Tonate virus infection in French Guiana: clinical aspects and seroepidemiologic study.

Two recent cases of human infection with Tonate virus, one of which was a fatal case of encephalitis, have renewed interest in these viruses in French Guiana. The clinical aspects of confirmed and probable cases of infection with this virus indicate that it has pathogenic properties in humans similar to those of other viruses of the Venezuelan equine encephalitis complex. To determine the prevalence of antibodies to Tonate virus in the various ethnic groups and areas of French Guiana, 3,516 human sera were tested with a hemagglutination inhibition test. Of these, 11.9% were positive for the virus, but significant differences in seroprevalence were found by age, with an increase with age. After adjustment for age, significant differences were found between places of residence. The prevalence of antibody to Tonate virus was higher in savannah areas, especially in the Bas Maroni (odds ratio [OR] = 22.2, 95% confidence interval [CI] = 15.2-32.4) and Bas Oyapock areas (OR = 13.4; 95% CI = 9.8-18.4). The ethnic differences observed in this study were due mainly to differences in place of residence, except that whites were significantly less frequently infected than other ethnic groups. This study indicates that Tonate virus infection is highly prevalent in French Guiana, especially in savannah areas.

Genetic reassortment of Rift Valley fever virus in nature.

Rift Valley fever virus (RVFV), a phlebovirus of the Bunyaviridae family, is an arthropod-borne virus which emerges periodically throughout Africa, emphasizing that it poses a major threat for animal and human populations. To assess the genetic variability of RVFV, several isolates from diverse localities of Africa were investigated by means of reverse transcription-PCR followed by direct sequencing of a region of the small (S), medium (M), and large (L) genomic segments. Phylogenetic analysis showed the existence of three major lineages corresponding to geographic variants from West Africa, Egypt, and Central-East Africa. However, incongruences detected between the L, M, and S phylogenies suggested that genetic exchange via reassortment occurred between strains from different lineages. This hypothesis, depicted by parallel phylogenies, was further confirmed by statistical tests. Our findings, which strongly suggest exchanges between strains from areas of endemicity in West and East Africa, strengthen the potential existence of a sylvatic cycle in the tropical rain forest. This also emphasizes the risk of generating uncontrolled chimeric viruses by using live attenuated vaccines in areas of endemicity.

[Present status of an arbovirus infection: yellow fever, its natural history of hemorrhagic fever, Rift Valley fever]. / Une arbovirose d'actualité: la fièvre jaune, son histoire naturelle face à une fièvre hémorragique, la fièvre de la vallée du Rift.

In the early 20th century, when it was discovered that the yellow fever virus was transmitted in its urban cycle by Aedes aegypti, measures of control were introduced leading to its disappearance. Progressive neglect of the disease, however, led to a new outbreak in 1927 during which the etiological agent was isolated; some years later a vaccine was discovered and yellow fever disappeared again. In the 1960s, rare cases of encephalitis were observed in young children after vaccination and the administration of the vaccine was forbidden for children under 10 years. Five years later, a new outbreak of yellow fever in Diourbel, Senegal, was linked to the presence of Aedes aegypti. In the late 1970s, the idea of a selvatic cycle for yellow fever arose. Thanks to new investigative techniques in Senegal and Côte d'Ivoire, the yellow fever virus was isolated from the reservoir of virus and vectors. The isolated virus was identified in monkeys and several vectors: Aedes furcifer, Aedes taylori, Aedes luteocephalus. Most importantly, the virus was isolated in male mosquitoes. Until recently, the only known cycle had been that of Haddow in East Africa. The virus circulate in the canopea between monkeys and Aedes africanus. These monkeys infect Aedes bromeliae when they come to eat in banana plantations. This cycle does not occur in West Africa. Vertical transmission is the main method of maintenance of the virus through the dry season. "Reservoirs of virus" are often mentioned in medical literature, monkeys having a short viremia whereas mosquitoes remain infected throughout their life cycle. In such a selvatic cycle, circulation can reach very high levels and no child would be able to escape an infecting bite and yet no clinical cases of yellow fever have been reported. The virulence--as it affects man--of the yellow fever virus in its wild cycle is very low. In areas where the virus can circulate in epidemic form, two types of circulation can be distinguished. Intermediate yellow fever--a term coined to define epidemia which do not correspond exactly to urban yellow fever. The cycle involves men and monkeys through wild vectors as Aedes furcifer but also through Aedes aegypti and the mortality rate is much lower than for urban epidemics. In urban yellow fever, man is the only vertebrate host involved in the circulation of the virus, the vector being generally Aedes aegypti. This vector maintains a selective pressure, increasing the transmission of virus capable of producing high viremia in man. In the selvatic cycles, two cycles can be distinguished: one of maintenance which does not increase the quantity of virus in circulation and one of amplification which does increase this quantity. As we shall see, it develops into an epizootic form but also in an epidemic form in man. When the decrease in yellow fevers across Africa is considered, it appears that all major epidemics occur in West Africa inspite of the presence of wild cycles of the yellow fever virus in Central and East Africa. For the rare epidemics that have occurred there, the vector has never been Aedes aegypti. In a recent outbreak in Kenya, the vector was Aedes bromeliae. The examination of part of the gene encoding for envelope protein showed the presence of two geographical types corresponding to West-Africa and Central East-Africa. Clinically speaking, yellow fever is an haemorrhagic fever with hepatitis similar to other haemorrhagic fevers such as Rift Valley fever. When, in 1987, an outbreak of haemorrhagic fever occurred in southern Mauritania, for several days it was thought to be yellow fever. Four days later, the diagnosis was corrected by isolating and identifying the virus as that of Rift Valley fever (RVFV). RVFV causes several pathogenic syndromes in human beings: acute febrile illness, haemorrhagic fever, haemorrhagic fever with hepatitis, nervous syndromes or ocular disease. Mortality rate was high for haemorrhagic fever with hepatitis, reaching 36%. (ABST

