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During a parasite survey in Brazilian amphibians from São Paulo state, Brazil, Gorgoderina parvicava Travassos, 1922 was found in the urinary bladder (11 adult worms) and (five juvenile worms) in the kidneys of the pepper-frog Leptodactylus labyrinthicus (Spix, 1824). Parasites were examined by microscopy and 28S rDNA and COI gene were sequenced and analyzed for the molecular study. The phylogenetic reconstructions resulted in identical topologies with highly supported values in the nodes in most clades using Maximum likelihood and Bayesian inference methods and positioned G. parvicava in the subclade formed by species of subfamily Gorgoderinae parasitizing the urinary bladder of amphibians. Molecular phylogenetic data showed that this species is related to other species of Gorgoderina. In addition, new molecular data and the analyses of genetic distances provide extra comparative data, which can be applied in further investigations on the taxonomic status and the diversity among Gorgoderina spp. and host-parasite relationships.

Two species of digenean trematodes of the family Brachycladiidae were obtained from two male dwarf sperm whales Kogia sima that stranded along the island of Kyushu, southern Japan in 2017. From the liver of the first animal, a single, large gravid specimen of a digenean species was collected. The morphological features were consistent with those of the genus Brachycladium. The worm had a large body and was characterized by anterior caeca without lateral diverticula, the shape of testes, ovary, and eggs. Molecular analyses using gene sequences of the 28S rRNA and the mitochondrial NADH dehydrogenase subunit 3 also supported the inclusion of this specimen into the genus Brachycladium. The identity of this worm is undetermined due to the lack of information on the genus and is reported as Brachycladium sp. From the cranial sinuses of the second animal, 33 specimens of digeneans were collected that were morphologically identified as Nasitrema gondo. This report documents a new host record for N. gondo, and the sequence information is provided for this digenean for the first time. This is the second record of digenean parasites for the family Kogiidae, and the first record with morphological and molecular information. The possibility of digenean infection in the liver and cranial sinus should be kept in mind during the necropsy of stranded kogiids., (Copyright © 2023. Published by Elsevier Ltd.)

Capsalids are monopisthocotylean monogenean parasites found on the skin and gills of fish. Capsalines (subfamily Capsalinae) are large-sized capsalids, parasitic on highly prized gamefish, and species of Tristoma parasitise only the gills of swordfish (Xiphias gladius). We obtained specimens of Tristoma integrum Diesing, 1850 from swordfish collected off Algeria in the Mediterranean Sea. Here, we describe the specimens, including the key systematics characters of dorsolateral body sclerites. One specimen was used for a next generation sequencing analysis but a part of it, including the sclerites, was mounted on a permanent slide, drawn, and deposited in a curated collection. We characterised the complete mitogenome, the ribosomal cluster (including 18S and 28S) and additional genes such as Elongation factor 1 alpha (EF1α) and Histone 3. We also retrieved molecular information from the host tissue present in the gut of the monogenean and provide the sequence of the complete rRNA cluster of the host, X. gladius. The mitogenome of T. integrum is 13 968 bp in length and codes for 12 protein, 2 rRNA and 22 tRNA. Phylogenies of capsalids were generated from 28S sequences and concatenated mitochondrial protein-coding genes, respectively. In the 28S phylogeny, most subfamilies based on morphology were not found to be monophyletic, but the Capsalinae were monophyletic. In both phylogenies, the closest member to Tristoma spp. was a member of the Capsaloides. In an Appendix, we report the complex nomenclatural history of Tristoma Cuvier, 1817 and its species., (© R. Gastineau et al., published by EDP Sciences, 2023.)

Paradiplozoon cirrhini n. sp. (Monogenea, Diplozoidae) is described from the gills of mud carp, Cirrhinus molitorella (Valenciennes, 1844) (Cyprinidae, Labeoninae), collected in Wuzhou, Guangxi Province, and Conghua, Guangdong Province as part of an ongoing survey of the diplozoid fauna in the Pearl River basin of China. The new Paradiplozoon species is distinguished from congeners by the structure of median plate and its outgrowth sclerites. The ITS2 sequences of the new species differ from all known available diplozoid sequences by 22.04%-38.34%. The new species is the first diplozoid species parasitic on Labeoninae in China. Molecular phylogenetic analyses using rRNA ITS2 placed Paradiplozoon cirrhini n. sp. in a sister position to the other Chinese Paradiplozoon, implying that Labeoninae represents an early and potentially ancestral host group for China Paradiplozoon. We also provided ITS2 sequences for four other diplozoids species, namely P. megalobramae Khotenovsky, 1982, P. saurogobionis (Jiang, et al., 1985) Jiang, Wu & Wang, 1989, Sindiplozoon hunanensis Yao & Wang, 1997, and Sindiplozoon sp., and validated their phylogenetic position. The results confirm that all diplozoid species are spilt into two major clades and show monophyly of Sindiplozoon but paraphyly of Paradiplozoon., (© J. Huang et al., published by EDP Sciences, 2023.)

Fish are often speckled with "black spots" caused by metacercarial trematode infection, inducing a host response. Cryptocotyle spp. (Opisthorchiidae) are among the parasites responsible for this phenomenon. So far, the impact on human health is still unknown. In addition, few publications dealing with black spot recovery, identification, distribution and diversity among commercially important fish are available. Moreover, "black spots" have been observed by fishermen on marine fish, revealing an appreciable but unquantified presence in consumed fish. An epidemiological survey of 1,586 fish from seven commercial species (herring, sprat, whiting, pout, dab, flounder, and plaice) was conducted in the Eastern English Channel and the North Sea in January 2019 and 2020. Encysted metacercariae were found in 325 out of 1,586 fish, with a total prevalence of 20.5%. Intensity of infection varied from 1 to 1,104 parasites. The recorded encysted metacercariae were identified either by microscopic examination or with molecular tools. Partial sequences of the mtDNA cox1 gene and of the rDNA ITS region were obtained. Two species of Cryptocotyle, Cryptocotyle lingua (Creplin, 1825) and Cryptocotyle concava (Creplin, 1825) were found. Metacercariae belonging to other trematode families were also identified. Molecular phylogenetic analysis and haplotype network construction were performed to confirm the identification and to study the potential presence of different populations of Cryptocotyle spp. This survey enabled us to describe the distribution of two species of Cryptocotyle in the English Channel and North Sea ecosystems. The observed differences in infestation levels between fish species and geographical areas will contribute to better understanding of the ecology of these parasites., (© M. Duflot et al., published by EDP Sciences, 2023.)

Monogenean parasites are often co-introduced with their fish hosts into novel areas. This study confirmed co-introduction of two dactylogyrids, Dactylogyrus squameus Gusev, 1955 and Bivaginogyrus obscurus (Gusev, 1955), and a newly described gyrodactylid species, Gyrodactylus pseudorasborae n. sp. into Europe along with their fish host, the invasive topmouth gudgeon Pseudorasbora parva (Temminck & Schlegel) from East Asia. All three species were observed in the lower Dnieper and middle Danube basin regions and had slightly larger haptoral hard parts than the same parasites in their native range. While dactylogyrids occurred sporadically, we recorded regular infection by G. pseudorasborae n. sp. at relatively high prevalence and abundance. This latter species was observed in both the native and non-native range of topmouth gudgeon, and resembles Gyrodactylus parvae You et al., 2008 recently described from P. parva in China. Both species were distinguished based on genetic analysis of their ITS rDNA sequence (6.6% difference), and morphometric differences in the marginal hooks and male copulatory organ. Phylogenetic analysis of dactylogyrid monogeneans showed that B. obscurus clustered with Dactylogyrus species parasitising Gobionidae and Xenocyprididae, including D. squameus, supporting recent suggestions of a paraphyletic origin of the Dactylogyrus genus. In addition to co-introduced parasites, topmouth gudgeon was infected with a local generalist, G. prostae Ergens, 1964, increasing the number of monogeneans acquired in Europe to three species. Nevertheless, monogenean infections were generally lower in non-native host populations, potentially giving an advantage to invading topmouth gudgeon., (© M. Ondračková et al., published by EDP Sciences, 2023.)

