Plerchis heterorchis N. sp. (Digenea: Acanthocolpidae Luhe, 1909) from fishes Lutjanus johnii and Otolithus argenteus of Karachi coast, Pakistan

A new species of trematodes Pleorchis heterorchis is described from the fishes Lutjanus johnii and Otolithus argenteus of Karachi coast. The new species is characterized by having a lanceolate body with a notch at the middle of the posterior end of the body. Body surface is smooth, ventral sucker rounded, situated at the anterior middle region of the body, pre-pharynx is well developed, widened posteriorly, pharynx muscular, oesophagus short, intestine H-shaped with anterior arms much shorter than the posterior, intestinal bifurcation almost in the middle of fore body, anterior caeca wide and short extending as far as anterior limit of pharynx. Posteriorly caeca reach to posterior end of the body with no lateral out pocketing. Testes 44 in number, intercecal arranged in 2 parallel rows, sub-globular, entire to slightly irregular, almost of same sizes extending immediately from posterior of the ovary to anterior of excretory vesicle. Cirrus pouch overlaps the ventral sucker, extends into hind body, terminating above the ovary, containing bipartite seminal vesicle, pars prostatica and ejaculatory duct. Genital pore behind the intestinal bifurcation and pre-acetabular. Ovary pre-testicular, consists of 16 follicles of varying sizes. Vitellaria lateral, follicular, extending from post bifurcal to posterior extremity. Excretory vesicle reaches to the posterior level of last pair of testes.

Stephanostomum tantabiddii n. sp. (Digenea: Acanthocolpidae) from Carangoides fulvoguttatus (Forsskal, 1775) (Perciformes: Carangidae) from Ningaloo Reef, Western Australia

A new species, Stephanostomum tantabiddii n. sp., is described from the yellowspotted trevally Carangoides fulvoguttatus from Ningaloo Reef, Western Australia. It has 38 - 45 circum-oral spines and the vitellarium reaches to no less than 17% of the hindbody length from the ventral sucker. It differs from other species of Stephanostomum with these characteristics by various combinations of the ventral hiatus of the circum-oral spine rows, the relatively long pars prostatica and short ejaculatory duct, the elongate body and the wide gaps between the gonads.

Ankistromeces mariae n. g., n. sp (Digenea : Sanguinicolidae) from Meuschenia freycineti (Monacanthidae) off Tasmania

Ankistromeces mariae n. g., n. sp. is described from Meuschenia freycineti (Monacanthidae), the six-spined leatherjacket, from off northern Tasmania. The new genus differs from the 21 other sanguinicolid genera in the combination of the anteriorly intercaecal and posteriorly post-caecal single testis, the presence of a cirrus-sac, the absence of an auxiliary external seminal vesicle, separate genital pores, the typically post-ovarian uterus and the H-shaped intestine. A. mariae is the first sanguinicolid to be reported from a monacanthid fish.

Prosorhynchoides lamprelli n. sp (Digenea : Bucephalidae) from the brassy trevally, Caranx papuensis (Teleostei : Carangidae), from off Lizard Island on the Great Barrier Reef, Australia

Prosorhynchoides lamprelli n. sp. ( Digenea: Bucephalidae) is described from the intestine of the brassy trevally, Caranx papuensis (Carangidae) from off Lizard Island, Great Barrier Reef, Australia. The new species is differentiated from other species of Prosorhynchoides Dollfus, 1929 by the shape and distribution of its vitelline follicles, the shape and extent of its uterus and the configuration of its digestive system. This is the first bucephalid to be described from Caranx papuensis; we have not encountered this species from other carangids or from over 1,500 individuals of other teleosts species we have found to be infected with bucephalids from the Great Barrier Reef.

