Two new species of <Emphasis Type="Italic">Haplorchoides</Emphasis> Chen, 1949 (Digenea: Heterophyidae) infecting an Australian siluriform fish, <Emphasis Type="Italic">Neoarius graeffei</Emphasis> Kner &amp; Steindachner

Combined morphological and molecular analyses are used to characterise two new species of Haplorchoides Chen, 1949 (Digenea: Heterophyidae) from an Australian siluriform fish. Haplorchoides maiwariensis n. sp. and H. daguilarensis n. sp. are described from the intestine of the Blue salmon catfish, Neoarius graeffei (Kner & Steindachner) (Siluriformes: Ariidae) from tidal reaches of the Brisbane River and from a freshwater creek off Lake Wivenhoe, Kipper Creek, Australia. The two new species most obviously differ from all previously described species of Haplorchoides in possessing clearly and reliably tripartite seminal vesicles. The two new species differ in the size and distribution of vitelline follicles, the size of the pigment granules and the form of the ventral sucker. Previously described species of Haplorchoides have been reported from Africa and Asia, principally from bagrid, schilbeid, silurid and sisorid catfishes; these are the first species reported from an ariid catfish. Complete ITS2 and partial 28S ribosomal DNA data were generated for both new species. The two species differ from each other by eight base pairs in the ITS2 region, and by 13 bp for the 28S region. The 28S rDNA sequence of H. daguilarensis agrees with a previously reported sequence from an unidentified species of Haplorchoides collected from N. graeffei in Lake Wivenhoe, Australia; we identify this previous report as relating to H. daguilarensis.     

<Emphasis Type="Italic">Lepotrema</Emphasis> Ozaki, 1932 (Lepocreadiidae: Digenea) from Indo-Pacific fishes,with the description of eight new species,characterised by morphometric and molecular features

We review species of the genus Lepotrema Ozaki, 1932 from marine fishes in the Indo-West Pacific. Prior to the present study six species were recognised. Here we propose eight new species on the basis of combined morphological and molecular analysis: Lepotrema acanthochromidis n. sp. ex Acanthochromis polyacanthus from the Great Barrier Reef (GBR); Lepotrema hemitaurichthydis n. sp. ex Hemitaurichthys polylepis and H. thompsoni from Palau and French Polynesia; Lepotrema melichthydis n. sp. ex Melichthys vidua from Palau and the GBR; Lepotrema amansis n. sp. ex Amanses scopas from the GBR; Lepotrema cirripectis n. sp. ex Cirripectes filamentosus, C. chelomatus and C. stigmaticus from the GBR; Lepotrema justinei n. sp. ex Sufflamen fraenatum from New Caledonia; Lepotrema moretonense n. sp. ex Prionurus microlepidotus, P. maculatus and Selenotoca multifasciata from Moreton Bay; and Lepotrema amblyglyphidodonis n. sp. ex Amblyglyphidodon curacao and Amphipron akyndynos from the GBR. We also report new host records and provide novel molecular data for two known species: Lepotrema adlardi Bray, Cribb & Barker, 1993 and Lepotrema monile Bray & Cribb, 1998. Two new combinations are formed, Lepotrema cylindricum (Wang, 1989) n. comb. (for Preptetos cylindricus) and Lepotrema navodonis (Shen, 1986) n. comb. (for Lepocreadium navodoni). With the exception of a handful of ambiguous records, the evidence is compelling that the host-specificity of species in this genus is overwhelmingly oioxenous or stenoxenous. This renders the host distribution in three orders and ten families especially difficult to explain as many seemingly suitable hosts are not infected. Multi-loci molecular data (ITS2 rDNA, 28S rDNA and cox1 mtDNA) demonstrate that Lepotrema is a good generic concept, but limited variability in sequence data and differences in phylogenies produced for different gene regions make relationships within the genus difficult to define.     

