Evidence for circulation of Rift Valley fever virus in wildlife and domestic animals in a forest environment in Gabon, Central Africa.

Rift Valley fever (RVF) is a mosquito-borne viral zoonosis caused by the Rift Valley fever virus (RVFV) that can infect domestic and wild animals. Although the RVFV transmission cycle has been well documented across Africa in savanna ecosystems, little is known about its transmission in tropical rainforest settings, particularly in Central Africa. We therefore conducted a survey in northeastern Gabon to assess RVFV circulation among wild and domestic animals. Among 163 wildlife samples tested using RVFV-specific RT-qPCR, four ruminants belonging to subfamily Cephalophinae were detected positive. The phylogenetic analysis revealed that the four RVFV sequences clustered together with a virus isolated in Namibia within the well-structured Egyptian clade. A cross-sectional survey conducted on sheep, goats and dogs living in villages within the same area determined the IgG RVFV-specific antibody prevalence using cELISA. Out of the 306 small ruminants tested (214 goats, 92 sheep), an overall antibody prevalence of 15.4% (95% CI [11.5-19.9]) was observed with a higher rate in goats than in sheep (20.1% versus 3.3%). RVFV-specific antibodies were detected in a single dog out of the 26 tested. Neither age, sex of domestic animals nor season was found to be significant risk factors of RVFV occurrence. Our findings highlight sylvatic circulation of RVFV for the first time in Gabon. These results stress the need to develop adequate surveillance plan measures to better control the public health threat of RVFV.

An experimental game to assess hunter's participation in zoonotic diseases surveillance.

BACKGROUND: Strengthening the surveillance of zoonotic diseases emergence in the wild meat value chains is a critical component of the prevention of future health crises. Community hunters could act as first-line observers in zoonotic pathogens surveillance systems in wildlife, by reporting early signs of the possible presence of a disease in the game animals they observe and manipulate on a regular basis. METHODS: An experimental game was developed and implemented in a forested area of Gabon, in central Africa. Our objective was to improve our understanding of community hunters' decision-making when finding signs of zoonotic diseases in game animals: would they report or dissimulate these findings to a health agency? 88 hunters, divided into 9 groups of 5 to 13 participants, participated in the game, which was run over 21 rounds. In each round the players participated in a simulated hunting trip during which they had a chance of capturing a wild animal displaying clinical signs of a zoonotic disease. When signs were visible, players had to decide whether to sell/consume the animal or to report it. The last option implied a lowered revenue from the hunt but an increased probability of early detection of zoonotic diseases with benefits for the entire group of hunters. RESULTS: The results showed that false alerts-i.e. a suspect case not caused by a zoonotic disease-led to a decrease in the number of reports in the next round (Odds Ratio [OR]: 0.46, 95% Confidence Interval [CI]: 0.36-0.8, p < 0.01). Hunters who had an agricultural activity in addition to hunting reported suspect cases more often than others (OR: 2.05, 95% CI: 1.09-3.88, p < 0.03). The number of suspect case reports increased with the rank of the game round (Incremental OR: 1.11, CI: 1.06-1.17, p < 0.01) suggesting an increase in participants' inclination to report throughout the game. CONCLUSION: Using experimental games presents an added value for improving the understanding of people's decisions to participate in health surveillance systems.

Exploratory analysis of the knowledge, attitudes and perceptions of healthcare workers about arboviruses in the context of surveillance in the Republic of Guinea.

BACKGROUND: The escalating risk and contemporary occurrences of arbovirus infections prompt a critical inquiry into the ability of nations to execute efficient surveillance systems capable to detect, prevent and respond to arbovirus outbreaks. Healthcare workers (HCWs) are the major actors in the surveillance of infectious diseases with epidemic potential. The objective of this study was to evaluate the knowledge, attitudes and perceptions of HCWs regarding arboviruses in the public health facilities of Conakry, Guinea. METHODS: A cross-sectional survey was conducted during the from December 27, 2022, to January 31, 2023, encompassing from public health facilities in Conakry. The data collection process encompassed various aspects, including the characteristics of health facilities, socio-demographic and professional attributes of HCWs, the information received concerning arboviruses and the sources of information, as well as a series of inquiries designed to evaluate their knowledge, attitudes and perceptions. Subsequently, scores were computed for knowledge, attitude and perception. To identify the factors influencing the knowledge, attitudes, and perceptions of HCWs regarding arboviruses, Decision Tree and Inference Conditional Tree models were used. RESULTS: A total of 352 HCWs participated in the study, comprising 219 from national hospitals, 72 from municipal hospitals and 61 from primary health centers. More than half of the respondents (54.3%) had never received information on arboviruses. Only 1% of the respondents had good knowledge about arboviruses, 95.7% had a negative attitude about arboviruses. Moreover, nearly 60% of the respondents had a moderate perception and 24.1% had a good perception. The analysis revealed significant associations between the knowledge and attitudes of respondents concerning arboviruses and their years of professional experience and age. CONCLUSION: This study highlights the imperative requirement for comprehensive training targeting HCWs to enhance their capacity for early case detection within healthcare facilities. Additionally, there is a crucial need for analogous studies adopting a mixed-methods approach across all healthcare regions in Guinea.