The gill monogenean Sparicotyle chrysophrii (Van Beneden & Hesse, 1863) Mamaev, 1984 is a specific and common parasite of wild and cultured gilthead sea bream Sparus aurata Linnaeus, 1758, able to cause disease and mortality in aquaculture systems. Few molecular studies have been carried out on this monogenean, and its population structure and genetic diversity are barely known. This study provides the first contribution to the population genetic variation of S. chrysophrii, based on two molecular markers - the structural ribosomal RNA (rRNA) for the large subunit (28S) and the cytochrome c oxidase subunit I (COI) gene. Samples were collected from the gills of farmed and wild S. aurata from Italy and the Spanish Mediterranean. The analysis included previously published sequences. The 28S rDNA analysis was consistent with previous studies of specimens isolated from S. aurata and confirmed the presence of only one species on the gills of this host in the Mediterranean Sea. The COI sequences analysis suggested that the samples isolated in a previous study from a different host species, wild Boops boops (Linnaeus, 1758) in the Adriatic Sea, may represent a new undescribed sister species of S. chrysophrii. The low nucleotide diversity of S. chrysophrii isolated only from S. aurata versus the high haplotype diversity revealed small differences between haplotypes. The haplotypes shared between wild and farmed hosts from Spain provided the first molecular evidence of the possible transfer of S. chrysophrii between wild and farmed populations of S. aurata. The mtDNA COI analysis did not show a clear genetic structure, probably the result of several factors including coevolution, wild and farmed host interactions, and host population structure in space and time., (© 2022. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

Background: Glutamate carboxypeptidase 2 (GCP2) belongs to the M28B metalloprotease subfamily encompassing a variety of zinc-dependent exopeptidases that can be found in many eukaryotes, including unicellular organisms. Limited information exists on the physiological functions of GCP2 orthologs in mammalian tissues outside of the brain and intestine, and such data are completely absent for non-mammalian species. Here, we investigate GCP2 orthologs found in trematodes, not only as putative instrumental molecules for defining their basal function(s) but also as drug targets., Methods: Identified genes encoding M28B proteases Schistosoma mansoni and Fasciola hepatica genomes were analyzed and annotated. Homology modeling was used to create three-dimensional models of SmM28B and FhM28B proteins using published X-ray structures as the template. For S. mansoni, RT-qPCR was used to evaluate gene expression profiles, and, by RNAi, we exploited the possible impact of knockdown on the viability of worms. Enzymes from both parasite species were cloned for recombinant expression. Polyclonal antibodies raised against purified recombinant enzymes and RNA probes were used for localization studies in both parasite species., Results: Single genes encoding M28B metalloproteases were identified in the genomes of S. mansoni and F. hepatica. Homology models revealed the conserved three-dimensional fold as well as the organization of the di-zinc active site. Putative peptidase activities of purified recombinant proteins were assayed using peptidic libraries, yet no specific substrate was identified, pointing towards the likely stringent substrate specificity of the enzymes. The orthologs were found to be localized in reproductive, digestive, nervous, and sensory organs as well as parenchymal cells. Knockdown of gene expression by RNAi silencing revealed that the genes studied were non-essential for trematode survival under laboratory conditions, reflecting similar findings for GCP2 KO mice., Conclusions: Our study offers the first insight to our knowledge into M28B protease orthologs found in trematodes. Conservation of their three-dimensional structure, as well as tissue expression pattern, suggests that trematode GCP2 orthologs may have functions similar to their mammalian counterparts and can thus serve as valuable models for future studies aimed at clarifying the physiological role(s) of GCP2 and related subfamily proteases., (© 2022. The Author(s).)

Leucochloridium spp. have been established in Taiwan since the 1930s, with five species listed: Leucochloridium turdi, Leucochloridium passeri, Leucochloridium muscularae, Leucochloridium sime, and Leucochloridium taiwanese listed until now. Because of the similar morphology among the adults of the Leucochloridiidae family, the taxonomic status of most of them should be reconsidered. Broodsacs are the most distinct characteristic shared by Leucochloridium flukes, and L. passeri is, by far, the only one whose broodsacs have been described. In this study, broodsacs collected from an infected Succinea sp. (amber snails) in Taiwan were morphologically compared to previous descriptions and sequenced using both nuclear and mitochondrial genetic markers for identification. Our samples resembled broodsacs of L. passeri previously described in northern Taiwan and those recently collected in Okinawa, Japan. The conspecific status of the broodsacs collected in Taiwan and Okinawa was supported by the identical DNA sequences identified in this study. Broodsacs have rarely been reported in tropical and subtropical regions, although some Leucochloridium spp. have been recorded. In such cases, genetic markers will be crucial to link Leucochloridium broodsacs in snails (intermediate host) with their corresponding adult stage in birds (definitive host)., (Copyright © 2022 Elsevier B.V. All rights reserved.)

Ligophorus kaohsianghsieni (Gusev, ) Gusev, was collected from the so-iuy mullet Planiliza haematocheilus (Temminck & Schlegel, 1845) from the Black Sea and the Sea of Japan. DNA sequences data for L. kaohsianghsieni , as well as its morphological characters from the Sea of Japan were obtained for the first time. Significant morphometric and genetic diversity between specimens of L. kaohsianghsieni from the Black-Azov Sea region and the Sea of Japan were not found. For the first time, the molecular phylogeny of L. kaohsianghsieni based on three fragments of the nuclear DNA ribosomal cluster (18S, internal transcribed spacer 1 and 28S) was reconstructed. Molecular analysis of Ligophorus species from the Atlantic and Pacific Oceans revealed a significant phylogenetic distance between L. kaohsianghsieni and two others species ( Ligophorus pilengas and Ligophorus llewellyni ) from the same host ( P. haematocheilus ) and region. This result indicates the lack of correspondence between the phylogenetic and geographical closeness of the hosts and the relation of their parasites from the genus Ligophorus .

A parasitological survey of terrestrial slugs and snails was conducted at popular dog walking locations across the city of Nottingham, with the intensions of finding gastropods infected with parasites of medical (or veterinary) importance such as lungworm (metastrongyloid nematodes) and trematodes. A total of 800 gastropods were collected from 16 sites over a 225 km 2 area. The extracted nematodes and trematodes were identified by molecular barcoding. Of the 800 gastropods collected, 227 were infected (172 had nematode infections, 37 had trematode infections and 18 had both nematode and trematode infections). Of the nematode infected gastropods genotyped, seven species were identified, Agfa flexilis , Angiostoma gandavense , Angiostoma margaretae , Cosmocerca longicauda , Phasmarhabditis hermaphrodita , Phasmarhabditis neopapillosa and an unknown Cosmocercidae species. Of the trematode infected gastropods genotyped, four species were identified, Brachylaima arcuate , Brachylaima fuscata , Brachylaima mesostoma and an unknown Plagiorchioidea species. No lungworm species were found within the city of Nottingham. To our knowledge, this study represents the first survey of gastropod-associated nematodes and trematodes in the East midlands of the United Kingdom.

The Asian freshwater snail Melanoides tuberculata has been established since the 1960s in the Americas, where it transmits cercariae of a small number of digenetic trematode species from its native range. In 2021-2022, 24 M. tuberculata were discovered shedding transversotrematid cercariae in Puerto Rico, where parasites of this snail have not been previously studied. Adult transversotrematids (in some cases, gravid) were found on field-caught fish and on fish exposed to shedding snails, including on fish species native to Puerto Rico. Adults and cercariae were identified as Transversotrema patialense (Soparkar, 1924), a species native to the Indomalayan region. Morphological identification was supported with 28S rDNA sequences closely matching that from unidentified transversotrematid cercariae in Thailand. The absence of T. patialense in snails collected prior to 2021, increasing prevalence of infection in snails collected thereafter, and lack of variation in parasite DNA sequences (28S, internal transcribed spacer 2, cytochrome c oxidase I) from three isolates are consistent with a recently introduced and possibly expanding parasite population. Transversotrema patialense has been recorded outside its native range before, but most studies (including a prior record in the Americas) reported the parasite from captive hosts from commercial sources such as pet shops. The present results thus provide the first demonstration of natural transmission of T. patialense in the Americas. Phylogenetic analysis of 28S but not of ITS2 show the transversotrematid genus Transversotrema Witenberg, 1944 is paraphyletic, with Crusziella Cribb, Bray and Barker 1992 nested within it., Competing Interests: Declaration of competing interest The authors declare no conflict of interest, financial or otherwise., (Copyright © 2022 Elsevier Inc. All rights reserved.)

