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Assessment of an rDNA probe filter hybridization assay for the detection of Rift Valley fever virus RNA in human serum samples from the Mauritanian epidemic.

The Rift Valley fever virus (RVFV) epidemic that occurred in southern Mauritania during the 1987 rainy season provided a unique opportunity to test and evaluate a recently developed, M-segment-specific, nucleic acid filter hybridization assay on a large collection of infected human serum samples. It afforded the opportunity to compare the procedure with two other methods for detecting virus: virus isolation and antigen detection by ELISA. The filter hybridization procedure employed a polyethylene-glycol-precipitation and proteinase-K-digestion sample treatment step developed specifically for preparing serum samples for hybridization. The procedure was less sensitive for detecting RVFV in the Mauritanian human viremic samples than in sera from experimentally infected monkeys used to evaluate this procedure. It was also less sensitive than an antigen detection procedure used to test the Mauritanian samples. However, we were able to detect virus RNA in a significant proportion of the virus-isolation-positive samples. Advances in sample preparation, labelling and detection procedures, and hybridization methods will improve the sensitivity, precision and ease of use of this assay and increase its value as a diagnostic tool.

Antibodies, Viral↗

The 2000 epidemic of Rift Valley fever in Saudi Arabia: mosquito vector studies.

In mid-September 2000, Rift Valley fever (RVF) virus was diagnosed as the cause of infection in humans and livestock in Jizan Region, Saudi Arabia. This is the first time that this arbovirus has been found outside Africa and Madagascar. Collections of mosquitoes (Diptera: Culicidae) were therefore undertaken (from 25 September to 10 October) at eight sites during the epidemic to obtain mosquitoes for attempted RVF virus isolation. Among 23 699 mosquito females tested, six isolations of RVF virus were made from 15 428 Culex (Culex) tritaeniorhynchus Giles and seven from 8091 Aedes (Aedimorphus) vexans arabiensis Patton [corrected]. Minimum mosquito infection rates per 1000 at sites with infected mosquitoes were 0.3-13.8 Cx. tritaeniorhynchus and 1.94-9.03 Ae. v. arabiensis. Viral activity moved northwards as collecting was in progress and collectors 'caught up' with the virus at the two most northerly sites on the last two trapping evenings. Other species occurred in small numbers and were identified but not tested. Both Cx. tritaeniorhynchus and Ae. v. arabiensis were susceptible to RVF virus and transmitted between hamsters, and an additional quantitative test with Cx. tritaeniorhynchus showed that 71-73% of mosquitoes became infected after ingesting 6.9-7.9 log10 FFU/mL of virus; transmission rates were 10% (post-infection day 14) and 26% (post-infection day 20). It was concluded that both species were vectors on grounds of abundance, distribution, preference for humans and sheep, the virus isolations and vector competence tests.

Animals↗

Rift Valley fever outbreak, Mauritania, 1998: seroepidemiologic, virologic, entomologic, and zoologic investigations.

A Rift Valley fever outbreak occurred in Mauritania in 1998. Seroepidemiologic and virologic investigation showed active circulation of the Rift Valley fever virus, with 13 strains isolated, and 16% (range 1.5%-38%) immunoglobulin (Ig) M-positivity in sera from 90 humans and 343 animals (sheep, goats, camels, cattle, and donkeys). One human case was fatal.

Animals↗

Serological evidence in sheep suggesting phlebovirus circulation in a Rift Valley fever enzootic area in Burkina Faso.

Within the Phlebovirus serogroup, Rift Valley fever (RVF) virus is endemo-enzootic in the African sahelian zone. Recently an RVF epizootic in West Africa prompted a serosurvey in the major sheep and cattle raising areas. Because of the close antigenic relationship between the phleboviruses it appeared of interest to evaluate the prevalence of the other phleboviruses also. In 1987, 482 sheep serum samples were collected in 2 different ecological zones of Burkina Faso and tested for the presence of phlebovirus antibodies. A sensitive but non-specific immunofluorescent antibody test and a specific enzyme-linked immunosorbent assay (ELISA) were used, with the following African phlebovirus antigens: Rift Valley fever (RVF), Arumowot, Gabek Forest, Gordil, Saint Floris and Odrenisrou. A total of 15.8% of the sera sampled had anti-RVF antibody in the ELISA. RVF virus appeared to be more active in drier areas such as the sahelian region, known to be an enzootic area for the disease. Antibodies to other phleboviruses were found in 11.8% of the samples, independent of RVF virus activity. It is assumed that sheep can be infected by different phleboviruses.