Seroprevalence of IgG Antibodies Directed against Dengue, Chikungunya and West Nile Viruses and Associated Risk Factors in Madagascar, 2011 to 2013.

Arboviruses have been shown to circulate in Madagascar, including West Nile, dengue, and chikungunya viruses, though the extent of their circulation remains poorly documented. We estimated the seroprevalence of these three arboviruses in Madagascar and determined risk factors associated with seropositivity. Serum samples obtained from 1680 individuals surrounding the Sentinel Health Centers network in all regions of the country were analyzed using ELISA and hemagglutination inhibition assays for dengue, chikungunya, and West Nile viruses IgG antibodies, and multivariate logistic regression models were run. Overall, 6.5% [IC 95% 3.2-9.9] were seropositive for dengue virus, predominantly of Dengue serotype 1, 13.7% [IC 95% 6.5-20.9] for chikungunya virus, and 12.7% [IC 95% 9.0-16.5] for West Nile virus. There was no association with age, showing that dengue and chikungunya viruses were likely recently introduced. Eastern and Northern parts were more affected by dengue and chikungunya viruses, while West Nile virus seemed to circulate in all parts of the country. Dengue and chikungunya seropositivity were notably associated with high levels of vegetation, as well as frequent work in the forest, and West Nile seropositivity with the presence of cultivated areas, as well as standard of living. This analysis gives a new insight into arboviruses circulation and transmission patterns in Madagascar.

Current evidences of the efficacy of mosquito mass-trapping interventions to reduce Aedes aegypti and Aedes albopictus populations and Aedes-borne virus transmission.

BACKGROUND: Over the past decades, several viral diseases transmitted by Aedes mosquitoes-dengue, chikungunya, Zika-have spread outside of tropical areas. To limit the transmission of these viruses and preserve human health, the use of mosquito traps has been developed as a complement or alternative to other vector control techniques. The objective of this work was to perform a systematic review of the existing scientific literature to assess the efficacy of interventions based on adult mosquito trap to control Aedes population densities and the diseases they transmit worldwide. METHODS AND FINDINGS: Following the Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) guidelines, a systematic review was conducted using the PubMed and Scopus databases. Among the 19 selected papers, lethal ovitraps were used in 16 studies, host-seeking female traps in 3 studies. Furthermore, 16 studies focused on the control of Ae. aegypti. Our review showed great heterogeneity in the indicators used to assess trap efficacy: e.g., the number of host-seeking females, the number of gravid females, the proportion of positive containers, the viral infection rate in female mosquitoes or serological studies in residents. Regardless of the type of studied traps, the results of various studies support the efficacy of mass trapping in combination with classical integrated vector control in reducing Aedes density. More studies with standardized methodology, and indicators are urgently needed to provide more accurate estimates of their efficacy. CONCLUSIONS: This review highlights gaps in the demonstration of the efficacy of mass trapping of mosquitoes in reducing viral transmission and disease. Thus, further large-scale cluster randomized controlled trials conducted in endemic areas and including epidemiological outcomes are needed to establish scientific evidence for the reduction of viral transmission risk by mass trapping targeting gravid and/or host-seeking female mosquitoes.

The COVID-19 pandemic should not jeopardize dengue control.