Division of labour through the formation of morphologically and functionally distinct castes is a recurring theme in the evolution of animal sociality. The mechanisms driving the differentiation of individuals into distinct castes remain poorly understood, especially for animals forming clonal colonies. We test the association between microbiomes and caste formation within the social trematode Philophthalmus attenuatus, using a metabarcoding approach targeting the bacterial 16S SSU rRNA gene. Clonal colonies of this trematode within snail hosts comprise large reproductive individuals which produce dispersal stages, and small, non-reproducing soldiers which defend the colony against invaders. In colonies extracted directly from field-collected snails, reproductives harboured more diverse bacterial communities than soldiers, and reproductives and soldiers harboured distinct bacterial communities, at all taxonomic levels considered. No single bacterial taxon showed high enough prevalence in either soldiers or reproductives to be singled out as a key driver, indicating that the whole microbial community contributes to these differences. Other colonies were experimentally exposed to antibiotics to alter their bacterial communities, and sampled shortly after treatment and weeks later after allowing for turnover of colony members. At those time points, bacterial communities of the two castes still differed across all antibiotic treatments; however, the caste ratio within colonies changed: after antibiotic disruption and turnover of individuals, new individuals were more likely to become reproductives than in undisturbed control colonies. Our results reveal that each caste has a distinct microbiome; whether the social context affects the microbiota, or whether microbes contribute to modulating the phenotype of individuals, remains to be determined., (© 2022 The Authors. Molecular Ecology published by John Wiley & Sons Ltd.)

A survey was conducted in the lowland areas of Binh Dinh and Phu Yen provinces in south central Vietnam to study the diversity of digenean cercariae within freshwater snails. These regions are known as highly endemic areas for the transmission of liver flukes, Fasciola spp. and Opisthorchis viverrini. In total, 13 snail species were found, and 3 of these were dominant, comprising 51.9% of all collected snails, i.e., Bithynia funiculata, Melanoides tuberculata, and Radix auriculata. Cercariae were found in 5 snail species, and among these, Indoplanorbis exustus had the highest overall prevalence of infection (38.5%). No cercarial infections were found in viviparid and ampullariid snails. Morphological identification resulted in 10 cercariae types that were illustrated and measured. Among them, xiphidiocercariae and echinostomata were the most common, causing 70.55% of all cercariae infections. Bithynia species had the most diverse digenean fauna. Molecular data revealed the presence of cercariae of 13 digenean species, including O. viverrini. The 18S rDNA sequences of echinostomata and xiphidiocercariae type 3, which were shed from different snail hosts, were similar to 2 and 3 digenean species, respectively. In conclusion, our findings highlight the diversity of digenean fauna in Binh Dinh and Phu Yen provinces., (© American Society of Parasitologists 2022.)

Plagiorchis elegans (Trematoda: Digenea) is mainly parasitic in the intestines of vertebrate animals, including humans, causing irreversible pathological damage and herd-spherical influences. However, little information is available about its molecular epidemiology, population genetics, and phylogeny. In the present study, we sequenced, assembled, and annotated the complete mitochondrial (mt) genome of P. elegans. Combining with the available mitochondrial data of subclass Digenea, phylogenetic analysis was performed based on Bayesian inference (BI). These results showed that the complete length of P. elegans is 13,862 bp, including 12 protein-coding genes (PCGs), 2 ribosomal RNAs (rRNAs), 22 transfer RNAs (tRNAs), and one non-coding region (NCR). There was an obvious A + T content from 61.0% to 71.3% and the values of the Ka/Ks ratio ranged from 0.119 (cox1) to 1.053 (nad6). In the BI analysis, different from previous studies, phylogenetic analysis showed genus Glypthelmins was paraphyletic rather than monophyletic and had a closer relationship with Plagiorchis and Orientocreadium. Additionally, the BI tree also presented that the genus Echinostoma was monophyletic. Our results provided molecular data in the family Plagiorchiidae proposing new insight within Xiphidiata and Echinostomata., 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 © 2022 Elsevier Inc. All rights reserved.)

Most trematodes and some cestodes have obligate life history features that include an asexual developmental stage that can produce genetically-identical individuals (clonemates) followed by an adult stage with sexual reproduction. These life history features can influence the evolutionary mechanism of inbreeding in parasites, especially among self-compatible hermaphroditic endoparasites whose mating opportunities are restricted to within hosts. As clonemate mating in hermaphroditic species produces a genetic inbreeding signature identical to that of self-mating, it is important to understand how clonemates are transmitted through their life stages. A handful of prior studies compared clone richness (number of clones) across life cycle stages to infer transmission processes and to characterize clone abundance distributions (CADs) among hosts. Here we illustrate the use of the proportion of clonemate dyads (PC) within hosts to describe the CADs. PC has several advantages as an ecological metric in that it is unbiased by sample size, takes into account relative parasite burdens, and has a direct transmission interpretation, i.e., the probability of cotransmitting clonemates. Moreover, PC is also an evolutionary metric as it can be used to estimate a potential clonemate mating rate. We demonstrate the use of PC in comparing CADs within and across 2 trematode developmental stages in the lancet fluke Dicrocoelium dendriticum. Also, we show how genetic estimates of apparent selfing (true selfing plus clonemate mating) at larval and juvenile stages can be compared to PC estimated at the adult stage to assess the contribution of clonemate mating to apparent selfing. The eco-evolutionary links presented are generalizable to assess sibling cotransmission as well. Thus, the framework presented herein will facilitate future field-based studies on the transmission and mating systems of parasitic flatworms., (© American Society of Parasitologists 2022.)

The Nile perch (Lates niloticus) is a notorious invasive species. The introductions of Nile perch into several lakes and rivers in the Lake Victoria region led to the impoverishment of trophic food webs, particularly well documented in Lake Victoria. Additionally, its parasites were co-introduced, including Dolicirroplectanum lacustre (Monogenea, Diplectanidae). Dolicirroplectanum lacustre is the single monogenean gill parasite of latid fishes (Lates spp.) inhabiting several major African freshwater systems. We examined the intra-specific diversification of D. lacustre from Lates niloticus in Lake Albert, Uganda (native range) and Lake Victoria (introduced range) by assessing morphological and genetic differentiation, and microhabitat preference. We expected reduced morphological and genetic diversity for D. lacustre in Lake Victoria compared with Lake Albert, as a result of the historical introductions. We found that D. lacustre displayed high morphological variability within and between African freshwaters, with two morphotypes identified, as in former studies. The single shared morphotype between Lake Albert and Lake Victoria displayed similar levels of haplotype and nucleotide diversity between the lakes. Mitonuclear discordance within the morphotypes of D. lacustre indicates an incomplete reproductive barrier between the morphotypes. The diversification in the mitochondrial gene portion is directly linked with the morphotypes, while the nuclear gene portions indicate conspecificity. Based on our results, we reported reduced genetic and morphological diversity, potentially being a result of a founder effect in Lake Victoria., (Copyright © 2022 Australian Society for Parasitology. Published by Elsevier Ltd. All rights reserved.)

The complete circular mitogenome of Paragonimus skrjabini miyazakii (Platyhelminthes: Paragonimidae) from Japan, obtained by PacBio long-read sequencing, was 17 591 bp and contained 12 protein-coding genes (PCGs), 2 mitoribosomal RNA and 22 transfer RNA genes. The atp 8 gene was absent, and there was a 40 bp overlap between nad 4L and nad 4. The long non-coding region (4.3 kb) included distinct types of long and short repeat units. The pattern of base usage for PCGs and the mtDNA coding region overall in Asian and American Paragonimus species ( P. s. miyazakii , P. heterotremus , P. ohirai and P. kellicotti ) and the Indian form of P. westermani was T > G > A > C . On the other hand, East-Asian P. westermani used T > G > C > A . Five Asian and American Paragonimus species and P. westermani had TTT/Phe, TTG/Leu and GTT/Val as the most frequently used codons, whereas the least-used codons were different in each species and between regional forms of P. westermani . The phylogenetic tree reconstructed from a concatenated alignment of amino acids of 12 PCGs from 36 strains/26 species/5 families of trematodes confirmed that the Paragonimidae is monophyletic, with 100% nodal support. Paragonimus skrjabini miyazakii was resolved as a sister to P. heterotremus. The P. westermani clade was clearly separate from remaining congeners. The latter clade was comprised of 2 subclades, one of the East-Asian and the other of the Indian Type 1 samples. Additional mitogenomes in the Paragonimidae are needed for genomic characterization and are useful for diagnostics, identification and genetic/ phylogenetic/ epidemiological/ evolutionary studies of the Paragonimidae.

