Syst Parasitol https://doi.org/10.1007/s11230-018-9803-3 Lepocreadiidae Odhner, 1905 and Aephnidiogenidae Yamaguti, 1934 (Digenea: Lepocreadioidea) of ﬁshes from Moreton Bay, Queensland, Australia, with the erection of a new family and genus . . Rodney A. Bray Thomas H. Cribb Scott C. Cutmore Received: 13 March 2018 / Accepted: 19 May 2018 The Author(s) 2018 Abstract Digeneans of the lepocreadioid families monotypic Gibsonivermis Bray, Cribb & Barker, Lepocreadiidae Odhner, 1905 and Aephnidiogenidae 1997 is isolated from all other lepocreadioids and Yamaguti, 1934 from Moreton Bay, off southern supports the erection of Gibsonivermidae n. fam., Queensland, Australia, are recorded, along with the which is deﬁned morphologically, based particularly erection of a new family, Gibsonivermidae. Molecular on the uniquely elongated male terminal genitalia, the data were generated for all representatives of these distribution of the uterus in the forebody and the families collected during this study and a phylogram presence of a uroproct. Mobahincia teirae n. g., n. sp. for members of the superfamily was generated based is reported from Platax teira (Forsska˚l) in Moreton on the partial 28S rDNA dataset, placing these species Bay and off Heron Island and New Caledonia. in context with those previously sequenced. This Recognition of this new genus is based on molecular phylogenetic analysis demonstrates that the results and the combination of caeca abutting the posterior body wall and the lack of an anterior body scoop or ﬂanges. The following lepocreadioid species This article was registered in the Ofﬁcial Register of are reported from Moreton Bay for the ﬁrst time: Zoological Nomenclature (ZooBank) as urn:lsid:zoobank.org: Bianium arabicum Sey, 1996 in Lagocephalus lunaris pub:907DBCEC-B908-4135-90AA-3950B0F75DD8. This (Bloch & Schneider), Diploproctodaeum cf. monstro- article was published as an Online First article on the online sum Bray, Cribb & Justine, 2010 in Arothron hispidus publication date shown on this page. The article should be cited by using the doi number. This is the Version of Record. (Linnaeus), Multitestis magnacetabulum Mamaev, 1970 and Neomultitestis aspidogastriformis Bray & This article is part of the Topical Collection Digenea. Cribb, 2003 in Platax teira and Opechona austrobacil- laris Bray & Cribb, 1998 in Pomatomus saltatrix Electronic supplementary material The online version of (Linnaeus). Bianium plicitum (Linton, 1928) is this article (https://doi.org/10.1007/s11230-018-9803-3) con- reported from Torquigener squamicauda (Ogilby) tains supplementary material, which is available to authorized users. for the ﬁrst time. Sequences of newly collected specimens of Austroholorchis sprenti (Gibson, 1987) R. A. Bray (&) indicate that the species forms a clade with other Department of Life Sciences, Natural History Museum, Cromwell Road, London SW7 5BD, UK members of the Aephnidiogenidae, agreeing with its e-mail: email@example.com morphology. The phylogenetic status of all newly sequenced species is discussed. T. H. Cribb S. C. Cutmore School of Biological Sciences, The University of Queensland, St Lucia, QLD 4072, Australia 123 Syst Parasitol Introduction described by Cribb & Bray (2010). Those collected were ﬁxed by pipetting into near-boiling saline and During January and July 2016, workshops were held at immediately preserved in formalin or 70% ethanol. the Moreton Bay Research Station at Dunwich on Whole-mounts were stained with Mayer’s paracar- North Stradbroke Island, off southern Queensland, mine or Mayer’s haematoxylin, dehydrated in a graded Australia, as part of a collaborative study of the ethanol series, cleared in beechwood creosote or metazoan parasite fauna of the ﬁshes, particularly the methyl salicylate and mounted in Canada balsam. commercially important ﬁshes, of Moreton Bay. The Measurements were made through a drawing tube on present work is a report on some of the digeneans an Olympus BH-2 microscope, using a Digicad Plus found, framed as an overview of our knowledge of the digitising tablet and Carl Zeiss KS100 software closely related families Lepocreadiidae Odhner, 1905 adapted by Imaging Associates, and are quoted in and Aephnidiogenidae Yamaguti, 1934 in Moreton micrometres, with the range and the mean in paren- Bay. The lepocreadioid fauna of Australian and other theses. The following abbreviations are used: Indo-Paciﬁc ﬁshes has been ‘subjected to recent NHMUK, Natural History Museum, London, UK; sustained study’ (Cribb & Bray, 2011). This has been MNHN, Museum National d’Histoire Naturelle, Paris, documented in some 31 articles (see Bray et al., 2009, France; QM, Queensland Museum Collection, Bris- and references therein; Bray et al., 2010b; Bray et al., bane, Australia. 2010a); however, much work remains to be done. Some genera are large and/or complex and require Molecular sequencing and phylogenetic analysis molecular data to elucidate their status. Specimens for molecular analysis were processed Bray & Cribb (2012) divided members of the according to the protocols used by Sun et al. (2014). Lepocreadiidae Odhner, 1905 as recognised by Bray The complete ITS2 rDNA region was ampliﬁed and (2005) into three families based on a molecular sequenced using the primers 3S (Morgan & Blair, 1995) phylogeny. These three families, the Lepocreadiidae, andITS2.2(Cribbetal., 1998) and the partial D1-D3 28S Lepidapedidae Yamaguti, 1958 and Aephnidio- rDNA region using LSU5 (Littlewood, 1994), 300F genidae Yamaguti, 1934, had previously been consid- (Littlewood et al., 2000), ECD2 (Littlewood et al., 1997) ered subfamilies of the Lepocreadiidae (see Bray, and 1500R (Snyder & Tkach, 2001). Geneious version 2005). In this paper, we analyse species of two of these 10.2.3 (Kearse et al., 2012) was used to assemble and edit three families found in Moreton Bay. A new family contiguous sequences and the start and end of the ITS2 and a new genus and species are erected. In addition, rDNA region were determined by annotation through the this report summarises information from earlier stud- ITS2 Database (Keller et al., 2009; Ankenbrand et al., ies in the region. Collections representing specimens 2015) using the ‘Metazoa’ model. of two lepocreadiid genera (Lepotrema Ozaki, 1932 The partial 28S rDNA sequences generated during and Preptetos Pritchard, 1960) and one lepidapedid this study were aligned with sequences of related genus (Postlepidapedon Zdzitowiecki, 1993) will be species of the Lepocreadioidea Odhner, 1905 from incorporated in genus-speciﬁc studies later and are GenBank using MUSCLE version 3.7 (Edgar 2004) thus not reported here. Novel 28S and ITS2 rDNA run on the CIPRES portal (Miller et al., 2010), with sequences are reported for all new collections, which ClustalW sequence weighting and UPGMA clustering enable the placement of many of the Moreton Bay for iterations 1 and 2. The resultant alignment was species in a wider phylogenetic context. reﬁned by eye using MESQUITE (Maddison & Maddison, 2017). The ends of each sequence were trimmed, and ambiguously aligned regions were Materials and methods identiﬁed and masked manually (those constituting more than three bases and present in greater than 5% of Specimen collection and morphological analysis the sequences in the dataset). Fish were collected by line-ﬁshing, spear-ﬁshing, Bayesian