Avian hepevirus infections were detected in chickens suffering from big liver and spleen disease or hepatitis–splenomegaly syndrome in Australia, the USA and Europe. Available data indicate their genetic relationship to mammalian hepatitis E virus (HEV). In the present study, the near-complete genomic sequences of an Australian and a European isolate of avian hepatitis E virus (avian HEV) are reported for the first time. Furthermore, the phylogenetic relationship to other avian HEVs is determined. Sequence analyses of these isolates identified major genetic differences among avian HEVs. Most of them are located within the open reading frame (ORF)1 region, although only a few lie within conserved motifs of predicted domains. Non-silent mutations in the ORF2 region suggest the presence of potentially different epitopes among avian HEV isolates. Finally, phylogenetic analysis confirmed the distant relationship to mammalian HEV and additionally suggested that the avian HEVs can be separated into three different genotypes: 1 (Australia), 2 (USA) and 3 (Europe), indicating a geographical distribution pattern.

Biochemical profiles, restriction endonuclease analysis (REA) and ribotyping were used to investigate Pasteurella multocida isolates from outbreaks of fowl cholera on 7 turkey farms in New South Wales. While only a single isolate was available from 5 of the farms, multiple isolates, 4 and 12 respectively, were available from the other 2 farms. The available field evidence suggested that 8 outbreaks had occurred with one farm suffering 2 outbreaks. The isolates obtained were all confirmed as Pasteurella multocida. Biochemical profiles allocated the isolates to 4 groups, 3 being variants of P multocida subsp multocida and the fourth being P multocida subsp septica. REA performed with HpaII established 7 groups. Ribotyping using the HpaII digests probed with the 16S rRNA operon of Haemophilus paragallinarum recognised the same 7 groups as REA. Unlike the biochemical profiles, both REA and ribotyping provided a fine subdivision that identified outbreaks as either related or unrelated. The REA and ribotyping patterns as well as biochemical profiles were stable for all isolates from the outbreaks in which multiple isolates were obtained from either the same bird or from different birds. REA and ribotyping were found to be superior to biotyping methods for the investigation of fowl cholera outbreaks.

In broiler breeder flocks in one broiler integration in Hungary, a new syndrome appeared in January 2005 with initially four successive post-peak flocks experiencing significant decreases in egg production. Clinically birds became depressed and there was a small increase in the mortality rate. Postmortem examinations revealed enlarged livers in up to 19% of birds dying, and enlarged spleens in some. Also observed were birds with either clotted blood or serosanguineous fluid in the abdomen and subcapsular haemorrhages of the liver. Histopathology and polymerase chain reaction excluded tumours and the presence of common tumour-associated viruses. Chronic bacterial infections (especially causing hepatitis, peritonitis and airsacculitis) were common but many enlarged livers had no obvious bacterial involvement. After a 9-month period during which a majority of flocks became affected, no newly affected flocks occurred. Investigations showed that all tested affected flocks were seropositive in the big liver and spleen (BLS) Agar Gel Immuno Diffusion (AGID) test. Subsequent flocks without post-peak egg-production drops were shown to be seronegative in the BLS AGID test, as were all the parent flocks contributing to the affected flocks. Liver samples and cloacal swabs were positive by polymerase chain reaction (aHEV helicase target), and calicivirus-like particles were demonstrated in bile samples from affected birds. These observations are similar to hepatitis-splenomegaly syndrome as described in North America and BLS syndrome as described in Australia. Histopathological features were a non-specific chronic hepatitis similar to those described in BLS and hepatitis-splenomegaly syndrome. Immunohistochemistry using a BLS-specific monoclonal antibody confirmed the presence of avian hepatitis E virus antigen in livers and spleen.

An Australian field isolate of Mycoplasma synoviae (MS), 89079/7NS, was exposed to the mutagen N-nitro-N'-methyl-N-nitrosoguanidine. Fifteen clones from the exposed culture were characterized for temperature sensitivity. Four clones labelled B, D, G, and H were temperature sensitive and were further characterized for their ability to colonize chickens and elicit an immune response. Serum antibody responses to MS were detected 3 wk after infection, by eyedrop, in 10 of 10 birds inoculated with 86079/7NS and clones B and G and in 9 of 10 birds inoculated with clone H. No MS antibody response was observed in any bird inoculated with clone D. MS was recovered from the upper trachea of 10 of 10 birds inoculated with clones B, G, and H at 2 wk after infection. No MS was isolated from birds inoculated with clone D. Clone H, designated MS-H, was selected as a potential vaccine candidate.

Standardized challenge viruses are essential for the evaluation of Marek's disease (MD) vaccines with many MD challenge preparations consisting of lymphocytes or whole blood from infected birds. Virus present in these preparations is difficult to quantify by tissue culture assays and, therefore, the infectious bird dose and long-term storage viability cannot be assured. We report on the properties of two low-passage virulent Australian MD viruses, the Woodlands No. 1 strain and strain MPF 57. Both strains were isolated in chicken embryo kidney cultures and adapted to grow in chicken embryo fibroblast cultures for a maximum of 14 passages. Both strains could be readily assayed in tissue culture and produced titres of 10 3 to 10 4 50% tissue culture infectious doses per ml (TCID 50 ). Birds inoculated at three different doses were observed over 10 weeks, and tissues examined for gross and histological lesions, bursarbody weight ratios and the presence of viraemia. Tissue culture-grown preparations of both strains were only slightly less virulent than the original lymphocyte challenge material and produced similar pathological responses and around 80% death or gross lesions. From bursarbody weight ratios strain MPF 57 appeared to be more virulent than the Woodlands No. 1 strain.

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A diverse group of Mycoplasma synoviae strains from various hosts, pathological processes, and geographic areas collected over about 25 years were analyzed by restriction endonuclease analysis. The results suggest that restriction endonuclease analysis of M. synoviae strains may be a useful strain identification tool to study epidemiological problems.