Mussel production is expanding worldwide, and in Brazil the main species currently produced is the mussel Perna perna. Bucephalid trematodes have been recorded in P. perna but their larval identification is problematic. In this context, the aims of this paper were to evaluate the prevalence of bucephalids in P. perna, perform taxonomic and phylogenetic trematode studies, and analyze potential histopathological alterations in the infected host. Mussels obtained by fishers from Guanabara Bay, Rio de Janeiro, Brazil were weighed and measured, and internal organ tissues and parasites were collected. Of the 69 analyzed mussels, 24.6 % (17/69) were parasitized by bucephalid larvae. Sporocysts were located mainly in host mantle. Mussels presented sporocysts and cercaria within the connective tissue of mantle, all without associated inflammatory reactions. Parasite loads varied from less than 5 % to > 50 % of parasitized tissue. Histopathological examinations indicated that male or female gonads were not observed in 77 % (10/13) of parasitized mussels and in 4 % (2/56) identified as non-parasitized in the histology but previously classified as parasitized in the stereomicroscopic analysis. Thus, the absence of gonads may be associated with parasitism. Prosorhynchoides sp. is reported herein for the first time in mussels sampled on the coast of Rio de Janeiro, with genetic and histological data reported for the intermediate host, sporocysts and cercariae. New 28S rDNA, 18S rDNA and ITS1, 5.8S and ITS2 sequences are provided., 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 © 2022 The Authors. Published by Elsevier Inc. All rights reserved.)

Marker-assisted selective breeding of fish with higher levels of resistance towards specific pathogens may improve fish health, but the impact of host genotype on susceptibility to multiple pathogen infections is still poorly investigated. This study examined the resistance in rainbow trout Oncorhynchus mykiss towards infection with the eye fluke Diplostomum pseudospathaceum. We used genetically selected rainbow trout, carrying SNPs associated with resistance towards the parasitic ciliate Ichthyophthirius multifiliis, and exposed the fish to eye fluke cercariae. We showed that fish partly resistant to I. multifiliis were more susceptible to eye fluke invasion. The expression of immune relevant genes (encoding innate and adaptive factors) was also affected as these genotypes responded less strongly to a secondary fluke infection. The complexity of genome architecture in disease resistance towards multiple pathogens is discussed., Competing Interests: The authors have declared that no competing interests exist.

Metacercariae of Tylodelphys sp. were found in the abdominal cavity of the Chinese sleeper (Perccottus glenii) collected in Liaoning Province and Inner Mongolia Autonomous Region of China. The sequences of the mitochondrial cox1 gene and ribosomal ITS1-5.8S rDNA-ITS2 region were obtained and used for molecular identification and phylogenetic assessment of this parasite species. Results of phylogenetic analyses based on ITS and cox1 markers showed that the metacercariae of Tylodelphys sp. ex P. glenii from China were conspecific with specimens of Tylodelphys sp. collected by Sokolov et al. (2013) from the same fish-host species captured earlier in West Siberia, Russia. The examined Tylodelphys sp. ex. P. glenii is the only member of the genus whose metacercariae parasitise the abdominal cavity of fish in northern Eurasia. Tylodelphys sp. ex P. glenii clustered with T. darbyi, T. immer, T. podicipina, and Tylodelphys sp. of Soldánová et al., 2017 based on mitochondrial DNA markers, and with T. darbyi, T. immer, T. kuerepus, and T. schreuringi using nuclear DNA markers.

Renschetrematidae is a very small family of digeneans parasitic in bats in southern and Southeast Asia. According to the original descriptions and the latest revision, its representatives are characterized by the presence of several unusual characters. Among them are the dorsal position of the genital pores, separate male and female genital pores, and the presence of an accessory sac (stylet pouch) associated with terminal genitalia and containing a stylet-like structure. Prior to our study, the phylogenetic relationships of the Renschetrematidae were unknown and DNA sequence data were absent from any of its representatives. In this work, we present the description of a new species of Renschetrema from bats in the Philippines. We also evaluate the phylogenetic affinities of the Renschetrematidae using newly obtained partial sequences of the 28S rRNA gene from Renschetrema specimens collected in the Philippines and Southeast China. The new species differs from previously described species by the position and orientations of the stylet pouch, presence of accessory spines around genital atrium, position of gonads, body shape and relative size of pharynx and oral sucker. Our phylogenetic analysis supports the status of the Renschetrematidae as an independent family within the superfamily Microphalloidea. In the phylogenetic tree, the Renschetrematidae appeared as an independent family-level lineage, basal to the remaining taxa within the Microphalloidea. Detailed examination of our specimens revealed significant errors in the current diagnoses of the genus Renschetrema and the Renschetrematidae. Amended diagnoses of the genus and the family are provided., (© 2022. The Author(s), under exclusive licence to Springer Nature B.V.)

We report a species of diplostomid fluke recovered from 3 carcasses of wild Korean raccoon dog, Nyctereutes procyonoides koreensis, in Korea. A total of 107 diplostomid flukes were recovered from the small intestines of Korean raccoon dogs, which were obtained from the Gangwon Wildlife Medical Rescue Center. Worms fixed with 10% neutral formalin were subjected to microscopic observation and those fixed in 70% ethanol were used for molecular genomic analysis. The worm was divided into 2 separate parts, forebody and hindbody, with a total length of 3,020-4,090 (3,855) µm and a width of 1,210-1,770 (1,562) µm. The boat-shaped forebody has a pair of characteristic tentacular appendage, 2 suckers, holdfast organ, and vitelline follicles. The oval to cylindrical hindbody has reproductive organs. The ovary was round or elliptical and located in the anterior of the testes. Two large testes were slightly segmented and tandemly arranged, occupying almost half of hindbody. The short uterus contained a relatively small number of unembryonated eggs sized 130-140×85-96 µm. The partial sequence of 18S rRNA of this fluke was consistent with Alaria alata. Based on the morphological and molecular characteristics, the diplostomid flukes recovered from the small intestine of Korean raccoon dogs were identified as A. alata (Digenea: Diplostomidae).

A study published in 2016 reported on an undescribed species of Apatemon (Strigeidae) from New Zealand that was previously well known from its larval stages. Only a single specimen from a mallard duck was available at the time, which was described and given the provisional name Apatemon sp. "jamiesoni". Specimens also obtained from a spotted shag were not in good enough condition to form the basis of a new species description. A black-backed gull has since been discovered with specimens of this strigeid, their identity confirmed by genetic similarity, allowing formal description and naming of this species. This paper provides a description of the new specimens from the black-backed gull, along with a comparison with the specimens from other bird hosts, reprises some data from Blasco-Costa et al. (Parasitol Res 115:271-289, 2016) and presents formally the name Apatemon jamiesoni n. sp. This species differs from all other species of Apatemon in its small size, particularly that of the ovary and testes. It is most similar to A. jamesi from which it differs in the size of the oral and ventral suckers., (© 2022. The Author(s).)

The best way to study digenean diversity combines molecular genetic methods, life-cycle studies and elaborate morphological descriptions. This approach has been barely used for one of the most widespread digenean taxa parasitizing fish – the superfamily Hemiuroidea. Here, we applied the integrative approach to the hemiuroideans from the family Derogenidae parasitizing fish at the White and Barents Seas. Analysis of 28S, 18S, 5.8S rDNA, ITS2 and cox1 gene sequences from sexually adult worms (maritae) showed genetic heterogeneity for 2 derogenid species known from this area: Derogenes varicus and Progonus muelleri . Thus, 2 pairs of genetic lineages were found: DV1 and DV2, PM1 and PM2, respectively. Data from other regions indicate that 2 more lineages of D. varicus probably exist. Based on previous records from the White and Barents Seas, we hypothesized that the cercariae found in the moonsnails (family Naticidae) belong to the Derogenidae and may help to differentiate these lineages as species. According to our results, Cercaria appendiculata from Cryptonatica affinis matched DV1, similar nameless cercariae from Euspira pallida and Amauropsis islandica matched DV2, and Cercaria octocauda from C. affinis matched PM1. We provide new data on the structure of these cercariae and discuss the life-cycle pattern of the studied digeneans.

