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.

The mitochondrial genome of Angiostrongylus mackerrasae is distinct from A. cantonensis and A. malaysiensis.

The native rat lungworm (Angiostrongylus mackerrasae) and the invasive rat lungworm (Angiostrongylus cantonensis) occur in eastern Australia. The species identity of A. mackerrasae remained unquestioned until relatively recently, when compilation of mtDNA data indicated that A. mackerrasae sensu Aghazadeh et al. (2015b) clusters within A. cantonensis based on their mitochondrial genomes (mtDNA). To re-evaluate the species identity of A. mackerrasae, we sought material that would be morphologically conspecific with A. mackerrasae. We combined morphological and molecular approaches to confirm or refute the specific status of A. mackerrasae. Nematodes conspecific with A. mackerrasae from Rattus fuscipes and Rattus rattus were collected in Queensland, Australia. Morphologically identified A. mackerrasae voucher specimens were characterized using amplification of cox1 followed by the generation of reference complete mtDNA. The morphologically distinct A. cantonensis, A. mackerrasae and A. malaysiensis are genetically distinguishable forming a monophyletic mtDNA lineage. We conclude that A. mackerrasae sensu Aghazadeh et al. (2015b) is a misidentified specimen of A. cantonensis. The availability of the mtDNA genome of A. mackerrasae enables its unequivocal genetic identification and differentiation from other Angiostrongylus species.

THADEUA GREENI-ASSOCIATED DERMATITIS IN AN EASTERN BETTONG (BETTONGIA GAIMARDI).

An adult female bettong (Bettongia gaimardi) presented with extensive alopecia and dermatitis affecting the ventral and lateral aspects of the neck and thorax. Microscopic examination of skin scrapings collected from the affected area revealed large numbers of the dermanyssid mite Thadeua greeni. A histopathologic diagnosis of chronic proliferative and hyperkeratotic perivascular dermatitis with intralesional mites was returned. Treatment with a combination of topical fipronil and parenteral ivermectin weekly for 3 wk resulted in the resolution of clinical signs and apparent elimination of the mite.

First report of parasitism by Ophidascaris robertsi (Nematoda) in a sugar glider (Petaurus breviceps, Marsupialia).

Third-stage larvae of Ophidascarsis robertsi (Nematoda: Ascaridoidea) were found on necropsy in a female sugar glider, Petaurus breviceps (Marsupialia: Petauridae), two in heart chambers and one free in the peritoneal cavity. The animal was bred in captivity and had previous contact with Australian pythons captured in nature, which could be the source of the infection. The histopathologic diagnosis was intraluminal and perivascular pulmonary hemorrhage possibly due to the parasitosis. It is the first report of parasitism by O. robertsi in a sugar glider.

Redescription of species of Gongylonema Molin, 1857 (Nematoda: Spiruroidea: Gongylonematidae) parasitic in some Australian vertebrate hosts and description of three new species.

Gongylonematid nematodes (Nematoda: Gongylonematidae) parasitic in some Australian vertebrates are described from the monotypic genus Gongylonema (Gongylonema) (Molin, 1857). Three previously incompletely described species from a megapod and murid rodents are re-described from limited material. Three additional species are described from murid rodents and macropodid, potoroid and phalangerid marsupials. A key to species is provided.

Cryptic species diversity in ticks that transmit disease in Australia.

