Molecular species delimitation of marine trematodes over wide geographical ranges: Schikhobalotrema spp. (Digenea: Haplosplanchnidae) in needlefishes (Belonidae) from the Pacific Ocean and Gulf of Mexico.

Geographical distribution plays a major role in our understanding of marine biodiversity. Some marine fish trematodes have been shown to have highly restricted geographical distributions, while some are known to occur over very wide ranges; however, very few of these wide distributions have been demonstrated genetically. Here, we analyse species of the genus Schikhobalotrema (Haplosplanchnidae) parasitizing beloniforms from the tropical west Pacific, the eastern Pacific and the Gulf of Mexico (GoM). We test the boundaries of these trematodes by integrating molecular and morphological data, host association, habitat of the hosts and geographical distribution, following a recently proposed and standardized delineation method for the recognition of marine trematode species. Based on the new collections, Schikhobalotrema huffmani is here synonymized with the type-species of the genus, Schikhobalotrema acutum; Sch. acutum is now considered to be widely distributed, from the GoM to the western Pacific. Additionally, we describe a new species, Schikhobalotrema minutum n. sp., from Strongylura notata and Strongylura marina (Belonidae) from La Carbonera coastal lagoon, northern Yucatán, GoM. We briefly discuss the role of host association and historical biogeography of the hosts as drivers of species diversification of Schikhobalotrema infecting beloniforms.

Hidden in the fog: morphological and molecular characterisation of Derogenes varicus sensu stricto (Trematoda, Derogenidae) from Sweden and Norway, and redescription of two poorly known Derogenes species.

Derogenes varicus (Müller, 1784) is widely reported as a trematode with exceptionally low host specificity and a wide, bipolar distribution. However, several recent studies have suggested that D. varicus represents a species complex and based on molecular evidence, four genetic lineages (labeled as "DV1-4") have been designated within the D. varicus species complex. This possibility requires improved (ideally molecular) characterisation of specimens from the type-host (Salmo salar) and type-locality (off Denmark). During examination of trematode parasites of fish from Scandinavian and Arctic waters (Sweden and Norway), we found specimens of D. varicus in the stomach of Merlangius merlangus off the coast of Sweden, and in Gadus morhua off the coast of Sweden and Norway; we compared them to D. varicus from the type-host, the Atlantic salmon Salmo salar from Norway, to verify their conspecificity. Newly generated sequences (28S rDNA, ITS2 and cox1) of Scandinavian and Arctic specimens consistent with D. varicus all formed a single clade, DV1. 28S sequences of D. varicus from S. salar from Norway, i.e., close to the Danish type locality, clustered within the DV1 clade along with sequences of D. varicus from various hosts including Limanda limanda, G. morhua and Myoxocephalus scorpius from the White Sea and the Barents Sea (Russia), without any host-related structuring. We thus consider that the lineage DV1 represents D. varicus sensu stricto. Additionally, specimens from M. merlangus had a similar morphology and anatomy to those of D. varicus from L. limanda, G. morhua and M. scorpius from T. Odhner's collection, supporting the presence of a single species in the DV1 lineage designated herein as D. varicus sensu stricto. We redescribe D. varicus sensu stricto, add new morphological characters and provide morphometric data. We infer that D. varicus types DV2-4 all relate to separate species. We also revise type-specimens of Derogenes minor Looss, 1901 from the A. Looss collection in the Swedish Museum of Natural History and provide redescriptions of it and of the type-species of the genus, Derogenes ruber Lühe, 1900. In light of their morphological distinctiveness relative to D. varicus sensu stricto, we reinstate D. parvus Szidat, 1950 and D. fuhrmanni Mola, 1912.

