Electrophoretic comparison of the genomes of North American bluetongue viruses, one Australian bluetongue virus, and three other related orbiviruses

The genomes of U.S. bluetongue viruses, an Australian bluetongue virus, and three other related orbiviruses were analyzed by polyacrylamide gel electrophoresis. The genomes were comprised of ten segments of double-stranded (ds) RNA. Estimates of the molecular weights of the dsRNA segments revealed that the U.S. bluetongue serotypes were remarkably similar. Although the dsRNA profiles of the viruses exhibited common segments, each virus had a distinct dsRNA profile. The usefulness of the genome analysis as a diagnostic tool for identification and for epidemiologic studies is discussed.     

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 ?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 survey of ruminants in some Caribbean and South American countries for type-specific antibody to bluetongue and epizootic haemorrhagic disease viruses

The results of a serological survey of ruminant livestock in some countries of the Caribbean and South America for type-specific antibody to bluetongue virus are reported. Using the microneutralisation test with the international serotypes 1 to 22 of bluetongue virus, antibodies to several types were detected. Analysis of the data indicated that in 1981-82 bluetongue virus types 6, 14 and 17, or viruses closely related to them, were infecting ruminants in this region of the world. Antibody to the related virus of epizootic haemorrhagic disease (serotype 1) was also detected in cattle. The difficulty in interpreting the epidemiological significance of data generated by a serological survey of this kind is discussed.     

Serogrouping of United States and some African serotypes of bluetongue virus using RT-PCR

The diagnostic potential of RT-PCR for detection of bluetongue virus (BTV) ribonucleic acid (RNA) sequence in cell culture and tissue samples from infected ruminants from United States, Sudan, South Africa and Senegal, was evaluated. The non structural protein 1 (NS1) gene of North American BTV serotype 11 was targeted for PCR amplification. The United States BTV serotypes 2, 10, 11, 13 and 17 and the Sudanese BTV serotypes 1, 2, 4 and 16 and BTV serotype 4 from South Africa and BTV serotype 2 from Senegal were studied. RNAs from all BTV field isolates used in this study, propagated in cell cultures, were detected by the described RT-PCR-based assay. The first specific 790bp BTV PCR products were amplified using a pair of outer primers (BTV1 and BTV2). Specificity of the PCR products was confirmed by a nested amplification of a 520bp PCR product using a pair of internal (nested) primers (BTV3 and BTV4). The BTV PCR products were visualized on ethidium bromide-stained agarose gels. Amplification products were not detected when the RT-PCR-based assay was applied to RNAs from closely related orbiviruses including, epizootic hemorrhagic disease virus (EHDV) prototypes serotypes 1, 2, 4; RNA from Sudanese isolate of palyam orbiviruses serogroup and total nucleic acid extracts from uninfected Vero cells. Application of the nested BTV RT-PCR to clinical samples resulted in amplification of BTV RNA from blood and serum samples from goats experimentally infected with BTV4 and from naturally infected sheep, goats, cattle and deer. The results of this study indicated that this RT-PCR assay could be applied for rapid detection of BTV, in cell culture and clinical samples from susceptible ruminants during an outbreak of the disease, in the United States and African.     

A serological comparison of the australian isolate of bluetongue virus type 20 (CSIRO 19) with bluetongue group viruses

Bluetongue virus serotype 20 (BTV20) (CSIRO 19 isolate) was compared with 17 other BTV serotypes using various serum neutralization (type antigen) tests to determine whether any serological relationships existed. Plaque-reduction neutralization tests employing 50% and 80% end-points could not clearly differentiate BTV20 from BTV4. Plaque-inhibition tests and quantal microtitre neutralization tests also showed a relationship between BTV20 and BTV4. Antisera against BTV20 and a Cyprus isolate of BTV4 (A SOT 1) showed a low level of cross-neutralization against BTV17. Investigation of plaque-reduction neutralization of virus—antiserum mixtures, by the calculation of regression curves and comparison of the area under the curves, showed that the BTV4 isolates studied could not be differentiated, and that BTV4 typing antiserum could not distinguish between BTV4 and BTV20, but that BTV20 antiserum could distinguish between BTV20 and BTV4. BTV20 did not show any significant type relationships with any of the other BTV types 1 to 17 using any of the neutralization tests. Our results suggest that BTV20 is closely related to, although not identical with, BTV4 and could be grouped as a subtype of BTV4. BTV17 appears to be distantly related to BTV20 and BTV4, but is clearly a distinct type.     