Larval didymozoids (Trematoda: Digenea) were discovered parasitizing the hemocoel of the heteropod Firoloida desmarestia (redia mean intensity = 13) and the chaetognaths Flaccisagitta enflata and Flaccisagitta hexaptera (metacercaria mean intensity = 1) during a 2014-2016 systematic study of parasites of zooplankton collected in the central and southern regions of the Gulf of California, Mexico. Didymozoid infection route during the early life cycle was inferred combining morphological (light microscopy) and molecular (mitochondrial cytochrome c oxidase subunit I gene, cox1) evidence. Didymozoid rediae parasitizing F. desmarestia were observed, just after field collection of the host, containing hundredths of completely developed cystophorous cercariae, releasing them though the birth pore at approximately one cercaria every 12 s. Cercariae lost their tails developing into a 'young metacercaria' in 1 d at 22 °C without need of an intermediate host. Molecular analysis of cox1 showed that rediae found in F. desmarestia belong to two distinct didymozoid species (Didymozoidae sp. 1 and sp. 2). Metacercariae parasitizing chaetognaths were morphologically identified as Didymozoidae type Monilicaecum and cox1 sequences showed that metacercariae of chaetognaths matched with these two Didymozoidae sp. 1, and sp. 2 species found parasitizing F. desmarestia, plus a third distinct Didymozoidae sp. 3. These are the first DNA sequences of cox1 gene from didymozoid larvae for any zooplankton taxonomic group in the world. We concluded that F. desmarestia is the first intermediate host of rediae and cercariae, and the chaetognaths are the second intermediate hosts where non-encysted metacercariae were found. The definitive host is still unknown because cox1 sequences of present study did not genetically match with any available cox1 sequence of adult didymozoid. Our results demonstrate a potential overlap in the distribution of two carnivorous zooplankton taxonomic groups that are intermediate hosts of didymozoids in the pelagic habitat. The didymozoid specimens were not identified to species level because any of the cox1 sequences generated here matched with the sequences of adult didymozoids currently available in GenBank and Bold System databases. This study provides baseline information for the future morphological and molecular understanding of the Didymozoidae larvae that has been previously based on the recognition of the 12 known morphotypes., (© 2022. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

Parasites might represent a helpful tool in understanding the historical dispersion and phylogeography of their hosts. In order to reveal whether the migration routes and diversification of hosts can be traceable in the genetic structures of their parasites, we investigated the diversity of paramphistomoid trematodes of Pelophylax frogs in 2 geographically distant European regions. Water frogs belonging to the genus Pelophylax represent a striking example of a species with a high variety of ecological adaptations and a rich evolutionary history. The parasites were collected from 2 Balkan endemic species, P. epeiroticus and P. kurtmuelleri , and 2 species in Slovakia, P. esculentus and P. ridibundus . While in Slovakia, Pelophylax frogs harboured 2 species, the diplodiscid Diplodiscus subclavatus and the cladorchiid Opisthodiscus diplodiscoides , only the former was recorded in the south-western Balkans. Remarkably high genetic diversity (16 unique mitochondrial cox 1 haplotypes, recognized among 60 novel sequences) was observed in D. subclavatus , and subsequent phylogenetic analyses revealed a strong population-genetic structure associated with geographical distribution. We also evidenced the existence of 2 divergent D. subclavatus cox 1 haplogroups in the south-western Balkans, which might be associated with the historical diversification of endemic water frogs in the regional glacial microrefugia.

Platynosomum is a digenean trematode causing hepatobiliary disease in cats in tropical and subtropical regions. The presence of Platynosomum species in Sri Lanka has not been previously reported or investigated. In the current study, we report a clinical case of a cat suffering from hepatic and biliary complications. Fine-needle aspiration of the biliary extract revealed a large number of parasite eggs, and the morphological and molecular identification of eggs was carried out. Molecular phylogenetics was performed using the nuclear ribosomal Internal Transcribed Spacer 2 (ITS2) and a portion of a mitochondrially encoded gene; Cytochrome C Oxidase subunit 1 (COX1). Through a combination of history, clinical signs, blood reports, ultrasound scanning, light microscopy of eggs from biliary aspirate and molecular studies, the disease was confirmed as parasitism caused by a Platynosomum like species. The Platynosomum species in Sri Lanka is phylogenetically related to Platynosomum illiciens reported from Costa Rica, Central America. This case emphasizes the importance of future studies in Sri Lanka regarding the prevalence and distribution of Platynosomum among cat populations. Further, the inclusion of feline platynosomiasis in the differential diagnoses list for hepatobiliary diseases is required., (Copyright © 2022 Elsevier B.V. All rights reserved.)

Biological category is effective to indicate the evolution of organism populations between past and present. Conventional taxonomy of human parasites mainly depends on important morphological features, which suffers from a problem of categorizing related-genera species with similar morphological characteristics. With recent advances in molecular biological technologies, the effective applications of mitochondrial and ribosomal biomarkers and sequencing greatly improve the development of the taxonomic rank of human parasites. Worldwide, the classification of human parasites have been continuously revised and improved. Hereby, we re-categorize parasitic Protozoa, Trematoda, Cestoda and Nematoda, so as to provide insights into the researches on molecular systematics and genetic evolution of human parasites.

White grub metacercariae were found in the livers and kidneys of diseased specimens of an introduced channel catfish, Ictalurus punctatus (Rafinesque), in Vietnam. Based on morphological features and 28S rDNA sequence analysis, the isolated metacercariae were identified as Dollfustrema bagarii (Digenea: Bucephalidae) Moravec & Sey. Histopathological examination shows that encysted metacercariae can change the tissue structure of the infected organs and is often accompanied by haemorrhaging and the presence of eosinophilic granular cell infiltration. Degenerative changes were also observed., (© 2022 John Wiley & Sons Ltd.)

The capsalid monogeneans are important pathogens that generally infect marine fishes and have a substantial impact on fish welfare in aquaculture systems worldwide. However, the current mitogenome information on capsalids has received little attention, limiting the understanding of their evolution and phylogenetic relationships with other monogeneans. This paper reports the complete mitochondrial genomes of Capsala katsuwoni and Capsala martinieri for the first time, which we obtained using a next-generation sequencing method. The mitogenomes of C. katsuwoni and C. martinieri are 13,265 and 13,984 bp in length, respectively. Both species contain the typical 12 protein-coding genes, 2 ribosomal RNA genes, 22 transfer RNA genes, and a control region. The genome compositions show a moderate A+T bias (66.5% and 63.9% for C. katsuwoni and C. martinieri , respectively) and exhibit a negative AT skew but a positive GC skew in both species. One gene block rearrangement was found in C. katsuwoni in comparison with other capsalid species. Instead of being basal to the Gyrodactylidea and Dactylogyridea or being clustered with Dactylogyridea, all species of Capsalidea are grouped into a monophyletic clade. Our results clarify the gene rearrangement process and evolutionary status of Capsalidae and lay a foundation for further phylogenetic studies of monogeneans.

A new cryptogonimid trematode, Siphoderina hustoni n. sp., is reported, collected off Lizard Island, Queensland, Australia, from the Maori snapper Lutjanus rivulatus (Cuvier). The new species is moderately distinctive within the genus. It is larger and more elongate than most other species of Siphoderina Manter, 1934, has the shortest forebody of any, a relatively large ventral sucker, a long post-testicular zone, and is perhaps most recognisable for the substantial space in the midbody between the ventral sucker and ovary devoid of uterine coils and vitelline follicles, the former being restricted to largely posterior to the ovary and the latter distributed from the level of the anterior testis to the level of the ovary. In phylogenetic analyses of 28S ribosomal DNA, the new species resolved with the other nine species of Siphoderina for which sequence data are available, all of which are from Queensland waters and from lutjanid and haemulid fishes. Molecular barcode data were also generated, for the ITS2 ribosomal DNA and cox1 mitochondrial DNA markers. The new species is the first cryptogonimid known from L. rivulatus and the first metazoan parasite reported from that fish in Australian waters., (© 2022. The Author(s).)

