Neuro-angiostrongylosis in wild Black and Grey-headed flying foxes (Pteropus spp).

OBJECTIVE: To identify nematodes seen in histological sections of brains of flying foxes (fruit bats) and describe the associated clinical disease and pathology. PROCEDURES: Gross and histological examination of brains from 86 free-living flying foxes with neurological disease was done as part of an ongoing surveillance program for Australian bat lyssavirus. Worms were recovered, or if seen in histological sections, extracted by maceration of half the brain and identified by microscopic examination. Histological archives were also reviewed. RESULTS: There was histological evidence of angiostrongylosis in 16 of 86 recently submitted flying foxes with neurological disease and in one archival case from 1992. In 10 flying foxes, worms were definitively identified as Angiostrongylus cantonensis fifth-stage larvae. A worm fragment and third stage larvae were identified as Angiostrongylus sp, presumably A cantonensis, in a further three cases. The clinical picture was dominated by paresis, particularly of the hindlimbs, and depression, with flying foxes surviving up to 22 days in the care of wildlife volunteers. Brains containing fifth-stage larvae showed a moderate to severe eosinophilic and granulomatous meningoencephalitis (n = 14), whereas there was virtually no inflammation of the brains of bats which died when infected with only smaller, third-stage larvae (n = 3). There was no histological evidence of pulmonary involvement. CONCLUSION: This is the first report of the recovery and identification of A cantonensis from free-living Australian wildlife. While angiostrongylosis is a common cause of paresis in flying foxes, the initial clinical course cannot be differentiated from Australian bat lyssavirus infection, and wildlife carers should be urged not to attempt to rehabilitate flying foxes with neurological disease.

Microsporidiosis in a Gouldian finch (Erythrura [Chloebia] gouldiae).

A 12-day-old nestling Gouldian finch (Erythrura [Chloebia] gouldiae) was presented for investigation of a mortality problem in nestling finches raised by Bengalese finch foster parents. On histological examination, large numbers of spores consistent with a microsporidian organism were present within the small intestinal mucosa. Electron microscopy and molecular studies (sequencing the 5' end of the ssu rRNA gene) further defined the organism as Encephalitozoon hellem. Sequence homology with other eukaryotes was determined using a BLASTN search from the NCBI GenBank database. The finch isolate sequences showed greater than 99% homology with those of previously reported human and avian isolates.

A description of Lecithocladium invasor n.sp. (Digenea: Hemiuridae) and the pathology associated with two species of Hemiuridae in acanthurid fish.

Lecithocladium invasor n.sp. is described from the oesophagus of Naso annulatus, N. tuberosus and N. vlamingii on the Great Barrier Reef, Australia. The worms penetrate the oesophageal mucosa and induce chronic transmural nodular granulomas, which expand the full thickness of the oesophageal wall and protrude both into the oesophageal lumen and from the serosal surface. We observed two major types of lesions: large ulcerated, active granulomas, consisting of a central cavity containing a single or multiple live worms; and many smaller chronic fibrous submucosal nodules. Small, identifiable but attenuated, worms and degenerate worm fragments were identified within some chronic nodules. Co-infection of the posterior oesophagus of the same Naso species with Lecithocladium chingi was common. L. chingi is redescribed from N. annulatus, N. brevirostris, N. tuberosus and N. vlamingii. Unlike L. invasor n.sp., L. chingi was not associated with significant lesions. The different pathenogenicity of the two species in acanthurid fish is discussed.

The spread of Angiostrongylus cantonensis in Australia.

Until the recent establishment of Angiostrongylus cantonensis in North America, Australia was the only developed region endemic for this parasite. Almost 50 years ago the life cycle was elucidated there, in the city of Brisbane, and the first human infections probably occurred in 1959. From the 1970s, increasing numbers of autochthonous infections have been reported along the central east coast of the continent (southeast Queensland and northern New South Wales), involving humans, rats, dogs, horses, flying foxes and marsupials. Ten years ago, the parasite was discovered in Sydney, almost 1,000 km to the south, in dogs. In that city, it has since been diagnosed as a cause of neurological disease in increasing numbers of dogs, flying foxes, marsupials and zoo primates. Presumably, these infections resulted from the ingestion of snails or slugs, and it seems that virtually all species of native and exotic terrestrial molluscs can serve as intermediate hosts. It is not known how the parasite was introduced to this continent, or how it has spread over such an extensive territory, although eventually its range could encompass the entire east coast, and potentially other regions. It is also not known if the almost identical, native species, A. mackerrasae, is able to infect people (or other non-rodent hosts). All worms recovered to date, from one fatal human case, and from many animal infections, have been confirmed as A. cantonensis.

Neuro-angiostrongyliasis: unresolved issues.

Angiostrongylus cantonensis, the rat lungworm, probably evolved with its hosts, members of the genus Rattus and closely related species, in south-east Asia. Since its first discovery in rats in China and in a case of human infection in Taiwan, the parasite has been found to infect humans and other mammals across a wide and ever-increasing territory, which now encompasses much of south-east Asia, Melanesia, Polynesia and eastern Australia. It has also established a foothold in Africa, India, the Caribbean and south-eastern USA. This dispersal has been a direct result of human activity, and in some cases has been linked with the spread of the African giant land snail, Achatina fulica. However, this snail is not critical to the extension of the parasite's range, as numerous other indigenous molluscan species serve as adequate intermediate hosts; the importance of Achatina to the life cycle may have been over-emphasized. In Australia, the parasite is established along parts of the east coast, and the presence of an indigenous close relative, Angiostrongylus mackerrasae, suggests a long association of the parasite with its local rat hosts, a situation analogous to that of Angiostrongylus malaysiensis in south-east Asia. These three Angiostrongylus species share virtually the same life cycle, but only A. cantonensis has been confirmed to be a human pathogen.

