Evidence that host ecology drives first intermediate host use in the Didymozoidae (Trematoda: Hemiuroidea): an asexual infection in a vermetid (Gastropoda).

The Didymozoidae (Trematoda: Hemiuroidea) is among the most speciose trematode families, known from a wide range of marine teleost fishes. Despite their richness, however, didymozoid life cycles are unusually poorly known; only two first intermediate hosts are known, a marine bivalve (Anadara trapezia) and a pelagic gastropod (Firoloida desmarestia). This study uses multi-locus molecular sequence data to identify a novel first intermediate host for the family, a sessile gastropod of the genus Thylacodes Guettard (Vermetidae). The didymozoid infection is not identified to species but, based on molecular phylogenetic analyses, it is close to Saccularina magnacetabula Louvard et al., 2022, which uses a bivalve as a first intermediate host. The distribution of known first intermediate hosts of didymozoids (a bivalve, a holoplanktonic gastropod and a sessile gastropod that feeds with the use of mucus nets) suggests that first intermediate host use within the Didymozoidae has been opportunistically driven by the trophic ecology of potential mollusc hosts and has involved significant host-switching events.

Gerricola queenslandensis n. g., n. sp., a new monorchiid trematode from the eastern Australian coast and its life cycle partially elucidated.

Of over 250 species of Monorchiidae Odhner, 1911, just four are known from gerreid fishes. Here, we report adult specimens of a new species infecting Gerres oyena (Forsskål) and Gerres subfasciatus Cuvier from off Heron Island and North Stradbroke Island, Queensland, Australia. The species is morphologically most similar to the concept of Lasiotocus Looss, 1907, which currently comprises eight species, in the possession of an unspined genital atrium, bipartite terminal organ, round oral sucker and unlobed ovary. However, phylogenetic analyses of the 28S ribosomal DNA gene region shows the species to be distantly related to the two sequenced species of Lasiotocus - Lasiotocus mulli (Stossich, 1883) Odhner, 1911 and Lasiotocus trachinoti Overstreet & Brown, 1970 - and that it clearly requires a distinct genus; thus, we propose Gerricola queenslandensis n. g., n. sp. Morphologically, G. queenslandensis n. g., n. sp. differs significantly from L. mulli and L. trachinoti only in the possession of distinctly longer caeca, which terminate in the post-testicular region, and in the absence of a distinct gap in the terminal organ spines. The remaining species of Lasiotocus possess caeca that also terminate in the post-testicular region, which might warrant their transfer to Gerricola n. g. However, doubt about their monophyly due to a combination of significant morphological variation, a lack of information on some features and infection of a wide range of hosts, lead us to retain these taxa as species of Lasiotocus until molecular sequence data are available to better inform their phylogenetic and taxonomic positions. Sporocysts and cercariae of G. queenslandensis n. g., n. sp. were found in a lucinid bivalve, Codakia paytenorum (Iredale), from Heron Island. Sexual adult and intramolluscan stages were genetically matched with the ITS2 ribosomal DNA and cox1 mitochondrial DNA regions. This is the second record of the Lucinidae as a first intermediate host for the Monorchiidae. Additionally, we report sporocysts and cercariae of another monorchiid infection in a tellinid bivalve, Jactellina clathrata (Deshayes), from Heron Island. Molecular sequence data for this species do not match any sequenced species and phylogenetic analyses do not suggest any generic position.

Two new species of threadlike blood flukes (Aporocotylidae), with a molecular revision of the genera Ankistromeces Nolan & Cribb, 2004 and Phthinomita Nolan & Cribb, 2006.