inference and maximum likelihood anal- seine netting and from the commercial tunnel-net yses of the 28S dataset were conducted to explore ﬁshery in Moreton Bay, Queensland, Australia. Fish relationships among these taxa. Bayesian inference were euthanised and examined for trematodes, as analysis was performed using MrBayes version 3.2.6 123 Syst Parasitol (Ronquist et al., 2012) and maximum likelihood Representative DNA sequences: ITS2 rDNA, four analysis using RAxML version 8.2.10 (Stamatakis, identical replicates (two in GenBank MH157055- 2014), both run on the CIPRES portal. The best MH157056); 28S rDNA, one sequence (GenBank nucleotide substitution model was estimated using MH157066). jModelTest version 2.1.10 (Darriba et al., 2012). Both New measurements: Supplementary Table S1. the Akaike Information Criterion (AIC) and Bayesian Information Criterion (BIC) predicted the GTR?I?C Remarks model as the best estimator; Bayesian inference and maximum likelihood analyses were conducted using The new specimens (Fig. 1A) are morphologically the closest approximation to this model. Nodal support identical to those reported from Moreton Bay by Bray in the maximum likelihood analysis was estimated by & Cribb (1998) from Whitley’s toadﬁsh Torquigener performing 100 bootstrap pseudoreplicates. Bayesian whitleyi (Paradice) and T. pleurogramma.New ITS2 inference analysis was run over 10,000,000 genera- rDNA sequences of specimens from T. squamicauda and tions (ngen = 10,000,000) with two runs each T. pleurogramma were identical. Analysis of the 28S containing four simultaneous Markov Chain Monte data showed that this species forms a strongly supported Carlo (MCMC) chains (nchains = 4) and every 1,000th clade with similar lepocreadiid species from tetraodon- tree saved. Bayesian inference analysis used the tiforms (other species of Bianium, Pelopscreadium following parameters: nst = 6, rates = invgamma, Dronen, Blend, Khalifa, Mohamadain & Karer, 2016, ngammacat = 4, and the priors parameters of the Diplocreadium Park, 1939, Diploproctodaeum La Rue, combined dataset were set to ratepr = variable. 1926 and Lobatocreadium Madhavi, 1972); nodal Samples of substitution model parameters and tree support for relationships within this clade was weak and branch lengths were summarised using the (Fig. 2). The two species of Bianium included in the parameters: sump burnin = 3,000 and sumt burnin = phylogenetic analyses are paraphyletic with respect to 3,000. Species of the families Cryptogonimidae Ward, species of Diplocreadium, Diploproctodaeum and Lo- 1917 and Apocreadiidae Skrjabin, 1942 were desig- batocreadium. The status of these specimens from nated as functional outgroup taxa, sensu Bray et al. Moreton Bay as identical to Bianum plicitum as (2009). described by Linton (1928) is yet to be tested by DNA sequence comparison, and we think it highly likely that Family Lepocreadiidae Odhner, 1905 forms from eastern Australian waters are not conspeciﬁc Subfamily Lepocreadiinae Odhner, 1905 with the original specimens from off north-eastern USA. Genus Bianium Stunkard, 1930 Bianium arabicum Sey, 1996 Bianium plicitum (Linton, 1928) Stunkard, 1931 Type-host: Lagocephalus lunaris (Bloch & Schneider) Syn. Psilostomum plicitum Linton, 1928 (Tetraodontiformes: Tetraodontidae), lunartail puffer. Type-host: Larus argentatus Pontoppidan (Charadri- Type-locality: Off Kuwait, Arabian Gulf. iformes: Laridae), herring gull. New records: Type-locality: Woods Hole, Massachusetts, USA. Host: Lagocephalus lunaris. New records: Locality: Off Wynnum North, Moreton Bay (2723 S, Hosts: Torquigener squamicauda (Ogilby), brush-tail 15311 E). toadﬁsh; T. pleurogramma (Regan), weeping toado Site in host: Intestine. (Tetraodontiformes: Tetraodontidae). Voucher material: Two specimens in the QM Localities:Ex T. squamicauda, Moreton Banks, G237254–5, one in the NHMUK 2018.3.26.2. 0 0 Moreton Bay (2724 S, 15320 E); ex T. pleuro- Representative DNA sequences: ITS2 rDNA, one gramma, off Amity, Moreton Bay (2724 S, sequence (GenBank MH157054); 28S rDNA, one 15326 E). sequence (GenBank MH157076). Site in host: Intestine. New measurements: Supplementary Table S1. Voucher material: Three specimens in the QM G237251–3, one in the NHMUK 2018.3.26.1. 123 Syst Parasitol Fig. 1 A, Bianium plicitum (Linton, 1928) ex Torquigener squamicauda, ventral view, uterus in outline; B, Bianium arabicum Sey, 1996 ex Lagocephalus lunaris, ventral view, uterus in outline; C, Mobahincia teirae n. g., n. sp. ex Platax teira, Moreton Bay, ventral view, uterus in outline; D, Mobahincia teirae n. g., n. sp. ex Platax teira, off Heron Island, ventral view, uterus in outline. Scale-bars:A, B, 500 lm; C, D, 200 lm 123 Syst Parasitol Remarks New records: Host: Trachinotus coppingeri Gu¨nther (Perciformes: In the original description of B. arabicum, Sey (1996) Carangidae), swallowtail dart. stated ‘Along the lateral sides of body longitudinal Locality: Off Green Island, Moreton Bay (2725 S, folds present, bending ventrally and ending at poste- 15314 E). rior extremity’. The longitudinal folds (ﬂanges) do not Site in host: Intestine. appear to reach the full length of the body in our Voucher material: Six specimens in the QM specimens (Fig. 1B). When describing specimens G237275–80. considered to be this species from the silverstripe Representative DNA sequences: ITS2 rDNA, three blaasop Lagocephalus sceleratus (Gmelin) off New replicates (one in GenBank MH157057); 28S rDNA, Caledonia, Bray et al. (2010a) said ‘it has full-length one sequence (GenBank MH157067). lateral folds of the body (or scoop-sides), although the full extent is not always visible on both sides of the Remarks worm’. These authors pointed out the similarity of these worms to those described from L. lunaris by Bray & Gibson (1990) redescribed the original Hafeezullah (1970) under the name B. plicitum specimens of Stephanostomum trachinoti Fischthal (Linton, 1928) from off Chennai (as Madras) in the & Thomas, 1968 and its synonym Opechona pseu- Bay of Bengal and by Shen & Tong (1990) under the dobacillaris Fischthal & Thomas, 1970, and placed name B. dayawanense Shen & Tong, 1990 from Daya the species in Clavogalea. Bray & Cribb (1998) Bay, China [in this case the host is quoted as L. lunaris redescribed the worm based on new material from the spadiceus (Richardson)]. The lateral ﬂanges of the large-spotted dart Trachinotus botla (Shaw) off Heron Bay of Bengal worms are similar to those seen in our Island (southern Great Barrier Reef) and T. coppingeri specimens and the dimensions are close to those found Gu¨nther off northern New South Wales and in by Sey (1996) and Bray et al. (2010a) (Supplementary Moreton Bay. Our newly collected material appears Table S1). The Chinese worms tend to be larger and identical to these descriptions. New 28S rDNA data the ﬂanges are illustrated as distinct ﬂaps reaching were identical to sequences reported by Bray et al. only to the ventral sucker level. It is not possible to be (2009) based on specimens from T. coppingeri certain of the status of all these forms, but it appears collected off Heron Island. Phylogenetic analysis of that the Bay of Bengal specimens are more similar to the 28S dataset indicates that, of taxa available for the worms here considered B. arabicum. Thisisthe analysis, this species is most closely related to ﬁrst report of B. arabicum