Ticks are important vectors of a broad range of pathogens in Australia. Many tick species are morphologically similar and are therefore difficult to identify using morphology alone, particularly when collected in the larval and nymphal life stages. We report here the application of molecular methods to examine the species diversity of ixodid ticks at two sites in southern New South Wales, Australia. Our taxon sampling included six morphologically characterised adult stage voucher specimens of Ixodes trichosuri, Ixodes tasmani, Ixodes fecialis and Ixodes holocyclus (the paralysis tick) and ~250 field collected specimens that were in the larva or nymph stage and thus not morphologically identifiable. One nuclear and two mitochondrial amplicons were sequenced using a combination of Sanger and Illumina MiSeq sequencing. Phylogenetic relationships were estimated using both maximum likelihood and Bayesian methods. Two clades with strong bootstrap and Bayesian support were observed across trees estimated from each of three markers and from an analysis of the concatenated sequences. One voucher specimen of I. trichosuri was located in one of these clades, while the other I. trichosuri voucher specimen was in a second clade with the remaining three identified species, suggesting these morphologically similar ticks may represent different cryptic species. Unidentified specimens were found across both clades, and molecular divergence of many of these is equal to or greater than that observed between identified species, suggesting additional unidentified species may exist. Further studies are required to understand the taxonomic status of ticks in Australia, and how this species diversity impacts disease risk for livestock, domestic animals, wildlife and humans.

Spatial scaling laws do not structure strongyloid nematode communities in macropodid hosts.

The characteristics of species within a community can influence the number of species that can coexist within that community. In particular, body size can constrain how many individuals can 'fit' into a community, and overlap in resource use between species depends on differences in their body sizes. Here, using data from 18 communities of strongyloid nematodes living in the stomachs of macropodid marsupials, we test key predictions derived from spatial scaling laws regarding the minimum similarity in body size between coexisting species believed to control how many species can coexist in a community. These communities are ideal systems for such a test: they consist of huge numbers of individuals from numerous species, all belonging to the same family (Chabertiidae) and living in the same host organ. Within these communities, we found that mean abundance correlated negatively with body size across all nematode species, whether body size was measured as length or volume. However, we found no support for the predictions of spatial scaling laws. First, the size ratios of pairs of adjacent-sized species did not decrease as a function of the size of the largest species in a pair. The few significant relationships observed were all positive, suggesting that the relative difference in size between adjacent species in the size hierarchy may in fact increase toward the upper end of the size spectrum. Second, the frequency distributions of body sizes were predominantly right-skewed amongst the communities investigated: within the size spectrum observed in a nematode community, small-bodied species greatly outnumber large-bodied ones, in sharp contrast to the predictions of spatial scaling laws. Nematode body size may thus determine the abundance achieved by a species but not how many species can coexist; the limiting similarity between coexisting species must depend on other biological traits.

Helminth parasites of Australasian monotremes and marsupials.

This work includes all published records, to April 2015, of the helminths occurring in Australasian monotremes and marsupials, with due regard for synonymy and an attempt to include life history studies, pathological observations and epidemiology. It also contains all unpublished records known to us and referrable, by accession numbers, to curated collections in Australia and overseas. Information is presented by host family, genus, species, sub-species or chromosome race and includes the names of all host species from which no parasites have been recorded. Most records pertain to free-living and wild animals; where they do not, they have been annotated appropriately. Unpublished information known to the authors has been included in annotations to entries, where appropriate. Parasites are arranged as follows: Trematoda, Cestoda, Nematoda, Acanthocephala, and their systematic position is indicated by abbreviations placed before the name. The authority for each parasite record is given after the author's name, as a number in parentheses, and this refers to the numbered (1-664) list of references.        A parasite-host list is presented alphabetically, irrespective of taxonomic affiliation together with the host species in which they are known to occur. Hosts are arranged initially by family and alphabetically within each family.

More parasitic myositis cases in humans in Australia, and the definition of genetic markers for the causative agents as a basis for molecular diagnosis.

Since 1998, there have been six reported human cases of myositis in Australia, attributable to infection with the nematode Haycocknema perplexum. However, an unequivocal diagnosis of H. perplexum infection and associated disease has been seriously compromised by a lack of molecular markers for this nematode. Here, we report new cases of disseminated myositis in two male patients from the states of Queensland and Tasmania in Australia, respectively; genetically characterize the causative agent from each case; and, also establish a PCR-based sequencing approach as a tool to support the diagnosis of future cases and to underpin epidemiological studies.

Australian ecosystems, capricious food chains and parasitic consequences for people.

