Draft genome sequence of Moroccan camelpox vaccine strain.

Prevention and control of camelpox can be achieved by efficient vaccination. A limited number of homologous attenuated vaccines have been commercialized. In this study, we report the draft genome sequence of camelpox virus vaccine strain "CAMPOX vaccine" after 175 passages of attenuation in Vero cells.

The prevalence, virulence, and serogroups of Dichelobacter nodosus and prevalence of Fusobacterium necrophorum in footrot lesions of sheep and cattle in Morocco.

Background and Aim: Footrot is a contagious disease of ruminants leading to severe economic losses. This study aimed to estimate the prevalence, virulence, and serogroups of Dichelobacter nodosus and the prevalence of Fusobacterium necrophorum in footrot lesions of sheep and cattle. Materials and Methods: A total of 106 pathogenic lesion samples were taken from 74 sheep and 32 cattle exhibiting typical footrot lesions and were analyzed for the presence of D. nodosus and F. necrophorum by real-time polymerase chain reaction (PCR). Both virulence and serogroup were estimated for D. nodosus positive samples. Results: Among the 106 samples, 89 were positive by PCR for F. necrophorum, D. nodosus, or both. Dichelobacter nodosus was detected at a rate of 78.3% versus 28.3% for F. necrophorum. Virulent D. nodosus strains were detected in 67.5% of positive samples, with a higher rate in sheep (73.4%) than in cattle (47.4%). Benign D. nodosus strains were detected in 57.8% of samples, with a lower prevalence rate in sheep (50%) than in cattle (84.2%). The positive samples of D. nodosus revealed the presence of three dominant serogroups (D, H, I) and three minor serogroups (G, C, A) by serogroup-specific multiplex PCR. Conclusion: The findings provided information on the prevalence of D. nodosus and F. necrophorum strains in footrot lesions of sheep and cattle in some regions of Morocco, which will be useful for developing an effective autovaccine for the prevention of this disease in cattle and sheep in these regions.

Long term immunity against Peste Des Petits Ruminants mediated by a recombinant Newcastle disease virus vaccine.

Peste des Petits Ruminants (PPR) is a highly contagious and often fatal disease of sheep and goats. Conventional live vaccines have been successfully used in endemic countries however, there are not completely safe and not allowing differentiation between vaccinated and infected animals (DIVA). In this study, a recombinant Newcastle disease virus (NDV) expressing the hemagglutinin of PPRV (NDV-PPRVH) was evaluated on small ruminants by serology response in sheep and goats, experimental infection in goats and immunity duration in sheep. The NDV-PPRVH vaccine injected twice at 28 days' interval, provided full protection against challenge with a virulent PPR strain in the most sensitive species and induced significant neutralizing antibodies. Immunological response in goats was slightly higher than sheep and the vaccine injected at 108.0 50 % egg infective dose/mL allowed anti-PPRV antibodies that lasted at least 12 months as shown by antibody response monitoring in sheep. The NDV vector presented a limited replication in the host and vaccinated animals remained negative when tested by cELISA based on PPRV nucleoprotein allowing DIVA. This recombinant vaccine appears to be a promising candidate in a free at risk countries and may be an important component of the global strategy for PPR eradication.

Safety and efficacy of a Bluetongue inactivated vaccine (serotypes 1 and 4) in sheep.

A new inactivated vaccine against Bluetongue virus (BTV) serotypes 1 and 4, was developed from field isolates. Safety and efficacy of the vaccine were evaluated in sheep by serological monitoring and virus nucleic acid detection after experimental infection of vaccinated animals. Seroconversion was observed in vaccinated animals at day 14 post vaccination (pv) with neutralizing antibody titer of 1.9 and 1.8 for serotypes 1 and 4, respectively. The titer increase significantly after the booster reaching 2.7 and persist one year >1.5 for both serotypes. After challenge with virulent isolates, vireamia was recorded in control animals, as evident by q-PCR with threshold cycles (Ct) ranging from 24 to 31 and peaked at day 10 post challenge, while no vireamia was detected in vaccinated animals. Vaccinated sheep were fully protected against the disease and infection.