Opisthorchiasis and related infections are persistent and substantial public health burdens from eastern Europe to southeastern and northern Asia. Snails of the family Bithyniidae act as first intermediate hosts not only for the trematodes of the family Opisthorchiidae but also for those of the family Notocotylidae. There are certain morphological similarities between the aforementioned trematode cercariae. In order to find natural local foci of opisthorchiasis, which are established primarily according to the presence of infected bithyniid snails at the area under examination, it is crucial to correctly identify the rediae and cercariae of the trematodes. The aim of our investigation was to evaluate the role of bithyniid snails in the transmission of Opisthorchiidae and Notocotylidae in ecosystems in the south of Western Siberia. We have been studying the process of bithyniid snail dissemination in Western Siberia and examining their infection by trematodes from 1994 until now. A total of 16,213 bithyniid snails in 23 water bodies (in 28 localities) of four major basins situated in the Novosibirsk region were inspected for trematode infestation. Long-term research has been conducted in the Kargat river estuary and the Ob river floodplain for 15 and 25 years, respectively. In both water bodies, the prevalence of notocotylid and opisthorchiid trematodes was positively correlated with the global land-ocean temperature index. Trematode parthenitae were identified if there were mature cercariae capable of leaving the shell of the host snail on their own. Identification to the species of opisthorchiid cercariae was verified using molecular genetic analysis methods targeting ITS2 locus. Opisthorchis felineus and Metorchis bilis, two opisthorchiid species that are potentially perilous to human health, were found in bithyniids in the Novosibirsk region. The mean prevalence of infection with notocotylid trematodes in bithyniid snails was higher than the corresponding prevalence of infection with opisthorchiid trematodes. The results of this study can be used to identify and predict natural foci of epidemiologically and/or epizootically dangerous diseases., (© 2022. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

Host-parasite coevolution is one of the main topics of the evolutionary biology of host-parasite associations. The majority of monogeneans parasitizing fish exhibit a high degree of host specificity. As a result, their evolutionary history might be intertwined with that of their fish hosts. The Cichlidae represent a diverse group of secondary freshwater fish with disjunctive distribution. Host-specific dactylogyrid monogeneans commonly parasitize cichlid fish. Their high diversity is associated with the main areas of cichlid distribution, i.e., Neotropical America and Africa. Nevertheless, the parasite fauna of cichlids from Neotropical America is still underexplored. A total of 31 cichlid species were examined for the presence of monogeneans, with 20 of them being parasitized. On these cichlids, 30 monogeneans belonging to the genera Gussevia, Trinidactylus, and Scadicleithrum were identified, 17 of them potentially representing new species for science. Phylogenetic analyses revealed three monophyletic groups of Neotropic cichlid monogeneans. Genus Gussevia was monophyletic, while Sciadicleithrum resulted polyphyletic. Sciedicleithrum from South America and Sciadicleithrum from Mexico represented two divergent lineages. The plesiomorphic Neotropical cichlid host group for dactylogyrid monogeneans was Cichlini, from which the representatives of other Neotropical cichlid tribes were colonised. Cophylogenetic analyses revealed a statistically significant cophylogenetic signal in the investigated host-parasite system, with host switch and duplication representing the main coevolutionary events for monogeneans parasitizing Neotropical cichlids. This scenario is in accordance with previous studies focussed on dactylogyridean monogeneans parasitizing freshwater fish in Europe and Africa., (Copyright © 2022. Published by Elsevier Ltd.)

Plesiochorus Looss, 1901 is a genus of Gorgoderidae infecting the urinary bladders of marine turtles globally. Currently, just two morphologically similar species are recognised, Plesiochorus cymbiformis (Rudolphi, 1819) Looss, 1901 and Plesiochorus elongatus Pigulevsky, 1953, which have been distinguished by molecular data and subtle morphological differences. Here we describe a new species, Plesiochorus irwinorum n. sp., infecting hawksbill turtles (Eretmochelys imbricata (L.)), which is primarily distinguished from the other two species of Plesiochorus on the basis of ITS2, cox1 and 28S sequence data. Morphometric data for specimens examined during this study overlap between P. cymbiformis and P. irwinorum n. sp. for every measured feature, rendering them functionally cryptic. However, principal components analysis clearly distinguishes the two species. Additionally, we report new specimens of P. cymbiformis, and provide new sequence data for specimens from Australian loggerhead (Caretta caretta (L.)) and hawksbill turtles. There is little understanding of the host-specificity or geographical distribution of the three species of Plesiochorus, and it remains possible that some of the previously reported sequences have been attributed to the wrong species., (© 2022. The Author(s).)

The Levant Basin is in many ways the world's most invaded marine ecosystem owing to the existence of the man-made Suez Canal. The invasion of free-living organisms through this pathway is increasingly documented and monitored in the past two decades, and their ecological impact recognized. Nonetheless, while tremendous scientific effort is invested in documenting introduced fishes, co-introduction events of these fishes and their parasites have drawn relatively little interest. In our research, we examined the presence of gill parasites (Monogenea) on the invasive narrow barred Spanish mackerel Scomberomorus commerson which has been known in the Mediterranean Sea for 80 years. The gills of S. commerson supported numerous, relatively large monogeneans (Monogenea: Gastrocotylinea), reaching prevalence levels of 100% with a mean intensity of ~ 80 worms per host. Using an integrated molecular and morphological approach, four gastrocotylinean species were identified: Gotocotyla acanthura, Cathucotyle cathuaui, Pricea multae, and Pseudothoracocotyla ovalis. Two species, C. cathuaui and P. ovalis, are reported here for the first time from the Mediterranean. Sequences of the 28S rRNA gene of G. acanthura from native hosts, Pomatomus saltatrix and Trachinotus ovatus, differed from individuals collected from S. commerson by 1.8%. We therefore suggest that the taxonomic status and distribution of G. acanthura should be revisited, and we recommend an integrated approach as essential to accurately detect co-introductions., (© 2022. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

Prosthogonimus cuneatus and Prosthogonimus pellucidus (Trematoda: Prosthogonimidae) are common flukes of poultry and other birds which can cause severe impacts on animal health and losses to the poultry industry. However, there are limited studies on the molecular epidemiology, population genetics, and systematics of Prosthogonimus species. In the present study, the complete mitochondrial (mt) genomes of P. cuneatus and P. pellucidus were determined to be 14,829 bp and 15,013 bp in length, respectively. Both mt genomes contain 12 protein-coding genes (PCGs) (cox1-3, nad1-6, nad4L, cytb, and atp6), 22 transfer RNA genes, two ribosomal RNA genes, and one non-coding region. Our comparative analysis shows that the atp6 genes of P. cuneatus and P. pellucidus are longer than any previously published atp6 genes of other trematodes. The lengths of the atp6 genes of P. cuneatus and P. pellucidus in this study seem unusual, and should therefore be studied further. The mt genes of P. cuneatus and P. pellucidus are transcribed in the same direction, and the gene arrangements are identical to those of Plagiorchis maculosus, Tamerlania zarudnyi, and Tanaisia sp., but different from those of Eurytrema pancreaticum, Dicrocoelium chinensis, and Brachycladium goliath. The mt genome A + T contents of P. cuneatus and P. pellucidus are 64.47% and 65.34%, respectively. In the 12 PCGs, ATG is the most common initiation codon, whereas TAG is the most common termination codon. The sequence identity of the same 12 PCGs among the eight trematodes (P. cuneatus, P. pellucidus, Pl. maculosus, D. chinensis, E. pancreaticum, B. goliath, T. zarudnyi, Tanaisia sp.) of Xiphidiata are 55.5%-81.7% at the nucleotide level and 43.9%-82.5% at the amino acid level. The nucleotide similarities among the complete mt genomes of the eight trematodes range from 54.1%-81.5%. Phylogenetic analysis based on the aligned concatenated amino acid sequences of the 12 PCGs shows that P. cuneatus and P. pellucidus cluster together and are sister to T. zarudnyi and Tanaisia sp., and this clade is more closely related to E. pancreaticum, Dicrocoelium spp. and Lyperosomum longicauda in the family Dicrocoeliidae, than it is to species in the families Plagiorchiidae and Brachycladiidae. These are the first reported complete mt genomes of Prosthogonimidae, and these data will provide additional molecular resources for further studies of Prosthogonimidae taxonomy, population genetics, and systematics., (Copyright © 2022. Published by Elsevier B.V.)