Animals↗

Studies on Rift Valley fever in some African murids (Rodentia: Muridae).

Brains, spleens and livers of 2214 murids, 27 shrews and 7 dormice, trapped at 7 sites in Rhodesia, were tested in 277 pools for the presence of Rift Valley Fever virus. There were no isolations of Rift Valley Fever, but 69 isolations of an unidentified virus were obtained. Sixteen out of 867 sera had low-titre haemagglutination-inhibition activity against Rift Valley Fever antigen, but only one out of 1260 sera had neutralizing antibody. The evidence suggests that murids fail to encounter infection in nature and are unlikely to play a role in circulation and dissemination of Rift Valley Fever virus. Four out of seven widely distributed species of muried, Rhabdomys pumilio, Saccostomys campestris, Aethomys chrysophilus and Lemniscomys griselda, were shown to be capable of circulating amounts of virus likely to be infective for mosquitoes.

Animals↗

Immunogenicity of combination DNA vaccines for Rift Valley fever virus, tick-borne encephalitis virus, Hantaan virus, and Crimean Congo hemorrhagic fever virus.

DNA vaccines for Rift Valley fever virus (RVFV), Crimean Congo hemorrhagic fever virus (CCHFV), tick-borne encephalitis virus (TBEV), and Hantaan virus (HTNV), were tested in mice alone or in various combinations. The bunyavirus vaccines (RVFV, CCHFV, and HTNV) expressed Gn and Gc genes, and the flavivirus vaccine (TBEV) expressed the preM and E genes. All vaccines were delivered by gene gun. The TBEV DNA vaccine and the RVFV DNA vaccine elicited similar levels of antibodies and protected mice from challenge when delivered alone or in combination with other DNAs. Although in general, the HTNV and CCHFV DNA vaccines were not very immunogenic in mice, there were no major differences in performance when given alone or in combination with the other vaccines.

Animals↗

First isolation of the Rift Valley fever virus from Culex poicilipes (Diptera: Culicidae) in nature.

Following the reemergence of Rift Valley fever (RVF) virus in southeastern Mauritania in 1998, an entomological survey was undertaken in the boundary area in Senegal to assess the extent of the virus circulation. During this study, RVF virus (36 strains) was isolated for the first time from Culex poicilipes in nature. The possible role of Cx. poicilipes as an RVF vector is discussed regarding its biology and ecology.

Animals↗

[Rift Valley Fever and phleboviroses in the Central African Republic].

During 1984 and 1985, six Rift Valley Fever virus strains (RVF) were isolated in Central African Republic, among them five from human samples. Three strains were isolated in 1985 at the end of the rainy season, from sera of patients dead with severe jaundice with haemorrhagic syndrome, what could evoke a little outbreak. At this occasion, these RVF strains and the other strains of phleboviruses previously isolated in CAR, were antigenically compared by Immuno-fluorescent assay (IFA) and Complement Fixation Test (CF), using mice immuno ascitic fluids prepared against each strain. A lot of cross reactions were noted between the different strains, but all the RVF strains seem to have the same antigenic outline.

Animals↗

Analytical study of a Rift Valley fever epidemic.

Three epidemiological investigations were carried out during and after the 1987 Rift Valley fever (RVF) epidemic in South Mauritania: a case control study, a cross-sectional study in the town of Rosso and a cross-sectional study of villages and encampments around Rosso. The case control study showed an association between epizootic and epidemic outbreaks. The study in Rosso town showed housing and district effects. Also, a medical survey of hospital staff showed the absence of interhuman contamination. The cross-sectional study of villages and encampments around Rosso showed heterogeneity corresponding to a village and ethnic effect. The human epidemic was linked to epizootic disease, but the "vectors" of the disease are not as yet known.

Animals↗

Rift Valley fever virus: some ultrastructural observations on material from the outbreak in Egypt 1977.

Rift Valley fever virus isolates from the 1977 outbreak in Egypt were studied at an ultrastructural level. The particles measured 90 to 110 nm in diam. using negative staining and sectioning techniques, with a core component of 80 to 85 nm. The surface of the virions was calculated to be covered by approx. 160 sub-units. The particles were found in smooth endoplasmic reticular systems, which were made up of either multi-tubular complexes, or of a single large vacuole. The majority of these membrane systems were found to be unassociated with Golgi apparatus. Inclusion bodies were found within the host cell nuclei (made up of rods and fine granules) and in the cytoplasm (aggregates of fine or coarse granules). The possible relationship of these structures to virus replication is discussed.