Characteristic Australian ecosystems or environments contain numerous food chains some of which may become capriciously side-tracked or appropriated by humans, with parasitic consequences for people in Australia and overseas. Twelve of 13 arboviruses affecting humans are of wildlife origin and all are transmitted by mosquitoes. In this case, transmission is thus associated with aquatic environments, many artificial. Zoonotic trematode (brachylaimiasis) and cestode (rodentoleposis) infections have been reported from semi-arid environments. Scabies and angiostrongylosis are associated with work, recreational and home environments. Four species of Rickettsia endemic in wildlife are acquired by humans from fleas, mites and ticks in bush and semi-urban environments. The enigmatic and life-threatening muspiceoid nematode, Haycocknema perplexum, is known from people associated with the natural environment in Tasmania; whether it comes from vertebrates, invertebrates, plants, soil or water is unknown. Food chains occurring in a range of Australian ecosystems and environments, some associated with feeding arthropods, others with accidental ingestion of invertebrates, may result in human exposure and infection. A range of organisms normally occurring in wildlife, domestic animals or the environment may be involved in causing human disease.

Species of Angiostrongylus (Nematoda: Metastrongyloidea) in wildlife: A review.

Twenty-one species of Angiostrongylus plus Angiostrongylus sp. (Nematoda: Metastrongyloidea) are known currently in wildlife. These occur naturally in rodents, tupaiids, mephitids, mustelids, procyonids, felids, and canids, and aberrantly in a range of avian, marsupial and eutherian hosts including humans. Adults inhabit the pulmonary arteries and right atrium, ventricle and vena cava, bronchioles of the lung or arteries of the caecum and mesentery. All species pass first-stage larvae in the faeces of the host and all utilise slugs and/or aquatic or terrestrial snails as intermediate hosts. Gastropods are infected by ingestion or penetration of first-stage larvae; definitive hosts by ingestion of gastropods or gastropod slime. Transmission of at least one species may involve ingestion of paratenic hosts. Five developmental pathways are identified in these life cycles. Thirteen species, including Angiostrongylus sp., are known primarily from the original descriptions suggesting limited geographic distributions. The remaining species are widespread either globally or regionally, and are continuing to spread. Small experimental doses of infective larvae (ca. 20) given to normal or aberrant hosts are tolerated, although generally eliciting a granulomatous histopathological response; large doses (100-500 larvae) often result in clinical signs and/or death. Two species, A. cantonensis and A. costaricensis, are established zoonoses causing neurological and abdominal angiostrongliasis respectively. The zoonotic potential of A. mackerrasae, A. malaysiensis and A. siamensis particularly warrant investigation. Angiostrongylus cantonensis occurs in domestic animals, mammalian and avian wildlife and humans in the metropolitan areas of Brisbane and Sydney, Australia, where it has been suggested that tawny frogmouths and brushtail possums may serve as biosentinels. A major conservation issue is the devastating role A. cantonensis may play around zoos and fauna parks where captive rearing of endangered species programmes may exist and where Rattus spp. are invariably a problem.

Biodiversity and parasites of wildlife: helminths of Australasian marsupials.

Despite current attempts to document the extent of biodiversity on Earth, significant problems exist in fully documenting the helminth parasites of wildlife. Using the Australasian marsupials as an example, we examine some of these difficulties, including challenges in collecting uncommon host species, the ongoing description of new species of marsupials, the presence of cryptic species, and the decline in taxonomic expertise in Australia. Although optimistic global predictions have been made concerning the rate of discovery and description of new species of animals, these predictions may not apply in the case of specific groups of animals such as the Australasian marsupials.

Pentastomids of wild snakes in the Australian tropics.

