of the diversity and ecological distribution of Clinostomum species in South American freshwater environments.

Digenetic trematodes of the genus Clinostomum are cosmopolitan parasites infecting fishes, amphibians, reptiles, and snails as intermediate hosts. Despite the broad geographical distribution of this genus, debate about the number of species and how they vary in host use has persisted. To better understand patterns of infection among host species and across life stages, we used large-scale field surveys and molecular tools to examine five species of amphibians and seven species of fishes from 125 California ponds. Among the 12,360 examined hosts, infection was rare, with an overall prevalence of 1.7% in amphibians and 9.2% in fishes. Molecular evidence indicated that both groups were infected with Clinostomum marginatum . Using generalized linear mixed effects models, host species identity and host life stage had a strong influence on infection status, such that Lepomis cyanellus (green sunfish) (49.3%) and Taricha granulosa (rough skinned newt) (9.2%) supported the highest overall prevalence values, whereas adult amphibians tended to have a higher prevalence of infection relative to juveniles (13.3% and 2.5%, respectively). Experimentally, we tested the susceptibility of two amphibian hosts ( Pseudacris regilla [Pacific chorus frog] and Anaxyrus boreas [western toad]) to varying levels of cercariae exposure and measured metacercariae growth over time. Pseudacris regilla was 1.3× more susceptible to infection, while infection success increased with cercariae exposure dose for both species. On average, metacarcariae size increased by 650% over 20 days. Our study highlights the importance of integrating field surveys, genetic tools, and experimental approaches to better understand the ecology of host–parasite interactions.

Trematode metacercariae are the most abundant and frequently encountered helminths in freshwater fish. Yet, accurate species identification remains challenging, potentially leading to an underestimation of trematode diversity. Using data from parasitological examinations of 1,030 fish (47 species) collected from diverse freshwater habitats in Lithuania (2022–2024), we assessed the contemporary diversity of trematode metacercariae, host associations, microhabitat preferences, and changes in diversity patterns and transmission dynamics. Through integrated morphological and molecular techniques, we identified metacercariae belonging to 51 species from eight families, more than doubling previously reported diversity (25 species). While trematode family composition remained largely unchanged – the Diplostomidae and Strigeidae remained the most diverse families – notable differences were observed at the species level. Metacercariae of the Echinochasmidae and Echinostomatidae were detected for the first time, while previously reported Clinostomidae were absent. Fish of the Leuciscidae hosted the highest trematode diversity. Host specificity of metacercariae was generally low, with most species being euryxenous. At the microhabitat level, eyes harboured the highest number of species, while muscles showed the highest metacercarial density. Notably, we detected species first genetically characterised in North America (Echinoparyphium sp. 2 and Ichthyocotylurus sp. 2) and species potentially belonging to the genus Neogogatea, previously known only from Asia and North America, highlighting potential invasion risks and suggesting that European trematode diversity remains substantially underestimated. Future efforts should obtain molecular data from correctly identified adult specimens to resolve the identity of species currently identified only to the genus or family level, thereby enabling assessment of their geographical distributions and ecological roles.

The genus Clinostomoides Dollfus, 1950 was erected to accommodate a single worm from Ardea goliath sampled in the Belgian Congo. The specimen was distinguished from other clinostomids by its large size and posterior genitalia. In the following years, metacercariae of Clinostomoides brieni, have been described in Clarias spp. in southern and western Africa. A few authors have referred to Clinostomum brieni, but all such usages appear to be lapsus calami, and the validity of Clinostomoides remains widely accepted. In this study our aim was: position C. brieni among the growing clinostomids molecular database, and redescribe the species with emphasis on characters that have emerged as important in recent work. We sequenced two nuclear (partial 18S and ITS) and one mitochondrial marker (partial cytochrome c oxidase I) and studied morphology in metacercariae from hosts and localities likely to harbour the type species (Clarias spp., Democratic Republic of the Congo, South Africa). Phylogenetic analysis shows C. brieni belongs within Clinostomum Leidy, 1856. We therefore transfer C. brieni to Clinostomum, amend the diagnosis for the genus Clinostomum and provide a critical analysis of other species in Clinostomoides, all of which we consider species inquirendae, as they rest on comparisons of different developmental stages.

