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Bovine Ephemeral Fever (BEF) is an arthropod-borne virus (arbovirus) that presents a significant challenge to the cattle industry due to its economic impact, primarily through the loss of milk production in dairy cows. Vaccination is the predominant strategy for managing the disease. We recently showed a vaccine effectiveness (VE) of 60 % of a vaccine based on the Australian 919 BEFV isolate, with a natural challenge occurring shortly after the administration of the second dose of the vaccine. Still, there is a lack of data regarding the duration of protective immunity after vaccination and its potential enhancement after the administration of three and four vaccine doses. To answer these questions we conducted a retrospective cohort study of 850 cows (7 herds), analyzing the influence of different vaccination regimens on VE and a serosurvey of 71 cows to test the longevity of BEFV-specific serum-neutralizing antibodies (SNAb). We adopted a quantitative methodology for BEF diagnosis with the use of commercially validated precision dairy monitoring technologies for milk reduction identification. Survival analysis was used to analyze the vaccine dose effectiveness. A Cox regression mixed-effect model (COXME) was fitted to the data. The analysis demonstrated the following VE compared to zero vaccine doses: 82 % (p-value<0.001) for four doses, 66 % (p-value<0.026) for three doses and 39 % (p-value = 0.3) for two doses. Corroborating with the VE results, the four-dose regimen exhibited the highest geometric mean titer (GMT) value (4.45, CI95% = 3.99, 4.91), followed by the three-dose regimen (3.53, CI95% = 3.08,3.98), and the two-dose regimen (2.17, CI95% = 1.77,2.57). In light of these findings, we recommend vaccinating calves as early as four to six months old with two doses spaced one month apart, followed by a third and even fourth dose administered between six to 12 months later, ideally close to the onset of the high-risk season. •Bovine ephemeral fever (BEF) is a viral disease, causing vast losses worldwide•Long-term effectiveness of the 919 attenuated BEF virus vaccine is not known•Dose-dependent vaccine effectiveness (VE) data is lacking•This retrospective cohort study shows a significant dose-dependent increase in VE•VE results corroborate with a similar dose-dependent increase in antibody response

Bovine ephemeral fever (BEF) is caused by the arthropod-borne bovine ephemeral fever virus (BEFV), which is a member of the family Rhabdoviridae and the genus Ephemerovirus . BEFV causes an acute febrile infection in cattle and water buffalo. In this study, a recombinant Newcastle disease virus (NDV) expressing the glycoprotein (G) of BEFV (rL-BEFV-G) was constructed, and its biological characteristics in vitro and in vivo , pathogenicity, and immune response in mice and cattle were evaluated. BEFV G enabled NDV to spread from cell to cell. rL-BEFV-G remained nonvirulent in poultry and mice compared with vector LaSota virus. rL-BEFV-G triggered a high titer of neutralizing antibodies against BEFV in mice and cattle. These results suggest that rL-BEFV-G might be a suitable candidate vaccine against BEF.

Background Bovine ephemeral fever virus (BEFV), the causative agent of bovine ephemeral fever, is an economically important pathogen of cattle and water buffalo. MicroRNAs (miRNAs) are endogenous 21-23 nt small non-coding RNA molecules that binding to a multiple of target mRNAs and functioning in the regulation of viral replication including the miRNA-mediated antiviral defense. However, the reciprocal interaction between bovine ephemeral fever virus replication and host miRNAs still remain poorly understood. The aim of our study herein was to investigate the exact function of miR-3470b and its molecular mechanisms during BEFV infection. Results In this study, we found a set of microRNAs induced by BEFV infection using small RNA deep sequencing, and further identified BEFV infection could significantly up-regulate the miR-3470b expression in Baby Hamster Syrian Kidney cells (BHK-21) after 24 h and 48 h post-infection (pi) compared to normal BHK-21 cells without BEFV infection. Additionally, the target association between miR-3470b and mitochondrial antiviral signaling protein (MAVS) was predicted by target gene prediction tools and further validated using a dual-luciferase reporter assay, and the expression of MAVS mRNA and protein levels was negatively associated with miR-3470b levels. Furthermore, the miR-3470b mimic transfection significantly contributed to increase the BEFV N mRNA, G protein level and viral titer, respectively, whereas the miR-3470b inhibitor had the opposite effect on BEFV replication. Moreover, the overexpression of MAVS or silencing of miR-3470b by its inhibitors suppressed BEFV replication, and knockdown of MAVS by small interfering RNA also promoted the replication of BEFV. Conclusions Our findings is the first to reveal that miR-3470b as a novel host factor regulates BEFV replication via directly targeting the MAVS gene in BHK-21 cells and may provide a potential strategy for developing effective antiviral therapy.

