D. H. Cybinski

Regulation of Expression of Genes Encoding Phosphate Transporters in Barley Roots

A family of genes that encode phosphate transporters has been isolated from the barley genome. Three of these genes, HVPT1, HVPT2 and HVPT3 are expressed in the roots. The polypeptides encoded by HVPT1, IIVPT2 and HVPT3 are similar in structure and show significant homology to phosphate transporters that have been isolated from dicotyledonous species. There is a high level of homology between HVPT1 and HVPT2 and these two genes are closely linked, lying on the same 20kB fragment of the genome. Although homologous in conserved regions and having a similar topology to HVPT1 and HVPT2, the polypeptide encoded by HVPT3 differs significantly from HVPT1 and HVPT2.

Serological studies of Australian and Papua New Guinean cattle and Australian sheep for the presence of antibodies against bluetongue group viruses

Following isolation of a virus (CSIRO19) from insects in Australia and its identification as bluetongue virus serotype 20 (BTV20), a nationwide survey of antibodies in cattle and sheep sera was undertaken. Initial studies using the serum neutralization (SN) test showed that the distribution of BTV20 antibodies in cattle was confined to the northern part of Australia. Group-reactive antibody tests (agar gel diffusion precipitin, AGDP, and complement-fixation, CF) showed group-reactive cattle sera south of the BTV20 zone (northern Australia), and southwards from Queensland to New South Wales. Very few group-reactive sheep sera (45 out of 16213) were found and these were of doubtful epidemiological significance. Some of these BTV group-reactive, BTV20-negative, sera were tested in SN tests against BTV1 to 17 and Ibaraki (IBA) virus. The results indicated that BTV1, or a closely related orbivirus, was active in cattle in Queensland, northern Western Australia, and New South Wales, and that antibody to BTV15 was present in some of the cattle sera in northern Western Australia and the Northern Territory. Antibody to IBA virus was present in some cattle sera in Queensland, northern Western Australia and New South Wales. SN antibody titres greater than or equal to 60 were also found to a number of other BTV serotypes in cattle sera in northern Western Australia and Queensland (principally, BTV2 and BTV7). Low level reactions were commonly observed against these and a number of other BTV serotypes, often in the same serum samples. Further, 22% of the group-reactive cattle sera did not react with any of the viruses in the SN tests. Such results were difficult to interpret in terms of known Australian BTV or BTV-related isolates.

Serological and biochemical factors in bovine ephemeral fever

Clinical signs of ephemeral fever, which were observed in individual cattle during two successive epidemics in 1973 and 1976, were related to biochemical, cellular and serological changes in the blood. The rise in peripheral blood neutrophil counts in samples collected from 12 sentinel cattle on a daily basis before, during and after natural disease in the two epidemics to mean peaks of 9.6-12.5 X 10(9) per litre, and fall in counts of lymphocytes to a trough of 5-7 X 10(9) per litre was found to occur on the same day as the fever peak. A fall in serum calcium levels from a normal mean of 2.55 mmol/l to 2.0 mmol/l occurred on the day clinical signs were most pronounced. Serum magnesium levels were affected to only a minor degree. Plasma fibrinogen rose from a normal mean of 5.0 milligrams to a peak of 18 milligrams on the second day of disease and fell towards normal in the week after recovery. Neutralizing antibodies to bovine ephemeral fever virus were detected up to 63 days prior to clinical disease, and the rise of antibody after recovery was secondary in pattern. Serological evidence of a prior infection with an antigenically related virus, Kimberley virus, was found in these animals. In more severe clinical cases of ephemeral fever serum calcium levels were as low as 1.95 mmol/l. Treatment of cattle showing clinical signs of the disease with phenylbutazone and calcium borogluconate was favourable.

Antigenic variation of the bovine ephemeral fever virus glycoprotein

SummaryGlycoprotein-specific monoclonal antibodies (MAbs) were used to select escape mutants of bovine ephemeral fever (BEF) virus to determine the escape frequency for different epitopes and to construct an epitope map. At least six antigenic sites were detected by this method and escape frequencies between 10−2 and 10−8 were recorded. One new non-conformational site was defined by a MAb, 5A5, which neutralized Berrimah and Kimberley viruses as well as three BEF virus strains. Batch to batch variation was detected in the BB7721 strain of BEF virus when tested for MAb neutralization. Eighteen strains of BEF virus, isolated from blood and insects from a variety of locations in Australia over a period of 33 years, were examined using MAbs and at least one epitope could not be detected in strains isolated since 1975. Implications for vaccine development are discussed.

Serological studies of two additional Australian blue-tongue virus isolates CSIRO 154 and CSIRO 156

Abstract Serological surveys revealed that some cattle in northern Australia possessed bluetongue virus (BTV) group-reactive (agar gel diffusion precipitin, AGDP, and complement-fixing, CF) antibodies, but not serum neutralizing (SN) antibodies, to BTV20, a new type previously found in Australia. Attempts were made during 1979 to isolate viruses causing these reactions. There was one isolate of a virus (CSIRO 154) and eight isolates of another virus (CSIRO 156) made from the blood of healthy cattle in the Northern Territory. These viruses could not be distinguished from BTV20 by AGDP, CF or fluorescent-abtibody tests and hence were designated members of the bluetongue serogroup. Serotyping was carried out using the plaque-inhibition and plaque-reduction SN tests. CSIRO 156 virus could not be distinguished from BTV1 by any of the SN tests and it was concluded that it was an Australian isolate of the BTV1 serotype. CSIRO 154 virus was found to be related to, but not identical with, BTV6. It is probably not one of the known 20 BTV serotypes and may represent a new BTV serotype. None of the three Australian BTV isolates is known to cause clinical disease in sheep or cattle under natural conditions, and biochemical comparisons with the African BTV serotypes may show differences not revealed by these serological studies.