The concurrent circulation of dengue and coronavirus disease 2019 (COVID-19) may produce many unfavourable outcomes-such as co-infections; delays in diagnosis, treatment, and mitigation measures; overwhelming of the healthcare system; underreporting of cases; deterioration in surveillance and control interventions; and exacerbation of social inequalities. Indeed, lockdown is greatly compromising the effectiveness of vector control, especially social mobilization campaigns and preventive insecticide spraying in private spaces (indoor and peridomestic spraying). Thus, failure to appropriately implement the full range of vector control interventions can lead to a reduction in their overall effectiveness and an increasing risk of vector-borne diseases circulating. Consequently, the health community and policy makers should develop proactive policies and allocate adequate resources to prevent and manage the resurgence of dengue and other vector-borne diseases in the new era of COVID-19.

Human Exposure to Hantaviruses Associated with Rodents of the Murinae Subfamily, Madagascar.

We conducted a national human serologic study of a hantavirus detected in Madagascar rodents using a commercial kit and a new ELISA targeting the virus. Our results suggest a conservative estimate of 2.7% (46/1,680) IgG seroprevalence. A second single-district study using the new ELISA revealed a higher prevalence (7.2%; 10/139).

Geographical distribution and relative risk of Anjozorobe virus (Thailand orthohantavirus) infection in black rats (Rattus rattus) in Madagascar.

BACKGROUND: Hantavirus infection is a zoonotic disease that is associated with hemorrhagic fever with renal syndrome and cardiopulmonary syndrome in human. Anjozorobe virus, a representative virus of Thailand orthohantavirus (THAIV), was recently discovered from rodents in Anjozorobe-Angavo forest in Madagascar. To assess the circulation of hantavirus at the national level, we carried out a survey of small terrestrial mammals from representative regions of the island and identified environmental factors associated with hantavirus infection. As we were ultimately interested in the potential for human exposure, we focused our research in the peridomestic area. METHODS: Sampling was achieved in twenty districts of Madagascar, with a rural and urban zone in each district. Animals were trapped from a range of habitats and examined for hantavirus RNA by nested RT-PCR. We also investigated the relationship between hantavirus infection probability in rats and possible risk factors by using Generalized Linear Mixed Models. RESULTS: Overall, 1242 specimens from seven species were collected (Rattus rattus, Rattus norvegicus, Mus musculus, Suncus murinus, Setifer setosus, Tenrec ecaudatus, Hemicentetes semispinosus). Overall, 12.4% (111/897) of Rattus rattus and 1.6% (2/125) of Mus musculus were tested positive for THAIV. Rats captured within houses were less likely to be infected than rats captured in other habitats, whilst rats from sites characterized by high precipitation and relatively low seasonality were more likely to be infected than those from other areas. Older animals were more likely to be infected, with infection probability showing a strong increase with weight. CONCLUSIONS: We report widespread distribution of THAIV in the peridomestic rats of Madagascar, with highest prevalence for those living in humid areas. Although the potential risk of infection to human may also be widespread, our results provide a first indication of specific zone with high transmission. Gathered data will be helpful to implement policies for control and prevention of human risk infection.

Prevalence of chronic hepatitis B virus infection and infrastructure for its diagnosis in Madagascar: implication for the WHO's elimination strategy.

BACKGROUND: WHO developed a global strategy to eliminate hepatitis B by 2030 and set target to treat 80% of people with chronic hepatitis B virus (HBV) infection eligible for antiviral treatment. As a first step to achieve this goal, it is essential to conduct a situation analysis that is fundamental to designing national hepatitis plans. We therefore estimated the prevalence of chronic HBV infection, and described the existing infrastructure for HBV diagnosis in Madagascar. METHODS: We conducted a stratified multi-stage serosurvey of hepatitis B surface antigen (HBsAg) in adults aged ≥18 years using 28 sentinel surveillance sites located throughout the country. We obtained the list of facilities performing HBV testing from the Ministry of Health, and contacted the person responsible at each facility. RESULTS: A total of 1778 adults were recruited from the 28 study areas. The overall weighted seroprevalence of HBsAg was 6.9% (95% CI: 5.6-8.6). Populations with a low socio-economic status and those living in rural areas had a significantly higher seroprevalence of HBsAg. The ratio of facilities equipped to perform HBsAg tests per 100,000 inhabitants was 1.02 in the capital city of Antananarivo and 0.21 outside the capital. There were no facilities with the capacity to perform HBV DNA testing or transient elastography to measure liver fibrosis. There are only five hepatologists in Madagascar. CONCLUSION: Madagascar has a high-intermediate level of endemicity for HBV infection with a severely limited capacity for its diagnosis and treatment. Higher HBsAg prevalence in rural or underprivileged populations underlines the importance of a public health approach to decentralize the management of chronic HBV carriers in Madagascar by using simple and low-cost diagnostic tools.