Animals↗

Isolation of Rift Valley fever virus from human peripheral blood mononuclear cells: Mauritanian epidemic.

During the Mauritanian Rift Valley fever (RVF) epidemic of 1987, peripheral blood mononuclear cells (PBMC) were studied from 78 sick patients. RVF virus (RVFV) was isolated in 5 cases, on Aedes pseudoscutellaris AP61, from both PBMC and serum. Among the 78 cases studied, RVF was proven in 19 cases (24.3%) by specific IgM detection, and in 12 cases (15.3%) by virus isolation from serum, of which 3 also exhibited anti-RVF IgM. Among the 5 PBMC-positive RVFV cases, 2 strains were isolated in the presence of specific IgM from patients presenting with neurologic signs. These observations raised the question as to the role of specific IgM in cellular infection, and suggest that, in certain cases, mononuclear cells may promote RVFV dissemination into brain cells. Further investigations need to be undertaken to determine the RVFV receptor expressed on PBMC membranes.

Antibodies, Viral↗

Clinical, virological and serological response of the West African dwarf sheep to experimental infection with different strains of Rift Valley fever virus.

West African dwarf sheep were inoculated with three different strains of Rift Valley fever virus (RVFV). Using infective mouse serum as the source of virus classical RVFV disease characterised by sudden onset, a sharp but transient febrile response, viraemia, abortions and the development of specific RVFV antibodies in surviving animals was observed. The severity of clinical response was, however, dependent on the strain of virus used, with animals inoculated with Smithburn's neuroadapted strain showing a milder response than those inoculated with either the Nigerian or Lunyo strain. The inoculation of sheep with RVFV infective mouse brain material of the three different strains resulted in a mild febrile response with low level viraemia. Immune sera from sheep inoculated with both the Nigerian and Smithburn's neurotropic strains did not neutralise the Lunyo virus strain in a mouse intracerebral neutralisation test; the reverse, however, was not the case. The findings indicate that the West African dwarf sheep is highly susceptible to RVFV infection and that previous reports of only a mild clinical response following inoculation with the Nigerian strain were due to infective mouse brain rather than infective mouse serum.

Animals↗

The effect of laboratory colonization on the vector-pathogen interactions of Egyptian Culex pipiens and Rift Valley fever virus.

Field and laboratory findings implicated Culex pipiens as a vector of Rift Valley fever (RVF) virus during the 1977-1978 epizootics/epidemics in Egypt. This study evaluated changes in infection and transmission rates, and viral titers in F1 through F16 generation Cx. pipiens mosquitoes orally infected with RVF virus. Infection and transmission rates of RVF virus by this species changed significantly during the colonization process. However, the ultimate viral titers of either the transmitting or the infected nontransmitting mosquitoes were not affected by the colonization process. Following ingestion of virus, Cx. pipiens could be separated into three distinct subpopulations, an uninfected group and two types of infected mosquitoes--transmitters and nontransmitters. Transmitters contained significantly more virus (approximately 100-fold) than nontransmitters. These results demonstrated that not every infected female mosquito should be considered a competent vector, even if the species (population) is known to be a primary vector. Transmission was also accomplished by probing mosquitoes which were unsuccessful in obtaining a blood meal. These data document the long-held suspicion that vector competence studies based upon laboratory-colonized specimens may not represent the field situation.

Animals↗

Efficacy of a Rift Valley fever virus vaccine against an aerosol infection in rats.

The formalin-inactivated Rift Valley fever virus (RVFV) vaccine, TSI-GSD-200, was administered subcutaneously to highly susceptible adult Wistar-Furth rats (LD50-1 p.f.u., ZH501 strain). Vaccine was administered on days 0, 7 and 28, the same time course used for at-risk personnel. Six months postimmunization, when the serum plaque-reduction neutralization titre (PRNT)80 had declined to low or undetectable levels, rats were challenged with 4.4 log10 p.f.u. of the virulent ZH501 strain in a nose-only dynamic aerosol apparatus. Ninety-seven per cent (33/34) of the non-vaccinated control rats died. In contrast, only 32% (33/105) of the vaccinated animals died. In vaccinated rats that succumbed, there was a doubling of the mean time to death and the cause of death shifted from hepatitis to encephalitis. Rats with a PRNT80 of greater than or equal to 1:40 were protected from clinical disease and histological evidence of hepatic or encephalitic lesions. While the precise mechanisms of immunity against aerosol challenge remain unresolved, here the serum PRNT titre correlated with protection.