Ankistromeces Nolan & Cribb, 2004 and Phthinomita Nolan & Cribb, 2006 are sister genera of threadlike blood flukes (Trematoda: Aporocotylidae) infecting teleost fishes of the tropical Indo-west Pacific. Here, we report new collections of these genera from Australia, Indonesia, and Japan. A new species of Ankistromeces, Ankistromeces kawamurai n. sp., is described from Siganus spinus (Linnaeus) off Okinawa, Japan, and a new species of Phthinomita, Phthinomita abdita n. sp., from Choerodon cephalotes (Castelnau), in Moreton Bay, Australia; the new species are morphologically cryptic within their respective genera and are delineated by molecular and ecological data. Ankistromeces olsoni Nolan & Cribb, 2006 is reported from Siganus fuscescens (Houttuyn) off Heron Island (southern Great Barrier Reef), Lizard Island (northern Great Barrier Reef), and Okinawa and Wakayama Prefectures, Japan and from Siganus spinus (Linnaeus) from off Bali, Indonesia. Ankistromeces mariae Nolan & Cribb, 2004 is re-reported from the type-host, Meuschenia freycineti (Quoy & Gaimard), from a new location, Gypsy Bay, Tasmania. Phthinomita poulini Nolan & Cribb, 2006 is re-reported from its type-locality, Lizard Island, from a range of mullids, including five new host species, and its range is extended to include Moreton Bay. Phthinomita symplocos Nolan & Cribb, 2006 is reported from Bali and P. hallae Nolan & Cribb, 2006, P. jonesi Nolan & Cribb, 2006, P. littlewoodi Nolan & Cribb, 2006, and P. munozae Nolan & Cribb, 2006 are each re-reported from their type-host and type-localities. New cox1 mtDNA data were generated for all known species of these two genera from new and archival material. Analyses of these data enabled an evaluation of all known Phthinomita species; P. robertsthomsoni Nolan & Cribb, 2006 is synonymised with P. adlardi Nolan & Cribb, 2006, and P. brooksi Nolan & Cribb, 2006 is synonymised with P. sasali Nolan & Cribb, 2006. We highlight the failure of ITS2 data to delineate closely related aporocotylid species. In contrast, cox1 sequence data are proving reliable and effective in this context and we recommend their incorporation in future studies of blood fluke taxonomy.

Pronocephaloid cercariae (Platyhelminthes: Trematoda) from gastropods of the Queensland coast, Australia.

The superfamily Pronocephaloidea Looss, 1899 comprises digeneans occurring in the gut and respiratory organs of fishes, turtles, marine iguanas, birds and mammals. Although many life cycles are known for species of the Notocotylidae Lühe, 1909 maturing in birds and mammals, relatively few are known for the remaining pronocephaloid lineages. We report the cercariae of five pronocephaloid species from marine gastropods of the Queensland coast, Australia. From Lizard Island, northern Great Barrier Reef, we report three cercariae, two from Rhinoclavis vertagus (Cerithiidae) and one from Nassarius coronatus (Nassariidae). From Moreton Bay, southern Queensland, an additional two cercariae are reported from two genotypes of the gastropod worm shell Thylacodes sp. (Vermetidae). Phylogenetic analysis using 28S rRNA gene sequences shows all five species are nested within the Pronocephaloidea, but not matching or particularly close to any previously sequenced taxon. In combination, phylogenetic and ecological evidence suggests that most of these species will prove to be pronocephalids parasitic in marine turtles. The Vermetidae is a new host family for the Pronocephaloidea.

Isorchis cannoni n. sp. (Digenea: Atractotrematidae) from Great Barrier Reef rabbitfishes and the molecular elucidation of its life cycle.

We describe Isorchis cannoni n. sp. from the rabbitfishes Siganus fuscescens (Houttuyn) and Siganus lineatus (Valenciennes) (Siganidae) collected off Heron Island, southern Great Barrier Reef, Australia and, using molecular data, demonstrate that 'Cercariae queenslandae II' of Cannon (1978) from the gastropod Clypeomorus batillariaeformis Habe & Kosuge (Cerithiidae) is the larval form of this new species. The cercariae of I. cannoni n. sp. develop in rediae, encyst in the environment after emergence, and are inferred to then be consumed by grazing rabbitfish. Additionally, we provide a new report of Isorchis currani Andres, Pulis & Overstreet, 2016 from the type host, Selenotoca multifasciata (Richardson) (Scatophagidae) collected in Moreton Bay, south-east Queensland, Australia, greatly expanding the known geographical range of this species. Molecular sequence data (ITS1, ITS2 and 28S rDNA) generated for I. cannoni n. sp. and the new specimens of I. currani, confirm the identification of I. currani and demonstrate a distinct genotype for I. cannoni n. sp. relative to other species of Isorchis Durio & Manter, 1969, for which molecular data are available. Isorchis cannoni n. sp. is morphologically distinct from all other species in the genus, and is further distinguished by utilizing species of Siganidae as definitive hosts, rather than species of Chanidae or Scatophagidae. Because haploporid and atractotrematid cercariae have well-developed reproductive organs, we find cercariae of these closely related families morphologically distinguishable in the same way as adult trematodes: atractotrematids have two symmetrical testes and haploporids have a single testis or, rarely, two tandem or oblique testes.