Reconstruction of Rift Valley fever transmission dynamics in Madagascar: estimation of force of infection from seroprevalence surveys using Bayesian modelling.

The force of infection (FOI) is one of the key parameters describing the dynamics of transmission of vector-borne diseases. Following the occurrence of two major outbreaks of Rift Valley fever (RVF) in Madagascar in 1990-91 and 2008-09, recent studies suggest that the pattern of RVF virus (RVFV) transmission differed among the four main eco-regions (East, Highlands, North-West and South-West). Using Bayesian hierarchical models fitted to serological data from cattle of known age collected during two surveys (2008 and 2014), we estimated RVF FOI and described its variations over time and space in Madagascar. We show that the patterns of RVFV transmission strongly differed among the eco-regions. In the North-West and Highlands regions, these patterns were synchronous with a high intensity in mid-2007/mid-2008. In the East and South-West, the peaks of transmission were later, between mid-2008 and mid-2010. In the warm and humid northwestern eco-region favorable to mosquito populations, RVFV is probably transmitted all year-long at low-level during inter-epizootic period allowing its maintenance and being regularly introduced in the Highlands through ruminant trade. The RVF surveillance of animals of the northwestern region could be used as an early warning indicator of an increased risk of RVF outbreak in Madagascar.

Correction: Integrated Analysis of Environment, Cattle and Human Serological Data: Risks and Mechanisms of Transmission of Rift Valley Fever in Madagascar.

[This corrects the article DOI: 10.1371/journal.pntd.0004827.].

Integrated Analysis of Environment, Cattle and Human Serological Data: Risks and Mechanisms of Transmission of Rift Valley Fever in Madagascar.

BACKGROUND: Rift Valley fever (RVF) is a vector-borne disease affecting ruminants and humans. Madagascar was heavily affected by RVF in 2008-2009, with evidence of a large and heterogeneous spread of the disease. The identification of at-risk environments is essential to optimize the available resources by targeting RVF surveillance in Madagascar. Herein, the objectives of our study were: (i) to identify the environmental factors and areas favorable to RVF transmission to both cattle and human and (ii) to identify human behaviors favoring human infections in Malagasy contexts. METHODOLOGY/PRINCIPAL FINDINGS: First, we characterized the environments of Malagasy communes using a Multiple Factor Analysis (MFA). Then, we analyzed cattle and human serological data collected at national level using Generalized Linear Mixed Models, with the individual serological status (cattle or human) as the response, and MFA factors, as well as other potential risk factors (cattle density, human behavior) as explanatory variables. Cattle and human seroprevalence rates were positively associated to humid environments (p<0.001). Areas with high cattle density were at risk (p<0.01; OR = 2.6). Furthermore, our analysis showed that frequent contact with raw milk contributed to explain human infection (OR = 1.6). Finally, our study highlighted the eastern-coast, western and north-western parts as high-risk areas for RVF transmission in cattle. CONCLUSIONS/SIGNIFICANCE: Our integrated approach analyzing environmental, cattle and human datasets allow us to bring new insight on RVF transmission patterns in Madagascar. The association between cattle seroprevalence, humid environments and high cattle density suggests that concomitant vectorial and direct transmissions are critical to maintain RVF enzootic transmission. Additionally, in the at-risk humid environment of the western, north-western and the eastern-coast areas, suitable to Culex and Anopheles mosquitoes, vectorial transmission probably occurs in both cattle and human. The relative contribution of vectorial or direct transmissions could be further assessed by mathematic modelling.

Genetic diversity of hepatitis B virus (HBV) in Madagascar.

Hepatitis B virus (HBV) is a DNA virus belonging to Hepadnaviridae family. Chronic infection with HBV is one major risk factor of hepatic disease. In Madagascar, former studies classified the country as part of high endemic area, as HBV prevalence can reach 23% in general population. However, this prevalence differs largely between urban and rural areas and is estimated to be, respectively, 5% and 26%. The aims of the present study were to describe the genetic diversity of HBV strains from different regions of Madagascar, and to describe the viral gene flow throughout the country by using phylogenetic analysis. This is the first large-scale molecular and phylogenetic study analyzing HBV sequences from 28 different Malagasy areas, never sampled in the past. In this study, the most prevalent genotype/sub-genotypes was E. Migration analysis showed a gene flow from zone 3 (rural) to zone 2 (suburban), and a greater gene flow from the middle part of Madagascar to the north than to the south. It is important to study the HBV infections in Madagascar and to monitor the potential spread of this viral strain inside this country. J. Med. Virol. 88:2138-2144, 2016. © 2016 Wiley Periodicals, Inc.