Gorgocephalus yaaji n. sp (Digenea : Gorgocephalidae) from the brassy chub Kyphosus vaigiensis (Perciformes : Kyphosidae) off Lizard Island, northern Great Barrier Reef and further records of G-kyphosi

A new species Gorgocephalus yaaji is described in the intestine of Kyphosus vaigiensis from the waters off Lizard Island, Queensland, Australia. It differs from Gorgocephalus kyphosi by its broader body shape, the extension of the vitellarium into the forebody, a relatively longer forebody, cirrus-sac and post-caecal region, and a shorter distance between the ventral sucker and the ovary. It differs from Gorgocephalus manteri in its size, its tandem testes, and the ratios of width, ventral sucker to ovary distance and ovary to testes distance to body-length. Gorgocephalus kyphosi is reported in the pyloric caeca of K. vaigiensis from waters off Moorea, French Polynesia, and Lizard Island, Queensland, Australia. Measurements and an illustration are given of the latter species.

Chaulioleptos haywardi n. gen., n. sp (Digenea : Sanguinicolidae) from Filimanus heptadactyla (Perciformes : Polynemidae) of Moreton Bay, Australia

Here we describe the first Species of sanguinicolid blood fluke (Trematoda: Digenea) from a polynemid fish. Chaulioleptos haywardi n. gen., n. sp. is described from Filimanus heptadacryla Cuvier, 1829 (Perciformes: Polynemidae), the sevenfinger threadfin from Sandgate, Moreton Bay (southeast Queensland, Australia). Chaidioleptos haywardi differs from existing sanguinicolid genera in the combined possession of the following 7 characters: 2 testes, an entirely postovarian uterus, a uterine chamber, separate genital pores, an H-shaped intestine with abbreviated anterior caeca, tegumental spines in incomplete ventromarginal transverse rows that are continuous along the length of the body, and vitelline follicles that are tightly compacted and subsequently appear to form a solid branching mass occupying the area anterior to intestinal bifurcation and extending posteriorly to the level of the posterior margin of the anterior testis. Chaulioleptos haywardi is most closely related to Paracardicola Martin, 1960 and Adelomyllos Nolan and Cribb, 2004.

Sanguinicola maritimus n. sp (Digenea : Sanguinicolidae) from Labridae (Teleostei : Perciformes) of southern Australian waters

A new species of Sanguinicola Plehn, 1905 is described from the marine teleosts Notolabrus parilus (Richardson) and N. tetricus (Richardson) (Perciformes: Labridae) from Western Australian and Tasmanian waters. This host distribution is strikingly anomalous; however, the present material fulfils the morphological criteria of Sanguinicola. S. maritimus n. sp. differs from previously described species in having the combination of a body 1,432-1,701 mu m long, the oesophagus 18.3-21.7% of the body length, the testis occupying 42.8-52.3% of the body length, an oviducal seminal receptacle and Mehlis' gland present, ovoid eggs, and vitelline follicles that extend anteriorly past the nerve commissure, laterally past the lateral nerve chords and posteriorly to the anterior margin of the cirrus-sac. S. maritimus also lacks a protrusible anterior proboscis. It also differs in the combination of host and geographical location, being the first Sanguinicola species from a marine teleost and the first from Australian waters.

Overstreetia olsoni n.sp. (Digenea: Zoogonidae) from the Capricorn silverside Atherinomorus capricornensis (Woodland) (Atherindae) off Heron Island, southern Great Barrier Reef

A new species, Overstreetia olsoni, is described from Atherinomorus capricornensis off Heron Island on the Great Barrier Reef off Queensland, Australia. It differs from its only known congener in that the enlarged spines of the circum-oral rows are smaller (

Stephanostomum talakitok n. sp (Digenea : Acanthocolpidae) from the golden trevalley, Gnathanodon speciosus (Perciformes : Carangidae), from Ningaloo Reef, Western Australia

A new species, Stephanostomum talakitok n. sp., is described from the golden trevally Gnathanodon speciosus, Ningaloo Reef, Western Australia. It has 36 (34-40) circum-oral spines and the vitellarium reaches to no less than 10-17% of the hindbody length from the ventral sucker. It differs from other species of Stephanostomum with these characteristics by combinations of the gradual diminution of the circum-oral spine size to a small mid-ventral spine, the contiguous gonads with no intervening vitelline follicles, the sucker-ratio and various other ratios, including the distance between the ventral sucker and the ovary and the distance the cirrus-sac reaches into the hindbody, both as a proportion of body length.