We describe a new species of Plagioporus Stafford, 1904 infecting the intestine of two catostomids in the eastern USA. We emend Plagioporus to account for Nearctic congeners having ceca terminating at the level of the testes (previously diagnosed as having ceca terminating in the post-testicular space only) and testes in the posterior body extremity (a feature not previously considered as having generic importance). Of the accepted Nearctic species, Plagioporus wataugaensis n. sp. resembles Plagioporus serotinus Stafford, 1904, Plagioporus hypentelii Hendrix, 1973, and Plagioporus hageli Fayton and Andres, 2016 but differs from them by the distribution of the vitellarium and proportional length and relative extent of the excretory vesicle. Plagioporus wataugaensis has vitelline fields that are discontinuous at the level of the ventral sucker (vs. continuous in P. serotinus and P. hypentelii) and follicles that surround the ceca (vs. wholly ventral to the ceca in P. hageli) and that span the midline dorsal to the testes (vs. slightly overlapping the lateral margins of the testes). The excretory vesicle of P. wataugaensis is wholly post-testicular and short (6-9% of the body length) (vs. reaching the level of the posterior testis, 14-24% of the body length). Phylogenetic analyses of the 28S, ITS1, 5.8S, and ITS2 rDNA recovered P. wataugaensis sister to Plagioporus sinitsini Mueller, 1934. A key to the Nearctic Plagioporus spp. is provided. We regard Plagioporus shawi (McIntosh, 1939) Margolis, 1970, Plagioporus serratus Miller, 1940, and Plagioporus loboides (Curran, Overstreet, and Tkach, 2007) Fayton and Andres, 2016 as incertae sedis., (Published by Elsevier B.V.)

We obtained new data on the complete mitochondrial DNA (mtDNA) and the ribosomal operon of the trematode Carassotrema koreanum (Digenea: Haploporata: Haploporidae), an intestinal parasite of Carassius auratus , using next-generation sequencing. The mtDNA of C . koreanum contained 13,965 bp, including 12 protein-coding genes, two ribosomal genes, 22 transport RNA (tRNA) genes and a non-coding region. The ribosomal operon of C . koreanum was 10,644 bp in length, including ETS1 (1449 bp), 18S ribosomal RNA (rRNA) gene (1988 bp), ITS1 ribosomal DNA (rDNA) (558 bp), 5.8S rRNA gene (157 bp), ITS2 rDNA (274 bp), 28S rRNA gene (4152 bp) and ETS2 (2066 bp). Phylogenetic analysis based on mtDNA protein-coding regions showed that C . koreanum was closely related to Parasaccocoelium mugili , a species from the same suborder Haploporata. Bayesian phylogenetic tree topology was the most reliable and confirmed the validity of the Haploporata. The results of sequence cluster analysis based on codon usage bias demonstrated some agreement with the results of the phylogenetic analysis. In particular, Schistosoma spp. were differentiated from the other members of Digenea and the members of Pronocephalata were localized within the same cluster. Carassotrema koreanum and P . mugili fell within different clusters. The grouping of C . koreanum and P . mugili within the same cluster was obtained on the basis of frequencies of 13 specified codons, of which three codon pairs were degenerate. A similarity was found between two haploporid species and two Dicrocoelium spp. in the presence of TTG start codon of the mitochondrial nad 5 gene. Our results confirmed the taxonomical status of the Haploporata identified in the previous studies and revealed some characteristic features of the codon usage in its representatives.

The assassin snail genus Anentome is widespread in Southeast Asia, and is distributed all over the world via the aquarium trade. One species of genus Anentome , Anentome helena , is known to act as intermediate host of parasitic trematodes. This study investigates the taxonomic diversity of larval trematodes infecting A . helena and Anentome wykoffi in Thailand. Larval trematodes were identified by combining morphological and DNA sequence data (cytochrome c oxidase I and internal transcribed spacer 2). Species delimitation methods were used to explore larval trematode species boundaries. A total of 1107 specimens of Anentome sp. were collected from 25 localities in Thailand. Sixty-two specimens of A . helena ( n = 33) and A . wykoffi ( n = 29) were infected by zoogonid cercariae, heterophyid metacercariae and echinostome metacercariae, with an overall prevalence of 5.6% (62/1107) and population-level prevalences in the range of 0.0-22.3%. DNA sequence data confirmed that the larval trematodes belong to the families Echinostomatidae, Heterophyidae and Zoogonidae. As such, this is the first report of zoogonid cercariae and heterophyid metacercariae in A . helena , and echinostome metacercariae in A . wykoffi . Moreover, this study provides evidence of tentative species-level differentiation between Thai Echinostoma sp. and Cambodian Echinostoma mekongi , as well as within Echinostoma caproni , Echinostoma trivolvis and Echinostoma revolutum .

Digenetic trematodes are important parasites of humans and animals. They have complex life cycles and typically infect a gastropod as the first intermediate host. Bithynia siamensis goniomphalos , the first intermediate host of the liver fluke, Opisthorchis viverrini , harbours a wide variety of other trematode species. Morphological details of cercariae of 20 trematode taxa from B . s . goniomphalos , collected mainly in Thailand from 2009 to 2014, were provided in an earlier paper. Correct identification to the species or genus level based on morphology of these cercariae is generally not possible. Therefore, we used molecular data to improve identification and to investigate the diversity of the species of trematodes infecting B . s . goniomphalos . We were successful in extracting, amplifying and sequencing portions of the 28S ribosomal RNA (rRNA) gene for 19 of these 20 types of cercaria, and the internal transcribed spacer 2 region for 18 types. BLAST searches in GenBank and phylogenetic trees inferred from the 28S rRNA sequences identified members of at least nine superfamilies and 12 families. Only a few cercariae could be assigned confidently to genus or species on the basis of the sequence data. Matching sequence data from named adult trematodes will be required for definitive identification. There is clearly a great diversity of trematode species utilizing B . s . goniomphalos in Thailand.

On the basis of the morphological characterization of Chelatrema neilgherriensis Manjula & Janardanan, 2006 recovered from the freshwater fish Barilius gatensis (Valenciennes, 1844) in the Wayanad region of the Western Ghats, the diagnostic features of the genus Chelatrema Gupta & Kumari, 1973 have been modified. Based on the phylogenetic analysis of C. neilgherriensis and comparative morphology studies relative to members of other families of Gorgoderoidea Looss, 1901, this genus is placed in a new family Chelatrematidae n. fam. The studies revealed the molecular and morphological closeness of Chelatrema with Paracreptatrematina limi Amin & Myer, 1982, and the latter is transferred to this new family. Hence the new family Chelatrematidae n. fam. comprises the genera Chelatrema and Paracreptatrematina .

We revise monotypic LeuceruthrusMarshall and Gilbert, 1905 (Azygiidae Lühe, 1909) by emending its generic diagnosis, redescribing its type species (Leuceruthrus micropteriMarshall and Gilbert, 1905), reassigning 2 species (Leuceruthrus stephanocauda [Faust, 1921] n. comb., Leuceruthrus ocalana [Smith, 1935] n. comb.), describing 2 new congeners (Leuceruthrus ksepkai n. sp. and Leuceruthrus blaisei n. sp.), and providing a phylogenetic analysis based on the internal transcribed spacer 2 (ITS2). Leuceruthrus is unique by having oblique preovarian testes, a vitellarium that does not extend anteriad into the forebody, and a uterus that is intercecal and between the ovary and ventral sucker. We describe the cercaria of Leuceruthrus cf. stephanocauda from cercariae shed from Elimia cf. carinifera and Elimia cf. modesta from Big Canoe Creek, Alabama. Leuceruthrus ksepkai n. sp. is described from cercariae shed from rasp elimia, Elimia floridensis (Reeve, 1860) from Holmes Creek, Florida, and Elimia sp. 1 from the Chocktawhatchee River, Florida. It differs from its congeners by the combination of having broadly rounded furcae with slight marginal pigmentation in live cercariae, no spines on the tail stem, distinct anterior and posterior ridges that flank the tail stem portion accommodating the withdrawn distome, minute protuberances occupying the lateral margin of the tail stem for its entire length, and protuberances that encircle the anterior third of the posterior tail stem (immediately posterior to the tail stem portion containing the withdrawn distome). Leuceruthrus blaisei n. sp. infects Elimia sp. and is the only known congener having proportionally small furcae (>tail stem maximum width) with a nipple-like distal projection and numerous minute projections on the tail stem surface. Our phylogenetic analysis included all of our new sequences plus all publicly available ITS2 sequences for Leuceruthrus spp. and Proteromera spp., and supported the monophyly of Leuceruthrus. It recovered L. ksepkai and L. cf. ksepkai in a polytomy, and Leuceruthrus blaisei as monophyletic (identical sequences), with L. micropteri and L. cf. stephanocauda recovered as sister taxa. The present study comprises the first systematic treatment of Leuceruthrus in over a century, the first description of a new species of Leuceruthrus in 117 yr, and the first taxonomic characterization of a species of Leuceruthrus from the Mobile River Basin. It also increases the number of accepted congeners from 1 to 5 and provides new host records (E. cf. carinifera and E. floridensis) for Leuceruthrus spp., (© American Society of Parasitologists 2022.)