Title: Caché dans le brouillard : caractérisation morphologique et moléculaire de Derogenes varicus sensu stricto (Trematoda, Derogenidae) de Suède et de Norvège, et redescription de deux espèces de Derogenes peu connues. Abstract: Derogenes varicus (Müller, 1784) est largement signalé comme un trématode avec une spécificité d'hôte exceptionnellement faible et une large distribution bipolaire. Cependant, plusieurs études récentes ont suggéré que D. varicus représente un complexe d'espèces. Cette possibilité nécessite une meilleure caractérisation (idéalement moléculaire) des spécimens de l'hôte-type (Salmo salar) et de la localité-type (au large du Danemark). Lors de l'examen des trématodes parasites des poissons des eaux scandinaves et arctiques (Suède et Norvège), nous avons trouvé des spécimens de D. varicus dans l'estomac de Merlangius merlangus au large de la Suède, et dans Gadus morhua au large de la Suède et de la Norvège ; nous les avons comparés à D. varicus de l'hôte-type, le saumon atlantique Salmo salar de Norvège, pour vérifier leur conspécificité. Les séquences nouvellement générées (ADNr 28S, ITS2 et cox1) de spécimens scandinaves compatibles avec D. varicus ont toutes formé un seul clade, DV1. Les séquences 28S de D. varicus de S. salar de Norvège, c'est-à-dire proches de la localité-type au Danemark, se regroupent dans le clade DV1 avec des séquences de D. varicus de divers hôtes dont Limanda limanda, G. morhua et Myoxocephalus scorpius de la mer Blanche et la mer de Barents (Russie), sans aucune structuration liée à l'hôte. Nous considérons donc que la lignée DV1 représente D. varicus sensu stricto. De plus, les spécimens de M. merlangus avaient une morphologie et une anatomie similaires à celles de D. varicus de L. limanda, G. morhua et M. scorpius de la collection de T. Odhner, confirmant la présence d'une seule espèce dans la lignée DV1 désignée ici comme D. varicus sensu stricto. Nous redécrivons D. varicus sensu stricto, ajoutons de nouveaux caractères morphologiques et fournissons des données morphométriques. Nous en déduisons que les types DV2-4 de D. varicus se rapportent tous à des espèces distinctes. Nous révisons également les spécimens-types de Derogenes minor Looss, 1901 de la collection d'A. Looss du Muséum Suédois d'Histoire Naturelle, et fournissons des redescriptions de cette espèce et de l'espèce-type du genre, Derogenes ruber Lühe, 1900. À la lumière de leur distinction morphologique par rapport à D. varicus sensu stricto, nous rétablissons D. parvus Szidat, 1950 et Derogenes fuhrmanni Mola, 1912.

First report of ryegrass cyst nematode, Heterodera mani, in Tasmania, Australia.

Cyst nematodes of the genus Heterodera are a major group of sedentary plant parasites causing a significant economic impact, restricting production and market access globally (Moens et al. 2018). The ryegrass cyst nematode Heterodera mani is in the Avenae group and is found predominantly in pastures and grasslands in Europe, California, and South Africa. It was first described by Mathews (1971) from Northern Ireland. Known hosts are grasses (family Poaceae), principally Lolium perenne (perennial ryegrass), but also Dactylis glomerata (cat grass) and Festuca pratensis (meadow fescue) (Subbotin et al. 2010). Mowat (1974) reported that H. mani causes negligible damage to the yield of L. perenne in pot trials; however, Maas & Brinkman (1982) determined that it may cause significant damage to spring and autumn-sown perennial ryegrass in field conditions. During a routine examination for potato cyst nematode from a farm near Mawbanna in north-west Tasmania, Australia, several pale to dark brown Heterodera cysts were extracted that were lemon shaped with the presence of a small vulval cone at the posterior end and a distinct neck. The J2 (n=20) stylet length ranged from 24-26 µm with round knobs deeply concave anteriorly, hyaline tail length was 37-42 µm, true tail length ranged from 59-68 µm and total body length varied from 526-559 µm. All the above characters match those described for H. mani (Subbotin et al. 2010). To verify this identification, DNA was extracted from five individual J2 juveniles from a single cyst using QIAamp DNA micro kit (Qiagen®), and two gene regions amplified: internal transcribed spacer region of ribosomal RNA (ITS-rRNA) with primer pair AB28 and TW81 and cytochrome oxidase 1 (CO1) with primer pair JB3 and JB5 (Bowles et al. 1992; Curran et al. 1994; Derycke et al. 2005). One PCR reaction contained 10 µM (1 µl each) of each primer, 12.5 µl of OneTaq® DNA Polymerase and 5 µl of DNA template with a final volume of 25 µl. PCR products were sent for purification and Sanger sequencing at Macrogen (Seoul, Rep. of Korea). All resulting sequences were trimmed, aligned, and analysed using Geneious Prime® 2022.0.1 (www.geneious.com). Five ITS sequences (accessions ON402852-ON402856) and five CO1 sequences (accessions ON402857-ON402861) were submitted to GenBank. These ITS sequences were very similar to each other and exhibited 99.16-100% similarity with that of H. mani isolate from Hamminkeln, Germany (AY148377) (Subbotin et al. 2018). The CO1 sequences exhibited 98.96-100% similarity with that of H. mani isolate from Washington, USA (MG523097) (Subbotin et al. 2003). Obtained sequences were mapped to reference sequences downloaded from NCBI GenBank and maximum likelihood phylogenetic trees were calculated. Due to the lack of further living nematode material, pot experiments were not performed. Such experiments are not feasible in Tasmania currently and transfer of live nematode material to the Australian mainland presents logistic and legal issues. However, morphological and molecular evidence for species determination of H. mani was unequivocal and contributes to the list of cyst nematode species present in Australia. This is the first detection of H. mani in Australia and is a range extension of the species from North America, Africa, and Europe to Australia. The nematode may cause damage to perennial ryegrass in Australia, however, impact on yield still needs to be investigated.