First report of a bucephalid digenean from an apogonid teleost: Prosorhynchoides apogonis n. sp from Cheilodipterus macrodon on the southern Great Barrier Reef, Australia

Prosorhynchoides apogonis n. sp. (Digenea: Bucephalidae) is described from the intestine of the apogonid Cheilodipterus macrodon on the southern Great Barrier Reef, Australia. The new species is differentiated from other species of Prosorhynchoides Dollfus, 1929 by the configuration of its digestive system, shape and distribution of its vitelline follicles, and the shape and extent of its uterus. This is the first bucephalid to be described from the teleost family Apogonidae, and our records suggest that this species is strongly host-specific, at least to the genus Cheilodipterus. The host family is consistent with the pattern of Prosorhynchoides being reported from a very wide range of piscivorous fish families. As a result of comparisons made in the description of this species, 18 species formerly included in Bucephaloides Hopkins, 1954, Bucephalopsis Diesing, 1855 and Neobucephalopsis Dayal, 1948 are transferred to Prosorhynchoides, 16 as new combinations and two as replacement names to prevent secondary homonymy.

The use and implications of ribosomal DNA sequencing for the discrimination of digenean species

In just over a decade, the use of molecular approaches for the recognition of parasites has become commonplace. For trematodes, the internal transcribed spacer region of ribosomal DNA (ITS rDNA) has become the default region of choice. Here, we review the findings of 63 studies that report ITS rDNA sequence data for about 155 digenean species from 19 families, and then review the levels of variation that have been reported and how the variation has been interpreted. Overall, complete ITS sequences (or ITS1 or ITS2 regions alone) usually distinguish trematode species clearly, including combinations for which morphology gives ambiguous results. Closely related species may have few base differences and in at least one convincing case the ITS2 sequences of two good species are identical. In some cases, the ITS1 region gives greater resolution than the ITS2 because of the presence of variable repeat units that are generally lacking in the ITS2. Intraspecific variation is usually low and frequently apparently absent. Information on geographical variation of digeneans is limited but at least some of the reported variation probably reflects the presence of multiple species. Despite the accepted dogma that concerted evolution makes the individual representative of the entire species, a significant number of studies have reported at least some intraspecific variation. The significance of such variation is difficult to assess a posteriori, but it seems likely that identification and sequencing errors account for some of it and failure to recognise separate species may also be significant. Some reported variation clearly requires further analysis. The use of a yardstick to determine when separate species should be recognised is flawed. Instead, we argue that consistent genetic differences that are associated with consistent morphological or biological traits should be considered the marker for separate species. We propose a generalised approach to the use of rDNA to distinguish trematode species.

An exceptionally rich complex of Sanguinicolidae von Graff, 1907 (Platyhelminthes : Trematoda) from Siganidae, Labridae and Mullidae (Teleostei : Perciformes) from the Indo-West Pacific Region