from Moreton Bay. Preptetos trulla (Linton, 1907), Prodistomum keyam Analysis of the 28S data showed that this species Bray & Cribb, 1996, Opechona austrobacillaris Bray forms a strongly-supported clade with similar lepocre- & Cribb, 1998 and Opechona kahawai Bray & Cribb, adiid species from tetraodontiforms; nodal support for 2003. These ﬁve species formed a clade in the relationships within this clade were weak (Fig. 2) and, phylogenetic analysis with C. trachinoti as sister to a as discussed above, the two species of Bianium did not clade of the other four species; however, nodal support form a clade. for this topology was poor (Fig. 2). Genus Clavogalea Bray, 1985 Genus Diplocreadium Park, 1939 Clavogalea trachinoti (Fischthal & Thomas, 1968) Diplocreadium tangaloomaense Bray, Cribb & Bray & Gibson, 1990 Barker, 1996 (emend.) Syns Stephanostomum trachinoti Fischthal & Thomas, 1968; Opechona pseudobacillaris Fischthal & Tho- Type-host: Paramonacanthus japonicus (Tilesius) mas, 1970 (Tetraodontiformes: Monacanthidae), hairﬁnned leatherjacket. Type-host: Trachinotus goreensis Cuvier (Perci- Type-locality: Off Tangalooma, Moreton Bay, formes: Carangidae), longﬁn pompano. Queensland. Type-locality: Off Iture, Elmina, Ghana. 123 Syst Parasitol Fig. 2 Relationships between members of the seven families of the superfamily Lepocreadioidea based on maximum likelihood analysis of the partial 28S rDNA dataset. Species from Moreton Bay are shown in bold and clades representing the Enenteridae, Gorgocephalidae, Gyliauchenidae and Lepidapedidae are collapsed for brevity. Maximum likelihood bootstrap support values are shown above the nodes and Bayesian inference posterior probabilities below. Support values\80 and\0.80 are not shown. Outgroup taxa are species of the Apocreadiidae and Cryptogonimidae. Abbreviations: Aephnidiog., Aephnidiogenidae; G, Gibsonivermidae; Out., outgroup taxa Remark Diploproctodaeum monstrosum Bray, Cribb & Justine, 2010 This species has not been detected since its original description and no material is available for molecular Type-host: Arothron stellatus (Anonymous) (Te- characterisation. traodontiformes: Tetraodontidae), stellate puffer. 123 Syst Parasitol Type-locality: Off Mermaid Beach, Lizard Island, Type-locality: Off Mud Island, Moreton Bay, Queensland, Australia. Queensland. Diploproctodaeum cf. monstrosum Remark New records This species has not been detected since its original Host: Arothron hispidus (Linnaeus), white-spotted description and no material is available for molecular puffer. characterisation. Locality: Off Peel Island, Moreton Bay (2730 S, 15320 E). Genus Lepocreadioides Yamaguti, 1936 Site: Intestine. Syn. Bicaudum Bilqees, 1971 Voucher material: Three specimens in the QM G237281–3. Lepocreadioides orientalis Park, 1939 Representative DNA sequences: ITS2 rDNA, two replicates (one in GenBank MH157059); 28S rDNA, Type-host: Cynoglossus joyneri Gu¨nther, red tongue- one sequence (GenBank MH157069). sole (Pleuronectiformes: Cynoglossidae). Type-locality: Off Simmi Island, North Tyoˆsen, Remarks Korea. Bray et al. (2010a) reported this species in Arothron Remark stellatus and A. mappa from off Lizard Island. They pointed out that the sequence of ‘Diploproctodaeum sp.’ This species has not been re-collected from Moreton from A. stellatus off Lizard Island (GenBank FJ788474), Bay since the report from the fourlined tonguesole used in the study of Bray et al. (2009), referred to this Cynoglossus bilineatus (Lace´pe`de) by Bray & Cribb species. 28S sequence data generated from the new (1998) and no material is available for molecular Moreton Bay material differs from that sequence by 5 characterisation. bases. No morphological differences could be detected between the two collections, but only a relatively small Genus Mobahincia n. g. number of specimens has been collected and the rather amorphous structure of these worms makes morpholog- Diagnosis ical comparisons difﬁcult. Given that the two sites are Body broader anteriorly, tapering posteriorly. Tegu- only approximately 1,650 km apart, a 5 bp difference in ment spined. Eye-spot pigment scattered at pharyngeal the 28S rDNA raises the possibility of the presence or level. Oral sucker transversely oval, subterminal. more than one species. However, we consider the current Ventral sucker rounded, smaller than oral sucker, in evidence insufﬁcient to suggest that specimens from anterior quarter of body-length. Prepharynx short. Moreton Bay represent a species distinct from that Pharynx oval. Oesophagus not detected. Intestinal occurring on the northern Great Barrier Reef but consider bifurcation dorsal to anterior part of ventral sucker or the issue worthy of further consideration in the light of just in forebody. Caeca straight, reaching to posterior additional genetic data from more variable gene regions extremity where they abut body wall at base on small (Blasco-Costa et al., 2016). For the present, the designa- indentations; ani possibly present. Testes two, oval, tion D. cf. monstrosum seems the best way to draw entire, tandem contiguous, in mid-hindbody. External attention to these issues. seminal vesicle large, saccular, dorsal to uterus. Cirrus-sac claviform. Internal seminal vesicle large, Diploproctodaeum yosogi Bray, Cribb & Barker, oval, curved. Pars prostatica oval vesicular, lined with 1996 anuclear cell-like bodies. Ejaculatory duct thick- walled muscular, long, complexly folded. Genital Type-host: Paramonacanthus japonicus (Tilesius) atrium small. Genital pore sinistral to antero-sinistral (Tetraodontiformes: Monacanthidae), hairﬁnned to ventral sucker. Ovary multilobate, immediately pre- leatherjacket. testicular. Seminal receptacle canalicular. Mehlis’ 123 Syst Parasitol gland dorsal to ovary. Uterus between ovary and genus the testes are symmetrical. We conclude that ventral sucker, intracaecal. Eggs tanned, operculate. the relationships of this form are best expressed by Vitellarium in follicular ﬁelds at ventral sucker level the erection of a new genus. and in hindbody. Parasites in intestine of ephippid ﬁshes. Mobahincia teirae n. sp. Type-species: Mobahincia teirae n. sp. Etymology: The generic name is a feminine noun Type-host: Platax teira (Forsskal) (Perciformes: derived from the localities at which this genus has Ephippidae), longﬁn batﬁsh. been found: Moreton Bay (Moba), Heron Island (hi), Type-locality: Four Beacons, Moreton Bay (2710 S, New Caledonia (nc). 