A molecular characterization of marsupial filarioid nematodes of the genus Breinlia.

Here we present the genetic relationships of 26 specimens of the genus Breinlia (Nematoda: Filarioidea) from a range of Australian marsupials using markers in the small subunit of nuclear ribosomal RNA and mitochondrial cytochrome c oxidase subunit 1 (cox1) genes and compare them with morphological determinations. The molecular data support the validity of most of the morpho-species included in the study and provide provisional insights into the phylogeny of the genus in Australian mammals, with dasyuroid marsupials appearing to be the original hosts. The recent discovery of Breinlia annulipapillata in the eye of a human brings this genus of parasites into the group of emerging infectious parasitic diseases.

Ocular Filariasis in Human Caused by Breinlia (Johnstonema) annulipapillata Nematode, Australia.

We report a human case of ocular filariasis, caused by a species of Breinlia nematode, from Queensland, Australia. Morphological and molecular evidence indicated that the nematode Breinlia (Johnstonema) annulipapillata, or a closely related taxon, likely transmitted from a macropodid marsupial host was involved, which might represent an accidental finding or an emerging zoonosis.

Eutetrarhynchus pacificus n. sp. (Cestoda: Trypanorhyncha) from Raja inornata Jordan & Gilbert (Batoidea: Rajiformes) off the coast of California with comments on congeners.

Eutetrarhynchus pacificus n. sp. is described from the spiral valve of Raja inornata Jordan & Gilbert off the coast of California, USA. The new species is distinguished from E. ruficollis (Eysenhardt, 1829) and E. leucomelanus (Shipley & Hornell, 1906) in having acraspedote rather than craspedote segments and a saccate rather than a branched uterus. It is distinguished from E. platycephali Palm, 2004 in lacking an enlarged hook in the eighth row of the basal armature and from E. beveridgei Schaeffner, 2013, which has a basal swelling and a distinctive basal armature. A partial redescription of E. ruficollis, the type-species of the genus, is provided based on available museum specimens, highlighting the need for a comprehensive redescription of this species to better define the characteristics of the genus. The presence of an undescribed species in museum collections is also noted. Based on the 28S ribosomal gene, the new species clustered with Dollfusiella in a molecular phylogenetic tree. The delimitation of Eutetrarhynchus and its relationship with Dollfusiella is discussed.

Redescription of Rugopharynx australis (Mönnig, 1926) and the description of R. moennigi n. sp. (Nematoda: Strongyloidea) from kangaroos (Marsupialia: Macropodidae) in Australia.

Rugopharynx australis (Mönnig, 1926) (Nematoda: Strongyloidea) is redescribed based on specimens from the type host, Osphranter rufus (Desmarest), together with matching DNA sequence data. Additional hosts were Macropus giganteus Shaw and Osphranter robustus (Gould) with single occurrences in M. fuliginosus (Desmarest), Notamacropus dorsalis (Gray), Lagorchestes conspicillatus Gould and Petrogale xanthopus Gray. Rugopharynx moennigi n. sp., formerly included within R. australis, is distinguished by shorter but overlapping spicule lengths and in the morphology of the gubernaculum as well as by molecular data. Rugopharynx moennigi n. sp. appears to be primarily parasitic in M. fuliginosus throughout its geographical range, but also infects M. giganteus, O. robustus and O. rufus in areas of host sympatry.

Four new species of Rugopharynx Mönnig, 1927 (Nematoda: Strongyloidea) parasitic in the stomachs of macropodid marsupials from Australia.

