R.A. Swan

Quantitative studies of ovine haemonchosis. I. Relationship between faecal egg counts and total worm counts

The successful operation of parasite-control programmes based on monitoring of faecal egg counts stimulated an examination of the quantitative relationship between egg counts and worm counts of Haemonchus contortus in sheep in the Goondiwindi area of south-east Queensland. Sixty-one commercial Merino sheep were selected from flocks where outbreaks of haemonchosis had occurred. Faecal samples were collected prior to euthanasia and eggs and worms subsequently counted and differentiated. A strong relationship (r2 = 0.83, ∗∗∗P < 0.001) was found between faecal egg counts and the total number of adult H. contortus. The size of the worm population, the time of year, and the ratio of male to female adult worms were minor sources of variation. The relationship was considered to be of value in the implementation of programmes for the diagnosis and control of haemonchosis in sheep flocks in the area.

A Novel Virus Detected in Papillomas and Carcinomas of the Endangered Western Barred Bandicoot (Perameles bougainville) Exhibits Genomic Features of both the Papillomaviridae and Polyomaviridae

ABSTRACT Conservation efforts to prevent the extinction of the endangered western barred bandicoot (Perameles bougainville) are currently hindered by a progressively debilitating cutaneous and mucocutaneous papillomatosis and carcinomatosis syndrome observed in captive and wild populations. In this study, we detected a novel virus, designated the bandicoot papillomatosis carcinomatosis virus type 1 (BPCV1), in lesional tissue from affected western barred bandicoots using multiply primed rolling-circle amplification and PCR with the cutaneotropic papillomavirus primer pairs FAP59/FAP64 and AR-L1F8/AR-L1R9. Sequencing of the BPCV1 genome revealed a novel prototype virus exhibiting genomic properties of both the Papillomaviridae and the Polyomaviridae. Papillomaviral properties included a large genome size (∼7.3 kb) and the presence of open reading frames (ORFs) encoding canonical L1 and L2 structural proteins. The genomic organization in which structural and nonstructural proteins were encoded on different strands of the double-stranded genome and the presence of ORFs encoding the nonstructural proteins large T and small t antigens were, on the other hand, typical polyomaviral features. BPCV1 may represent the first member of a novel virus family, descended from a common ancestor of the papillomaviruses and polyomaviruses recognized today. Alternatively, it may represent the product of ancient recombination between members of these two virus families. The discovery of this virus could have implications for the current taxonomic classification of Papillomaviridae and Polyomaviridae and can provide further insight into the evolution of these ancient virus families.

Wide range of Chlamydiales types detected in native Australian mammals

The Chlamydiales are a unique order of intracellular bacterial pathogens that cause significant disease of birds and animals, including humans. The recent development of a Chlamydiales-specific 16S rDNA polymerase chain reaction (PCR) assay has enabled the identification of Chlamydiales DNA from an increasing range of hosts and environmental sources. Whereas the Australian marsupial, the koala, has previously been shown to harbour several Chlamydiales types, no other Australian marsupials have been analysed. We therefore used a 16S rDNA PCR assay combined with direct sequencing to determine the presence and genotype of Chlamydiales in five wild Australian mammals (gliders, possums, bilbies, bandicoots, potoroos). We detected eight previously observed Chlamydiales genotypes as well as 10 new Chlamydiales sequences from these five Australian mammals. In addition to PCR analysis we used antigen specific staining and in vitro culture in HEp-2 cell monolayers to confirm some of the identifications. A strong association between ocular PCR positivity and the presence of clinical disease (conjunctivitis, proliferation of the eyelid) was observed in two of the species studied, gliders and bandicoots, whereas little clinical disease was observed in the other animals studied. These findings provide further evidence that novel Chlamydiales infections occur in a wide range of hosts and that, in some of these, the chlamydial infections may contribute to clinical disease.