Monorchis lewisi n. sp. (Trematoda: Monorchiidae) from the surf bream, Acanthopagrus australis (Sparidae), in Moreton Bay, Australia.

We describe Monorchis lewisi n. sp. (Monorchiidae) from the surf bream, Acanthopagrus australis (Günther, 1859) (Sparidae), in Moreton Bay, eastern Australia. The new species differs from most existing species of Monorchis Monticelli, 1893 in its possession of an elongate I-shaped excretory vesicle, and from other congeners in the relative configuration of the gut and suckers. Ovipusillus mayu Dove & Cribb, 1998 is re-reported from Gnathanodon speciosus (Forsskål, 1775) (Carangidae) from Moreton Bay. We report new second internal transcribed spacer (ITS2) and 28S rDNA sequence data for both species. Bayesian inference and Maximum Likelihood analyses of the 28S rDNA dataset suggest that existing subfamily and genus concepts within the family require substantial revision.

Expansion of the concept of the Opistholebetinae Fukui, 1929 (Digenea: Opecoelidae Ozaki, 1925), with Magnaosimum brooksae n. g., n. sp. from Tripodichthys angustifrons (Hollard) (Tetraodontiformes: Triacanthidae) in Moreton Bay, Australia.

The Opecoelidae Ozaki, 1925, the largest trematode family, currently lacks an adequate subfamilial organisation. In particular, recent analyses have shown that the sequenced representatives of the Opistholebetinae Fukui, 1929 are nested among taxa currently recognised in the Plagioporinae Manter, 1947, which itself is polyphyletic. The concept of the Opistholebetinae persists because its 25 or so species are united by a distinctive morphology and a narrow host range; species are known only from tetraodontiform fishes, specifically of the Tetraodontidae and Diodontidae, and possess a muscular post-oral ring, unusual pigmented granules throughout the body and a ventral sucker positioned more posteriorly than is typical for other opecoelids. Here we report on Magnaosimum brooksae n. g., n. sp. from a tetraodontiform, Tripodichthys angustifrons (Hollard) (Triacanthidae), collected in Moreton Bay, Australia. The new species has a longer forebody than hindbody, but lacks a post-oral ring and pigmented granules. It resolves to the plagioporine clade containing the Opistholebetinae, but does not form a monophyletic grouping with those taxa. We propose to expand the concept of the Opistholebetinae to include Magnaosimum, as well as Gaevskajatrema Gibson & Bray, 1982, Macvicaria Gibson & Bray, 1982, Pachycreadium Manter, 1954, Peracreadium Nicoll, 1909, Pinguitrema Siddiqi & Cable, 1960, Propycnadenoides Fishcthal & Kuntz, 1964, Pseudopycnadena Saad-Fares & Maillard, 1986, Pycnadena Linton, 1911 and Pycnadenoides Yamaguti, 1938.

The ghost of parasites past: eggs of the blood fluke Cardicola chaetodontis (Aporocotylidae) trapped in the heart and gills of butterflyfishes (Perciformes: Chaetodontidae) of the Great Barrier Reef.

We explored the distribution of Cardicola chaetodontis in chaetodontid fishes from the Great Barrier Reef. We found just four infections of adult worms in 238 individuals of 26 chaetodontid species. By contrast, eggs were present in hearts of 75 fishes (31·5%) and 19 of 26 chaetodontid species (all Chaetodon species). In 10 cases eggs contained moving miracidia; all the others were dead and degenerating. Eggs were sought in the gills of 51 individual fish. There were 17 cases of eggs being present in gills while present in the heart, but also 13 cases where eggs were absent from gills but present in the heart, suggesting that eggs remain longer in heart tissue than in gills. ITS2 rDNA sequences from two adult worms and eggs extracted from gills of five fishes (all different species) were identical to previously reported sequences of C. chaetodontis except for a single base-pair difference in two samples. We conclude that aporocotylid eggs trapped in fish heart tissues may inform understanding of the distributions and host ranges of aporocotylids, especially where adult prevalence is low. The low host-specificity of C. chaetodontis contrasts with higher specificity of trematodes of chaetodontids that have trophic transmission.