Four new species of <Emphasis Type="Italic">Paradiscogaster</Emphasis> Yamaguti, 1934 (Digenea: Faustulidae) from batfishes (Perciformes: Ephippidae) on the Great Barrier Reef,Australia

Examination of three species of batfishes (Teleostei: Epphippidae) from off Lizard and Heron Islands on the Great Barrier Reef led to the discovery of specimens of the trematode genus Paradiscogaster Yamaguti, 1934 (Digenea: Faustulidae). Morphological analysis demonstrated that the new specimens represented four morphotypes which we interpret to be new species: Paradiscogaster martini n. sp., P. vichovae n. sp. and P. brayi n. sp. from Platax orbicularis (Forsskål) and P. pinnatus (Linnaeus) off Lizard Island, and P. nitschkei n. sp. from P. teira (Forsskål) off Heron Island. Published material was re-examined and the specimens identified as P. chaetodontis okinawensis Yamaguti, 1971 from P. pinnatus from Okinawa, Japan, actually represent the new species P. brayi n. sp., demonstrating that some species of Paradiscogaster have wide geographical distributions. ITS2 rDNA data for the four morphotypes differ by 4–39 base pairs confirming the delineation of the four species proposed. A feature of this study is the recognition of Platax spp. as an important host group for Paradiscogaster, with the new species placing them as the second richest host group for these parasites after the Chaetodontidae.     

Morphology and molecules reveal the alien <Emphasis Type="Italic">Posthodiplostomum centrarchi</Emphasis> Hoffman, 1958 as the third species of <Emphasis Type="Italic">Posthodiplostomum</Emphasis> Dubois, 1936 (Digenea: Diplostomidae) in Europe

Metacercariae of two species of Posthodiplostomum Dubois, 1936 (Digenea: Diplostomidae) were subjected to morphological and molecular studies: P. brevicaudatum (von Nordmann, 1832) from Gasterosteus aculeatus (L.) (Gasterosteiformes: Gasterosteidae), Bulgaria (morphology, cox1 and ITS1-5.8S-ITS2) and Perca fluviatilis L. (Perciformes: Percidae), Czech Republic (morphology, cox1, ITS1-5.8S-ITS2 and 28S); and P. centrarchi Hoffman, 1958 from Lepomis gibbosus (L.) (Perciformes: Centrarchidae), Bulgaria (morphology, cox1 and ITS1-5.8S-ITS2) and Slovakia (cox1 and ITS1-5.8S-ITS2). In addition, cercariae of P. cuticola (von Nordmann, 1832) from Planorbis planorbis (L.) (Mollusca: Planorbidae), Lithuania (morphology and cox1) and metacercariae of Ornithodiplostomum scardinii (Schulman in Dubinin, 1952) from Scardinius erythrophthalmus (L.) (Cypriniformes: Cyprinidae), Czech Republic, were examined (morphology, cox1, ITS1-5.8S-ITS2 and 28S). These represent the first molecular data for species of Posthodiplostomum and Ornithodiplostomum Dubois, 1936 from the Palaearctic. Phylogenetic analyses based on cox1 and ITS1-5.8S-ITS2, using O. scardinii as the outgroup and including the three newly-sequenced Posthodiplostomum spp. from Europe and eight published unidentified (presumably species-level) lineages of Posthodiplostomum from Canada confirmed the distinct status of the three European species (contrary to the generally accepted opinion that only P. brevicaudatum and P. cuticola occur in the Palaearctic). The subspecies Posthodiplostomum minimum centrarchi Hoffmann, 1958, originally described from North America, is elevated to the species level as Posthodiplostomum centrarchi Hoffman, 1958. The undescribed “Posthodiplostomum sp. 3” of Locke et al. (2010) from centrarchid fishes in Canada has identical sequences with the European isolates of P. centrarchi and is recognised as belonging to the same species. The latter parasite, occurring in the alien pumpkinseed sunfish Lepomis gibbosus in Europe, is also supposed to be alien for this continent. It is speculated that it colonised Europe long ago and is currently widespread (recorded in Bulgaria, Slovakia and Spain); based on the cox1 sequence of an adult digenean isolate from the Ebro Delta, Spain, only the grey heron (Ardea cinerea L.) (Ciconiiformes: Ardeidae) is known to be its definitive host in Europe.     