Relative brain size and morphology of some South African bats

Measures of relative brain size and brain macromorphology are described for four species of Microchiroptera, two from the Vespertilionidae and two from the Rhinolophidae, and two species from the Pteropodidae (Megachiroptera). Four brain parameters (brain length, hemisphere length, brain width and brain height) were measured, and converted to indices of proportion to allow interspecific comparison. The megachiropteran species have relatively larger brains than the microchiropteran species, this being primarily a result of the greater development of the cerebral hemispheres in the former group. Of the microchiropteran species, Miniopterus schreibersii has the highest value for comparative brain size and its brain shows several characteristics of the more encephalized Megachiroptera such as reduced exposure of the mesencephalic tectum, and the presence of prominent cerebral sulci. The possibility that the differences in brain size and development between the Megachiroptera and the Microchiroptera, may be related to diet, or that they may be of phylogenetic origin, is discussed.     

Bluetongue and related viruses in Nigeria: Experimental infection of West African dwarf sheep with Nigeria strains of the viruses of epizootic haemorrhagic disease of deer and bluetongue

West African dwarf sheep were inoculated by the subcutaneous route with epizootic haemorrhagic disease of deer (EHD) virus or bluetongue (BT) virus. No clinical disease was observed following primary EHD or BT infection, or subsequent challenge with either homologous or heterologous virus. However, viraemia was detected in non-immune sheep exposed to BT virus, but not in BT- or EHD-immune sheep challenged with either virus, or in non-immune sheep exposed to primary EHD virus infection. Complement fixing antibodies developed against both EHD and BT viruses following the primary infection with either virus, or subsequent challenge with homologous or heterologous virus. Following a primary infection, virus-neutralising (VN) antibodies developed only against the inoculated virus, while the detection of VN antibodied to both viruses followed the challenge of an EHD- or BT-immuned sheep with either the homologous or heterologous virus. These findings further support previous reports of a relationship between EHD and BT viruses. between EHD and BT viruses.     

Rapid methods for comparing the double-stranded RNA genome profiles of bluetongue virus

Various double-stranded RNA extraction procedures, gel electrophoresis systems, and methods to detect the RNA bands in the gel were investigated to find the most rapid methods to obtain the genome profiles of bluetongue virus in small volumes (1–25 ml) of infected cell culture fluids. Rapid double-stranded RNA extraction procedures coupled with staining the acrylamide gel slabs with ethidium bromide or silver nitrate resulted in well-defined genome profiles from bluetongue virus infected cell cultures in 6–48 h. Radioactive labelling of viral RNA with ³²P was time consuming, cumbersome and expensive. These techniques detect less than 0.5 μg of double-stranded RNA which can be obtained from one 1-ml well of a 24-well cluster plate of bluetongue virus infected cell monolayers. The methods were therefore suitable for rapid comparisons of the electropherotypes of multiple virus isolates.     

A comparison of some serological tests for bluetongue virus infection.

The plaque neutralization, complement fixation, and agar gel precipitin tests were compared by measuring bluetongue virus antibody in 137 serums from experimental animals (cattle and sheep) and suspected field reactors (cattle and deer). In general, the tests agreed well with each other. Plaque neutralization titers began earlier than the other two and went much higher than the complement fixation titers. Plaque neutralization titers usually peaked between two and three weeks after exposure and complement fixation titers from four to six weeks. The greater sensitivity of the plaque neutralization test allowed the detection of all complement fixation and agar gel precipitin reactors whereas occasionally the latter two tests failed to detect plaque neutralization reactors.     

Serological survey of ruminant livestock in some countries of the Caribbean region and South America for antibody to bluetongue virus

A serological survey of 6250 sera from cattle, sheep and goats in seven Caribbean and two South American countries showed that antibody to bluetongue virus was widely distributed in each species throughout the survey area. Overall prevalences of antibody were 70 per cent in cattle, 67 per cent in sheep and 76 per cent in goats as assessed by an immunodiffusion test. Within countries the percentage prevalences were Jamaica 77, St Kitts/Nevis 70, Antigua 76, St Lucia 82, Barbados 61, Grenada 88, Trinidad and Tobago 79, Guyana 52 and Surinam 84. No clinical cases of bluetongue have been confirmed in the area surveyed and there are no virus isolates available to indicate which serotype(s) of virus is/are causing the infection(s).     