15321 E). ZooBank registration: To comply with the regulations Other localities: Off Heron Island (2327 S, set out in article 8.5 of the amended 2012 version of 15155 E); Noumea Fish Market, New Caledonia. the International Code of Zoological Nomenclature Site in host: Intestine. (ICZN, 2012), details of the new genus have been Type-material: Holotype QM G237256 and 12 para- submitted to ZooBank. The Life Science Identiﬁer types QM G237257–60, NHMUK 2018.3.26.5–8. (LSID) for Mobahincia n. g. is urn:lsid: Voucher material: Off Heron Island: QM G237261; zoobank.org:act:8543D3CA-81FC-43A7-9ACB-6BA off New Caledonia: MNHN JNC2872F. 5DE6D6BA4. Representative DNA sequences: ITS2 rDNA, ﬁve replicates (one in GenBank MH157058); 28S rDNA, Remarks one sequence (GenBank MH157068). ZooBank registration: To comply with the regulations The species on which this new genus is based set out in article 8.5 of the amended 2012 version of appears morphologically closely related to mem- the International Code of Zoological Nomenclature bers of Diploproctodaeum and Bianium in having (ICZN, 2012), details of the new species have been its caeca abutting the posterior body wall, giving submitted to ZooBank. The Life Science Identiﬁer the appearance of ani, the usual condition in (LSID) for Mobahincia teirae n. sp. is urn:lsid:- species of the latter genera; however, there is no zoobank.org:act:7FD7D6C8-D114-4C5E-BEDA- indication of an anterior scoop as is usually present E25CB81979E9. in these taxa. Molecular evidence suggests unam- Etymology: The speciﬁc epithet is derived from that of biguously that the new genus is not closely related the host species. to members of these two genera. The exact relationship of this species is not well resolved in Description (Fig. 1C, D) the phylogram derived from the 28S analyses of the currently available lepocreadiid sequences (many [Based on 7 ovigerous and seven non-ovigerous relationships within the family have poor support), specimens from Moreton Bay, 1 specimen from off but it is clear that it does not resolve within the Heron Island and 1 specimen from off New Caledonia; well-supported clade which includes Diploprocto- measurements given in Table 1.] Body broader ante- daeum and Bianium species (Fig. 2). Following the riorly, tapering posteriorly. Body spines small on key to the Lepocreadiidae produced by Bray anterior ‘shoulders’, much more robust along remain- (2005), the species appears closest to members of der of body, reach close to posterior extremity. Eye- Lobatocreadium or Pseudocreadium Layman, spot pigment scattered at pharyngeal level. Oral sucker 1930; the new genus differs from both in the transversely oval, subterminal. Ventral sucker presence of long caeca abutting the body-wall and rounded, smaller than oral sucker, in anterior quarter the terminal excretory pore. The vitellarium is of body-length. Prepharynx short, mainly in posterior more extensive in both species of Lobatocreadium concavity of oral sucker. Pharynx oval. Oesophagus and Pseudocreadium, andinmembers of thelatter not detected. Intestinal bifurcation dorsal to anterior 123 Syst Parasitol Table 1 Measurements and ratios of Mobahincia teirae ex Platax teira Locality Moreton Bay (n = 7) New Caledonia Heron Island (n = 1) (n = 1) Range Mean Body 685–1,018 9 344–427 834 9 383 750 9 395 1,013 9 415 Forebody length 186–234 204 214 221 Pre-oral lobe length 0–5 3 6 5 Oral sucker 104–141 9 148–190 125 9 170 127 9 179 115 9 170 Prepharynx length 0–35 5 0 8 Pharynx 82–100 9 80–105 90 9 89 107 9 97 84 9 86 Oesophagus length 0 18 24 Distance from intestinal bifurcation to ventral sucker 0–16 4 0 0 (IB-VS) Distance from vitellarium to ventral sucker 0 0 19 0 Ventral sucker 79–112 9 90–122 96 9 103 89 9 98 93 9 99 Cirrus-sac 129–189 9 71–90 158 9 79 121 9 44 175 9 56 Distance from external seminal vesicle to ventral sucker 85–118 102 70 169 Distance from ventral sucker to ovary (VS-Ov) 20–51 34 17 61 Ovary 84–115 9 112–169 102 9 134) 64 9 110 132 9 133 Distance from ovary to anterior testis 0 0 0 0 Anterior testis 118–144 9 123–147 128 9 131 112 9 141 163 9 137 Distance between testes 0 0 0 0 Posterior testis 106–214 9 107–137 157 9 121 125 9 144 209 9 129 Post-testicular distance 112–190 150 128 190 Post-caecal distance 0–25 5 0 0 Eggs 58–70 9 26–41 62 9 34 63 9 23 69 9 33 Width (%) 41.9–59.0 46.5 52.7 41.0 Forebody (%) 21.8–27.2 24.7 28.5 21.8 Sucker length ratio 1:0.67–0.90 1:0.77 1:0.70 1:0.81 Sucker width ratio 1:0.58–0.64 1:0.60 1:0.55 1:0.58 Oral sucker: pharynx width 1:1.72–2.09 1:1.92 1:1.84 1:1.98 Ventral sucker to ovary (%) 2.63–5.02 4.04 2.32 6.06 External seminal vesicle to ventral sucker as % of VS-Ov 283–418 351 403 274 Post-testicular distance (%) 16–20 18 17.1 18.8 Prepharynx (%) 0–16.4 2.34 0 3.51 Oesophagus (%) 0 0 2.37 2.37 Distance IB-VS (%) 0–1.76 0.50 0 0 Vitellarium to ventral sucker distance (%) 0 0 2.47 0 Ovary to anterior testis (%) 0 0 0 0 Distance between testes (%) 0 0 0 0 Cirrus-sac length (%) 16.4–22.0 19.0 16.2 17.2 Pre-vitelline distance 186–234 204 195 221 Pre-vitelline distance (%) 21.8–27.2 24.7 26.0 21.8 Oesophagus length as % of forebody length 0 0 8.33 10.9 Distance IB-VS as % of forebody length 0–7.31 2.02 0 0 Vitellarium to ventral sucker distance as % of 0 0 8.66 0 forebody length Note: (%), percent of body-length where not otherwise noted; IB-VS, intestinal bifurcation to ventral sucker distance. Where length is followed by width, the two are separated by an ‘9’ 123 Syst Parasitol part of ventral sucker or just in forebody. Caeca Genus Multitestis Manter, 1931 straight, reach to posterior extremity where they abut body wall at base on small indentations; ani possibly Multitestis magnacetabulum Mamaev, 1970 present. Testes 2, oval, entire, tandem contiguous, in mid- Type-host: Platax orbicularis (Forsskal) (ﬁrst host hindbody. External seminal vesicle large, saccular, listed) (Perciformes: Ephippidae), orbicular batﬁsh. dorsal to uterus. Cirrus-sac claviform. Internal seminal Type-locality: Gulf of Tonkin. vesicle large, oval, curved. Pars prostatica oval New records: vesicular, lined with anuclear cell-like bodies. Ejac- Host: Platax teira (Forsska˚l) (Perciformes: Ephippi- ulatory duct thick-walled, muscular, long, complexly dae), longﬁn batﬁsh. folded. Genital atrium small. Genital pore closely Locality: Four Beacons, Moreton Bay (2710 S, sinistral to antero-sinistral to ventral sucker. 15321 E). Ovary multilobate (about 14–20 lobes), immediately Site in host: Intestine. pre-testicular. Seminal receptacle saccular, dorsal to Voucher material: Six voucher specimens QM anterior testis. Laurer’s canal not detected. Mehlis’ gland G237262–7, three NHMUK 2018.3.26.9–11. dorsal to ovary. Uterus between ovary and ventral sucker, Representative DNA sequences: ITS2 rDNA, two intracaecal. Eggs tanned, operculate. Vitellarium follic- replicates (one in GenBank MH157061); 28S rDNA, ular, in extensive dorsal and ventral ﬁelds, from level of one sequence (GenBank MH157071). ventral sucker to posterior extremity; ﬁelds conﬂuent at New measurements: Supplementary Table S2. level of testes and in post-testicular region. Excretory pore terminal; excretory vesicle narrow Remarks posteriorly, widens abruptly and reaches at least to posterior testes. This is the ﬁrst record of this species from Moreton Bay. Bray & Cribb (2003a)reporteditfrom Platax teira off Remarks Heron Island and Bray et al. (2009) used sequences from that collection in their molecular study of the superfamily Several species of Diploproctodaeum are found in Lepocreadioidea. 