Seroprevalence of specific viral infections in confiscated orangutans (Pongo pygmaeus)

A serological survey of confiscated orangutans was conducted to determine the prevalence of specific viral infections cross reacting with human viruses. Antibodies specific for human hepatitis A (HAV) and B (HBV) viruses, herpes simplex viruses (HSV), and human T‐lymphotropic virus (HTLV types I and II), as well as for the simian type D retroviruses (SRV types 1 to 3) and simian immunodeficiency virus (SIV) were tested in samples from 143 orangutans. Results revealed a high prevalence of potential pathogens. The most prevalent viral infection found was HBV (59.4% prevalence) of which 89.4% of infected individuals seroconverted to the non‐infectious state and 10.6% remained as chronic carriers. Antibodies to HAV, HSV, HTLV‐1, and SRV were also detected but at a lower prevalence. There was no evidence of lentiviral infections in this group of animals. The results confirm the importance of quarantine and the need for diagnostic differentiation of virus infections to determine if they are of human origin or unique orangutan viruses.

Quantitative studies of ovine haemonchosis. 2. Relationship between total worm counts of Haemonchus contortus, haemoglobin values and bodyweight

The relationships between Haemonchus contortus burdens, haemoglobin levels and bodyweight, were examined in 50 commercial Merino sheep, 9-18 months of age, in south-east Queensland where haemonchosis was endemic. Worm counts and haemoglobin levels were predictable (P less than 0.001, r = 0.68) from the equations log worm count = B.wt. X 0.06942 + log worm count X (-2.18896) + 12.51557. Bodyweight was not predictable from either worm counts or haemoglobin levels. Although bodyweight was represented in both equations, it was only a small and insignificant contributor. When haemoglobin levels became moderately depressed (less than 10.5 g%) or severely depressed (less than 8.0 g%), then the number of worms associated with these haemoglobin levels varied with the bodyweight of the sheep. For sheep up to 20 kg, 10.5 g% Hb was associated with 112 worms and 8 g% with 355 worms. However, 355 worms caused only moderate depression of Hb levels in sheep over 50 kg, and 1259 worms were required to cause severe depression (less than 8.0 g%) in sheep over 50 kg. Three sheep were found to be severely anaemic in the presence of less than 100 worms. It was postulated that additional factors of time and nutrition may have affected these sheep. The sheep were of low bodyweight, in poor condition (low fat score) and were subject to high stocking rates on long grass pastures and high rainfall in autumn. Other internal parasite species present were Nematodirus spp. and Trichostrongylus spp. However, their numbers were low (57 and 218 worms, respectively) and did not contribute to anaemia. The correlation between H. contortus and Nematodirus spp. was found to be negative but non-significant (r = -0.26, P less than 0.01), while the correlation between H. contortus and Trichostrongylus spp. was positive and significant (r = 0.38, P less than 0.01).

OCULAR CHLAMYDIALES INFECTIONS OF WESTERN BARRED BANDICOOTS (PERAMELES BOUGAINVILLE) IN WESTERN AUSTRALIA

Abstract The western barred bandicoot (Perameles bougainville) is an endangered species, free ranging on only two islands off the coast of Western Australia (Dorre and Bernier Islands). Conservation efforts are currently directed at reintroducing these marsupials into predator-proof enclosures and habitats in historical distribution ranges on the mainland in the southwest of Western Australia and in South Australia. In September 2000, 19 western barred bandicoots were captured on Bernier Island for translocation, and 11 of these had evidence of at least one of the following eye conditions: corneal opacity, conjunctivitis, ocular discharge, and blepharitis. Five bandicoots were examined, and conjunctival and cloacal swabs were collected. Polymerase chain reaction for Chlamydiales was positive in four bandicoots. Four Chlamydiales types were identified by gene sequencing, including a strain of Chlamydia pecorum different from strains previously found in koalas and several new Chlamydiales genotypes. The bandicoots responded excellently to treatment with oxytetracyline weekly for 6 wk, and topical oxytetracycline and neomycin were administered topically to both eyes s.i.d. for 4 mo.