Gymnophallus deliciosus is a type species of the genus Gymnophallus. We collected this trematode species from gallbladder of Larus argentatus caught in the White Sea (Kandalaksha Bay, Chupa Inlet) and obtained the sequences of its nuclear 18S and 28S rDNA genes. Our recently obtained phylogenetic data support a sister group relationship of this species with G. choledochus. However, other species of the genus Gymnophallus-G. australis (KM246854) and G. minutus (KM268111)-do not branch with the group G. choledochus + G. deliciosus or with each other. Our study revealed that the genus Gynmnophallus is probably a polyphyletic taxon, and the genus affiliation of its representatives should be re-examined in future., (© 2022. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

A new species of lepocreadiid, Opechonoides opisthoporus n. sp., is described infecting 12 pomacentrid fish species from the Great Barrier Reef, Australia, with Abudefduf whitleyi Allen & Robertson as the type-host. This taxon differs from the only other known member of the genus, Opechonoides gure Yamaguti, 1940, in the sucker width ratio, cirrus-sac length, position of the testes, position of the pore of Laurer's canal, and relative post-testicular distance. The new species exhibits stenoxenic host-specificity, infecting pomacentrids from seven genera: Abudefduf Forsskål, Amphiprion Bloch & Schneider, Neoglyphidodon Allen, Neopomacentrus Allen, Plectroglyphidodon Fowler & Ball, Pomacentrus Lacépède and Stegastes Jenyns. Phylogenetic analyses of 28S rDNA sequence data demonstrate that O. opisthoporus n. sp. forms a strongly supported clade with Prodistomum orientale (Layman, 1930) Bray & Gibson, 1990. The life cycle of this new species is partly elucidated on the basis of ITS2 rDNA sequence data; intermediate hosts are shown to be three species of Ctenophora. New host records and molecular data are reported for Lepocreadium oyabitcha Machida, 1984 and Lepotrema amblyglyphidodonis Bray, Cutmore & Cribb, 2018, and new molecular data are provided for Lepotrema acanthochromidis Bray, Cutmore & Cribb, 2018 and Lepotrema adlardi (Bray, Cribb & Barker, 1993) Bray & Cribb, 1996. Novel cox1 mtDNA sequence data showed intraspecific geographical structuring between Heron Island and Lizard Island for L. acanthochromidis but not for L. adlardi or O. opisthoporus n. sp., (© 2022. The Author(s).)

The phylogenetic relationships established in the family Allocreadiidae by genetic methods suggest that the center of origin, distribution, and divergence of the studied family is, apparently, eastern regions of Laurasia (the territory of the modern Southeast Asia), from where these trematodes penetrated through the Amur paleomicrocontinent and Beringia to North America and further through Central America to South, as well as through Transbaikalia and Yakutia to the western part of Eurasia., (© 2022. Pleiades Publishing, Ltd.)

Specimens of Dolichoperoides macalpini (Nicoll, 1914) (Digenea: Dolichoperoididae) were collected from Australian venomous snakes (Elapidae): Notechis scutatus Peters, 1861 and Austrelaps superbus (Günther, 1858) from Tasmania and surrounding islands and N. s. occidentalis Glauert, 1948 from wetlands near Perth, Western Australia. Despite variation in morphological measurements, genetic analysis showed that the one species of digeneans infected the snakes from all locations. This study presents the first DNA sequences for D. macalpini (internal transcribed spacer, 18S, 28S), confirming its placement in a family separate from the Reniferidae and Telorchiidae. Analysis of the infection dynamics of infection in Western Australian snakes showed significant differences in levels of infection between wetland locations, season and year of collection. Infection of D. macalpini was reported in the gastrointestinal tract, including the mouth, in freshly euthanised snakes in Western Australia, and in the lung in Tasmanian snakes, consistent with earlier reports. Differences in morphology and site of infection are suggested to be due to a combination of season and maturity of the digenean, with infection potentially occurring early in the season, as the snakes emerge from torpor. The need for research on the seasonal dynamics of infection with this parasite is discussed., (© 2022. The Author(s).)

Dactylogyridae is one of the most studied families of parasitic flatworms with more than 1000 species and 166 genera described to date including ecto- and endoparasites. Dactylogyrid monogeneans were suggested as model organisms for host-parasite macroevolutionary and biogeographical studies due to the scientific and economic importance of some of their host lineages. Consequently, an array of phylogenetic research into different dactylogyrid lineages has been produced over the past years but the last family-wide study was published 16 years ago. Here, we provide a meta-analysis of the phylogenetic relationships of Dactylogyridae including representatives of all genera with available molecular data (n = 67). First, we investigate the systematic informativeness of morphological characters widely used to diagnose dactylogyrid genera through a parsimony analysis of the characters, character mapping, and phylogenetic comparative methods. Second, we provide an overview of the current state of the systematics of the family and its subfamilies, and summarise potentially poly- and paraphyletic genera. Third, we elaborate on the implications of taxonomic, citation, and confirmation bias in past studies. Fourth, we discuss host range, biogeographical, and freshwater-marine patterns. We found two well-supported macroclades which we assigned to the subfamilies Dactylogyrinae and Ancyrocephalinae. These subfamilies further include 16 well-supported clades with only a few synapomorphies that could be deduced from generic diagnoses in the literature. Furthermore, few morphological characters considered systematically informative at the genus level display a strong phylogenetic signal. However, the parsimony analysis suggests that these characters provide little information on the relationships between genera. We conclude that a strong taxonomic bias and low coverage of DNA sequences and regions limit knowledge on morphological and biogeographical evolutionary patterns that can be inferred from these results. We propose addressing potential citation and confirmation biases through a 'level playing field' multiple sequence alignment as provided by this study., (Copyright © 2022. Published by Elsevier Ltd.)

This paper presents a new haploporid digenean that expands the number of species of Saccoccoelioides to 27. The new species, Saccocoelioides kirchneri n. sp. was collected from the intestine of Cnesterodon decemmaculatus (Poeciliidae: Cyprinodontiformes) from Lago del Bosque, La Plata, Argentina. The new species possesses the diagnostic features for Saccocoelioides: a sac like ceca; the vitellarium confined in two irregular groups of follicles distributed between the ventral sucker and the anterior margin of the testis; and a uterus confined largely in the hind-body, but encroaching into the range of the ventral sucker. The new species is differentiated from the 26 congeners by the body size, pharynx size, ventral sucker size, posterior extent of ceca, posterior extent of uterus and egg size. S. kirchneri n. sp. also is supported by the molecular analysis.

The present paper describes a new species of Dermadena (Digenea) parasitizing Pseudobalistes naufragium in Puerto Pizarro, northern Peru, using light and scanning electronic microscopy (SEM). Additionally, molecular analysis was also performed to determine the phylogenetic affinities of Dermadena within the Lepocreadiidae. The new species is differentiated from Dermadena spatiosa, Dermadena stirlingi and Dermadena lactophrysi by presenting a curved and well-developed external seminal vesicle. Also, SEM revealed numerous dome-shaped tegument protuberances forming glandular papillae with transversal wrinkles arranged roughly in concentric rows around the acetabular region, varying in size from large at the middle of the body to small at the margin. In the molecular phylogeny, the new species formed a well-supported clade with sequences of species from the Lepocreadiidae, confirming that it belongs to this family.