A re-evaluation of diversity of the Aporocotylidae Odhner, 1912 in <Emphasis Type="Italic">Siganus fuscescens</Emphasis> (Houttuyn) (Perciformes: Siganidae) and associated species

The aporocotylid fauna of the mottled spinefoot, Siganus fuscescens (Houttuyn), from Moreton Bay, Queensland, Australia, was characterised using a combined morphological and molecular approach. Four aporocotylid species were identified, three belonging to the genus Ankistromeces Nolan & Cribb, 2006 and one to Cardicola Short, 1953. Specimens of Cardicola matched an undescribed species from the same host and locality; this species is described as Cardicola mogilae n. sp. Phylogenetic analyses of ITS2 and 28S data showed that C. mogilae n. sp. forms a strongly supported clade with other Cardicola species from siganid fishes. We record Ankistromeces olsoni Nolan & Cribb, 2006 in Moreton Bay for the first time, redescribe A. dunwichensis Nolan & Cribb, 2006 on the basis of new specimens and sequence data and re-report Ankistromeces sp. X from Moreton Bay based on molecular data. We review the status of the ten putative species of aporocotylids reported from siganids. Small variation in ITS2 rDNA sequences, in association with different geographic localities, was previously used to separate Cardicola lafii Nolan & Cribb, 2006 from C. parilus Nolan & Cribb, 2006, C. bartolii Nolan & Cribb, 2006 from C. watsonensis Nolan & Cribb, 2006, C. tantabiddii Nolan & Cribb, 2006 from Cardicola sp. 2, Ankistromeces sp. Y from A. olsoni and Ankistromeces sp. X from Ankistromeces sp. Z. These five combinations are reinterpreted as each representing a single species; Cardicola lafii is recognised as the senior synonym of C. parilus and C. bartolii as the senior synonym of C. watsonensis. This study thus suggests that six, rather than ten, species should be recognised as infecting S. fuscescens. This richness remains greater than is known for any other fish species and siganids are, so far, unique among fishes in harbouring two strongly radiated lineages of aporocotylids.     

Two monorchiid species from the freckled goatfish, <Emphasis Type="Italic">Upeneus tragula</Emphasis> Richardson (Perciformes: Mullidae), in Moreton Bay,Australia, including a proposal of a new genus

Two monorchiid species are reported from the freckled goatfish, Upeneus tragula Richardson, from Moreton Bay, Queensland, Australia. Specimens of a species new to science were most morphologically similar to species of the genus Timonia Bartoli & Prevot, 1966, but significant differences in the arrangement of the testes (symmetrical vs oblique) and morphology of the terminal organ (bipartite vs unipartite) necessitate the proposal of a new genus; Madhavia n. g. is proposed for M. fellaminutus n. sp. Specimens of the second species are identified as Parachrisomon delicatus (Manter & Pritchard, 1964) Madhavi, 2008, extending its known range from Hawaii to Australia. Complete ITS2 and partial 28S rDNA sequence data were generated for both species and analysed with those for other monorchiids available on GenBank. Phylogenetic analyses of the 28S rDNA dataset showed that both genera are distinct from other sequenced monorchiids, but overall the resolution between genera is poor and more sequence data are required to elucidate relationships within the family. We propose to transfer Timonia stunkardi (Ahmad, 1985) and Timonia vinodae (Ahmad, 1987) to the genus Neotimonia Madhavi, 2008, as Neotimonia stunkardi (Ahmad, 1985) n. comb. and Neotimonia vinodae (Ahmad, 1987) n. comb. Additionally, we were unable to locate any literature on Parachrisomon brotulidorum (Toman, 1973) Madhavi, 2008 and consider this species as nomen nudum.     

Taxonomy and phylogenetic position of <Emphasis Type="Italic">Phyllactinia takamatsui</Emphasis>, a newly described powdery mildew on cotoneaster,based on molecular and morphological data

The newly recognised powdery mildew species Phyllactinia takamatsui on Cotoneaster nummularius (Rosaceae) is described and illustrated. This species, collected in Kerman Province, Iran, is well characterised by its conidial morphology and rDNA ITS sequences clearly different from allied species. Conidia are broadly ellipsoid to subcylindrical, i.e. they are not clavate-spathuliform as in most Phyllactinia species. The rDNA ITS sequence analysis showed that this species is closely allied to other species described on hosts belonging to Rosaceae, such as Ph. mali and Ph. pyri-serotinae. The ITS sequence of P. takamatsui was 92 to 94 % similar to that of the closest known relatives. The new species is described in detail, illustrated and compared with other similar taxa.     