Pentastomids are endoparasites of the respiratory system of vertebrates, maturing primarily in carnivorous reptiles. Adult and larval pentastomids can cause severe pathology resulting in the death of their intermediate and definitive hosts. The study of pentastomids is a neglected field, impaired by risk of zoonoses, difficulties in species identification, and life cycle complexities. We surveyed wild snakes in the tropics of Australia to clarify which host species possess these parasites, and then sought to identify these pentastomids using a combination of morphological and molecular techniques. We detected pentastomid infections in 59% of the 81 snakes surveyed. The ubiquity of pentastomid infections in snakes of the Australian tropics sampled in this study is alarmingly high considering the often-adverse consequences of infection and the recognized zoonotic potential of these parasites. The pentastomids were of the genera Raillietiella and Waddycephalus and infected a range of host taxa, encompassing seven snake species from three snake families. All seven snake species represent new host records for pentastomids of the genera Raillietiella and/or Waddycephalus. The arboreal colubrid Dendrelaphis punctulatus and the terrestrial elapid Demansia vestigiata had particularly high infection prevalences (79% and 100% infected, respectively). Raillietiella orientalis infected 38% of the snakes surveyed, especially frog-eating species, implying a frog intermediate host for this parasite. Raillietiella orientalis was previously known only from Asian snakes and has invaded Australia via an unknown pathway. Our molecular data indicated that five species of Waddycephalus infect 28% of snakes in the surveyed area. Our morphological data indicate that features of pentastomid anatomy previously utilised to identify species of the genus Waddycephalus are unreliable for distinguishing species, highlighting the need for additional taxonomic work on this genus.

A further patient with parasitic myositis due to Haycocknema perplexum, a rare entity.

A new genus of nematode, Haycocknema perplexum, causing polymyositis in humans, was first described in two Australian patients from Tasmania in 1998. Three patients with myositis due to the same nematode were reported from northern Queensland in 2008. We report the sixth case from Australia, a 50-year-old man, also from Tasmania. He had a 2-year history of progressive weakness, weight loss of 10 kg and dysphagia. Muscle biopsy was initially interpreted as polymyositis with eosinophils. Maximum creatine kinase (CK) level was 5700 U/L and full blood examination was normal. He deteriorated after several months of treatment with prednisolone and methotrexate and review of the muscle biopsy showed intramyofibre parasites of H. perplexum. After 3 months of treatment with albendazole therapy, he made a very good clinical recovery and his CK decreased to 470 U/L. This uniquely Australian parasite can mimic polymyositis and leads to significant irreversible morbidity (two of the previous patients still have weakness and elevated CK after years) and even mortality (one died), if diagnosed late or after corticosteroids. Diagnosis can only be made by histopathology of muscle biopsy.

Using combined morphological, allometric and molecular approaches to identify species of the genus Raillietiella (Pentastomida).

Taxonomic studies of parasites can be severely compromised if the host species affects parasite morphology; an uncritical analysis might recognize multiple taxa simply because of phenotypically plastic responses of parasite morphology to host physiology. Pentastomids of the genus Raillietiella are endoparasitic crustaceans primarily infecting the respiratory system of carnivorous reptiles, but also recorded from bufonid anurans. The delineation of pentastomids at the generic level is clear, but the taxonomic status of many species is not. We collected raillietiellids from lungs of the invasive cane toad (Rhinella marina), the invasive Asian house gecko (Hemidactylus frenatus), and a native tree frog (Litoria caerulea) in tropical Australia, and employed a combination of genetic analyses, and traditional and novel morphological methods to clarify their identity. Conventional analyses of parasite morphology (which focus on raw values of morphological traits) revealed two discrete clusters in terms of pentastome hook size, implying two different species of pentastomes: one from toads and a tree frog (Raillietiella indica) and another from lizards (Raillietiella frenatus). However, these clusters disappeared in allometric analyses that took pentastome body size into account, suggesting that only a single pentastome taxon may be involved. Our molecular data revealed no genetic differences between parasites in toads versus lizards, confirming that there was only one species: R. frenatus. This pentastome (previously known only from lizards) clearly is also capable of maturing in anurans. Our analyses show that the morphological features used in pentastomid taxonomy change as the parasite transitions through developmental stages in the definitive host. To facilitate valid descriptions of new species of pentastomes, future taxonomic work should include both morphological measurements (incorporating quantitative measures of body size and hook bluntness) and molecular data.