New vectors of Rift Valley fever in West Africa.

After an outbreak of Rift Valley fever in Southern Mauritania in 1987, entomologic studies were conducted in a bordering region in Sénégal from 1991 to 1996 to identify the sylvatic vectors of Rift Valley fever virus. The virus was isolated from the floodwater mosquitoes Aedes vexans and Ae. ochraceus. In 1974 and 1983, the virus had been isolated from Ae. dalzieli. Although these vectors differ from the main vectors in East and South Africa, they use the same type of breeding sites and also feed on cattle and sheep. Although enzootic vectors have now been identified in West Africa, the factors causing outbreaks remain unclear.

Variability of the NS(S) protein among Rift Valley fever virus isolates.

Eighteen strains of Rift Valley fever (RVF) virus collected over a period of 38 years and isolated from diverse localities in Africa and from various hosts (human, animal and arthropod) were investigated by RT-PCR followed by sequencing of the NS(S) protein coding region. This region was chosen to analyse variability because, in contrast to the N protein, the NS(S) protein differs in various phleboviruses and there exists an RVF virus (clone 13) in which 70% of the NS(S) ORF is deleted, suggesting that this sequence is under a weak selective pressure. Sequence data indicated that percentage divergence among isolates ranged from 0 to 9.6% at the nucleotide level and from 0 to 9.5% at the amino acid level. Phylogenetic analysis based on the NS(S) gene revealed two major lineages: Egyptian and sub-Saharan. This led to the establishment of the relatedness between strains and insights into the NS(S) protein, the function of which is still undetermined. Alignment of the deduced amino acid sequences indicated that the cysteine residues are conserved, as are several motifs representing potential phosphorylation sites.

Extensive nucleotide changes and deletions within the envelope glycoprotein gene of Euro-African West Nile viruses.

We compared the sequence of an envelope protein gene fragment from 21 temporally distinct West Nile (WN) virus strains, isolated in nine African countries and in France. Alignment of nucleotide sequences defined two groups of viruses which diverged by up to 29%. The first group of subtypes is composed of nine WN strains from France and Africa. The Austral-Asian Kunjin virus was classified as a WN subtype in this first group. The second group includes 12 WN strains from Africa and Madagascar. Four strains harboured a 12 nucleotide in-frame deletion. The loss of the corresponding four amino acids resulted in the loss of the potential glycosylation site present in several WN strains. The distribution of virus subtypes into two lineages did not correlate with host preference or geographical origin. The isolation of closely related subtypes in distant countries is consistent with WN viruses being disseminated by migrating birds.

Enzootic activity of Rift Valley fever virus in Senegal.