Seroepidemiological Study of Interepidemic Rift Valley Fever Virus Infection Among Persons with Intense Ruminant Exposure in Madagascar and Kenya.

In this cross-sectional seroepidemiological study we sought to examine the evidence for circulation of Rift Valley fever virus (RVFV) among herders in Madagascar and Kenya. From July 2010 to June 2012, we enrolled 459 herders and 98 controls (without ruminant exposures) and studied their sera (immunoglobulin G [IgG] and IgM through enzyme-linked immunosorbent assay [ELISA] and plaque reduction neutralization test [PRNT] assays) for evidence of previous RVFV infection. Overall, 59 (12.9%) of 459 herders and 7 (7.1%) of the 98 controls were positive by the IgG ELISA assay. Of the 59 ELISA-positive herders, 23 (38.9%) were confirmed by the PRNT assay (21 from eastern Kenya). Two of the 21 PRNT-positive study subjects also had elevated IgM antibodies against RVFV suggesting recent infection. Multivariate modeling in this study revealed that being seminomadic (odds ratio [OR] = 6.4, 95% confidence interval [CI] = 2.1-15.4) was most strongly associated with antibodies against RVFV. Although we cannot know when these infections occurred, it seems likely that some interepidemic RVFV infections are occurring among herders. As there are disincentives regarding reporting RVFV outbreaks in livestock or wildlife, it may be prudent to conduct periodic, limited, active seroepidemiological surveillance for RVFV infections in herders, especially in eastern Kenya.

Evidence for circulation of the rift valley fever virus among livestock in the union of Comoros.

Rift Valley fever virus (RVFV) is an arthropod-borne phlebovirus reported to be circulating in most parts of Africa. Since 2009, RVFV has been suspected of continuously circulating in the Union of Comoros. To estimate the incidence of RVFV antibody acquisition in the Comorian ruminant population, 191 young goats and cattle were selected in six distinct zones and sampled periodically from April 2010 to August 2011. We found an estimated incidence of RVFV antibody acquisition of 17.5% (95% confidence interval (CI): [8.9-26.1]) with a significant difference between islands (8.2% in Grande Comore, 72.3% in Moheli and 5.8% in Anjouan). Simultaneously, a longitudinal entomological survey was conducted and ruminant trade-related information was collected. No RVFV RNA was detected out of the 1,568 blood-sucking caught insects, including three potential vectors of RVFV mosquito species. Our trade survey suggests that there is a continuous flow of live animals from eastern Africa to the Union of Comoros and movements of ruminants between the three Comoro islands. Finally, a cross-sectional study was performed in August 2011 at the end of the follow-up. We found an estimated RVFV antibody prevalence of 19.3% (95% CI: [15.6%-23.0%]). Our findings suggest a complex RVFV epidemiological cycle in the Union of Comoros with probable inter-islands differences in RVFV circulation patterns. Moheli, and potentially Anjouan, appear to be acting as endemic reservoir of infection whereas RVFV persistence in Grande Comore could be correlated with trade in live animals with the eastern coast of Africa. More data are needed to estimate the real impact of the disease on human health and on the national economy.

Detection of Bartonella quintana in African body and head lice.

Currently, the body louse is the only recognized vector of Bartonella quintana, an organism that causes trench fever. In this work, we investigated the prevalence of this bacterium in human lice in different African countries. We tested 616 head lice and 424 body lice from nine African countries using real-time polymerase chain reaction targeting intergenic spacer region 2 and specific B. quintana genes. Overall, B. quintana DNA was found in 54% and 2% of body and head lice, respectively. Our results also show that there are more body lice positive for B. quintana in poor countries, which was determined by the gross domestic product, than in wealthy areas (228/403 versus 0/21, P < 0.001). A similar finding was obtained for head lice (8/226 versus 2/390, P = 0.007). Our findings suggest that head lice in Africa may be infected by B. quintana when patients live in poor economic conditions and are also exposed to body lice.