Two new species of <Emphasis Type="Italic">Bacciger</Emphasis> Nicoll, 1914 (Trematoda: Faustulidae) in species of <Emphasis Type="Italic">Herklotsichthys</Emphasis> Whitley (Clupeidae) from Queensland waters

Two new species of Bacciger Nicoll, 1914 (Faustulidae) are described infecting clupeids collected from the waters off Queensland, Australia; Bacciger minor n. sp. is described from Herklotsichthys castelnaui (Ogilby) in Moreton Bay, southern Queensland and Bacciger major n. sp. is described from Herklotsichthys quadrimaculatus (Rüppell) collected off Lizard Island, on the northern Great Barrier Reef. The two species both differ from previously described species of Bacciger in the combination of their generally elongate bodies, an entire rather than deeply lobed ovary, vitelline follicles that reach to at least the intestinal bifurcation, instead of restricted to further posteriorly but principally distributed in the hindbody, and intestinal caeca extending posteriorly well past the ventral sucker. The two new species have non-overlapping size ranges and differ in their sucker ratios, the distribution of the vitelline follicles and in the shape of the cirrus-sac. ITS2 and 28S rDNA sequence data distinguish the two new species unambiguously. Phylogenetic analysis of available 28S data show they are most closely related to Pseudobacciger cheneyae Sun, Bray, Yong, Cutmore & Cribb, 2014, also recorded off Lizard Island. These are the first faustulids reported from species of Herklotsichthys Whitley, but overall members of the Clupeidae undoubtedly harbours the richest faustulid fauna of any fish family. Baccigeroides ovatus (Price, 1934) n. comb. is proposed for Bacciger ovatus (Price, 1934) Bray & Gibson, 1980 (syn. B. opisthonema Nahhas & Cable, 1964) based on the position of the genital pore being far anteriorly removed from the ventral sucker.     

<Emphasis Type="Italic">Mayarhynchus karlae</Emphasis> n. g., n. sp. (Acanthocephala: Neoechinorhynchidae), a parasite of cichlids (Perciformes: Cichlidae) in southeastern Mexico,with comments on the paraphyly of <Emphasis Type="Italic">Neoechynorhynchus</Emphasis> Stiles &amp; Hassall, 1905

Mayarhynchus n. g. (Acanthocephala: Neoechinorhynchidae) is erected for Mayarhynchus karlae n. g, n. sp. described from the intestine of four species of cichlid fishes distributed from southeastern Mexico. The new genus placed in the family Neoechinorhynchidae (Ward, 1917) Van Cleave, 1928, is readily distinguished from the other 17 genera in the family by having a small proboscis armed with 45–46 relatively weak rooted hooks arranged in nine longitudinal rows of five hooks each. In addition, Mayarhynchus n. g., n. sp. is diagnosed by the presence of a short trunk, body wall with five dorsal and one ventral giant hypodermal nuclei, proboscis receptacle nearly cylindrical with single layered wall, lemnisci broad and flat with large nuclei, testes in tandem, cement gland with eight large nuclei, and eggs elongate to oval. Partial sequences of the cytochrome c oxidase subunit 1 (cox1), internal transcribed spacers (ITS1 + 5.8S + ITS2), and the D2-D3 domains of the large subunit rRNA gene (28S) were obtained for five specimens of the new species and other species belonging to the Neoechinorhynchidae. Phylogenetic analyses confirmed that the new genus belongs to the Neoechynorhynchidae and indicated that the genus Neoechynorhynchus Stiles & Hassall, 1905 is not monophyletic. Comparison between three populations of the new species yielded nine variable sites for cox1, 11 for ITS and four for 28S.     