Safety and immunogenicity of the Rift Valley fever arMP-12 ΔNSm21/384 candidate vaccine in pregnant ewes.

Rift Valley fever (RVF) poses a threat to human and animal health as well as economic losses due to abortion, new-born teratogenic effect and mortality. Safe and effective vaccines are critically needed to prevent the disease in humans and livestock. The objective of this study was to assess safety and immunogenicity of the Rift Valley fever virus (RVFV) arMP-12DNSm21/384 attenuated vaccine in 32 pregnant ewes at different stages of pregnancy including 17 ewes vaccinated during the early stage (G1) of pregnancy (<35 days) and 15 ewes vaccinated during the last two stages (G2) of pregnancy (>35 days). Ewes were monitored for clinical observations, rectal temperature and abortions and lambs were monitored for general health and rectal temperature. Vaccinated ewes and lambs were periodically sampled for their neutralizing antibody response to RVFV vaccination. All ewes were positive for antibody two weeks post-vaccination and 79% of ewes were positive at delivery. None of the 32 ewes aborted during pregnancy and all ewes vaccinated during the G2 stages of pregnancy gave birth to healthy lambs. However, among the 17 ewes vaccinated during the G1 stage of pregnancy, 2 ewes gave birth to 2 lambs with fore limb malformations that died at 1-day of age. One ewe gave birth to 2 punny twins that died at 2 days of age. Another ewe, gave birth to one lamb with a deformed tail that died at 20 days of age. At post-mortem, tissues of dead lambs (spleen, lung, brain and long bone) were negative for RVFV by PCR assay. While the findings did not link the malformed lambs directly to infection by the vaccine virus, these results indicated that pregnant sheep should not be vaccinated with the RVFV arMP-12DNSm21/384 vaccine during the first month of gestation.

Immunological responses and potency of the EG95NC- recombinant sheep vaccine against cystic echinococcosis.

Cystic echinococcosis (CE) is a zoonotic disease caused by the cestode parasite Echinococcus granulosus. The disease has an important impact on human health as well as economic costs including the cost of treatment as well as loss of productivity for the livestock industry. In many parts of the world where the disease is endemic, sheep and other livestock play an important role in the parasite's transmission. A vaccine to protect livestock against CE can be effective in reducing transmission and economic costs of the disease. A recombinant antigen vaccine has been developed against infection with E. granulosus (EG95) which could potentially be used to reduce the level of E. granulosus transmission and decrease the incidence of human infections. Further development of the EG95 recombinant vaccine as a combined product with clostridial vaccine antigens is one potential strategy which could improve application of the hydatid vaccine by providing an indirect economic incentive to livestock owners to vaccinate against CE. In this study we investigated the efficacy of the EG95 recombinant vaccine produced in Morocco by vaccination of sheep, including a combined vaccine incorporating EG95 and clostridia antigens. Vaccination with EG95 either as a monovalent vaccine or combined with clostridia antigens, protected sheep against a challenge infection with E. granulosus eggs and induced a strong, long lasting, and specific antibody response against the EG95 antigen.

Peste des petits ruminants pathogenesis on experimental infected goats by the Moroccan 2015 isolate.

BACKGROUND: Peste des petits ruminants (PPR) is a viral disease of major economic importance on small ruminants. Goats are usually known to be more susceptible to the disease. Infection chronology, virus circulation, and the disease early detection need to be better understood. This study evaluates the tissue tropism and pathogenesis of PPR following experimental infection of goats using a lineage IV virus, the most dominant in the world originated from Asia. PPRV infection was experimentally induced in 4 six-month-old goats by intra-nasal and intravenous route of cell virus suspension and from infectious mashed tissue. The clinical signs were observed and goats were euthanized at predetermined clinical score level for post-mortem examinations and PPRV detection by RT-PCR. Clinical signs of infection were present, pyrexia, serous-mucopurulent nasal discharges, coughing, diarrhea and asthenia, for both cell virus suspension and infectious mashed tissue. PPRV genome was highly detected in swabs and tissues with clinical signs dominated by pulmonary attack and digestive symptoms secondary. RESULTS: Results of this study indicates that PPRV is an invasive infection in animals that in a short period, less than 10 days, invade all vital organs. On live animals, early diagnostic may be easily done on lacrimal and rectal swabs. CONCLUSION: The experimental PPRV-infection model using the cell virus suspension is suitable for vaccine evaluation as a standard model.