Four new species of Rugopharynx Mönnig, 1927 are described from macropodid marsupials in Australia, some identified initially using molecular methods. Rugopharynx thetidis n. sp. from Thylogale thetis (Lesson) was initially included within R. sigma Chilton, Beveridge & Andrews, 1993 but was identified as being distinctive using molecular methods and is differentiated morphologically from R. sigma by the more anterior position of the deirid and by the distinctive curvature of the spicule tips. Rugopharynx thylogale n. sp., also from Thylogale thetis, is distinguished by the extreme elongation of the dorsal lobe of the bursa and spicule length (1.07-1.23 mm). Rugopharynx solitarius n. sp., again from Thylogale thetis, was initially included within Rugopharynx zeta (Johnston & Mawson, 1939) but differs morphologically in the features of the bursa. Rugopharynx disiunctus n. sp. from Macropus fuliginosus (Desmarest) was initially included within Rugopharynx rho Beveridge & Chilton, 1999 but was identified as distinctive based on DNA sequence differences. The new species differs from R. rho in the pattern of striations on the buccal capsule and in the lengths of the spicules. Additional records of species of Rugopharyx in M. fuliginosus in Western Australia are provided.

New species of Wallabinema Beveridge, 1983 (Nematoda: Strongyloidea) and a redescription of W. gallardi (Johnston & Mawson, 1939) from Australian macropodid marsupials.

Two new species of Wallabinema Beveridge, 1983 are described from the sacculated fore-stomachs of macropodid marsupials in Australia. Wallabinema petrogale n. sp. from the rock wallabies Petrogale penicillata (J. Gray) and P. inornata Gould, from Queensland, differs from all congeners in having its four sub-median lips subdivided at the base. Wallabinema macropodis n. sp. from the black stripe wallaby Macropus dorsalis (J. Gray) and the red-necked pademelon Thylogale thetis (Lesson), also from Queensland, is most similar to congeners with the nerve-ring encircling the oesophageal isthmus (W. parvispiculare Beveridge, 1983, W. tasmaniense Beveridge, 1983 and W. thylogale Beveridge, 1983), but differs in the length of the spicules. Wallabinema gallardi (Johnston & Mawson, 1939) is redescribed from T. thetis in Queensland and New South Wales, with T. thetis considered to be the type-host.

Ascarophisnema hoiae n. sp. (Nematoda: Cystidicolidae), from the stomach of the trumpeter whiting, Sillago maculata Quoy & Gaimard (Perciformes: Sillaginidae) from Moreton Bay, Queensland, Australia.

Ascarophisnema hoiae n. sp. (Nematoda: Cystidicolidae) is described from the stomach of the trumpeter whiting, Sillago maculata Quoy & Gaimard (Perciformes: Sillaginidae) from Moreton Bay, Queensland, Australia. It differs morphologically from the only other valid congener, A. tridentatum Moravec & Justine, 2010 in the shape of the sub-labium and in the lengths of the spicules and the morphology of their distal tips (bifid). It represents the first record of this genus from Australia and appears to be highly oioxenous, having been found only in this host species among 133 other species of fish examined at the same locality.

New species of Macropostrongyloides Yamaguti, 1961 (Nematoda: Strongylida) and the redescription of Ma. baylisi (Wood, 1930) from Australian macropodid marsupials.

Specimens of four genetically distinct groups of Macropostrongyloides baylisi Wood, 1930 were analysed morphologically. Each genotype was found to represent a morphologically distinct species: Ma. baylisi from Osphranter robustus woodwardi (Thomas) and Osphranter robustus erubescens (Sclater); Ma. spearei n. sp. from Osphranter robustus robustus (Gould) and O. r. erubescens; Ma. mawsonae n. sp. from Macropus giganteus Shaw and Ma. woodi n. sp. from Osphranter rufus (Desmarest). The new species described here are differentiated primarily by several male-specific features that have been overlooked in previous taxonomic revisions. These features include striations on the terminal part of the spicule ala, the papillae surrounding the genital cone and the bursal striations. Furthermore, scanning electron photomicrographs have revealed greater details of previously undefined structures within the buccal cavity that warrant further investigations.

Genetic variation within the genus Macropostrongyloides (Nematoda: Strongyloidea) from Australian macropodid and vombatid marsupials.