Recombinant baculoviruses expressing the BEFV envelope glycoprotein G and non-structural glycoprotein G NS were constructed. The G protein expressed in insect cells was located on the cell surface and induced spontaneous cell fusion at mildly acidic pH. The expressed G protein reacted with MAbs to continuous and conformational neutralization sites (G1, G2, G3b and G4), but not to conformational site G3a. The expressed G NS protein was also located on the cell surface but did not exhibit fusogenic activity. The G NS protein reacted with polyclonal antiserum produced from vaccinia-virus-expressed recombinant G NS but did not react with G protein antibodies. A His 6 -tagged, soluble form of the G protein was expressed and purified by Ni 2+ –NTA chromatography. The purified G protein reacted with BEFV-neutralizing MAbs to all continuous and conformational antigenic sites. The highly protective characteristics of the native BEFV G protein suggest that the secreted, baculovirus-expressed product may be a useful vaccine antigen.

Bovine ephemeral fever (BEF) is an economically important viral vector-borne cattle disease. Several live-attenuated, inactivated and recombinant vaccines have been tested, demonstrating varying efficacy. However, to the best of our knowledge, duration of immunity conferred by an inactivated vaccine has never been reported. In the last decade, Israel has faced an increasing number of BEF outbreaks. The need for an effective vaccine compatible with strains circulating in the Middle East region led to the development of a MONTANIDE™ ISA 206 VG (water-in-oil-in-water), inactivated vaccine based on a local strain. We tested the safety, immunogenicity and duration of immunity conferred by this vaccine. The induced neutralizing antibody (NA) response was followed for 493 days in 40 cows vaccinated by different protocols. The vaccine did not cause adverse reactions or a decrease in milk production. All cows [except 2 (6.7%) which did not respond to vaccination] showed a significant rise in NA titer of up to 1:256 following the second, third or fourth booster vaccination. Neutralizing antibody levels declined gradually to 1:16 up to 120 days post vaccination. This decline continued in cows vaccinated only twice, whereas cows vaccinated 3 or 4 times showed stable titers of approximately 1:16 for up to 267 days post vaccination. At least three vaccinations with the inactivated BEF vaccine were needed to confer long-lasting immunity. These results may have significant implications for the choice of vaccination protocol with inactivated BEF vaccines. Complementary challenge data should however be added to the above results in order to determine what is the minimal NA response conferring protection from clinical disease.

Bovine ephemeral fever (or 3-day sickness) is an acute febrile illness of cattle and water buffaloes. Caused by an arthropod-borne rhabdovirus, bovine ephemeral fever virus (BEFV), the disease occurs seasonally over a vast expanse of the globe encompassing much of Africa, the Middle East, Asia and Australia. Although mortality rates are typically low, infection prevalence and morbidity rates during outbreaks are often very high, causing serious economic impacts through loss of milk production, poor cattle condition at sale and loss of traction power at harvest. There are also significant impacts on trade to regions in which the disease does not occur, including the Americas and most of Europe. In recent years, unusually severe outbreaks of bovine ephemeral fever have been reported from several regions in Asia and the Middle East, with mortality rates through disease or culling in excess of 10–20%. There are also concerns that, like other vector-borne diseases of livestock, the geographic distribution of bovine ephemeral fever could expand into regions that have historically been free of the disease. Here, we review current knowledge of the virus, including its molecular and antigenic structure, and the epidemiology of the disease across its entire geographic range. We also discuss the effectiveness of vaccination and other strategies to prevent or control infection.