Cerebrospinal angiostrongyliasis in five captive tamarins (Sanguinus spp).

Four cotton-top tamarins (Sanguinus oedipus oedipus) and one emperor tamarin (S imperator subgrisescens) housed in a zoo became depressed, anorexic, paraparetic and eventually paralysed. The animals died within 5 days to 18 months of the appearance of clinical signs. Histological examination showed nonsuppurative and eosinophilic meningoencephalitis, and metastrongyle nematode larvae were found within subarachnoid spaces of all animals and within the spinal cord of one. Intact larvae with features consistent with Angiostrongylus cantonensis were recovered from the brain of one animal. This parasite is the classical cause of eosinophilic meningoencephalitis in many parts of the world and the diagnosis can be strongly suspected on clinical grounds. In endemic areas like south-east Queensland, protection of captive animals against infection with A cantonensis is a difficult balance between providing a stimulating, natural setting and eliminating potentially infectious definitive, intermediate and paratenic hosts. This is the first report of cerebrospinal angiostrongyliasis in tamarins and nonhuman primates in Australia.

Intestinal mucus entrapment of Trichinella spiralis larvae induced by specific antibodies.

Entrapment of muscle larvae (ML) occurred in vitro when antibodies specific for Trichinella spiralis were added directly to intestinal mucus from normal non-immunized rats or when mucus was collected from pups suckling a T. spiralis-infected dam. Normal rat serum immunoglobulins failed to promote mucus entrapment and complement did not appear to play a part in the entrapment process. Differences were not observed in the efficiency of entrapment of ML by mucus harvested from different regions of the small intestine. Employing a panel of monoclonal antibodies (mAb) specific for excretory-secretory antigen (ESA), we attempted to dissociate antibody-mediated protection from mucus entrapment. We assessed mucus entrapment and rapid expulsion by these mAb in vivo, and observed protection in the absence of significant, immediate mucus entrapment in two cases. In addition, we measured mucus entrapment of ML in two in vitro assays. One assay employed intestinal mucus harvested from pups suckling dams that had been injected i.v. with a mAb. Results confirmed those obtained in vivo and indicated that antibodies were present in the intestinal lumina of passively immunized pups. In the second in vitro assay, mAb were added individually to mucus from pups suckling non-immunized dams. Results from these assays suggested that certain antibody isotypes may be processed in vivo in ways that influence, either positively or negatively, their abilities to cause mucus entrapment.

The role of the antibody Fc region in rapid expulsion of Trichinella spiralis in suckling rats.

When an IgG2c monoclonal antibody specific for Trichinella spiralis muscle stage larvae was cleaved with pepsin to yield F(ab')2 fragments, the latter retained their capacity to cause mucus entrapment and rapid expulsion of larvae from the intestines of suckling rats. When fed to pups, the F(ab')2 fragments of this antibody and the F(ab')2 fragments of a similarly prepared IgG2a antibody caused mucus entrapment of muscle larvae (ML), demonstrating that trapping is not dependent upon the Fc region of the antibody molecule. Despite the fact that these two antibodies had similar specificities and that their F(ab')2 fragments caused larval entrapment in mucus, F(ab')2 fragments of the IgG2a antibody failed to protect rat pups. Fragments of the IgG2c antibody caused rapid expulsion when injected into pups, but the distribution of larvae was dramatically different from when the fragments were delivered orally. These results indicate that entrapment of T. spiralis in mucus is not in itself the cause of the expulsion. The more likely possibility is that antibody impedes a function of Trichinella spiralis that is related to the capacity of the parasite to reside in its epithelial niche.

The role of mucus in antibody-mediated rapid expulsion of Trichinella spiralis in suckling rats.

Rat pups suckling dams parasitized by Trichinella spiralis express rapid expulsion, a protective response that is associated with the entrapment of infectious muscle larvae in intestinal mucus. Immunofluorescent studies revealed that antibodies were bound to the surfaces of the entrapped larvae. Mucus binding and rapid expulsion occurred in normal pups dosed with larvae coated with antibodies prepared from infected rat serum. Subsequent experiments revealed that entrapped larvae escaped from mucus after 2 hr in vitro incubation in saline. Escape correlated with the loss of the surface-bound antibodies, suggesting that mucus entrapment was reversible and dependent on antibody coating. Finally, when protective antibodies were injected 1, 2 or 6 hr after larvae were administered to pups, the parasites were forced to leave their epithelial niche and became enveloped in mucus. The above findings suggest that mucus trapping of T. spiralis larvae is dependent upon the coating of larvae by antibody, but that trapping is reversible, and is not in itself the pivotal event in rapid expulsion. The primary mechanism of rapid expulsion appears to be antibody-mediated inhibition of processes required for the parasite to maintain itself in the epithelium.