Enenterum kyphosi Yamaguti, 1970 and Enenterum petrae n. sp. (Digenea: Enenteridae) from kyphosid fishes (Centrarchiformes: Kyphosidae) collected in marine waters off eastern Australia.

Species of the digenean genus Enenterum Linton, 1910 (Lepocreadioidea: Enenteridae) are characterised primarily by their elaborate oral suckers, which are divided into varying numbers of anteriorly directed lobes, and their host-restriction to herbivorous marine fishes of the family Kyphosidae. We describe Enenterum petrae n. sp. from the brassy chub Kyphosus vaigiensis (Quoy Gaimard) collected off Lizard Island, Great Barrier Reef, Queensland, Australia. Enenterum petrae n. sp. is readily differentiated from congeners by its unique oral sucker morphology, in having a minute pharynx, and the combination of a genital cap and accessory sucker. We also provide the first record of Enenterum kyphosi Yamaguti, 1970 from Australia based on material obtained from the blue sea chub Kyphosus cinerascens (Forsskl) collected off Lizard Island and North Stradbroke Island, Queensland. Morphologically, our specimens of E. kyphosi agree closely with descriptions of this species from Hawaii and South Africa, and despite lack of molecular data from outside of Australian waters, we consider all three reports to represent a single, widespread species. The first ITS2 and COI mtDNA gene sequences for species of Enenterum are provided and molecular phylogenetic analyses of 28S rDNA gene sequences place these species in a strongly-supported clade with the type-species of the genus, Enenterum aureum Linton, 1910. The oral suckers of both E. kyphosi and E. petrae n. sp. can be interpreted as having varying numbers of lobes depending on the particular specimen and how the division between lobes is defined. Scanning electron microscopical images improves understanding of the morphology of the enenterid oral sucker, and permits speculation regarding the evolutionary history leading to its specialisation in this lineage.

Reliability and Utility of Standard Gene Sequence Barcodes for the Identification and Differentiation of Cyst Nematodes of the Genus Heterodera.

Difficulties inherent in the morphological identification of cyst nematodes of the genus Heterodera Schmidt, 1871, an important lineage of plant parasites, has led to broad adoption of molecular methods for diagnosing and differentiating species. The pool of publicly available sequence data has grown significantly over the past few decades, and over half of all known species of Heterodera have been characterized using one or more molecular markers commonly employed in DNA barcoding (18S, internal transcribed spacer [ITS], 28S, coxI). But how reliable are these data and how useful are these four markers for differentiating species? We downloaded all 18S, ITS, 28S, and coxI gene sequences available on the National Center for Biotechnology Information (NCBI) database, GenBank, for all species of Heterodera for which data were available. Using a combination of sequence comparison and tree-based phylogenetic methods, we evaluated this dataset for erroneous or otherwise problematic sequences and examined the utility of each molecular marker for the delineation of species. Although we find the rate of obviously erroneous sequences to be low, all four molecular markers failed to differentiate between at least one species pair. Our results suggest that while a combination of multiple markers is best for species identification, the coxI marker shows the most utility for species differentiation and should be favored over 18S, ITS, and 28S, where resources are limited. Presently, less than half the valid species of Heterodera have a sequence of coxI available, and only a third have more than one sequence of this marker.

Molecular characterisation and updated description of Neoechinorhynchus aldrichettae Edmonds, 1971 (Acanthocephala: Neoechinorhynchidae), based on material from Aldrichetta forsteri (Valenciennes) collected in Tasmania, Australia.

We report on Neoechinorhynchus aldrichettae Edmonds, 1971 (Acanthocephala: Neoechinorhynchidae), obtained from yellow-eye mullet Aldrichetta forsteri (Valenciennes) (Mugiliformes: Mugilidae) from the Huon River, Tasmania, Australia. We provide new 18S and 28S rDNA gene sequence data for N. aldrichettae, assess its phylogenetic position relative to other species of Neoechinorhynchus and provide an updated morphological account of this species including detail of features omitted in the type-description, specifically of the apical organ, a collar at the base of the neck and a para-receptacle structure associated with the proboscis receptacle. We determine that eggs in this species are ovoid, without polar prolongations of fertilisation membrane, which permits assignment of N. aldrichettae to the subgenus Neoechinorhynchus. Our phylogenetic analyses place N. (N.) aldrichettae in a clade with other species of Neoechinorhynchus which parasitise mullets in marine and estuarine waters. We find that, in terms of monophyletic clades, the current subgeneric classification system for Neoechinorhynchus is not reflected in our phylogenetic analyses.