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.

The separation of Clinostomum complanatum Rudolphi, 1814 and Clinostomum marginatum Rudolphi, 1819 has long been unclear. Recent data confirm the validity of the junior species, C. marginatum, by ∼1% differences in its 18S rDNA sequences. We collected adults and metacercariae of C. complanatum and C. marginatum and found reliable morphological differences in the genital complex at both developmental stages. In addition, we identified basic morphometrics (distance between suckers, body width) in metacercariae that may be useful for discriminating the species. The morphological differences were supported by the comparison of sequences of internal transcribed spacers of ribosomal DNA and of the mitochondrial gene cytochrome c oxidase I (COI) from 39 specimens. In 36 specimens, the average divergence between the species was 7.3% in ITS and 19.4% in COI sequences. Two specimens from North America and 1 from Europe had sequences that did not allow them to be clearly allied with either species.

Species of the genus Tylodelphys (Diplostomidae) have a cosmopolitan distribution. Metacercariae of these species infect the eye, brain, pericardial sac or body cavity of fish second intermediate hosts, and the adults are found in piscivorous birds of many orders. An unnamed species of Tylodelphys from the eyes of bullies ( Gobiomorphus cotidianus ) was characterized molecularly and morphologically as a metacercaria in a previous study, in which it was predicted that the adult of this species would be found in the Australasian crested grebe. Two specimens of this bird became available and specimens of the unnamed Tylodelphys species were, indeed, found in them, confirmed by identity of genetic sequence data. Found to differ morphologically from its congeners, the new species is here described as Tylodelphys darbyi n. sp. Three species are closest to the new species in morphology: Tylodelphys glossoides , T. immer and T. podicipina robrauschi. Compared with T. darbyi n. sp. these three species are slightly larger and possess longer eggs. Tylodelphys glossoides also differs in having a wider oral sucker and T. podicipina robrauschi in having comma- or kidney-shaped pseudosuckers and an ovary that reaches a larger size, along with higher upper limits for body width, hind body and sucker width, holdfast and oesophagus length, and pharynx, pseudosucker and testes length and width. Tylodelphys immer also differs from T. darbyi n. sp. in having a shorter ventral sucker and the largest pseudosuckers of any Tylodelphys species.

Metacercariae of species of Euclinostomum have been found encysted in kidney, liver, and muscles of several fish species, while adults occur in the mouth, pharynx, larynx, trachea, and upper esophagus of fish-eating birds. The aim of this work was to gather molecular and morphological data from the type species, Euclinostomum heterostomum, as a starting point for needed revisions of the genus. Metacercariae were collected from cichlids in Lake Kinneret, Israel, and all were identified as E. heterostomum based on morphology. This identification was further confirmed by principal components analyses that revealed no partitions in morphometric resemblance, indicating that the material represents a single species, and that the specimens were morphometrically similar to other descriptions of E. heterostomum. In phylogenetic analysis of concatenated partial internal transcribed spacer 1 and 2, 5.8S rDNA, and partial cytochrome c oxidase I barcode sequences, the isolates of Euclinostomum obtained in this study form a single, monophyletic group separate from recently published data from Euclinostomum sp. from Thailand and from Clinostomum species, which are also monophyletic. The morphological and molecular data reported in the present work can provide a useful point of reference for future studies.

A complete morphological description, supported by molecular data, of Clinostomum detruncatum metacercariae is reported in this study. The metacercariae were found infecting Synbranchus marmoratus from the Paraná River, municipality of Guaíra, Paraná State, Brazil. The taenioidean uterus is the main morphological characteristic of this species. Both neighbor-joining (NJ) and maximum likelihood (ML) trees showed 2 clear separate clusters grouping together the species from the Palearctic region (Clinostomum complanatum, Clinostomum cutaneum, Clinostomum phalacrocoracis, Clinostomum philippinense) and those from the Nearctic/Neotropical regions (Clinostomum marginatum, Clinostomum tataxumui, C. detruncatum, Clinostomum sp. 7). The latter clade is divided into 2 subclades grouping species from North America and Mexico (C. marginatum and C. tataxumui), and those from Brazil (C. detruncatum and Clinostomum sp. 7).