Polymorphic Sites in ITS1-5.8S rDNA-ITS2 Region in Hybridogenic Genus × <Emphasis Type="Italic">Elyhordeum</Emphasis> and Putative Interspecific Hybrids <Emphasis Type="Italic">Elymus</Emphasis> (Poaceae: Triticeae)

The genus Elymus L. is a complicated aggregate of ecological and geographical races, species, subspecies, varieties, and hybrids. We suggest that comparative analysis of intragenomic polymorphism of internal transcribed spacers ITS1 and ITS2 of 35S rRNA genes in the supposed hybrids and their possible “parents” can be one of the approaches to verification of hybrid origin of the samples collected in nature to confirm or reject the hypotheses about their possible “parents.” Polymorphic sites (PS) in ITS of 23 Elymus species, as well as in two supposed interspecific Elymus hybrids and in a supposed intergeneric hybrid between Elymus × Hordeum determined as × Elyhordeum sp., were analyzed in the work. We collected all hybrids in the Altai. There were 2 and 5 PS in two samples of E. dahuricus and 1 and 4 PS in two studied samples of E. schrenkianus in the ITS1-5.8S rDNA-ITS2 region. From 0 to 4 (modes 0 and 3) PS were detected in 32 samples relating to 21 tetraploid Elymus species. More PS (14) were found in the × Elyhordeum sp. sample. A large number of single nucleotide substitutions were found in 5.8S rRNA in × Elyhordeum. It was shown that about half of them do not change the secondary structure of the 5.8S rRNA molecule, so these molecules probably retain the ability to work as a component of large subunit of a ribosome. On the other hand, the absence or weakening of 5.8S rDNA homogenization in × Elyhordeum indirectly suggests that a significant part of 5.8S rDNA is not transcribed. Paradoxically, ITS sequences of × Elyhordeum sp. are less polymorphic than 5.8S rDNA. There are no ITS sequences derived from Hordeum among × Elyhordeum ITS sequenced by Sanger method. No traces of the H subgenome and a subgenome originating from Agropyron (P-subgenome) are seen in the Alt 10–278 plant genome (a chimera, combining the morphological traits of Elymus, Elytrigia, and Agropyron). In this plant, as well as in the supposed intersectional hybrid Alt 11–60 distinguished by a mosaic of the traits typical for the E. caninus × E. mutabilis species, only 4 and 5 PS, respectively, are detected when sequencing by Sanger method. The comparison of ITS sequences of the supposed Elymus Alt 10–278 hybrid and its probable “parents” demonstrates that one of the species of the Elymus macrourus kinship circle, as well as the Elytrigia geniculata, could be one of its ancestors. The comparison of the ITS sequence of the supposed parental species with ITS of Alt 11–60 samples and five PS of the supposed Alt 11–60 hybrid does not contradict the hypothesis that this is an intersectional hybrid of the first generation that emerged with the involvement of E. caninus and E. mutabilis common in the Altai.     

<Emphasis Type="Italic">Melampsora salicis-bakko</Emphasis>, a new species on willows in Japan evidenced by morphological and molecular phylogenetic analyses

Through the morphological and molecular examinations of Melampsora species on willows, we clarified the taxonomic identity of the rust specimens on Salix bakko, S. hultenii and S. leucopithecia from Japan and described the following rust fungus as a new species, Melampsora salicis-bakko. This rust fungus resembled M. caprearum in morphology of teliospores, but it differed from M. caprearum mainly in the density of spines on the urediniospores. Molecular phylogenetic analyses using the rDNA ITS region (complete ITS1, 5.8S rRNA gene and ITS2) revealed that M. salicis-bakko was monophyletic, and that this rust fungus was distinct from other Melampsora species, including M. caprearum.     