28S sequence data generated from new Platax spp., namely D. plataxi Mamaev, 1970, D. collections from Moreton Bay differed from the Heron rutellum (Mamaev, 1970) and D. tsubameuo Bray & Island specimens (GenBank FJ788485) by a single base. Cribb, 2003; all three species have caeca abutting the A single base difference is consistent with the minor body-wall and are often described as having ani geographical variation found between these locations for (Mamaev, 1970;Bray&Cribb, 2003a). Other lepocre- other trematodes (e.g. Cutmore et al., 2016;Brookset al., adiid species from Platax spp., such as Deraiotrema 2017); however, given that this single base difference (an platacis Machida, 1982, Neomultitestis palauensis A to T transversion) is within the in the ﬁrst 15 bases of Machida, 1982 and N. aspidogastriformis Bray & Cribb, the start of the sequence, and that this base position is an 2003 are also described as having ani or the appearance A in all other taxa included in the analysis, we predict of ani (Machida, 1982;Bray&Cribb, 2003a). that the difference in FJ788485 is a sequencing misread. Phylogenetic analysis of the 28S dataset showed This species has also been reported from the same host in that this species does not form a strongly-supported the waters off New Caledonia by Bray & Justine (2012). clade with any particular clade of lepocreadiids. The new species was sister to a clade including Neoprepte- Genus Neomultitestis Machida, 1982 tos arusettae Machida, 1982, Multitestis magnacetab- ulum Mamaev, 1970 and Neomultitestis aspidogas- Neomultitestis aspidogastriformis Bray & Cribb, triformis, the latter two of which are Platax-infecting 2003 species; however, nodal support for this relationship was poor. The new species was not closely related to Type-host: Platax teira (Forsska˚l) (Perciformes: species of other genera which have similar caecal Ephippidae), longﬁn batﬁsh. terminations, namely Diploproctodaeum, Bianium Type-locality: Off Heron Island, Queensland, Australia. and Pelopscreadium. New records 123 Syst Parasitol Host: Platax teira. Cribb, 1998). Our specimens from Moreton Bay are Locality: Four Beacons, Moreton Bay (2710 S, indistinguishable from those described by Bray & 15321 E). Cribb (1998), and we are conﬁdent that the new Site in host: Intestine. specimens are conspeciﬁc with those from off Fre- Voucher material: One voucher specimen lodged in mantle and Iluka. the QM G237268. New 28S sequence data generated for O. austrobacil- Representative DNA sequences: ITS2 rDNA, one laris differs from those of O. kahawai,from Arripis sp. sequence (GenBank MH157062); 28S rDNA, one off Tasmania, by just a single base. Unfortunately, no sequence (GenBank MH157072). ITS2 rDNA sequencedata(asuperiormarkerfor species New measurements: Supplementary Table S2. delineation) are available for the Tasmanian species. Bray & Cribb (2003b) distinguished these two species by Remarks the sucker-ratio and the pseudoesophagus/oesophagus length ratio, and by the forebody being proportionally Bray & Cribb (2003a) reported this species from much longer in O. kahawai (40–44 vs 28–35% of body P. teira off Heron Island, and Bray et al. (2009) used length) (Supplementary Table S2; Bray & Cribb, 1998). 28S rDNA sequences from that collection in their Given the minor genetic differences, the relationship molecular study of the superfamily Lepocreadioidea. between these two morphologically distinct forms This is the ﬁrst report of N. aspidogastriformis from warrants further study. Phylogenetic analysis of the Moreton Bay. New 28S data generated from Moreton 28S dataset showed these two species of Opechona to be Bay specimens were identical to those of this species most closely related to Prodistomum keyam; however, off Heron Island (GenBank FJ788489). nodal support for this clade was poor. Genus Opechona Looss, 1907 Genus Prodistomum Linton, 1910 Opechona austrobacillaris Bray & Cribb, 1998 Prodistomum keyam Bray & Cribb, 1996 Type-host: Pomatomus saltatrix (Linnaeus), tailor Type-host: Monodactylus argenteus (Linnaeus) (Per- (Perciformes: Pomatomidae). ciformes: Monodactylidae), silver moony. Type-locality: Off South Mole, Fremantle, Western Type-locality: Off Hope Island, Queensland, Australia. Australia. New material: New records: Host: Pomatomus saltatrix. Host: Monodactylus argenteus. Locality: Off Garden Island, Moreton Bay (2736 S, Locality: In Port of Brisbane Land Reclamation, 0 0 0 15320 E). Moreton Bay (2721 S, 15311 E); off Amity, More- 0 0 Site in host: Intestine. ton Bay (2724 S, 15326 E). Voucher material: Two specimens in the QM Site in host: Intestine. G237269–70, one in the NHMUK 2018.3.26.3. Voucher material: Four specimens in the QM Representative DNA sequences: ITS2 rDNA, two G237271–4, one in the NHMUK 2018.3.26.3. replicates (one in GenBank MH157063); 28S rDNA, Representative DNA sequences: ITS2 rDNA, three one sequence (GenBank MH157073). identical replicates (one in GenBank MH157064); 28S New measurements: Supplementary Table S2. rDNA, one sequence (GenBank MH157074). New measurements: Supplementary Table S2. Remarks Remarks This is the ﬁrst report of this species from Moreton Bay. Although the type-locality is off Western Aus- Bray & Cribb (1996) and Bray et al. (2009) reported tralia, the original description also reported and this host/species combination in Moreton Bay. Bray described this species from the eastern coast of et al. (2009) used sequences of this species from this Australia, off Iluka in New South Wales (Bray & host in Moreton Bay in their molecular study of the 123 Syst Parasitol superfamily Lepocreadioidea. Molecular data from considered Gibsonivermis a genus ‘incertae sedis new specimens collected in this study were identical to within the superfamily’ Lepocreadioidea and ‘too those (FJ788493) from Bray et al. (2009). Phyloge- enigmatic to allow conﬁdent placement’. Barker et al. netic analysis of the 28S dataset showed P. keyam to (1993) sequenced the D1 domain of the 28S ribosomal be most closely related to Opechona austrobacillaris RNA gene of the type-species of this new taxon under and O. kahawai, but with low support (Fig. 2). Bray & its old name Intusatrium berryi Gibson, 1987 but did Justine (2012) reported this species from the same host not apparently submit the sequence to GenBank (it is from the waters around New Caledonia. itemised in the paper). In the early days of the development of molecular studies, few digenean Family Gibsonivermidae n. fam. sequences were available. The tree produced by Barker et al. (1993) included two other lepocreadioids, Diagnosis Gyliauchen sp. (Gyliauchenidae) and Tetracerasta Body elongate-oval, ﬂattened. Tegument armed with blepta Watson, 1984 (Aephnidogenidae), which clus- small spines. Oral sucker subglobular, subterminal. tered with Gibsonivermis, but they stated that ‘evi- Ventral sucker rounded, pre-equatorial. Prepharynx dence for the monophyly of the two lepocreadiids distinct. Pharynx oval. Oesophagus distinct. Intestinal [Tetracerasta and Gibsonivermis] was weak’. The bifurcation in mid-forebody. Caeca form uroproct at molecular phylogeny inferred from 28S data reported here conﬁrms that Gibsonivermis does not belong to posterior extremity. Testes two, lobed to almost entire, tandem, slightly separated, in mid-hindbody. External any of the six accepted lepocreadioid families (i.e. seminal vesicle very elongate, tubular, coiled, reaches Lepocreadiidae, Aephnidiogenidae, Enenteridae, Gor- well into hindbody. Cirrus-sac long, attenuated, coiled gocephalidae, Gyliauchenidae, Lepidapedidae; see proximally. Internal seminal vesicle tubular, coiled. Bray & Cribb, 2012), constituents of which all form Pars prostatica long, narrow. Ejaculatory duct elon- strongly supported clades. It is distinct enough, both gate, muscular, expands distally. Genital atrium small. morphologically and genetically, to warrant the pro- Genital pore dextrally submedian, ventral to pharynx. posal of a new family. In the current analyses, the Ovary with 4–6 lobes, pretesticular, slightly separated Gibsonivermidae was sister to the Lepidapedidae, from