The genetic variation and taxonomic status of the four morphologically-defined species of Macropostrongyloides in Australian macropodid and vombatid marsupials were examined using sequence data of the ITS+ region (=first and second internal transcribed spacers, and the 5.8S rRNA gene) of the nuclear ribosomal DNA. The results of the phylogenetic analyses revealed that Ma. baylisi was a species complex consisting of four genetically distinct groups, some of which are host-specific. In addition, Ma. lasiorhini in the common wombat (Vombatus ursinus) did not form a monophyletic clade with Ma. lasiorhini from the southern hairy-nosed wombat (Lasiorhinus latifrons), suggesting the possibility of cryptic (genetically distinct but morphologically similar) species. There was also some genetic divergence between Ma. dissimilis in swamp wallabies (Wallabia bicolor) from different geographical regions. In contrast, there was no genetic divergence among specimens of Ma. yamagutii across its broad geographical range or between host species (i.e. Macropus fuliginosus and M. giganteus). Macropostrongyloides dissimilis represented the sister taxon to Ma. baylisi, Ma. yamagutii and Ma. lasiorhini. Further morphological and molecular studies are required to assess the species complex of Ma. baylisi.

Worm burdens and associated histopathological changes caused by gastrointestinal nematodes in alpacas from Australia.

In this study, 100 gastrointestinal tracts of Australian alpacas were examined to assess the worm burden and to identify the species of nematode present. Faecal samples were collected from 97 alpacas and processed for faecal egg counts (FECs). For identification of the species, both molecular (multiplexed-tandem polymerase chain reaction [MT-PCR]) and morphological techniques were used. Total worm counts (TWCs) revealed a mean burden of 1300 worms, with the highest burden of 29,000 worms. The average egg count was 501 eggs per gram of faeces (EPG), with the highest count of 3500 EPG. Nineteen different species of gastrointestinal nematodes (GINs) were identified, and Graphinema auchenia, Camelostrongylus mentulatus and Trichuris tenuis were recovered from Australian alpacas for the first time. Haemonchus contortus was the most prevalent nematode (81%) followed by C. mentulatus (60%). The majority of the nematodes found are shared with sheep, goats and cattle. Findings of this study provide useful insights into the spectrum of GINs and their burden in Australian alpacas.

Epidemiology of gastrointestinal nematodes of alpacas in Australia: II. A longitudinal study.

We conducted a longitudinal survey on 13 alpaca farms in four climatic zones of Australia to understand the epidemiology of gastrointestinal nematodes (GINs) of alpacas. A total of 1688 fresh faecal samples were collected from both sexes of alpacas from May 2015 to April 2016 and processed for faecal egg counts (FEC) and molecular identification of eggs using the multiplexed-tandem PCR assay. Based on egg morphology, the overall prevalence of GINs was 61% while that for strongyles was 53%. The overall mean FEC was 168 eggs per gram (EPG) of faeces, with the highest count of 15,540 EPG. Weaners had the highest prevalence (73%) and mean FEC (295 EPG) of GINs followed by tuis, crias and adults. Alpacas in the winter rainfall zone had the highest prevalence (68%) as well as FEC (266 EPG) followed by Mediterranean-type, non-seasonal and summer rainfall zones. Trichostrongylus spp. (83%, 89/107), Haemonchus spp. (71%, 76/107) and Camelostrongylus mentulatus (63%, 67/107) were the three most common GINs of alpacas across all climatic zones. The mixed-effects zero-inflated negative binomial regression model used in this study showed that it could help to design parasite control interventions targeted at both the herd level and the individual alpaca level. The findings of this study showed that the epidemiology of GINs of alpacas is very similar to those of cattle and sheep, and careful attention should be paid when designing control strategies for domestic ruminants co-grazing with alpacas.

Epidemiology of gastrointestinal nematodes of alpacas in Australia: I. A cross-sectional study.