In two areas of Senegal where previous evidence of Rift Valley fever (RVF) virus circulation was detected, Barkedji in the Sahelian bioclimatic zone and Kedougou in the Sudano-Guinean zone, a longitudinal study of the enzootic maintenance of RVF virus was undertaken from 1991 to 1993. Mosquitoes, sand flies, and ticks were collected and domestic ungulates were monitored with serologic surveys. Rift Valley fever virus was not isolated in Kedougou. In Barkedji, RVF virus was isolated from Aedes vexans and Ae. ochraceus mosquitoes collected in traps near ground pools and cattle droves and from one health sheep. Sand flies were not involved in the maintenance cycle. Seroconversions were recorded in three (1.9%) of 160 monitored sheep and goats. The interepizootic vectors appeared to belong to the Aedes subgenus Neomelaniconion in East Africa, and to the subgenus Aedimorphus in West Africa. Epizootics in East Africa are associated with an increase in rainfall. However, factors associated with epizootics remain unknown for West Africa.

Surveillance for yellow fever virus in eastern Senegal during 1993.

During the 1993 rainy season, 15,806 mosquitoes, including 14,304 Aedes ssp., were collected and tested for virus infection in 702 and 547 pools, respectively. Aedes furcifer (Edwards) was the most abundant species collected throughout the survey period. Yellow fever (YF) virus was detected in 187 pools: Ae. furcifer (123 isolates), Ae. taylori (Edwards) (41 isolates), and Ae. luteocephalus (Newstead) (23 isolates). A high prevalence of immunoglobulin (IgG) antibodies was found in human and simian populations. Results clearly indicated that increased sylvatic YF activity in eastern Senegal has the increased the risk of YF transmission among rural populations in West Africa. Our results showed that a minimal survey period may be effective in detecting the circulation of YF in the Kedougou area.

[Ngari virus (Bunyaviridae: Bunyavirus). First isolation from humans in Senegal, new mosquito vectors, its epidemiology]. / Le virus Ngari (Bunyaviridae: Bunyavirus). Premiers isolements chez l'homme au Sénégal, nouveaux vecteurs culicidiens, le point sur son épidémiologie.

Ngari virus (NRI) (Bunyaviridae, genus Bunyavirus) was isolated first from male Aedes simpsoni mosquitoes in Southeastern Senegal in 1979. Then, it was recovered from several mosquito species in Senegal, Burkina Faso, Central African Republic and Madagascar. A potential pathogenicity of NRI virus in humans was suspected when the virus was isolated from two patients in Dakar in October and November 1993. The large diversity of Culicidae vectors and feeding patterns showed a large heterogeneity of vertebrate hosts. The wide geographical distribution of NRI virus in different bioclimatic areas indicated an important adaptability of the virus. Ngari virus epidemiology will need further investigations in order to approach the real pathogenicity of such emerging virus.

Short report: Rift Valley fever in western Africa: isolations from Aedes mosquitoes during an interepizootic period.

Thirteen strains of Rift Valley fever virus were isolated from Aedes vexans and Ae. ochraceus mosquitoes collected in October and November 1993 in northern Senegal. Entomologic and serologic data show that the risk of a new epizootic is increasing in this region.

Isolations of West Nile and Bagaza viruses from mosquitoes (Diptera: Culicidae) in central Senegal (Ferlo).

During October-November 1990, 31,497 mosquitoes consisting of 25 different species were collected in Barkedji, Ferlo area (Senegal), and tested for virus infection. Viruse were isolated from 55 of 407 pools. Eighteen pools were found positive for both Bagaza virus (BGA) and West Nile virus (WN). One alphavirus (Babanki [BBK] and 72 flaviviruses (19 BGA, 53 WN) were isolated from Culex poicilipes Theobald (29 WN, 8 BGA), C. neavei Theobald (3 WN, 1 BGA), Mimomyia hispida Theobald (8 WN, 6 BGA, and 1 BBK), M. lacustris Edwards (4 WN, 1 BGA), M. splendens Theobald (6 WN, 2 BGA), Mimomyia. spp. (2 WN), and Aedeomyia africana Neveu-Lemaire (1 WN). These were the first isolations of arboviruses from A. africana and Mimomyia species. C. poicilipes and possibly Mimomyia spp. may be involved in an avian-mosquito cycle of West Nile virus transmission in Senegal.

Dengue 2 outbreak in southeastern Senegal during 1990: virus isolations from mosquitoes (Diptera: Culicidae).