anterior testis. Seminal vesicle between ovary with branch lengths between the two families similar and anterior testis. Uterus pre-ovarian, intercaecal; to those found between the Lepocreadiidae and lateral slings extend into forebody. Metraterm narrow. Aephnidiogenidae, and the Enenteridae and Gyli- Vitellarium follicular; ﬁelds reach from anterior auchenidae, indicative of a family level distinction. region of hindbody or ventral sucker to posterior Gibsonivermis berryi (Gibson, 1987) Bray, Cribb & extremity. Excretory vesicle I-shaped, reaches ante- Barker, 1997 has several features very unusual for rior testis. In intestine of marine teleosts. species within the superfamily, the most striking of Type-genus: Gibsonivermis Bray, Cribb & Barker, which is the form of the male terminal genitalia 1997. (Fig. 3A, B). The cirrus-sac is elongate, narrow, coiled ZooBank registration: To comply with the regulations proximally and contains a long tubular coiled internal set out in article 8.5 of the amended 2012 version of seminal vesicle, a long narrow pars prostatica and a the International Code of Zoological Nomenclature muscular ejaculatory duct which widens distally (ICZN, 2012), details of the new family have been (Gibson, 1987). The external seminal vesicle is long, submitted to ZooBank. The Life Science Identiﬁer tubular and coiled and merges into the internal seminal (LSID) for Gibsonivermidae n. fam. is urn:lsid:- vesicle. Gibson (1987) described a constriction of the zoobank.org:act:F50B45FB-6B41-44B4-8FD2- seminal vesicle as it enters the cirrus-sac but stated 5042D1AD938F. that it was only seen in sections. We have not been able to detect this constriction in whole-mounted worms. If Remarks it is always present, it is obscured by the folds of the seminal vesicle in the region dorsal to the ventral Bray et al. (1997), in proposing Gibsonivermis, stated sucker in all the specimens we examined. This folding that it is ‘not immediately clear to which subfamily also usually obscures the precise posterior extent of this genus belongs’, and Bray & Cribb (2012) the cirrus-sac wall. Other distinguishing features, 123 Syst Parasitol which are rare or absent in other lepocreadioids, on the southern Great Barrier Reef. Bray et al. (1999) include a uroproct and a signiﬁcant proportion of the summarised the knowledge of the parasites of the uterus in the forebody. At present, no other lepocre- Sillaginidae and found that no ‘lepocreadiids’ were adioids appear to have characters in any way resem- reported outside Australian waters, but that in this bling those of specimens of Gibsonivermis. region a few unusual, apparently endemic, forms The single species of Gibsonivermis is so far occurred, namely species of Gibsonivermis, Austro- known only from Moreton Bay and off Heron Island holorchis Bray & Cribb, 1997 and Lepidapedella Fig. 3 A, Gibsonivermis berryi (Gibson, 1987)ex Sillago ciliata. Holotype, ventral view, uterus in outline; B, Gibsonivermis berryi (Gibson, 1987)ex Sillago analis. Male terminal genitalia, with ventral sucker and gut in outline. Scale-bars: A, 1,000 lm; B, 500 lm 123 Syst Parasitol Remark Bray, Cribb & Pichelin, 1999. Austroholorchis is now known to be an aephnidiogenid (see below). Gibson (1987) and Bray et al. (1997) reported this Lepidapedella is an unusual worm, which likewise does not agree well with any lepocreadioid family, species from the golden-line whiting Sillago analis Whitley, S. ciliata and the trumpeter whiting S. but shows no morphological similarities to G. berryi, and was placed in the Lepidapedidae by Bray & maculata Quoy & Gaimard, (Perciformes: Sil- laginidae) from Moreton Bay. ITS2 rDNA data were Cribb (2012). The species of endemic Australian lepocreadioid genera which are not reported from found to be identical for specimens of this species sillaginids include the lepocreadiids Amphicreadium infecting S. ciliata from Moreton Bay and off Heron Bray & Cribb, 2001, Cliveus Bray & Cribb, 1997 and Island. Rugocavum Bray & Cribb, 1997, the lepidapedids Harveytrema Kruse, 1979 and Scaphatrema Bray & Family Aephnidiogenidae Yamaguti, 1934 Cribb, 1997, and the unassigned Paraneocreadium Genus Austroholorchis Bray & Cribb, 1997 Kruse, 1978 and Jericho Bray & Cribb, 1997 (Kruse, 1978, 1979; Bray & Cribb, 1997a, 2001, 2012). None Austroholorchis sprenti (Gibson, 1987) Bray & Cribb, 1997 of members of these genera exhibit any great similarity to G. berryi, although the single species Syn. Holorchis sprenti Gibson, 1987 of Paraneocreadium has some extension of the uterus into the forebody and the testes are lobed Type-host: Sillago maculata Quoy & Gaimard (Per- (Kruse, 1978, 1979; Bray & Cribb, 1997a). Consid- ciformes: Sillaginidae), trumpeter whiting. ering the recognition that Gibsonivermis warrants a Type-locality: Deception Bay, Moreton Bay. separate family-level status within the Lepocrea- New records: Host: Sillago ciliata Cuvier. dioidea, the phylogenetic status of these other distinctive, apparent ‘‘southern endemics’’, is of great Locality: Off Dunwich, Moreton Bay (2729 S, 15323 E). interest. Since 1999, only records of opecoelids and transversotrematids have been added to the known Voucher material: Seven specimens in the QM G237284–90. sillaginid digenean fauna (Aken’Ova, 2003; Ake- n’Ova et al., 2008; Cutmore et al., 2016). Representative DNA sequences: ITS2 rDNA, four replicates (one in GenBank MH157065); 28S rDNA, Genus Gibsonivermis Bray, Cribb & Barker, one sequence (GenBank MH157075). Remarks Gibsonivermis berryi (Gibson, 1987) Bray, Cribb & Gibson (1987) and Bray & Cribb (1997b) reported this Barker, 1997 Syn. Intusatrium berryi Gibson, 1987 species from Sillago analis, S. ciliata and S. maculata from Moreton Bay. Analyses of the 28S data generated Type-host: Sillago ciliata Cuvier (Perciformes: Sil- during this study indicate that this species forms a strongly-supported clade with all other included laginidae), sand whiting. Type-locality: Deception Bay, off Moreton Bay. aephnidiogenids. Within the aephnidiogenid clade, New record: A. sprenti formed a strongly-supported clade with Host: Sillago ciliata. species of Aephnidiogenes Nicoll, 1915, Holorchis Locality: Off Dunwich, Moreton Bay (2729 S, Stossich, 1901 and Neolepocreadium Thomas, 1960, 15323 E). sister to the two freshwater anguilliform-infecting Voucher specimens: Six specimens in the QM species Stegodexamene anguillae Watson, 1984 and Tetracerasta blepta Watson, 1984. G237291–6. Representative DNA sequences: ITS2 rDNA, two replicates (one in GenBank MH157060); 28S rDNA, one sequence (GenBank MH157070). 