This study involved a national cross-sectional survey of gastrointestinal nematodes (GINs) of alpacas in Australia. A total of 1545 fresh faecal samples were collected from both sexes of alpacas and processed for faecal egg counts (FEC) and molecular identification of nematodes using the multiplexed tandem PCR assay. Based on egg morphology, the overall prevalence of GINs was 66% while that for strongyles was 59%. The overall mean FEC was 276 eggs per gram (EPG) of faeces, with the highest count of 17,415 EPG. Male alpacas had a higher prevalence (68%, 334/490) as well as mean FEC (328 ± 60 EPG) of GINs than females (63%, 602/954; 227 ± 26, respectively). Weaners had the highest prevalence (80%) whereas tuis had the highest FEC (402 EPG) of nematodes. The highest prevalence (77%, 293/383) and FEC (630 EPG) of GINs were observed in the summer rainfall zone followed by the Mediterranean-type rainfall, non-seasonal rainfall and winter rainfall zones. The characterisation of nematode DNA isolated from faeces revealed the occurrence of seven different GINs, including Camelostrongylus mentulatus, Cooperia spp., Haemonchus spp., Oesophagostomum spp., Ostertagia ostertagi, Teladorsagia circumcincta and Trichostrongylus spp. Besides, Nematodirus spp. and Trichuris spp. were also found during FECs. The prevalence of Haemonchus spp. was highest in the summer rainfall zone while that of C. mentulatus was highest in the Mediterranean-type rainfall, non-seasonal rainfall and winter rainfall zones. The findings of this study revealed that alpacas harbour many of the same nematodes as sheep and cattle.

New species and new records of species of Cloacina von Linstow, 1898 (Nematoda: Strongylida) parasitic in the western scrub wallaby, Notamacropus irma (Jourdan) (Marsupialia: Macropodidae) from Western Australia.

The helminth parasites of the western scrub wallaby or black-glove wallaby, Notamacropus irma (Jourdan) which occurs in Western Australia are relatively poorly documented. Six new species of the strongyloid genus Cloacina von Linstow, 1898 (Strongylida: Chabertiidae) are described namely C. asymmetrica n. sp., C. brazellei n. sp., C. harriganae n. sp., C. hobbsi n. sp., C. middletoni n. sp. and C. woodi n. sp. A redescription of C. laius Beveridge, 1999 from the same host species is included. Molecular sequence data (ITS1 and ITS2 ribosomal DNA) were obtained for C. asymmetrica, C. brazellei, C. hobbsi, C. middletoni and from the previously described species C. themis Beveridge, 1998 occurring in the same host species. Phylogenetically, C. asymmetrica, C. hobbsi and C. middletoni formed a distinct clade, suggesting the possibility of within-host speciation. Cloacina themis clustered with a group of morphologically distinctive species in a separate clade and C. brazellei clustered in a third clade but with poor support. This pattern of congeners in a single host species occurring in multiple clades mirrors the situation in other kangaroos and wallabies. Species of Cloacina from N. irma reported thus far therefore consist of a series of species found only in this host, with two species (C. brazellei and C. laius) shared with the sympatric macropodid Setonix brachyurus (Quoy & Gaimard).

New genera, species and records of rhinebothriidean cestodes (Platyhelminthes) parasitic in Australian stingrays (Elasmobranchii: Batoidea).

Collections of rhinebothriidean cestodes (Platyhelminthes) from Australian batoid elasmobranchs revealed the presence of a number of new genera and species. Ruptobothrium louiseuzeti n. g., n sp. is described from the reticulate whipray, Himantura australis Last, Naylor & Manjaji-Matsumoto, from off the Northern Territory and Mixobothrium queenslandense n. g., n sp. is described from the green sawfish, Pristis zijsron Bleeker, from off north-eastern Queensland. Two new species of Rhabdotobothrium Euzet, 1953 are described: Rhabdotobothrium meridionale n. sp. from the southern eagle ray Myliobatis tenuicaudatus Hector from off South Australia and Rhabdotobothrium anoxypristidis n. sp. from the narrow sawfish, Anoxypristis cuspidatus (Latham) from off north Western Australia. A new species of Scalithrium Healy & Reyda, 2016, Scalithrium australiense n. sp., is described from the reticulate whipray, Himantura australis Last, Naylor & Manjaji-Matsumoto, from off northern Western Australia. Scalithrium smitii (Shinde, Deshmukh & Jadhav, 1981) n. comb. is reported from Australian waters for the first time in the black spotted stingray Maculabatis toshi (Whitley) from off northern Western Australia. New host and geographical records are provided for Stillabothrium jeanfortiae Forti, Aprill & Reyda, 2016 from the brown whipray Maculobatis toshi (Whitley) and the black-spotted whipray, Maculabatis cf. astra (Last, Manjaji-Matsumoto & Pogonoski) from Moreton Bay in southern Queensland.