Bovine ephemeral fever virus (BEFV) is an arthropod-borne rhabdovirus that causes disabling clinical signs and major economic losses in cattle and water buffalo. The disease is well documented in Asia, Africa, and the Middle East; however, the seroprevalence of BEFV in different regions and bovine breeds in the Kingdom of Saudi Arabia (KSA) is unknown. The aim of this study was to analyze risk factors which affect the prevalence of antibodies against BEFV in small herds of cattle in four geographical regions of KSA. A total of 1480 serum samples from non-BEFV vaccinated small herds of cattle were collected from the Eastern, Jizan, Qasim, and Riyadh regions (370 samples per region) during the summer of 2010. Serum neutralization test was used to detect antibodies against BEFV. There was a significant effect of region, breed, sex, and age on the seroprevalence of BEFV. Seropositive ratios were 18, 18, 26, and 12 % for the Eastern, Jizan, Qasim, and Riyadh regions, respectively (P = 0.00002); 23.2 % for dairy and 13.7 % for non-dairy breeds (P = 0.00004); 24.4 % for males and 14.6 % for females (P = 0.00004); and 15.4, 29.1, and 11.4 % for animals <1 year, 1–3 years, and >3 years, respectively (P < 0.001). Risk analysis showed a significant effect of different regions of KSA on the seroprevalence of BEFV. Host risk factors (age, sex, and breed) showed also a significant effect on the seroprevalence of BEFV. This indicates active circulation of this virus in small herds of cattle. Insect control strategies and BEFV vaccination programs during the spring are recommended to reduce the spread of BEFV and minimize subsequent economic losses as this is adopted in many enzootic countries.

The aim of this study was to investigate the immunogenicity of a plasmid deoxyribonucleic acid (DNA) vaccine encoding the G1 epitope of bovine ephemeral fever virus (BEFV) G glycoprotein in mice. A plasmid DNA carrying the G1 gene was constructed and designated as pcDNA3.1-G1. The expression of the target gene was confirmed in human embryonic kidney 293 (HEK 293) cells transfected with pcDNA3.1-G1 by indirect immunofluorescent staining. Immunisation experiments were intramuscularly carried out by vaccinating 6-week-old female mice in four groups, including the pcDNA3.1-G1 construct, pcDNA3.1 (+) plasmid alone, BEF-inactivated vaccine and phosphate-buffered saline (PBS) (1X) three times with 2-week intervals. Fourteen days after the last immunisation, the animals were bled and the resulting sera were tested for anti-G1-specific antibodies by immunoblotting analysis, indirect enzyme-linked immunosorbent assay (ELISA) and virus neutralisation (VN) test. Serological assays showed that the pcDNA3.1-G1 construct expressing G1 protein was able to elicit specific antibodies against this antigen. Virus neutralisation test showed that pcDNA3.1-G1 could induce anti-BEFV-neutralising antibodies in mice. Our findings indicated that a new dimension can be added to vaccine studies for bovine ephemeral fever (BEF) using eukaryotic expression plasmids encoding the G1 antigen in the future.

The South African vaccine strain of lumpy skin disease virus (type SA-Neethling) is currently being developed as a vector for recombinant vaccines of economically important livestock diseases throughout Africa. In this study, the feasibility of using the viral thymidine kinase gene as the site of insertion was investigated and recombinant viruses were evaluated in animal trials. Two separate recombinants were generated and selected for homogeneity expressing either the structural glycoprotein gene of bovine ephemeral fever virus (BEFV) or the two structural glycoprotein genes of Rift Valley fever virus (RVFV). Both recombinants incorporate the enhanced green fluorescent protein (EGFP) as a visual marker and the Escherichia coli guanine phosphoribosyl transferase (gpt) gene for dominant positive selection. The LSDV-RVFV recombinant construct (rLSDV-RVFV) protected mice against virulent RVFV challenge. In a small-scale BEFV-challenge cattle trial the rLSDV-BEFV construct failed to fully protect the cattle against virulent challenge, although both a humoral and cellular BEFV-specific immune response was elicited.

The prevalence of antibody to the viruses of bovine virus diarrhoea (BVD), bovine herpes virus type1 (BHV1), rift valley fever (RVF), bovine ephemeral fever (BEF) and bluetongue (BT) and to Leptospira sp. was determined in wildlife populations in Zimbabwe. Evidence of infection with BVD virus was found in 14 of the 16 species examined but was greatest in eland Taurotragus oryx, nyala Tragelaphus angasi and bushbuck Tragelaphus scriptus. Persistent infection with BVD virus was found in 1 of 303 antibody-free eland but not in the smaller sample of 102 antibody-free buffalo Syncerus caffer. Antibody to BHV1 was widespread, being found in 10 of 16 species with the highest prevalence being in buffalo and eland. Antibody to RVF was most prevalent in black rhino Diceros bicornis and white rhino Ceratotherium simum, buffalo and waterbuck Kobus ellipsiprymnus. Both BEF and BT were widespread in all the species examined. Evidence of infection with Leptospira sp. was found in 7 species. Infections were due to up to 3 of 8 different serovars.