Antigenic relationship of Ibaraki,bluetongue, and epizootic Hemorrhagic disease viruses

Ibaraki virus, which causes a bluetongue-like disease of cattle in Japan, was compared antigenically with the four serotypes of bluetongue virus (BTV) found in the U.S. and with the two serotypes of epizootic hemorrhagic disease virus (EHDV). No antigenic relationship was found between Ibaraki virus and BTV serotypes 10, 11, 13, and 17 in tests for group or serotype-specific antigens. However, Ibaraki virus and EHDV were related antigenically. The agar gel precipitin and indirect fluorescent antibody tests for group antigens showed two-way cross relationships between Ibaraki virus and EHDV serotypes 1 and 2. The more restrictive serotype-specific neutralization test revealed that antigenic relatedness was stronger between Ibaraki virus and the serotype 2 (Alberta strain) of EHDV than between Ibaraki virus and the serotype 1 (New Jersey strain) of EHDV.     

The helminth parasites of various artiodactylids from some South African nature reserves

The helminth species composition and helminth burdens of 4 grey duikers, 12 bushbuck, 2 nyala, 2 giraffe, a steenbok, an oribi, a waterbuck and a tsessebe from the Kruger National Park (KNP); of a steenbok and a greater kudu from the farm Riekerts Laager, Transvaal; of a single blue duiker from the Tsitsikama Forest National Park, and of a blue wildebeest, a red hartebeest, a gemsbok and 2 springbok from the Kalahari Gemsbok National Park (KGNP) were collected, counted and identified. New parasite records are: Agriostomum equidentatum from the gemsbok, Cooperia neitzi from the bushbuck, Cooperia sp. from the gemsbok and the red hartebeest, Cooperia yoshidai from the waterbuck and the tsessebe, Dictyocaulus viviparus from the bushbuck, Haemonchus bedfordi from the waterbuck, Haemonchus contortus from the gemsbok, Haemonchus krugeri from the steenbok from the KNP, Impalaia nudicollis from the gemsbok and the red hartebeest, Impalaia tuberculata from the oribi and the waterbuck, Impalaia spp. from the kudu, Longistrongylus meyeri from the steenbok from Riekerts Laager and the gemsbok, Longistrongylus sabie from the steenbok from the KNP, Longistrongylus schrenki from the tsessebe, Parabronema sp. from the tsessebe and the red hartebeest, Paracooperia serrata from the gemsbok and the steenbok from the KGNP, Pneumostrongylus calcaratus from the bushbuck, Strongyloides sp. from the gemsbok, Trichostrongylus sp. from the gemsbok, the red hartebeest and the steenbok from the KGNP, Trichostrongylus axei from the blue duiker, Trichostrongylus falculatus from the bushbuck and the oribi, Trichostrongylus instabilis from the bushbuck, the steenbok from the KNP and the oribi and Trichostrongylus thomasi from the grey duikers and tsessebe. Host specificity of the parasites was not marked and crossinfestation was common. This was not true for the giraffe, since none of the helminths of these animals were found in the antelope and vice versa.     

Climatic factors associated with the infection of herds of cattle with bluetongue viruses

The incidence of bluetongue virus infection of 15 cattle herds in Queensland, Australia, was determined by a serum neutralization test. The maximum temperature (°C), minimum temperature (°C) and rainfall (mm) data were obtained from the meteorological recording stations closest to each herd. Using unweighted least-squares regression analysis, the best statistical model explaining the most variability in the herd incidence rate included the ratio between the maximum and minimum temperature recorded at both 1 month and 6 months preceding seroconversion, and rainfall recorded at both 2 months and 6 months preceding seroconversion. More than 90% of the variability in the incidence of bluetongue virus infection in the herds was explained by the model, a considerable improvement on previous models that used prevalence data. The prospective nature of the study also supports a strong causal relationship between climatic factors and the occurrence of infection in cattle herds.Abbreviations SN serum neutralization - R _infa ^sup2 adjusted coefficient of multiple determination - AIC Akaike's information criterion - FPE Akaike's final prediction error - PRESS predicted sum of squares