During 1990, Dengue-2 (DEN-2) virus was isolated for the first time from mosquitoes (Aedes furcifer, six isolates; Ae. taylori, six isolates; Ae. luteocephalus, seven isolates) collected during an epidemic in which DEN-2 virus also was isolated from humans. Numerous isolations have been made previously from mosquitoes in the absence of human infection. In Senegal, DEN-2 virus appears to be maintained in an enzootic cycle and, therefore, plays an expanding role in human disease and increases the need for effective surveillance in mosquito populations.

First isolations of arboviruses from phlebotomine sand flies in West Africa.

For the first time in West Africa, arboviruses were isolated from phlebotomine sand fly pools. One strain of Chandipura virus (a Vesiculovirus), four strains of Saboya virus (a Flavivirus), and one strain of a not yet identified virus were isolated. Three hundred twenty-two pools were established from a population of 33,917 sand flies caught in CO2 light traps in the Ferlo Sahelian region of Senegal from November 1991 to December 1992. This is the first isolation of Chandipura virus from any arthropod in Africa. Saboya virus has already been isolated from small rodents in Senegal; thus, its transmission cycle probably involves rodentophilic sand flies. No strain of Rift Valley fever phlebovirus, which caused an epizootic in this region in 1987, was isolated. During the same time at the same site, 11 sand fly species were identified from 4,191 specimens caught on sticky traps, including Phlebotomus duboscqi, a leishmaniasis vector.

Geographic distribution and evolution of yellow fever viruses based on direct sequencing of genomic cDNA fragments.

We have compared the nucleotide sequence of an envelope protein gene fragment encoding amino acids 291 to 406 of 22 yellow fever (YF) virus strains of diverse geographic and host origins isolated over a 63 year time span. The nucleotide fragment of viral RNA was examined by direct sequencing of a PCR product derived from complementary DNA. Alignment with the prototype Asibi strain sequence showed divergence of 0 to 21.5% corresponding to a maximum of 5.2% divergence in the amino acid sequence. Taking 10% nucleotide divergence as a cut-off point, the 22 YF virus strains fell into three topotypes which corresponded to different geographical areas, namely West Africa, Central-East Africa, and South America. Two subgroups were defined in West Africa, a genotypic group circulating in the sylvatic zone of the western part of Africa, from western Ivory Coast-Mali to Senegal, and a group responsible for large outbreaks from eastern Ivory Coast-Burkina Faso to Cameroon. Strains from Central-East Africa showed a low ratio of transition:transversion of about 1 instead of 8 to 10 for other strains, when their nucleotide sequences were compared with those of other African strains. This may reflect a more distant relationship between the former strains and the others. No change was observed in the highly conserved amino acid domain encompassing the TGD sequence, an important determinant of flavivirus tropism and pathogenesis. Our results support earlier observations on the genetic relationships between YF isolates established by T1 oligonucleotide fingerprinting and offer a useful tool for the understanding of YF virus distribution and evolution.

Simian immunodeficiency viruses from central and western Africa: evidence for a new species-specific lentivirus in tantalus monkeys.

Although up to 50% of African green monkeys (AGMs) are infected by simian immunodeficiency viruses (SIV) in their natural habitat, they remain asymptomatic carriers of these lentiviruses. They provide an attractive model to study not only the origin but also the link among genetic variation, host-virus adaptation, and pathogenicity of primate lentiviruses. SIVagm have been isolated from three species of AGM: the vervet (Cercopithecus pygerythrus), the grivet (Cercopithecus aethiops), and the sabaeus (Cercopithecus sabaeus) monkey. We studied four new SIVagm isolates from a fourth AGM species, the tantalus monkey (Cercopithecus tantalus), caught in the Central African Republic, and four new isolates from feral sabaeus monkeys from Senegal. Antigenic properties and partial env sequences were used to evaluate the diversity among these isolates. Alignment of env sequences in SIVagm isolated from tantalus and sabaeus monkeys permitted detailed mapping of the variable and conserved domains in the external glycoprotein. Genetic distances indicated that SIVagm isolates from tantalus monkeys are the most divergent among SIVagm in feral AGMs in Africa. The fact that AGMs are infected by four distinct lentiviruses, each specific for a single AGM species, supports the hypothesis of a coevolution of these viruses and their natural hosts and suggests that SIV transmission is a rare event among separated AGM species in the wild.