123 Syst Parasitol Phylogenetic results species of several other trematode families have been shown to be genetically identical between the Great Alignment of the 28S rDNA dataset (Table 2) yielded Barrier Reef and Moreton Bay (Brooks et al., 2017; 1,299 characters (including indels). Deleted ambigu- Yong et al., 2018). More controversial is the close ously aligned regions amounted to 49 bases (less than molecular similarity of Opechona austrobacillaris 4% of the alignment), resulting in a ﬁnal dataset of from Moreton Bay and O. kahawai from Tasmanian 1,250 characters for phylogenetic analysis. Bayesian waters, which brings into question the status of these inference and maximum likelihood analyses of the forms. Although morphologically similar, they do 28S rDNA dataset resulted in phylograms with almost appear to be readily distinguishable. Prodistomum identical topologies (Fig. 2). Only the relationship keyam, O. austrobacillaris and O. kahawai are between the specimens of Diploproctodaeum mon- included in a moderately-supported clade, which is strosum and Diploproctodaeum cf. monstrosum and poorly resolved internally. Preptetos trulla (Linton, that between Lepidapedoides angustus Bray, Cribb & 1907) is in this clade and is clearly not placed in the Barker, 1996 and Preptetos caballeroi Pritchard 1960 correct genus given its distance from the type-species, were different. The topology was almost identical (but P. caballeroi Pritchard, 1960. Preptetos trulla, Prodis- expanded relative) to that found by Bray et al. (2009), tomum keyam, O. austrobacillaris and O. kahawai are in which all lepocreadioid taxa formed a strongly also similar morphologically. supported clade to the exclusion of cryptogonimid and Moreton Bay members of the similar, and contro- apocreadiid outgroup taxa. The now seven accepted versially separated, genera Bianium and Diploprocto- families each formed monophyletic clades, all of daeum formed a well-supported clade, but were which were strongly supported; nodal support for internally poorly resolved and clearly need greater relationships within the familial clades was lower, sampling, both of species and genes, for a convincing especially for those in the lepocreadiid clade. The arrangement to emerge. The morphological charac- type- and only species of the Gibsonivermidae formed teristics that are currently used to separate these genera a well-supported clade with the lepidapedids. Most are evidently unreliable. The ﬁve-base difference in genera for which there were more than one sequenced sequences between D. monstrosum (‘Diploprocto- species included formed monophyletic clades (Gor- daeum sp.’ in Bray et al., 2009) from off Lizard Island gocephalus Manter, 1966, Holorchis, Hypocreadium and D. cf. monstrosum from Moreton Bay indicates Ozaki, 1936, Lepidapedon Stafford, 1904, Opechona, that there are potentially two closely related species in Paragyliauchen Yamaguti, 1934 and Proenenterum Queensland waters. However, few specimens have Manter, 1954), but several formed notably poly- been collected from either location, and currently phyletic assemblages (Bianium, Diploproctodaeum there are not enough morphological or molecular data and Preptetos). to justify the proposal of a new species; this complex needs further study. Members of the Lepocreadiidae sensu stricto in Discussion Moreton Bay were divided into two major clades reﬂecting the ﬁndings reported by Bray et al. (2009), Our phylogenetic hypotheses are inferred from the who labelled the clades as VII and VIII. Clade VII phylogram generated from the 28S rDNA dataset, but includes what might be considered ‘typical’ lepocre- all are supported by morphology. This phylogram adiids, mostly occurring in shallow water and, as far as includes the sequences used by Bray et al. (2009) and is known, with a gastropod ﬁrst intermediate host. Bray & Cribb (2012) in their reviews of the phylogeny Clade VIII includes many species from reef ﬁshes, and systematics of lepocreadioids and allows us to set especially tetraodontiform ﬁshes, with just one the Moreton Bay worms in context. The uncontrover- resolved life-cycle which utilises a bivalve ﬁrst sial results in the tree are the ﬁnding of identical intermediate host (Hassanine, 2006). The distribution sequences for Neomultitestis aspidogastriformis and of these clades in terms of their assemblage and the Clavogalea trachinoti from off Heron Island and in nature of their hosts is worthy of further exploration, Moreton Bay and the near identical sequences for but a much wider understanding of the genetic Multitestis magnacetabulum from the same localities; 123 Syst Parasitol Table 2 Collection data and GenBank accession numbers for lepocreadioid species analysed in this study Species Host GenBank References ID Lepocreadioidea Aephnidiogenidae Yamaguti, 1934 Aephnidiogenes major Yamaguti, 1934 Diagramma pictum labiosum FJ788468 Bray et al. (2009) (Macleay) Austroholorchis sprenti (Gibson, 1987) Sillago ciliata Cuvier MH157075 Present study Holorchis castex Bray & Justine, 2007 Diagramma pictum pictum FJ788476 Bray et al. (2009) (Thunberg) Holorchis gigas Bray & Cribb, 2007 Plectorhinchus chrysotaenia FJ788477 Bray et al. (2009) (Bleeker) Neolepocreadium caballeroi Thomas, 1960 Trachinotus blochii (Lace´pe`de) FJ788488 Bray et al. (2009) Stegodexamene anguillae Macfarlane, 1951 Gobiomorphus cotidianus McDowall KF484005 Herrmann et al. (2014) Tetracerasta blepta Watson, 1984 Posticobia brazieri (Smith) FJ788494 Bray et al. (2009) Enenteridae Yamaguti, 1958 Enenterum aureum Linton, 1910 Kyphosus vaigiensis (Quoy & AY222232 Olson et al. (2003) Gaimard) Koseiria xishaensis Gu & Shen, 1983 Kyphosus vaigiensis AY222233 Olson et al. (2003) Proenenterum ericotylum Manter, 1954 Aplodactylus arctidens Richardson FJ788499 Bray et al. (2009) Proenenterum isocotylum Manter, 1954 Aplodactylus arctidens FJ788500 Bray et al. (2009) Gibsonivermidae n. fam. Gibsonivermis berryi (Gibson, 1987) Sillago ciliata MH157070 Present study Gorgocephalidae Manter, 1966 Gorgocephalus kyphosi Manter, 1966 Kyphosus vaigiensis AY222234 Olson et al. (2003) Gorgocephalus yaaji Bray & Cribb, 2005 Kyphosus cinerascens (Forsska˚l) KU951489 Huston et al. (2016) Gorgocephalus sp. Austrolittorina unifasciata (Gray) KU951485 Huston et al. (2016) Gyliauchenidae Fukui, 1929 Affecauda annulata Hall & Chambers, 1999 Naso tuberosus Lace´pe`de FJ788501 Bray et al. (2009) Paragyliauchen arusettae Machida, 1984 Pomacanthus sexstriatus (Cuvier) FJ788503 Bray et al. (2009) Paragyliauchen sp. Centropyge bicolor (Bloch) FJ788502 Bray et al. (2009) Petalocotyle adenometra Hall & Cribb, 2000 Prionurus microlepidotus Lace´pe`de FJ788504 Bray et al. (2009) Robphildollfusium fractum (Rudolphi, 1819) Sarpa salpa (Linnaeus) FJ788505 Bray et al. (2009) Lepocreadiidae Odhner, 1905 Bianium arabicum Sey, 1996 Lagocephalus lunaris (Bloch & MH157076 Present study Schneider) Bianium plicitum (Linton, 1928) Torquigener pleurogramma (Regan) MH157066 Present study Clavogalea trachinoti (Fischthal & Thomas, 1968) Trachinotus coppingeri Gu¨nther MH157067 Present study Diplocreadium tsontso Bray, Cribb & Barker, 1996 Balistoides conspicillum (Bloch & FJ788472 Bray et al. (2009) Schneider) Diploproctodaeum momoaafata Bray, Cribb & Barker, Ostracion cubicus Linnaeus FJ788474 Bray et al. (2009) Diploproctodaeum monstrosum Bray, Cribb & Justine, Arothron stellatus (Anonymous) FJ788473 Bray et al. (2009) Diploproctodaeum cf. monstrosum Arothron hispidus (Linnaeus) MH157069 Present study Echeneidocoelium indicum Simha & Pershad, 1964 Echeneis naucrates Linnaeus FJ788475 Bray et al. (2009) 123 Syst Parasitol Table 2 continued Species Host GenBank References ID Hypocreadium patellare Yamaguti, 1938 Balistoides viridescens (Bloch & FJ788478 Bray et al. (2009) Schneider) Hypocreadium picasso Bray, Cribb & Justine, 2009 Rhinecanthus aculeatus (Linnaeus) FJ788479 Bray et al. (2009) Hypocreadium toombo Bray & Justine, 2006 Pseudobalistes fuscus (Bloch & FJ788480 Bray et al. (2009) Schneider) Lepidapedoides angustus Bray, Cribb & Barker, 1996 Epinephelus cyanopodus FJ788482 Bray et al. (2009) (Richardson) Lepotrema