Anjozorobe hantavirus, a new genetic variant of Thailand virus detected in rodents from Madagascar.

Until now, there was only serological evidence that hantaviruses were circulating in rodents and infecting humans from Madagascar. To assess the presence of a hantavirus on the island, between October, 2008, and March, 2010, we sampled 585 rodents belonging to seven species in the Anjozorobe-Angavo forest corridor, 70 km north from the capital city Antananarivo. A hantavirus was detected from organs of the ubiquist roof rat (Rattus rattus) and of the endemic Major's tufted-tailed rat (Eliurus majori). Amazingly, sequence analysis of the S (small), M (medium), and L (large) coding DNA sequence of this virus showed that the Anjozorobe strain (proposed name) was a new genetic variant of Thailand virus (THAIV) that comprises other variants found in Southeast Asia. Because THAIV is suspected of causing hemorrhagic fever with renal syndrome in humans, ongoing studies are addressing the risk of infection by this new variant in the Malagasy population.

The role of wild mammals in the maintenance of Rift Valley fever virus.

Rift Valley fever virus (RVFV) is a zoonotic arbovirus affecting primarily domestic ruminants and humans. Numerous vector species are known or implicated in the transmission of RVFV. The role of mammals in the maintenance of RVFV, and the existence of a wild mammal reservoir in the epidemiologic cycle of RVFV, remain largely unknown. Our objective is to present a detailed review of studies undertaken on RVFV, often associated with wild mammals, with the aim of focusing future research on potential reservoirs of the virus. Natural and experimental infections related to RVFV in several mammalian orders, including Artiodactyla, Chiroptera, Rodentia, Primata (nonhuman), Perissodactyla, Carnivora, Proboscidea, Erinaceomorpha, and Lagomorpha, are reviewed; the first four orders have received the greatest attention. The possible role of wild ruminants, especially African buffalo (Syncerus caffer), is also discussed. Conflicting results have been published concerning rodents but, based on the literature, the likely candidate species include the African genera Arvicanthis and Micaelamys and the widely introduced roof rat (Rattus rattus). Members of the orders Chiroptera and Rodentia should receive greater attention associated with new research programs. For the other orders mentioned above, few data are available. We are unaware of any investigation concerning the orders Afrosoricida and Soricomorpha, which are represented in the geographic area of RVFV and can be abundant. As a first step to resolve the question of wild mammals as a reservoir of RVFV, serologic and virologic surveys should be promoted during epizootic periods to document infected wild animals and, in the case of positive results, extended to interepidemic periods to explore the role of wild animals as possible reservoirs.

Detection, isolation, and genetic characterization of Rift Valley fever virus from Anopheles (Anopheles) coustani, Anopheles (Anopheles) squamosus, and Culex (Culex) antennatus of the Haute Matsiatra region, Madagascar.

Following veterinary alerts of Rift Valley fever (RVF) in the districts of Fianarantsoa I and II in November 2008 and in the district of Ambalavao in April 2009, entomological and virological investigations were carried out to identify the mosquito species that could act as RVF virus (RVFV) vectors in the region. A total of 12,785 adult mosquitoes belonging to 5 genera and 21 species were collected. After identification, mosquitoes were pooled by species, sex, and female status (fed or unfed) and then stored at -80°C. Of 319 pools of unfed monospecific female mosquito tested by real-time RT-polymerase chain reaction, RVFV was detected in 1 pool of Anopheles coustani, 5 pools of An. squamosus, and 2 pools of Culex antennatus mosquitoes. The virus was isolated in mosquito cell lines from two of the five Real Time-RT-polymerase chain reaction (real time-RT-PCR) positive pools of An. squamosus mosquitoes. From the eight RVFV strains detected, partial S, M, and L genome segments sequences were obtained. The phylogenetic analysis of these sequences showed that the strains circulating in mosquitoes were genetically close to those that circulated in livestock and humans during RVF outbreaks in 2008 and 2009. This study, therefore, provides strong evidence that An. squamosus, An. coustani, and Cx. antennatus could play a role as vectors of the RVFV during the disease outbreaks in 2008-2009. Bioecological, genetic, and RVF transmission studies on these three mosquito species are needed to address this question and thus improve prevention and control of future RVF outbreaks in Madagascar, where these species are present.