STABLE ISOTOPE SIGNATURES OF AN ACANTHOCEPHALAN AND TREMATODE FROM THE HERBIVOROUS MARINE FISH KYPHOSUS BIGIBBUS (PERCIFORMES: KYPHOSIDAE).

Stable isotope analyses of carbon and nitrogen (δ13C and δ15N) are useful for elucidating consumer relationships of free-living organisms, as carbon isotopes indicate dietary carbon sources and incremental increases in nitrogen isotopic enrichment are correlated with increases in trophic position. However, host-parasite relationships are more difficult to interpret using isotopes, as data from different host-parasite systems rarely show any consistent pattern. This inconsistency of pattern reflects the complexity of host-parasite relationships, but also the scarcity of data from a diverse assemblage of host-parasite systems. We present stable isotope data from a host-parasite system including 2 ecologically contrasting helminths, an acanthocephalan (Filisoma filiformis) and a digenetic trematode (Enenterum sp.), which co-occur in the intestine of the same marine fish (Kyphosus bigibbus), the diet of which consists almost exclusively of macroalgae. We obtained δ13C and δ15N data from K. bigibbus muscle, stomach contents, and pooled infrapopulations of Enenterum sp. and F. filiformis. Consistent with other isotope studies including acanthocephalans, F. filiformis was depleted in δ13C and δ15N relative to K. bigibbus. Although Enenterum sp. exhibited values for δ13C similar to those for F. filiformis, they were enriched in δ15N relative to the acanthocephalan, with a signature similar to that of K. bigibbus. These findings are discussed within a host-ecosystem context, highlighting the importance of considering species-specific biology when interpreting host-parasite relationships using stable isotopes. Our study adds to the growing body of literature indicating that absorptive feeders, such as acanthocephalans, are typically depleted in δ13C and δ15N relative to their hosts, whereas trematodes, with a greater diversity of feeding opportunities, exhibit a wide variety of isotopic signatures across life stage and different host-parasite systems.

Gorgorhynchoides pseudocarangis n. sp. (Acanthocephala: Isthmosacanthidae) from Pseudocaranx dentex (Carangidae) in southeast Queensland, Australia, with comments on the Isthmosacanthidae.

Gorgorhynchoides pseudocarangis n. sp. (Isthmosacanthidae), is described from the intestine of the white trevally Pseudocaranx dentex (Bloch & Schneider) (Carangiformes: Carangidae) collected in Moreton Bay, Queensland, Australia. The new species has a proboscis armature of 27-28 rows of 16-17 hooks. It is most similar morphologically to Gorgorhynchoides bullocki Cable & Marafachisi, 1970 and Gorgorhynchoides gnathanodontos Smales, 2014 but differs from the former in having a longer proboscis with more rows of hooks, ventral hooks 6/7-12 with notched tips and trunk spines which do not extend onto the anterior bulbous swelling, and from the latter in having a longer proboscis, ventral hooks 6/7-12 with notched tips, more circles of trunk spines, larger eggs and a proboscis armature with all hooks lacking manubria. Previous molecular phylogenetic analyses have shown that the genus Serrasentis Van Cleave, 1923 is sister to Gorgorhynchoides Cable & Linderoth, 1963, although some have failed to resolve these two lineages in separate monophyletic clades. We performed novel single-gene and concatenated phylogenetic analyses using cox1 mtDNA, 18S and 28S rDNA gene-sequences, resolving Gorgorhynchoides and Serrasentis in monophyletic sister clades and demonstrating that Gorgorhynchoides pseudocarangis n. sp. is phylogenetically distinct from related species for which molecular sequence data are available. We view the previous amendment of the Isthmosacanthidae to include the genera Golvanorhynchus Noronha, Fabio & Pinto, 1987, Gorgorhynchoides, Isthmosacanthus Smales 2014 and Serrasentis, and the transfer of the family to the Polymorphida, as the most satisfactory classification at present, although additional molecular evidence would provide greater stability.

Correction to: Molecular characterisation of acanthocephalans from Australian marine teleosts: proposal of a new family, synonymy of another and transfer of taxa between orders.

Shortly after publication it was brought to authors' attention that two of the cox1 sequences reported in the study, those of Neoechinorhynchus tylosuri (MN692675) and Transvena annulospinosa (MN692690) were potentially erroneous. After investigation, it was determined that this was indeed the case and was caused by contamination of original sequencing results. They were found to be near-duplicates of other species from the same sequencing batch. These sequences have been removed from GenBank. Unfortunately, this means that no cox1 sequence data were provided for the above two species in the referenced study. The remaining cox1 sequences reported have been checked and are reliable. Furthermore, cox1 sequence data were not analysed as part of the study, and thus the above error does not affect the results or conclusions of the study. Corrections to the text in reference to the above are made in Table 1 (removal of the above GenBank accession numbers), on page 10 ("Sequence data for all three targeted markers were obtained for 13 (rather than 15) of the 17 acanthocephalan species studied") and on page 19 ("We generated new cox1 sequence data for all but three (rather than one) of the acanthocephalan species from our collection…").