Safety and immunogenicity of a live attenuated Rift Valley Fever recombinant arMP-12ΔNSm21/384 vaccine candidate for sheep, goats and calves.

Rift Valley fever (RVF) causes serious health and economic losses to the livestock industry as well as a significant cause of human disease. The prevention of RVF in Africa is a global priority, however, available vaccines have only been partially effective. Therefore, the objective of this study was to evaluate the safety and immunogenicity of a live, attenuated recombinant RVFV arMP-12ΔNSm21/384 nucleotide deletion vaccine candidate in domestic ruminants. Evaluation involved testing to determine the infectivity titer of the vaccine virus in Vero cells for industrial scale up vaccine production. Safety experiments were conducted to determine the potential of the vaccine virus to revert to virulence by serial passages in sheep, the possibility of virus spread from vaccinated sheep and calves to unvaccinated animals, and the potential health effects of administering overdoses of the vaccine to sheep, goats and calves. The immunogenicity of 3 doses of 104, 105 and 106 Tissue Culture Infectious Doses50% (TCID50) of the vaccine was assessed in 3 groups of 10 sheep and 3 groups of 10 goats, and doses of 105, 106 and 107 TCID50 was evaluated in 3 groups of 10 calves subcutaenous vaccintation. The results showed that the infectivity titer of the vaccine virus was 108.4 TCID50/ml, that the vaccine did not spread from vaccinated to un-vaccinated animals, there was no evidence of reversion to virulence in sheep and the vaccine overdoses did not cause any adverse effects. The immunogenicity among sheep, goats and calves indicated that doses of 104-106 TCID50 elicited detectable antibody by day 7 post-vaccination (PV) with antibody titers ranging from 0.6 log to 2.1 log on day 14 PV with sustained titers through day 28 PV. Overall, these findings indicated that the RVFV arMP-12ΔNSm21/384 vaccine is a promising candidate for the prevention of RVF among domestic ruminants.

Comparative evaluation of three capripoxvirus-vectored peste des petits ruminants vaccines.

Sheep and goat pox (SGP) with peste des petits ruminants (PPR) are transboundary viral diseases of small ruminants that cause huge economic losses. Recombinant vaccines that can protect from both infections have been reported as a promising solution for the future. SGP was used as a vector to express two structural proteins hemagglutinin or the fusion protein of PPRV. We compared immunity conferred by recombinant capripoxvirus vaccines expressing H or F or both HF. Safety and efficacy were evaluated in goats and sheep. Two vaccine doses were tested in sheep, 104.5TCDI50 in 1ml dose was retained for the further experiment. Results showed that the recombinant HF confers an earlier and stronger immunity against both SGP and PPR. This recombinant vaccine protect also against the disease in exposed and unexposed sheep. The potential Differentiating Infected from Vaccinated Animals of recombinant vaccines is of great advantage in any eradication program.

Vaccination Against Hydatidosis: Molecular Cloning and Optimal Expression of the EG95NC- Recombinant Antigen in Escherichia coli.

Cystic echinococcosis (CE) is a widely distributed zoonosis that is highly endemic in the Mediterranean basin. The disease represents a serious public health threat and causes economic losses. The parasite life-cycle involves dogs and ruminants as definitive and intermediate hosts; humans are accidently infected, causing serious clinical issues. Vaccination of ruminants and dog treatments represent the most efficient measures to prevent parasite transmission. The recombinant protein vaccine, EG95, has been used successfully in sheep vaccine trials against CE in several countries. In this study, we expressed the modified antigen, EG95NC-GST, in Escherichia coli for use as a vaccine against Echinococcus granulosus in ruminants. We tested three different media formulations for E. coli culture and established for each culture conditions for optimal levels of soluble EG95 expression. The results demonstrate that SOC and TB media provided high yields in cell density and EG95 protein expression. Purification of the recombinant protein with affinity chromatography (using FPLC) was also performed to increase the purity of the EG95NC--GST antigen.