Aerosols↗

Transstadial and horizontal transmission of Rift Valley fever virus in Hyalomma truncatum.

We exposed Hyalomma truncatum and Rhipicephalus appendiculatus to Rift Valley fever (RVF) virus in order to assess the possible role of these ticks as enzootic/epizootic RVF vectors. The virus replicated in H. truncatum after intracoelomic inoculation, and a minimum transmission rate of 17% was achieved after 15 days intrinsic incubation. The virus persisted at least 58 days in these ticks. Virus was also shown to pass transstadially from inoculated H. truncatum nymphs to adults, with peak viral titers reaching 10(3.5) plaque-forming units (PFU) in adult males after they were provided with bloodmeals. Virus was recovered from adult females 121 days after they were inoculated as nymphs. Viral titers peaked in inoculated male ticks after dropping off a host (mean titer = 10(4.3) PFU). RVF virus was not detected in pools of eggs and larval progeny from 11 infected female H. truncatum. H. truncatum larvae and nymphs did not become infected after ingesting greater than 10(2.0) PFU while feeding on a RVF viremic hamster. The number of infected specimens declined rapidly after RVF virus was inoculated into R. appendiculatus adults, and virus was undetectable 12 days post-inoculation.

Animals↗

Characterization of clone 13, a naturally attenuated avirulent isolate of Rift Valley fever virus, which is altered in the small segment.

The 74HB59 strain of Rift Valley fever (RVF) virus, isolated from a human case in the Central African Republic, was shown to be composed of a heterogeneous population of viruses when plaque-purified clones were analyzed for their reactivity with monoclonal antibodies (MAbs) directed against the nucleocapsid (N) protein or the nonstructural (NSs) protein. One of these clones, C13, was of particular interest in that it proved to be avirulent in mice and hamsters, and highly immunogenic. Although C13 showed normal reactivity with a large panel of MAbs directed at the glycoproteins, it failed to react with specific MAbs or polyclonal antibodies directed at the NSs protein and with a specific MAb recognizing the N protein of the Egyptian strains. Consequently, the small RNA segment, which encodes the N and NSs proteins in an ambisense strategy, was sequenced and compared with the existing sequence of the attenuated MP-12 RVF virus strain. We found that the NSs gene contained, in addition to two conservative coding changes, a large internal deletion of 549 nucleotides that removes 69% of the open reading frame but conserves in-frame the N and C termini of the predicted translation product. In addition, the sequence revealed that the N protein of C13 contained a single amino acid change. Clone C13 replicated normally in certain cell types in vitro and in Culex pipiens mosquitoes after intrathoracic inoculation, but established abortive infections in MRC-5 human fibroblasts.

Animals↗

Further evaluation of a mutagen-attenuated Rift Valley fever vaccine in sheep.

A previous study demonstrated that a mutagen-attenuated Rift Valley fever virus (RVFV) vaccine, RVF MP-12, was immunogenic and non-abortogenic when ewes, 90-110 days pregnant, were inoculated with 5 x 10(5) plaque-forming units (p.f.u.) of the virus strain. The ewes delivered live, healthy lambs that had no neutralizing antibody to RVFV until after they had ingested colostrum. To assess further the safety and protective capability of this candidate vaccine, six pregnant ewes were inoculated with 5 x 10(3) p.f.u. of RVF MP-12 and challenged with 5 x 10(5) p.f.u. of virulent ZH-501 strain of RVFV 30 days later. No viraemia was detected after vaccination or challenge and all six ewes delivered live, healthy lambs. Those lambs tested before their nursing did not have neutralizing antibody to RVFV but quickly acquired antibody titres of 1:320 to greater than or equal to 1:10,240 after ingesting colostrum. To test the safety of the RVF MP-12 immunogen in neonates, lambs less than or equal to 7 days old, born to unvaccinated ewes, were inoculated with 5 x 10(5) p.f.u. of RVF MP-12. With the exception of brief pyrexia in 18 of 26 lambs, and a transient low-titred viraemia in 16 of 26 lambs after inoculation, no untoward effects were observed. Serum-neutralizing antibody to RVFV was detected 5-7 days after inoculation. Lambs vaccinated with either 5 x 10(5) or 5 x 10(3) p.f.u. of RVF MP-12 were protected against virulent RVFV challenge at 14 days postvaccination.

Animals↗