New collections of blood flukes (Aporocotylidae) from fishes of the tropical Indo-west Pacific, including a new genus, two new species and molecular evidence that Elaphrobates chaetodontis (Yamaguti, 1970) is widespread in the region.

We report new collections of the Aporocotylidae from Australia, French Polynesia, and Japan. A new species of Cardicola Short, 1953 is described from Scomberomorus commerson (Lacépède) (Scombridae), off Lizard Island. Cardicola nolani n. sp. can be distinguished from its congeners based on the position of the oötype, the position of the male genital pore, and the absence of an oral sucker. A new species is described from Abalistes stellatus (Anonymous) (Balistidae), also from off Lizard Island. Phylogenetically the new species forms a strongly-supported clade with Cardicola yuelao Yong, Cutmore & Cribb, 2018, which also infects balistids. These two species are distinct from all other aporocotylids in the combination of exceptionally short anterior and long posterior caeca, a lanceolate body, a single testis, an entirely post-ovarian uterus and the position of the oötype; a new genus, Balistidicola, is proposed for them. Balistidicola corneri n. sp. and B. yuelao (Yong, Cutmore & Cribb, 2018) n. comb. are essentially morphologically cryptic, only distinguishable by the form of the spination (B. corneri has five spines per row and B. yuelao has six). Elaphrobates chaetodontis (Yamaguti, 1970) is reported from 21 species of butterflyfishes (Chaetodontidae) from nine locations in tropical Indo-west Pacific; cox1 sequence data demonstrate extensive geographical structuring in this species. Braya jexi Nolan & Cribb, 2006, Elaphrobates milleri (Nolan & Cribb, 2006), and P. corventum Overstreet & Køie, 1989 are each re-reported from their type-hosts, and Pearsonellum pygmaeus Nolan & Cribb, 2004 and Balistidicola yuelao are each reported from a new host.

A complex of the blood fluke genus Psettarium (Digenea: Aporocotylidae) infecting tetraodontiform fishes of east Queensland waters.

Seven species of Psettarium (Digenea: Aporocotylidae), including four new species, are reported from tetraodontiform fishes from off coastal east Queensland. Psettarium pandora n. sp. infects the yellow boxfish, Ostracion cubicus (Ostraciidae), the first known aporocotylid to infect this family of fishes. Three new species are reported from pufferfishes of the genus Arothron (Tetraodontidae): Psettarium yoshidai n. sp. infects the map puffer (Arothron mappa), Psettarium hustoni n. sp. infects the black-spotted puffer (A. nigropunctatus) and Psettarium martini n. sp. infects the starry puffer (A. stellatus). We also report three species of Psettarium from Australian waters for the first time. Paracardicola hawaiensis Martin, 1960, the sole species of Paracardicola, is redescribed based on specimens collected from the type-host, the stars-and-stripes puffer, Arothron hispidus. Paracardicola is synonymised with Psettarium and P. hawaiensis is recombined as Psettarium hawaiiense (Martin, 1960) n. comb. Psettarium pulchellum Yong, Cutmore, Bray, Miller, Semarariana, Palm & Cribb, 2016, described from the narrow-lined puffer (Arothron manilensis) from off Bali, Indonesia, is reported from the same fish species at two locations on the Queensland coast, significantly extending the range of this species. Psettarium nolani (Bray, Cribb & Littlewood, 2013), originally described from French Polynesia, is reported from A. hispidus, A. manilensis and A. stellatus, representing both new host and locality records for this species. Molecular phylogenetic analysis shows these species to all be closely related, such that they cannot be considered to represent separate genera despite their differing morphology. Analysis of 28S sequence data for Psettarium anthicum Bullard & Overstreet, 2006, a non-tetraodontiform-infecting species, shows it to be distantly related to all other species of Psettarium for which sequence data are available. The species is re-assigned to a new genus, Cardallagium n. gen., as Cardallagium anthicum (Bullard & Overstreet, 2006) n. comb. We think it likely that the host range of species of Psettarium is limited to tetraodontiform fishes. We assessed the infection biology of two species, P. nolani and P. hawaiiense n. comb. infecting A. hispidus, using histology to assess the pathways of egg release for these species. Eggs of both species were observed in both circulatory and visceral organs of infected hosts, often in high numbers. Eggs were seen trapped in the mucosal layer of the intestine and, in rare instances, causing lesions in the laminar epithelium, providing the strongest evidence yet that they pass through the gut wall and escape the host via the faeces. Lastly, we discuss the biogeographical implications of our findings, noting that some Psettarium species now show very wide geographical distributions.