Re-emergence of Peste des Petits Ruminants virus in 2015 in Morocco: Molecular characterization and experimental infection in Alpine goats.

Peste des Petits Ruminants (PPR) is a transboundary viral disease of small ruminants that causes huge economic losses in Africa, The Middle East and Asia. In Morocco, the first PPR outbreak was notified in 2008. Since then no cases were reported for seven years, probably due to three successive vaccination campaigns during 2008-2011 and close surveillance at the border areas. In June 2015, the disease re-emerged in Morocco, raising questions about the origin of the virus. The PPR virus was confirmed by qRT-PCR and virus was isolated from clinical samples on VeroNectin-4 cells. The disease was experimentally reproduced in Alpine goats using both sheep and goat derived outbreak isolates. Molecular characterization of the 2015 Moroccan PPR isolate confirmed the identity of the virus as lineage IV, closely related to the 2012 Algerian (KP793696) and 2012 Tunisian (KM068121) isolates and significantly distinct from the previous PPRV Morocco 2008 strain (HQ131927). Therefore this study confirms a new incursion of PPR virus in Morocco during 2015 and highlights the urgency of implementation of a common control strategy to combat PPR in Maghreb region in North Africa.

VeroNectin-4 is a highly sensitive cell line that can be used for the isolation and titration of Peste des Petits Ruminants virus.

Peste des Petits Ruminants virus (PPRV) is a member of the Morbillivirus subgroup of the family Paramyxoviridae, and is one of the most contagious diseases of small ruminants throughout Africa and the rest of the world. Different cell lines have previously been used to isolate PPRV but with limited success. Thus, to improve the isolation of Morbilliviruses, human, canine, and goat homologues of the lymphocyte receptor signaling lymphocyte activation molecule (SLAM) have been introduced into cells that can support virus replication. However, the amino acid sequence of SLAM varies between species, and often requires adaptation of a particular virus to different versions of the receptor. The protein sequence of Nectin-4 is highly conserved between different mammals, which eliminate the need for receptor adaptation by the virus. Cell lines expressing Nectin-4 have previously been used to propagate measles and canine distemper viruses. In this study, we compared infections in Vero cells expressing canine SLAM (VeroDogSLAM) to those in Vero cells expressing Nectin-4 (VeroNectin-4), following inoculations with wild-type strains of PPRV. Virus isolation using VeroNectin-4 cells was successful with 23% of swabbed samples obtained from live infected animals, and was 89% effective using post-mortem tissues of infected sheep. By contrast, only 4.5% efficiency was observed from swab samples and 67% efficiency was obtained in virus isolation from post-mortem tissues using VeroDogSLAM cells. The average incubation period for virus recovery from post-mortem tissues was 3.4 days using VeroNectin-4 cells, compared with 5.5 days when using VeroDogSLAM cells. The virus titers of PPRV obtained from VeroNectin-4 cells were also higher than those derived from VeroDogSLAM cells. A comparison of the growth kinetics for PPRV in the two cell lines confirmed the superiority of VeroNectin-4 cells for PPR diagnostic purposes and vaccine virus titration.

Equine encephalosis virus: evidence for circulation beyond southern Africa.

Prior to the recent outbreak of equine encephalosis in Israel in 2009, equine encephalosis virus (EEV) had only been isolated from equids in South Africa. In this study we show the first evidence for the circulation of EEV beyond South Africa in Ethiopia, Ghana and The Gambia, indicating that EEV is likely to be freely circulating and endemic in East and West Africa. Sequence analysis revealed that the EEV isolate circulating in The Gambia was closely related to an EEV isolate that was isolated from a horse from Israel during the EEV outbreak in 2009, indicating that the two viruses have a common ancestry. Interestingly horses in Morocco tested negative for EEV antibodies indicating that the Sahara desert may be acting as a geographical barrier to the spread to the virus to North African countries. This evidence for EEV circulation in countries in East and West Africa sheds light on how the virus may have reached Israel to cause the recent outbreak in 2009.