New species of Rhinebothrium Linton, 1890 (Cestoda: Rhinebothriidea) parasitic in Australian stingrays (Elasmobranchii: Batoidea).

Seven new species of the cestode genus Rhinebothrium Linton, 1890 are reported from the spiral intestines of batoid elasmobranchs from the coasts of Australia. The new species are: Rhinebothrium dasyatidis n. sp. from the smooth stingray Bathytoshia brevicaudata (Hutton) from Spencer Gulf, South Australia; Rhinebothrium bunburyense n. sp. from the southern eagle ray, Myliobatis tenuicaudatus Hector from off Bunbury, Western Australia; Rhinebothrium vandiemeni n. sp. from the reticulate whipray, Himantura australis Last, Naylor & Manjaji-Matsumoto from off Cape van Diemen, Northern Territory; Rhinebothrium fluviorum n. sp. from the estuary stingray, Hemitrygon fluviorum (Ogilby) from Moreton Bay, Queensland; Rhinebothrium urolophi n. sp. from the wide stingaree Urolophus expansus McCulloch from off Beachport, South Australia; Rhinebothrium nickoli n. sp. from the brown whipray Maculabatis toshi (Whitley) and the reticulate whipray, Himantura australis Last, Naylor & Manjaji-Matsumoto, from Nickol Bay, Western Australia and from the white-spotted guitarfish Rhynchobatus australiae (Whitley) from off Broome, Western Australia and Rhinebothrium fungiforme n. sp. from the estuary stingray, Hemitrygon fluviorum (Ogilby) from Fog Bay in the Northern Territory.

A new species of Kiluluma Skrjabin, 1916 (Nematoda: Strongyloidea) from the white rhinoceros, Ceratotherium simum (Burchell) from South Africa.

A new species of Kiluluma Skrjabin, 1916, Kiluluma ornata n. sp., is described from the intestine of the white rhinoceros Ceratotherium simum (Burchell) from South Africa. The new species is virtually identical with a species described as Kiluluma sp., but not named due to a paucity of material then available. The new species most closely resembles K. solitaria Thapar, 1924 and K. ceratotherii Beveridge & Jabbar, 2013, in the possession of an undulating anterior margin of the buccal capsule but differs in the leaf crown elements which have bulbous, lip-like expansions, which are lacking in K. solitaria and K. ceratotherii.

Tetraphyllidean and onchoproteocephalidean cestodes of elasmobranchs from Moreton Bay, Australia: description of two new species and new records for seven described species.

Parasitological examination of elasmobranchs of Moreton Bay, Queensland, Australia, resulted in the discovery of cestodes belonging to several armed genera of the Tetraphyllidea and Onchoproteocephalidea. Two new tetraphyllideans, Yorkeria moretonensis n. sp. and Yorkeria williamsi n. sp., are described from Chiloscyllium cf. punctatum (Hemiscylliidae). Yorkeria moretonensis n. sp. differs from its congeners in the possession of vitelline follicles that are discontinuous in the region of the ovary and in the length of its pedicels. Yorkeria williamsi n. sp. is most similar to Y. parva Southwell, 1927, but has larger, oval bothridia, longer pedicels and differences in the sizes of the scolex hooks. Yorkeria longstaffae Caira, Jensen & Rajan, 2007 is reported from Moreton Bay for the first time, and Spiniloculus mavensis Southwell, 1925 is re-reported from the type-locality and likely type-host (Moreton Bay and Chiloscyllium cf. punctatum, respectively), over 90 years after its original description. Six known onchoproteocephalideans, Acanthobothrium cannoni Campbell & Beveridge, 2002, A. chisholmae Campbell & Beveridge, 2002, A. ocallaghani Campbell & Beveridge, 2002, A. margieae Fyler, 2011, Megalonchos shawae Caira, Reyda & Mega, 2007 and M. sumansinghai Caira, Reyda & Mega, 2007, are reported from Moreton Bay for the first time, representing significant range extensions for all species.