We describe an unprecedented radiation of sanguinicolid blood flukes ( Digenea: Sanguinicolidae) from two species of Labridae (Choerodon venustus and C. cauteroma), seven species of Mullidae (Mulloidichthys vanicolensis, Parupeneus barberinoides, P. barberinus, P. bifasciatus, P. cyclostomus, P. indicus and P. multifasciatus) and ten species of Siganidae (Siganus argenteus, S. corallinus, S. doliatus, S. fuscescens, S. lineatus, S. margaritiferus, S. puellus, S. punctatus, S. virgatus and S. vulpinus) from sites off Australia and Palau. The flukes were morphologically similar in having the combination of a long thread-like body, tegumental spines in lateral transverse rows, a vestigial oral sucker bearing concentric rows of fine spines, an H-shaped intestine, a cirrussac, a notch level with the male genital pore, a lateral or post-ovarian uterus, a uterine chamber and separate genital pores. These species are divided into two genera on the basis of testis number. Sanguinicolids from Siganus fuscescens have a single large testis between the intestinal bifurcation and the ovary and are placed in Ankistromeces Nolan & Cribb, 2004. Species from the remaining nine species of Siganidae, Labridae and Mullidae are placed in Phthinomita n. g.; these species have two testes, the anterior testis being large and between the intestinal bifurcation and the ovary whereas the small posterior testis is at the posterior end of the body and appears rudimentary or degenerate and probably non-functional. The second internal transcribed spacer (ITS2) of ribosomal DNA ( rDNA) from 29 host/parasite/location combinations (h/p/l) was sequenced together with that of Ankistromeces mariae Nolan & Cribb, 2004 for comparison. From 135 samples we found 19 distinct genotypes which were interpreted as representing at least that many species. Replicate sequences were obtained for 25 of 30 h/p/l combinations ( including A. mariae); there was no intraspecific variation between replicates sequences for any of these. Interspecific variation ranged from 1 - 41 base differences (0.3 - 12.7% sequence divergence). The 19 putative species were difficult to recognise by morphological examination. We describe 13 new species; we do not describe (= name) six species characterised solely by molecular sequences and three putative species for which morphological data is available but for which molecular data is not. We have neither morphological nor molecular data for sanguinicolids harboured in five hosts species ( Siganus margaritiferus, S. puellus, Choerodon cauteroma, Parupeneus indicus and P. multifasciatus) in which we have seen infections. Where host species were infected in different localities they almost always harboured distinct species. Some host species ( for example, S. argenteus and S. lineatus from Lizard Island) harboured two or three species in a single geographical location. This suggests that, for parts of this system, parasite speciation has outstripped host speciation. Distance analysis of ITS2 showed species from each host family ( Siganidae, Mullidae and Labridae) did not form monophyletic clades to the exclusion of species from other host families. However, a host defined clade was formed by the sequences from sanguinicolids from S. fuscescens.

Microphallus fonti sp. n. (Digenea: Microphallidae) from the Red Swamp Crawfish in Southern United States

A new species of digenean, Microphallus fonti, is described from the red swamp crawfish in Louisiana, USA. It has a small pharynx and a rudimentary gut like M. opacus and possibly related species from crayfishes, but it differs from them by its relatively large male copulatory papilla and a conspicuous metraterm.

CYTOCHEMICAL DEMONSTRATION OF CHOLINERGIC, SEROTONINERGIC AND PEPTIDERGIC NERVE ELEMENTS IN GORGODERINA-VITELLILOBA (TREMATODA, DIGENEA)

Standard enzyme cytochemical and indirect immunocytochemical techniques have been used in conjunction with light and confocal scanning laser microscopy (CSLM) to visualize cholinergic, serotoninergic and peptidergic nerve elements in whole-mount preparations of the amphibian urinary-bladder fluke, Gorgoderina vitelliloba. Cholinesterase (ChE) activity was localized in paired anterior ganglia, a connecting dorsal commissure and in the origins of the ventral nerve cords. Cholinergic ganglia were also evident in shelled embryos in the uterus. Serotonin-immunoreactivity (IR) was more extensive than ChE activity and was identified in both the central and peripheral nervous systems. Serotoninergic nerve fibres were associated with the somatic musculature and female reproductive ducts. Antisera to nine mammalian peptides and one invertebrate (FMRFamide) peptide have been used to investigate the peptidergic nervous system in the parasite. Immunoreactivity was obtained to five peptides, namely pancreatic polypeptide (PP), peptide YY (PYY), neuropeptide Y (NPY), substance P (SP) and FMRFamide. Peptidergic nerve fibres were found to be more abundant than demonstrable cholinergic or serotoninergic nerve fibres. NPY-IR was identified only in the main components of the central nervous system. However, PP- and PYY-IR occurred in the anterior ganglia, dorsal commissure, main nerve cords and in numerous small varicose fibres that ramified throughout the worm. Additionally, PP-immunoreactive nerve fibres were found to innervate the musculature of the female reproductive tracts. Six sites of IR were found in the acetabulum, using antisera directed towards the C-terminal end of PP and PYY, and these matched with the distribution of six non-ciliated rosette-like papillae observed by scanning electron microscopy. SP- and FMRFamide-IR were identified in the CNS, and FMRFamide-immunopositive nerve fibres were also evident in association with the gonopore/cirrus region and with the terminal excretory pore. Results are discussed with respect to possible roles for each of the neurochemical types.