Metazoan Parasite Life Cycles: Significance for Fish Mariculture.

Marine fish are a nutritious and high-value food commodity, but many wild-capture fisheries are in decline. Thus, marine fish culture is expected to expand greatly in coming years. Management of disease is a major problem in this industry, and metazoan parasites are among the most significant disease agents in terms of economic loss and animal welfare. Current methods for controlling metazoan parasites are mostly reactionary and rely on chemical treatment. Such methods are ultimately unsustainable. Here, we summarise the life cycles of marine metazoan parasites and how this knowledge can be used for nonchemical management and control. To aid the sustainable growth of marine fish culture, we advocate for a renewed research focus on the basic life history characteristics of parasites.

Proposal of Spinulacorpus biforme (Smales, 2014) n. g., n. comb. and the Spinulacorpidae n. fam. to resolve paraphyly of the acanthocephalan family Rhadinorhynchidae Lühe, 1912.

Previous phylogenetic analyses of the Acanthocephala have demonstrated that the families Rhadinorhynchidae Lühe, 1912 and Transvenidae Pichelin & Cribb, 2001 are sister clades. However, a recent study found that the Rhadinorhynchidae is paraphyletic, due to the basal position of Rhadinorhynchus biformis Smales, 2014 relative to the Rhadinorhynchidae + Transvenidae. We reassess these relationships with new single-gene and concatenated phylogenetic analyses utilising cox1 mtDNA, 18S rDNA and 28S rDNA, including recently available sequences. Although topologies differed among single-gene analyses, the overall results support R. biformis as representative of a lineage distinct from those of the Rhadinorhynchidae and Transvenidae. Examination of additional specimens of R. biformis allowed us to identify morphological characters that further support this hypothesis. These results lead us to propose transfer of R. biformis to a new genus (Spinulacorpus n. g.) and family (Spinulacorpidae n. fam.) to resolve the paraphyly of the Rhadinorhynchidae.

Molecular characterisation of acanthocephalans from Australian marine teleosts: proposal of a new family, synonymy of another and transfer of taxa between orders.

We provide molecular data (cox1, 18S rDNA and 28S rDNA) for 17 acanthocephalan species and 20 host-parasite combinations from Australian marine teleosts collected from off Queensland, Australia. Fourteen of these acanthocephalans are characterised with molecular data for the first time and we provide the first molecular data for a species of each of the genera Heterosentis Van Cleave, 1931, Pyriproboscis Amin, Abdullah & Mhaisen, 2003 and Sclerocollum Schmidt & Paperna, 1978. Using 18S and 28S rDNA sequences, the phylogenetic position of each newly sequenced species is assessed with both single-gene and concatenated 18S+28S maximum likelihood and Bayesian inference analyses. Additional phylogenetic analyses focusing on the genus Rhadinorhynchus Lühe, 1912 and related lineages are included. Our phylogenetic results are broadly consistent with previous analyses, recovering previously identified inconsistencies but also providing new insights and necessitating taxonomic action. We do not find sufficient evidence to recognise the Gymnorhadinorhynchidae Braicovich, Lanfranchi, Farber, Marvaldi, Luque & Timi, 2014 as distinct from the Rhadinorhynchidae Lühe, 1912. The family Gymnorhadinorhynchidae and its sole genus, Gymnorhadinorhynchus Braicovich, Lanfranchi, Farber, Marvaldi, Luque & Timi, 2014, are here recognised as junior synonyms of Rhadinorhynchidae and Rhadinorhynchus, respectively. The two species currently assigned to Gymnorhadinorhynchus are recombined as Rhadinorhynchus decapteri (Braicovich, Lanfranchi, Farber, Marvaldi, Luque & Timi, 2014) n. comb. and Rhadinorhynchus mariserpentis (Steinauer, Garcia-Vedrenne, Weinstein & Kuris, 2019) n. comb. In all of our analyses, Rhadinorhynchus biformis Smales, 2014 is found basal to the Rhadinorhynchidae + Transvenidae Pichelin & Cribb, 2001, thus resulting in a paraphyletic Rhadinorhynchidae. It appears that R. biformis may require a new genus and family; however, morphological data for this species are currently insufficient to adequately distinguish it from related lineages, thus we defer the proposal of any new higher-rank names for this species. Species of the genus Sclerocollum, currently assigned to the Cavisomidae Meyer, 1932, are found nested within the family Transvenidae. We transfer the genus Sclerocollum to the Transvenidae and amend the diagnosis of the family accordingly. The genera Gorgorhynchoides Cable & Linderoth, 1963 and Serrasentis Van Cleave, 1923, currently assigned to the Rhadinorhynchidae, are supported as sister taxa and form a clade in the Polymorphida. We transfer these genera and Golvanorhynchus Noronha, Fabio & Pinto, 1978 to an emended concept of the Isthomosacanthidae Smales, 2012 and transfer this family to the Polymorphida. Lastly, Pyriproboscis heronensis (Pichelin, 1997) Amin, Abdullah & Mhaisen, 2003, currently assigned to the Pomphorhynchidae Yamaguti, 1939, falls under the Polymorphida in our analyses with some support for a sister relationship with the Centrorhynchidae Van Cleave, 1916. As this species clearly does not belong in the Pomphorhynchidae and is morphologically and molecularly distinct from the lineages of the Polymorphida, we propose the Pyriprobosicidae n. fam. to accommodate it.