New species of Cloacina von Linstow, 1898 (Nematoda: Strongyloidea) parasitic in the stomachs of wallaroos, Osphranter spp. (Marsupialia: Macropodidae) from northern Australia.

Three new species of the parasitic nematode genus Cloacina von Linstow, 1898 (Strongyloidea: Cloacininae) are described from the stomachs of wallaroos, Osphranter spp. (Marsupialia: Macropodidae), from northern Australia. Cloacina spearei n. sp. is described from O. robustus woodwardi (Thomas) and O. antilopinus (Gould) and is distinguished from congeners by the shape of the cephalic papillae, the shallow buccal capsule, the presence of an oesophageal denticle and the convoluted but non-recurrent vagina in the female. Cloacina longibursata n. sp. also from O. robustus woodwardi and O. antilopinus is distinguished from congeners by the elongate dorsal lobe of the bursa, with the origin of the lateral branchlets posterior to the principal bifurcation, in the features of the spicule tip, the lack of bosses lining the oesophagus and the absence of an oesophageal denticle. Cloacina crassicaudata n. sp., from the same two host species was formerly identified as C. cornuta (Davey & Wood, 1938). Differences in the cephalic cuticle (inflation lacking in the new species), the shape of the cephalic papillae, the dorsal oesophageal tooth and the spicule tips, as well as differences in the sequences of the internal transcribed spacers of the nuclear ribosomal DNA, indicate that this is an independent species. The geographical distribution of this species is disjunct with populations in both the Northern Territory and Queensland. Possible reasons for the disjunct distribution are discussed.

Lobocapillaria austropacifica n. g., n. sp. (Nematoda: Capillariidae) from the obtuse barracuda Sphyraena obtusata Cuvier (Sphyraenidae, Perciformes) off eastern Australia.

Based on light and scanning electron microscopical studies, a new nematode parasite, Lobocapillaria austropacifica n. sp. (Capillariidae), is described from the gall-bladder of the marine fish (obtuse barracuda) Sphyraena obtusata Cuvier (Perciformes: Sphyraenidae) from off the eastern Pacific coast of Australia, for which a new genus Lobocapillaria n. g. is established. This new genus is mainly characterised by a single row of stichocytes, the presence of two large, conspicuously elongated lateral caudal lobes and a pair of subventral papillae at their base in males, a flat spicule distended laterally towards its proximal end and provided with superficial rough transverse grooves, a spicular canal and a very long, aspinose spicular sheath with a conspicuous expansion near its proximal end when evaginated. Capillaria sphyraeni Parukhin, 1971 is transferred to Lobocapillaria as L. sphyreni (Parukhin, 1971) n. comb. A key to capillariid genera containing species parasitic in fishes is provided.

Phylogenetic relationships of species of the oesophageal parasitic nematode genera Cyclostrongylus and Spirostrongylus (Strongyloidea: Chabertiidae: Cloacininae) with their wallaby hosts (Marsupialia: Macropodidae).

A phylogeny for seven species of Cyclostrongylus and the monotypic genus Spirostrongylus (Nematoda: Chabertiidae), all highly host specific parasites of the oesophagi of wallabies (Marsupialia: Macropodidae), was constructed using sequence data for the first and second internal transcribed spacers (ITS-1 and ITS-2) of the nuclear ribosomal DNA. There was no evidence for co-speciation, or for the sympatric or synxenic speciation of Cyclostrongylus alatus and Cyclostrongylus perplexus, both of which are parasites of Macropus rufogriseus. Rather, host switching, correlating with geographical distributions, appeared to provide some explanation of the pattern of speciation observed.