Did biogeographical processes shape the monogenean community of butterflyfishes in the tropical Indo-west Pacific region?

Geographical distribution of parasite species can provide insights into the evolution and diversity of parasitic communities. Biogeography of marine parasites is poorly known, especially because it requires an understanding of host-parasite interactions, information that is rare, especially over large spatial scales. Here, we have studied the biogeographical patterns of dactylogyrid parasites of chaetodontids, one of the most well-studied fish families, in the tropical Indo-west Pacific region. Dactylogyrid parasites were collected from gills of 34 butterflyfish species (n=560) at nine localities within an approximate area of 62millionkm2. Thirteen dactylogyrid species were identified, with richness ranging from 6 to 12 species at individual localities. Most dactylogyrid communities were dominated by Haliotrema angelopterum or Haliotrema aurigae, for which relative abundance was negatively correlated (ρ=-0.59). Parasite richness and diversity were highest in French Polynesia and the Great Barrier Reef (Australia) and lowest in Palau. Three biogeographic regions were identified based on dactylogyrid dissimilarities: French Polynesia, characterised by the dominance of H. angelopterum, the western Pacific region dominated by H. aurigae, and Ningaloo Reef (Australia), dominated by Euryhaliotrema berenguelae. Structure of host assemblages was the main factor explaining the dissimilarity (turnover and nestedness components of the Bray-Curtis dissimilarity and overall Bray-Curtis dissimilarity) of parasite communities between localities, while environment was only significant in the turnover of parasite communities and overall dissimilarity. Spatial structure of localities explained only 10% of the turnover of parasite communities. The interaction of the three factors (host assemblages, environment and spatial structure), however, explained the highest amounts of variance of the dactylogyrid communities, indicating a strong colinearity between the factors. Our findings show that spatial arrangement of chaetodontid dactylogyrids in the tropical Indo-west Pacific is primarily characterised by the turnover of the main Haliotrema spp., which is mainly explained by the structure of host assemblages.

Three new species of blood flukes (Digenea: Aporocotylidae) infecting pufferfishes (Teleostei: Tetraodontidae) from off Bali, Indonesia.

We describe three new species of blood flukes (Aporocotylidae) and propose their classification within the genus Psettarium Goto & Ozaki, 1929. All three species were collected from the circulatory systems of pufferfishes caught off Bali, central Indonesia. Psettarium pulchellum n. sp. was found in the gills of both the narrow-lined puffer (Arothron manilensis de Procé) and the spiny blaasop (Tylerius spinosissimus Regan), while P. ogawai n. sp. and P. jimbaranense n. sp. were found in the gills of the reticulated puffer (Arothron reticularis Bloch & Schneider). The morphological characteristics of these taxa necessitated emendation of the diagnosis for the genus Psettarium, to accommodate the presence of an oral sucker, multiple or entirely post-caecal testes and a degenerate posterior testis. Features such as proportion of body length occupied by the oesophagus, and posterior caeca being ≥7× the length of anterior caeca, are no longer regarded as useful genus-level characters. Additionally, Sasala nolani is reassigned to this genus as Psettarium nolani n. comb. In phylogenetic analyses of the 28S and ITS2 rDNA regions, all three new taxa form a well-supported clade, together with Psettarium sinense and Psettarium nolani n. comb., the two other species of tetraodontid-infecting aporocotylids for which comparative rDNA data were available. The short branch lengths within this clade, despite dramatic morphological differences between the five species, suggest that rapid morphological diversification has occurred among the tetraodontid-infecting aporocotylids. The genus Psettarium has long been considered problematic. Further commentary is given on the history of this genus and how the issues presented might be resolved.