An identity crisis in the Indo-Pacific: molecular exploration of the genus Koseiria (Digenea: Enenteridae).

We explore the growing issue of cryptic speciation in the Digenea through study of museum material and newly collected specimens consistent with the enenterid genus Koseiria from five species of the Kyphosidae and Chaetodontoplus meredithi Kuiter (Pomacanthidae) collected in the Indo-Pacific. We use an integrated approach, employing traditional morphometrics, principal components analysis (PCA), and molecular data (ITS2 and 28S rDNA). Our results support recombination of Koseiria allanwilliamsi Bray & Cribb, 2002 as Proenenterum allanwilliamsi (Bray & Cribb, 2002) n. comb. and transfer of Koseiria huxleyi Bray & Cribb, 2001 to a new genus as Enenterageitus huxleyi (Bray & Cribb, 2002) n. comb. Molecular data indicate the presence of four further species consistent with Koseiria, one from Western Australia (sequence data only) and three from eastern Australia. All three eastern Australian species are morphologically consistent with Koseiria xishaensis Gu & Shen, 1983, but distinct from all other previously described species. Although K. xishaensis has been reported from Australia, we conclude that the similarity of the present forms to the original description of K. xishaensis means records of this species from Japan, Palau and Australia are unreliable. Because the eastern Australian forms cannot be reliably ascribed to K. xishaensis, we describe Koseiria argalea n. sp., Koseiria laiphopharophora n. sp., and Koseiria pyknophora n. sp., following application of PCAs and iterative refinement of species concepts and type series. These analyses did not allow convincing identification hypotheses for all specimens examined. In this genus, both morphological and molecular data, together with reliable host identifications, are essential for species recognition, and thus we refrain from attempting to name samples lacking molecular data. The issues presented by these taxa encapsulate those of trematodes in the region as a whole. Many records require dramatically improved supporting data, leading to substantial uncertainly in the identification of this fauna.

A new genus and species of the trematode family Gyliauchenidae Fukui, 1929 from an unexpected, but plausible, host, Kyphosus cornelii (Perciformes: Kyphosidae).

The Enenteridae Yamaguti, 1958 and Gyliauchenidae Fukui, 1929 exhibit an interesting pattern of host partitioning in herbivorous fishes of the Indo-West Pacific. Enenterids are known almost exclusively from fishes of the family Kyphosidae, a group of herbivorous marine fishes common on tropical and temperate reefs. In contrast, gyliauchenids are found in most of the remaining lineages of marine herbivorous fishes, but until the present study, had never been known from kyphosids. Here we report on the first species of gyliauchenid known from a kyphosid. Endochortophagus protoporus gen. nov., sp. nov. was recovered from the Western buffalo bream, Kyphosus cornelii (Whitley, 1944), collected off Western Australia. Kyphosus cornelii also hosts an enenterid, Koseiria allanwilliamsi Bray & Cribb, 2002, and is thus the first fish known in which enenterids and gyliauchenids co-occur. Molecular phylogenetic analyses place the new species close to those of Affecauda Hall & Chambers, 1999 and Flagellotrema Ozaki, 1936, but there is sufficient morphological evidence, combined with the unusual host, to consider it distinct from these genera. We discuss factors which may have contributed to the host partitioning pattern observed between enenterids and gyliauchenids.

Collastoma esotericum (Neodalyellida: Umagillidae), a new species of sipunculan-inhabiting rhabdocoel from Queensland, Australia.

