Widespread Reassortment Contributes to Antigenic Shift in Bluetongue Viruses from South Africa.

Bluetongue (BT), a viral disease of ruminants, is endemic throughout South Africa, where outbreaks of different serotypes occur. The predominant serotypes can differ annually due to herd immunity provided by annual vaccinations using a live attenuated vaccine (LAV). This has led to both wild-type and vaccine strains co-circulating in the field, potentially leading to novel viral strains due to reassortment and recombination. Little is known about the molecular evolution of the virus in the field in South Africa. The purpose of this study was to investigate the genetic diversity of field strains of BTV in South Africa and to provide an initial assessment of the evolutionary processes shaping BTV genetic diversity in the field. Complete genomes of 35 field viruses belonging to 11 serotypes, collected from different regions of the country between 2011 and 2017, were sequenced. The sequences were phylogenetically analysed in relation to all the BTV sequences available from GenBank, including the LAVs and reference strains, resulting in the analyses and reassortment detection of 305 BTVs. Phylogenomic analysis indicated a geographical selection of the genome segments, irrespective of the serotype. Based on the initial assessment of the current genomic clades that circulate in South Africa, the selection for specific clades is prevalent in directing genome segment reassortment, which seems to exclude the vaccine strains and in multiple cases involves Segment-2 resulting in antigenic shift.

Safety and efficacy of Rift Valley fever Smithburn and Clone 13 vaccines in calves.

Two modified live attenuated vaccines against the disease Rift Valley fever (RVF) have been tested for safety and efficacy in young calves. The RVF Smithburn vaccine produced in South Africa and used successfully to prevent and control the disease in endemic sub-Saharan countries was compared to the candidate vaccine RVF Clone 13. Five sero-negative calves per vaccine group were vaccinated with a single dose of each vaccine and tested for antibody response. All vaccinated calves were challenged with a highly virulent RVF virus together with five unvaccinated calves used as control of the challenge. Protection was confirmed in all vaccinated animals as they did not show any clinical signs typical of RVF. A good neutralizing antibody response was induced post-vaccination and no viraemia could be detected post-challenge in calves of both vaccine groups. All non-vaccinated control animals showed clinical symptoms of RVF, high viraemia and were euthanized. This study reported the first case of blindness in cattle resulting from virulent RVF virus infection in unvaccinated calves used as negative controls.

In vivo cross-protection to African horse sickness Serotypes 5 and 9 after vaccination with Serotypes 8 and 6.

The polyvalent African horsesickness (AHS) attenuated live virus (AHS-ALV) vaccine produced at Onderstepoort Biological Products incorporates 7 of the 9 known serotypes circulating in southern Africa. Serological cross-reaction has been shown in vitro to Serotypes 5 and 9 by Serotypes 8 and 6 respectively, but the degree of in vivo cross-protection between these serotypes in vaccinated horses has not previously been reported. Due to the increasing incidence of AHS Serotypes 5 and 9 in the field, over the last 3-4 seasons of AHS in South Africa, and the absence of Serotypes 5 and 9 in the AHS-ALV vaccine, it was necessary to conduct a vaccination-challenge study to determine in vivo cross-protection of vaccine-incorporated Serotypes 8 and 6 respectively. Groups of horses were vaccinated with either the polyvalent AHS-ALV vaccine or a monovalent Serotype 6 (vAHSV6) or 8 (vAHSV8) vaccine to determine the cross-protection of vaccinated horses following challenge with virulent AHS virus (AHSV) of either Serotype 5, 6, 8 or 9. Serial vaccination of naive horses with the polyvalent AHS-ALV vaccine generated a broad neutralizing antibody response to all vaccine strains as well as cross-neutralizing antibodies to Serotypes 5 and 9. Booster vaccination of horses with monovalent vaccine vAHSV6 or vAHSV8 induced an adequate protective immune response to challenge with homologous and heterologous virulent virus. In vivo cross-protection between AHSV6 and AHSV9 and AHSV8 and AHSV5 respectively, was demonstrated.

Evaluation of the efficacy and safety of the Rift Valley Fever Clone 13 vaccine in sheep.

The efficacy and safety of the naturally attenuated Rift Valley Fever (RVF) Clone 13 vaccine were evaluated in ovines in three different experiments involving 38 ewes at different stages of pregnancy, their offsprings and four rams. In Experiment 1, 4 rams and a total of 13 pregnant ewes were vaccinated and monitored during vaccination and after a challenge with a virulent RVF virus. The ewes were vaccinated at either 50 or 100 days of pregnancy and some were challenged after lambing. In Experiment 2, nine oestrus-synchronized ewes were vaccinated at 50 days of pregnancy and challenged at 100 days of pregnancy together with 5 unvaccinated ewes at the same stage of pregnancy. In Experiment 3, 16 oestrus-synchronized ewes were vaccinated with 3 different doses of the RVF Clone 13 vaccine and challenged together with unvaccinated pregnant ewes at either 30 or 50 days of pregnancy. The results from the three experiments indicated that the vaccine did not induce clinical manifestation of RVF such as abortion in pregnant ewes, teratogeny in their offsprings, or pyrexia in all vaccinated animals. Vaccination with RVF Clone 13 vaccine also prevented clinical RVF following virulent challenge at different stages of pregnancy while unvaccinated control ewes showed pyrexia, aborted or died of RVF. A vaccine dose-response effect was also observed.

Evaluation of the pathogenicity of African Horsesickness (AHS) isolates in vaccinated animals.

BACKGROUND: The polyvalent African Horsesickness (AHS) attenuated live vaccine (ALV) produced by Onderstepoort Biological Products (OBP) Ltd., South Africa, has been associated with some safety concerns and alleged cases of vaccine failure or vaccine-induced disease. The risk of reassortment and reversion to virulence is a common concern associated with the use of ALVs, and a phenomenon reported for viruses with segmented RNA genomes. The purpose of this study was to determine whether or not reassortment of AHS vaccine strains could result in reassortants and reversion to virulence and therefore cause AHS in susceptible horses. METHODS: Clinical or field isolates of AHS were obtained from horses with AHS symptoms or disease post vaccination. AHS-naïve horses were inoculated with these isolates and monitored for clinical reactions. Laboratory tests were performed at intervals to determine immune responses and viraemia. Viral RNA extraction and complete genome amplification of monovalent AHS-ALV vaccine strains and isolates collected post-vaccination was conducted. cDNA of the genome segments were run on PAGE to determine mobility patterns and genome segments 2, 3, 4, 5 and 6 sequenced for phylogenetic analysis. RESULTS: No clinical symptoms typical of AHS were observed in inoculated horses and all showed a good immune response. A comparison of mobility patterns of the amplified cDNA genome on PAGE allowed the identification and differentiation of reassortants, which were confirmed by sequence and phylogenetic analysis of the nucleotide sequences. CONCLUSION: This study, however, showed no indications that vaccine reassortants were pathogenic or lethal after inoculation in susceptible horses. Assumptions of virulence or reversion to virulence of vaccine reassortants post-vaccination in horses could not be substantiated.