Analysis of two genomic variants of orang-utan hepadnavirus and their relationship to other primate hepatitis B-like viruses

We recently described orang-utan hepadnavirus (OuHV) (Warren et al., Journal of Virology, 73, 7860-7865, 1999). Phylogenetic analyses indicated that the various isolates of OuHV can be divided into two genomic variants. Two representatives from each genomic cluster were analysed both molecularly and phylogenetically. Their genome organization was highly similar to other hepadnaviruses of apes and humans. The complete genome sequences of the two OuHV types had an overall 5% sequence difference. Research on 25 seropositive Bornean orang-utans showed that, of the 19 animals infected with one variant, 12 originated from East Kalimantan. Phylogenetic analysis was performed using the full-length genomes of various primate hepadnaviruses. The tree topology revealed one cluster of Old World hepadnaviruses that is divided into two subclusters, one consisting of the ape viruses, and the other comprising the human genotypes A-E. These data suggest that the great apes and gibbons have been infected with a common ancestor hepadnavirus.

Cutaneous Papillomatosis and Carcinomatosis in the Western Barred Bandicoot (Perameles bougainville)

A progressive wart-like syndrome in both captive and wild populations of the Western barred bandicoot (WBB) is hindering conservation efforts to prevent the extinction of this endangered marsupial. In this study, 42 WBBs exhibiting the papillomatosis and carcinomatosis syndrome were examined. The disease was characterized by multicentric proliferative lesions involving cutaneous and mucosal surfaces, which were seen clinically to increase in size with time. Grossly and histologically the smaller skin lesions resembled papillomas, whereas the larger lesions were most commonly observed to be squamous cell carcinomas. Large amphophilic intranuclear inclusion bodies were observed in hyperplastic conjunctival lesions of 8 WBBs under light microscopy. Conjunctival lesions from 2 WBBs examined using transmission electron microscopy contained a crystalline array of spherical electrondense particles of 45-nm diameter, within the nucleus of conjunctival epithelial cells, consistent with a papillomavirus or polyomavirus. Conjunctival samples from 3 bandicoots that contained intranuclear inclusion bodies also demonstrated a positive immunohistochemical reaction after indirect immunohis-tochemistry for papillomavirus structural antigens. Ultrastructural and/or immunohistochemical evidence of an etiologic agent was not identified in the nonconjunctival lesions examined. Here we describe the gross, histopathologic, ultrastructural, and immunohistochemical findings of a papillomatosis and carcinomatosis syndrome recently identified in the WBB.

Platynosomum fastosum in ex-captive orangutans from Indonesia.

The liver fluke Platynosomum fastosum was identified upon necropsy of three ex-captive orangutans (Pongo pygmaeus) which had been part of a rehabilitation program for reintroduction to the wild. This trematode has not been reported in orangutans previously and is commonly found in cats in Southeast Asia. Cross infection from cats via intermediate hosts, to orangutans kept in captivity as pets, could explain their presence in the latter. Although P. fastosum caused intrahepatic and bile duct damage, death of the hosts could not be attributed solely to the presence of the liver fluke infection.

Characterization of a simian T-lymphotropic virus from a wild-caught orang-utan (Pongo pygmaeus) from Kalimantan, Indonesia

In a recent serological survey among 143 ex-captive orang-utans two individuals were found that reacted positive in an ELISA detecting antibodies which cross-react with human T-lymphotropic virus type I (HTLV-I) antigens. Infection of both animals with an HTLV-I or simian T-lymphotropic virus (STLV)-like virus was confirmed by Western blot analysis. A third wild-caught animal, which was not part of the original serological survey, was also found to be infected with an HTLV-related virus in a diagnostic PCR assay and Western blot assay. Nucleotide sequence analysis of the 709 bp PCR fragment from the tax/rex region of the HTLV/STLV genome confirmed infection of orang-utans with an STLV similar to but clearly distinct from other Asian STLVs.