Homologous and heterologous antibody reactions in sera from cattle naturally infected with bovine ephemeral fever group viruses

Four viruses belonging to the bovine ephemeral fever (BEF) group have been isolated from bovine blood. Infection of cattle with BEF virus was associated with neutralizing antibody responses to BEF, Kimberley (KIM), Berrimah (BRH) and Adelaide River (ADE) viruses, with highest antibody titres to BEF and KIM viruses. Infection of one cow with KIM virus was associated with a homologous neutralizing antibody response and nil or minimal responses to the other three viruses. Infection of a steer with ADE virus was associated with a rise in neutralizing antibody levels to ADE virus and to KIM virus, but not to BEF or BRH viruses. Infection of a steer with BRH virus was associated with marked neutralizing antibody rises to BRH and BEF viruses and small rises to KIM and ADE viruses. An antibody rise to BEF virus did not necessarily indicate recent BEF virus infection, and should be considered of diagnostic value only when taken in conjunction with clinical signs of disease.

Antibodies against certain bluetongue and epizootic haemorrhagic disease viral serotypes in Indonesian ruminants.

The orbiviruses contain several important viruses of livestock including bluetongue (BT) and epizootic haemorrhagic disease of deer (EHD) which share some group antigens. Preliminary screening of sera for antibodies to orbiviruses by the agar gel immunodiffusion (AGID) test has previously revealed widespread infections with the BT group in Indonesia. However serum neutralization (SN) tests give a more accurate estimate of exposure to each serotype in the BT and EHD groups, and in this study were applied to sera that had reacted previously in the AGID test. Five different serotypes of BT and one serotype of EHD virus were studied. Reactors to BT serotype 20 were the most prevalent, followed by EHD type 5 and BT types 21, 12, 1 and 17. Antibodies against BT serotype 20 were present in cattle, buffaloes, goats and sheep, but were most common in buffaloes. Buffaloes showed the highest exposure to the BT serotypes tested. Antibody to EHD type 5 occurred most frequently in cattle. Antibodies against all BT and EHD serotypes tested were found in buffaloes and cattle while goats had antibodies against BT types 20, 21 and EHD type 5 and sheep had antibodies only against BT type 20.

Isolation of Tibrogargan virus, a new Australian rhabdovirus, from Culicoides brevitarsis.

CSIRO 132 virus, which is new to science in Australia, and probably the world, has been isolated from Culicoides brevitarsis. Electron micrographs show that it resembles a rhabdovirus. Antibodies to the new virus have been detected in water buffaloes and cattle, but not in 58 human beings, 14 camels, 21 dogs, 67 goats, 15 horses, 43 pigs, 154 sheep, 98 wallabies or 38 possums. The distribution of antibodies in cattle lies within the distribution range of C. brevitarsis. It has not so far been associated with disease. The name Tibrogargan is proposed for the new virus.

Seroepidemiology of arboviruses among seabirds and island residents of the Great Barrier Reef and Coral Sea

Duplicate neutralization tests were done on 401 avian and 101 human sera from island residents collected in the Coral Sea and on Australias Great Barrier Reef against 19 known arboviruses. Antibodies to a potentially harmful flavivirus, Gadgets Gully virus, were equally present (4%) in both avian and human sera. Antibodies to another flavivirus, Murray Valley Encephalitis, and an ungrouped isolate, CSIRO 1499, were also present in both populations with non-significantly different incidences. Antibodies to Upolu, Johnston Atoll, Lake Clarendon, Taggert, Saumarez Reef and CSIRO 264 viruses were restricted to seabirds. Island residents with antibodies to Ross River and Barmah Forest viruses are thought to have been exposed to these viruses on the mainland as antibody to both viruses was absent among seabirds. These results indicate that consideration should be given to tick-associated arboviruses as potential public health hazards on islands where both seabird and human activities interact.

Genomic Characterisation of Vinegar Hill Virus, An Australian Nairovirus Isolated in 1983 from Argas Robertsi Ticks Collected from Cattle Egrets

This report describes the near complete genomic sequence and subsequent analysis of Vinegar Hill virus (VINHV; tentative member of the genus Orthonairovirus, family Nairoviridae, order Bunyavirales). VINHV is the second nairovirus reported to be isolated on mainland Australia and the first to be sequenced and analysed. Our genetic analysis shows that VINHV belongs to the Dera Ghazi Khan genogroup, a group of viruses previously isolated in other parts of the world including Asia, South Africa, and the USA. We discuss possible routes of entry for nairoviruses into Australia and the need to understand the virome of Australian ticks in the context of new and emerging disease.