Molecular characterization of hard tick <Emphasis Type="Italic">Haemaphysalis longicornis</Emphasis> from China by sequences of the internal transcribed spacers of ribosomal DNA

In the present study, the entire first and second internal transcribed spacer (ITS-1 and ITS-2) regions of nuclear ribosomal DNA (rDNA) of Haemaphysalis longicornis from China were amplified by polymerase chain reaction. The 45 representative amplicons were sequenced, and sequence variation in the ITS was examined. The ITS sequences of H. longicornis were 3644 bp in size, including the part of 18S rDNA, 28S rDNA sequences and the complete ITS-1, 5.8S rDNA and ITS-2 sequences. Sequence analysis revealed that the ITS-1, 5.8S rDNA and ITS-2 of this hard tick were 1582, 152, and 1610 bp in size, respectively. The intra-specific sequence variations of ITS-1 and ITS-2 within H. longicornis were 0–2 and 0–2.2%; however, the inter-specific sequence differences among members of the genus Haemaphysalis were significantly higher, being 35.1–55.2 and 37–52% for ITS-1 and ITS-2, respectively. The molecular approach employed in this study provides the foundation for further studies of the genetic variation of H. longicornis from different hosts and geographical origins in China.     

Myrothecium-like (Ascomycota,Hypocreales) species from tropical areas: <Emphasis Type="Italic">Digitiseta</Emphasis> gen. nov. and additions to <Emphasis Type="Italic">Inaequalispora</Emphasis> and <Emphasis Type="Italic">Parvothecium</Emphasis>

Inaequalispora and Parvothecium are two myrothecium-like, closely related genera of Hypocreales. They are also morphologically similar, sharing sporodochial conidiomata, penicillate conidiophores, fusiform to ellipsoidal conidia accumulating in a green slimy drop, and hypha-like setoid extensions emerging through the conidial mass. During a revision of myrothecium-like isolates originating from rainforest areas of South America (Ecuador, Brazil) and Southeast Asia (Singapore), multilocus phylogenetic inferences (based on DNA sequence data of ITS, partial nuc 28S, and partial tef1a, rpb2 and tub2) and morphological studies concordantly revealed the occurrence of two undescribed species of Inaequalispora (I. longiseta sp. nov. and I. cylindrospora sp. nov.) and one undescribed species of Parvothecium (P. amazonensesp. nov.). Myrothecium setiramosum, M. dimorphum, and two undescribed taxa form the base of a new lineage, sister to the current Parvothecium lineage. This lineage is recognized as Digitiseta gen. nov., typified by D. setiramosa comb. nov. Digitiseta dimorpha comb. nov. is also proposed, and the new species D. parvodigitata sp. nov. and D. multidigitata sp. nov. are described.     

Two species of <Emphasis Type="Italic">Neometadena</Emphasis> Hafeezullah &amp; Siddiqi, 1970 (Digenea: Cryptogonimidae) from Moreton Bay,Australia, including the description of <Emphasis Type="Italic">Neometadena paucispina</Emphasis> n. sp. from Australian Lutjanidae

A survey of the trematode fauna of lutjanid fishes off the east coast of Queensland (QLD), Australia revealed the presence of two species of Neometadena Hafeezullah & Siddiqi, 1970 (Digenea: Cryptogonimidae). Neometadena paucispina n. sp. is described from the intestine and pyloric caeca of Lutjanus fulviflamma (Forsskål) and L. russellii (Bleeker) from Moreton Bay, in southeast QLD. Specimens of the type- and only other species, N. ovata (Yamaguti, 1952) Miller & Cribb, 2008, were recovered from L. carponotatus (Richardson), L. fulviflamma, L. fulvus (Forster), L. russellii, and L. vitta (Quoy & Gaimard) off Lizard Island, on the northern Great Barrier Reef (GBR). Neometadena paucispina is distinguished from N. ovata in having fewer oral spines (55–65 vs 67–80). Alignment of novel molecular data for these two taxa revealed that they differ consistently by 13 nucleotides (1.5%) over the partial large subunit (LSU), 34 nucleotides (6.6%) over the internal transcribed spacer 1 (ITS1), 0 nucleotides over the 5.8S, and 21 nucleotides (7.3%) over the ITS2 rDNA regions. Despite relatively large samples of L. carponotatus, L. fulviflamma and L. russellii from three distinct locations along the east coast of QLD (i.e. Moreton Bay in the south, Heron Island in central QLD and Lizard Island in northern QLD), these two species have been found at only one site each with neither species at Heron Island. These distributions are discussed in the context of the wide distribution of other cryptogonomid species in the same hosts elsewhere in the Indo-West Pacific.