clavatum Ozaki, 1932 Acanthochromis polyacanthus FJ788483 Bray et al. (2009) (Bleeker) Lobatocreadium exiguum (Manter, 1963) Pseudobalistes fuscus FJ788484 Bray et al. (2009) Mobahincia teirae n. g., n. sp. Platax teira (Forsskal) MH157068 Present study Multitestis magnacetabulum Mamaev, 1970 Platax teira MH157071 Present study Neohypocreadium dorsoporum Machida & Uchida, 1987 Chaetodon ﬂavirostris Gu¨nther FJ788487 Bray et al. (2009) Neomultitestis aspidogastriformis Bray & Cribb, 2003 Platax teira MH157072 Present study Neopreptetos arusettae Machida, 1982 Pomacanthus sexstriatus FJ788490 Bray et al. (2009) Opechona austrobacillaris Bray & Cribb, 1998 Pomatomus saltatrix Linnaeus MH157073 Present study Opechona kahawai Bray & Cribb, 2003 Arripis trutta (Forster) FJ788491 Bray et al. (2009) Pelopscreadium spongiosum (Bray & Cribb, 1998) Ostracion cubicus FJ788469 Bray et al. (2009) Preptetos caballeroi Pritchard, 1960 Naso vlamingii (Valenciennes) AY222236 Olson et al. (2003) Preptetos trulla (Linton, 1907) Ocyurus chrysurus (Bloch) AY222237 Olson et al. (2003) Prodistomum keyam Bray & Cribb, 1996 Monodactylus argenteus (Linnaeus) MH157074 Present study Lepidapedidae Yamaguti, 1958 Bulbocirrus aulostomi Yamaguti, 1965 Aulostomus chinensis (Linnaeus) FJ788470 Bray et al. (2009) Intusatrium robustum Durio & Manter, 1968 Bodianus perditio (Quoy & FJ788481 Bray et al. (2009) Gaimard) Lepidapedon beveridgei Campbell & Bray, 1993 Coryphaenoides armatus (Hector) AJ405263 Bray et al. (2009) Lepidapedon desclersae Bray & Gibson, 1995 Mora moro (Risso) AJ405264 Bray et al. (1999) Lepidapedon discoveryi Bray & Gibson, 1995 Coryphaenoides armatus AJ405265 Bray et al. (1999) Lepidapedon elongatum (Lebour, 1908) Gadus morhua Linnaeus AJ405266 Bray et al. (1999) Lepidapedon gaevskayae Campbell & Bray, 1993 Coryphaenoides armatus AJ405267 Bray et al. (1999) Lepidapedon rachion (Cobbold, 1858) Gadus morhua AJ405260 Bray et al. (1999) Lepidapedon sommervillae Bray & Gibson, 1995 Coryphaenoides guentheri (Vaillant) AJ405268 Bray et al. (1999) Lepidapedon zubchenkoi Campbell & Bray, 1993 Coryphaenoides leptolepis Gu¨nther AJ405269 Bray et al. (1999) Myzoxenus insolens (Crowcroft, 1945) Notolabrus tetricus (Richardson) FJ788486 Bray et al. (2009) Neolepidapedon smithi Bray & Gibson, 1989 Mora moro AJ405270 Bray et al. (1999) Postlepidapedon opisthobifurcatum (Zdzitowiecki, 1990) Muraenolepis marmorata Gu¨nther KY497957 Sokolov et al. (2018) Postlepidapedon uberis Bray, Cribb & Barker, 1997 Choerodon venustus (De Vis) FJ788492 Bray et al. (2009) Profundivermis intercalarius Bray & Gibson, 1991 Coryphaenoides armatus AJ405271 Bray et al. (1999) Outgroup taxa Apocreadiidae Skrjabin, 1942 Homalometron armatum (MacCallum, 1895) Lepomis microlophus (Gu¨nther) AY222241 Olson et al. (2003) Neoapocreadium splendens Cribb & Bray, 1999 Scolopsis monogramma (Cuvier) AY222242 Olson et al. (2003) Paraschistorchis zancli (Hanson, 1953) Zanclus cornutus (Linnaeus) AY222240 Olson et al. (2003) Cryptogonimidae Ward, 1917 123 Syst Parasitol Table 2 continued Species Host GenBank References ID Adlardia novaecaledoniae Miller, Bray, Goiran, Justine & Nemipterus furcosus (Valenciennes) FJ788496 Bray et al. (2009) Cribb, 2009 Caecincola parvulus Marshall & Gilbert, 1905 Micropterus salmoides (Lace´pe`de) AY222231 Olson et al. (2003) Australian temperate marine ﬁsh. Systematic Parasitology, structuring of this diverse family is needed before such 55, 127–133. an analysis can be completed. Aken’Ova, T. O., Cribb, T. H., & Bray, R. A. (2008). Eight new The new status of Gibsonivermis, as the type-genus species of Macvicaria Gibson and Bray, 1982 (Digenea: for a monotypic family, has been argued above. This Opecoelidae) mainly from endemic temperate marine ﬁshes of Australia. ZooKeys, 1, 23–58. new status is consistent with the observations of Cribb Ankenbrand, M. J., Keller, A., Wolf, M., Schultz, J., & Fo¨rster, & Bray (2011) that new trematode families are now F. (2015). ITS2 Database V: Twice as much. Molecular principally recognised from among known taxa rather Biology and Evolution, 32, 3030–3032. than as a result of completely new discoveries. We Barker, S. C., Blair, D., Garrett, A. R., & Cribb, T. H. (1993). Utility of the D1 domain of nuclear 28S rRNA for phylo- suspect that further genetic exploration of unique genetic inference in the Digenea. Systematic Parasitology, trematode taxa will likely lead to more families being 26, 181–188. proposed within the Lepocreadioidea. Blasco-Costa, I., Cutmore, S. C., Miller, T. L., & Nolan, M. J. (2016). Molecular approaches to trematode systematics: Acknowledgements We thank John Page and Dave ‘best practice’ and implications for future study. Systematic Thompson for their assistance in the collection of ﬁshes in Parasitology, 93, 295–306. Moreton Bay, and all members of the Marine Parasitology Bray, R. A. (2005). Family Lepocreadiidae Odhner, 1905. In: Research Group at the University of Queensland for assistance Jones, A., Bray, R. A. & Gibson, D. I. (Eds), Keys to the with dissections. Trematoda. Volume 2. Wallingford: CABI Publishing and The Natural History Museum, pp. 545–602. Bray, R. A., & Cribb, T. H. (1996). Two species of Prodistomum Funding RAB, THC and SCC acknowledge the Australian Linton, 1910 (Digenea: Lepocreadiidae) from marine Biological Resources Study (ABRS) for their ongoing support. This study was funded by the ABRS National Taxonomy ﬁshes of Australia. Systematic Parasitology, 35, 59–67. Research Grant RF215-40. Bray, R. A., & Cribb, T. H. (1997a). Lepocreadiid (Digenea) species from members of the marine teleost family Cheilodactylidae from south-western Australia, including Compliance with ethical standards four new genera and ﬁve new species. Systematic Para- sitology, 37, 27–45. Conﬂict of interest The authors declare that they have no Bray, R. A., & Cribb, T. H. (1997b). The subfamily Aephnid- conﬂict of interest. iogeninae Yamaguti, 1934 (Digenea: Lepocreadiidae), its status and that of the genera Aephnidiogenes Nicoll, 1915, Ethical approval All applicable institutional, national and Holorchis Stossich, 1901, Austroholorchis n. g., Pseu- international guidelines for the care and use of animals were daephnidiogenes Yamaguti, 1971, Pseudoholorchis Yam- followed. aguti, 1958 and Neolepocreadium Thomas, 1960. Systematic Parasitology, 36, 47–68. Open Access This article is distributed under the terms of the Bray, R. A., & Cribb, T. H. (1998). Lepocreadiidae (Digenea) of Creative Commons Attribution 4.0 International License (http:// Australian coastal ﬁshes: new species of Opechona Looss, creativecommons.org/licenses/by/4.0/), which permits unre- 1907, Lepotrema Ozaki, 1932 and Bianium Stunkard, 1930 stricted use, distribution, and reproduction in any medium, and comments on other species reported for the ﬁrst time or provided you give appropriate credit to the original poorly known in Australian waters. Systematic Parasitol- author(s) and the source, provide a link to the Creative Com- ogy, 41, 123–148. mons license, and indicate if changes were made. Bray, R. A., & Cribb, T. H. (2001). Amphicreadium n. g. (Di- genea: Lepocreadiidae) from monacanthid ﬁshes (Te- traodontiformes) from the coast of northern Tasmania. Systematic Parasitology, 49, 205–209. Bray, R. A., & Cribb, T. H. (2003a). 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