The Umagillidae Wahl, represent a group of endosymbiotic Platyhelminthes which inhabit two disparate invertebrate host groups, the echinoderms and sipunculans. Sipunculan-inhabiting umagillids are morphologically distinct from those inhabiting echinoderms and have traditionally been placed in a distinct genus and subfamily, Collastoma Dörler, and the Collastominae Wahl, respectively. Although molecular data are available for umagillid species inhabiting echinoids and holothurians, species inhabiting sipunculans have yet to be evaluated with molecular data. Collastoma esotericum n. sp. from the sipunculan Phascolosoma scolops (Selenka & de Man) collected in Moreton Bay, southeast Queensland, Australia, is described. Phylogenetic analyses based on 18S rRNA gene sequences placed C. esotericum in a clade with species of the Umagillidae and the Provorticinae Luther, with strong support. However, within this clade the relationship of C. esotericum to the Umagillidae and Provorticinae was not resolved.

Molecular systematics of the digenean community parasitising the cerithiid gastropod Clypeomorus batillariaeformis Habe & Kusage on the Great Barrier Reef.

A rich fauna of digenetic trematodes has been documented from the Great Barrier Reef (GBR), yet little is known of the complex life-cycles of these parasites which occur in this diverse marine ecosystem. At Heron Island, a small coral cay at the southern end of the GBR, the intertidal marine gastropod Clypeomorus batillariaeformis Habe & Kusage (Cerithiidae) is especially abundant. This gastropod serves as an intermediate host for 12 trematode species utilising both fish and avian definitive hosts. However, 11 of these species have been characterised solely with morphological data. Between 2015 and 2018 we collected 4870C. batillariaeformis from Heron Island to recollect these species with the goal of using molecular data to resolve their phylogenetic placement. We found eight of the 12 previously known species and two new forms, bringing the total number of digenean species known to parasitise C. batillariaeformis to 14. The families of this trematode community now include the Atractotrematidae Yamaguti, 1939, Bivesiculidae Yamaguti, 1934, Cyathocotylidae Mühling, 1898, Hemiuridae Looss, 1899, Heterophyidae Leiper, 1909, Himasthlidae Odhner, 1910, Microphallidae Ward, 1901, and Renicolidae Dollfus, 1939. Molecular data (ITS and 28S rDNA) were generated for all trematode species, and the phylogenetic position of each species was determined. The digenean community parasitising C. batillariaeformis includes several common species, as well as multiple species which are uncommon to rare. Although most of those trematodes in the community which exploit fishes as definitive hosts have remained common, the composition of those which utilise birds appears to have shifted over time.

The phylogenetic position of Choerodonicola Cribb, 2005 (Digenea: Opecoelidae) with a partial life-cycle for a new species from the blue-barred parrotfish Scarus ghobban Forsskål (Scaridae) in Moreton Bay, Australia.

Choerodonicola Cribb, 2005 is a minor genus of opecoelid trematodes defined for species with exceptionally small eggs but otherwise generalised morphology. Four species are currently recognised, all from fishes collected in Japanese waters but each from different perciform families: a labrid, a scarid, a sparid and pinguipeds. We report on a new species, Choerodonicola arothokoros n. sp., from the blue-barred parrotfish Scarus ghobban Forsskål (Scaridae) collected in subtropical waters of Moreton Bay, south-east Queensland, Australia. Using genetic sequence data for the ITS2 rDNA marker, we matched adult C. arothokoros to intramollsucan stages discovered in an intertidal gastropod Herpetopoma atratum (Gmelin) (Vetigastropoda: Chilodontidae) collected in close proximity to the fish hosts. Notably, the cercariae lack a penetration stylet and are among the smallest known in the Opecoelidae. We provide the first assessment of the phylogenetic position of Choerodonicola based on sequence data generated for the phylogenetically informative 18S and 28S rRNA coding regions, for C. arothokoros and also C. renko Machida, 2014, which we recollected from the yellowback seabream Dentex hypselosomus Bleeker from the fish market in Minabe, Wakayama Prefecture, Japan. In our analyses, species of Choerodonicola resolved to neither of the major marine Plagioporinae (sensu lato) clades, clustering instead with Trilobovarium parvvatis Martin, Cutmore & Cribb, 2017, Podocotyloides parupenei (Manter, 1963) Pritchard, 1966 and Macvicaria magellanica Laskowski, Jezewski & Zdzitowiecki, 2013. This clade is phylogenetically distinctive such that it has the potential to be recognised as a new opecoelid subfamily, but further investigation is required to establish the bounds for such a grouping and to determine the morphological and/or life-history patterns reflected by the phylogeny. Finally, we propose C. interruptus (Manter 1954) n. comb. for a species previously recognised in Plagioporus Stafford, 1904 and known only from Pseudolabrus miles (Schneider & Forster), a labrid endemic to New Zealand.

Molecular phylogeny of the Haplosplanchnata Olson, Cribb, Tkach, Bray and Littlewood, 2003, with a description of Schikhobalotrema huffmani n. sp.

We describe Schikhobalotrema huffmani n. sp. from Tylosurus crocodilus (Péron and Leseur) (Belonidae) collected off Lizard Island, Great Barrier Reef, Queensland, Australia and Tylosurus gavialoides (Castelnau) collected from Moreton Bay, Queensland. Schikhobalotrema huffmani n. sp., along with Schikhobalotrema ablennis (Abdul-Salam and Khalil, 1987) Madhavi, 2005, Schikhobalotrema acutum (Linton, 1910) Skrjabin and Guschanskaja, 1955 and Schikhobalotrema adacutum (Manter, 1937) Skrjabin and Guschanskaja, 1955 are distinguished from all other species of Schikhobalotrema Skrjabin and Guschanskaja, 1955 in having ventral suckers which bear lateral lobes and have longitudinal apertures. Schikhobalotrema huffmani n. sp. differs from S. ablennis in having an obvious post-vitelline region and a longer forebody. From S. acutum, S. huffmani n. sp. differs in having a prostatic bulb smaller than the pharynx and more anterior testis. From S. adacutum, S. huffmani n. sp. differs in having more prominent ventral sucker lobes, a conspicuous prostatic bulb and a longer forebody. We also report the first Australian record of Haplosplanchnus pachysomus (Eysenhardt, 1829) Looss, 1902, from Mugil cephalus Linnaeus (Mugilidae) collected in Moreton Bay. Molecular sequence data (ITS2, 18S and 28S rDNA) were generated for Schikhobalotrema huffmani n. sp., H. pachysomus and archived specimens of Hymenocotta mulli Manter, 1961. The new 18S and 28S molecular data were combined with published data of five other haplosplanchnid taxa to expand the phylogeny for the Haplosplanchnata. Bayesian inference and Maximum Likelihood analyses recovered identical tree topology and demonstrated the Haplosplanchnata as a well-supported monophyletic group. However, relationships at and below the subfamily level remain poorly resolved.

The life-cycle of Gorgocephalus yaaji Bray & Cribb, 2005 (Digenea: Gorgocephalidae) with a review of the first intermediate hosts for the superfamily Lepocreadioidea Odhner, 1905.

Using novel molecular and morphological data we elucidated the life-cycle of Gorgocephalus yaaji Bray & Cribb, 2005 from off Lizard Island, on the northern Great Barrier Reef, Australia. ITS2 rDNA sequences generated for larval trematodes from the infected snail species Echinolittorina austrotrochoides Reid (Littorinidae) were identical to those from adult G. yaaji from the fish Kyphosus cinerascens (Forsskål) (Kyphosidae). Cercariae develop in rediae in E. austrotrochoides, emerge from the snail, encyst on algae as metacercariae, and are inferred to then be consumed by the herbivorous definitive fish host, K. cinerascens. In addition, we generated the first ITS2 rDNA sequences for a gorgocephalid previously reported from the littorind gastropod Austrolittorina unifasciata Gray. Although infections previously reported from A. unifasciata were the first larval gorgocephalids characterised, this study is the first to connect an intramolluscan infection to a sexual adult. In light of the new life-cycle information, a review of mollusc associations for the digenean superfamily Lepocreadioidea was performed, highlighting gaps in the knowledge and revealing patterns of host-parasite association. We find that distinct patterns of first intermediate host association are discernible for three lepocreadioid lineages: the Aephnidiogenidae Yamaguti, 1934, Gorgocephalidae Manter, 1966, and the Lepocreadiidae Odhner, 1905. However, the evolutionary origin for these patterns of host association remains unclear.

Trematodes of fishes of the Indo-west Pacific: told and untold richness.

The Indo-west Pacific is a marine bioregion stretching from the east coast of Africa to Hawaii, French Polynesia and Easter Island. An assessment of the literature from the region found reports of 2,582 trematode species infecting 1,485 fish species. Reports are concentrated in larger fishes, undoubtedly reflecting the tendency for larger hosts to be infected by more species of parasites as well as a collecting bias. Many hundreds of fish species, including many from families known to be rich in trematodes, have yet to be reported as hosts. Despite some areas (the Great Barrier Reef, Hawaii and the waters off China, India and Japan) receiving sustained attention, none can be considered to be comprehensively known. Several regions, most importantly in East Africa, French Polynesia and the Coral Triangle, are especially poorly known. The fauna of the Indo-west Pacific has been reported so unevenly that we consider it impossible to predict the true trematode richness for the region. We conclude that the greatest gap in our understanding is of the geographical distribution of species in the Indo-west Pacific. This is highlighted by the fact that 87% of trematodes in the region have been reported no more than five times. The reliable recognition of species is a major problem in this field; molecular approaches offer prospects for resolution of species identification but have been little adopted to date.