Serological Evidence of West Nile and Usutu Viruses Circulation in Domestic and Wild Birds in Wetlands of Mali and Madagascar in 2008.

The geographical distribution and impact on animal and human health of both West Nile and Usutu viruses, two flaviviruses of the Japanese encephalitis complex, have been increasing during the past two decades. Both viruses circulate in Europe and Africa within a natural cycle between wild birds and mosquitoes, mainly from the Culex genus. We retrospectively analyzed sera from domestic and wild birds sampled in 2008 in two wetlands, namely the Inner Niger Delta, Mali, and the Lake Alaotra area, Madagascar. Sera were first tested using a commercial ID Screen West Nile Competition Multi-species ELISA kit. Then, positive sera and sera with insufficient volume for testing with ELISA were tested with a Microneutralization Test. In Mali, the observed seroprevalence in domestic birds was 28.5% [24.5; 32.8] 95%CI, 3.1 % [1.8; 5.2] 95%CI, 6.2% [3.4; 10.2] 95%CI and 9.8 % [7.3; 12.8] 95%CI, for West Nile virus (WNV), Usutu virus (USUV), undetermined flavivirus, and WNV/USUV respectively. Regarding domestic birds of Madagascar, the observed seroprevalence was 4.4 % [2.1; 7.9]95%CI for WNV, 0.9% [0.1; 3.1] 95%CI for USUV, 1.3% [0.5; 2.8] 95%CI for undetermined flavivirus, and null for WNV/USUV. Among the 150 wild birds sampled in Madagascar, two fulvous whistling-ducks (Dendrocygna bicolor) were positive for WNV and two for an undetermined flavivirus. One white-faced whistling-duck (Dendrocygna viduata) and one Hottentot teal (Spatula hottentota) were tested positive for USUV. African and European wetlands are linked by wild bird migrations. This first detection of USUV-as well as the confirmed circulation of WNV in domestic birds of two wetlands of Mali and Madagascar-emphasizes the need to improve the surveillance, knowledge of epidemiological patterns, and phylogenetic characteristics of flavivirus in Africa, particularly in areas prone to sustained, intense flavivirus transmission such as wetlands.

Landscape attributes driving avian influenza virus circulation in the Lake Alaotra region of Madagascar.

While the spatial pattern of the highly pathogenic avian influenza H5N1 virus has been studied throughout Southeast Asia, little is known on the spatial risk factors for avian influenza in Africa. In the present paper, we combined serological data from poultry and remotely sensed environmental factors in the Lake Alaotra region of Madagascar to explore for any association between avian influenza and landscape variables. Serological data from cross-sectional surveys carried out on poultry in 2008 and 2009 were examined together with a Landsat 7 satellite image analysed using supervised classification. The dominant landscape features in a 1-km buffer around farmhouses and distance to the closest water body were extracted. A total of 1,038 individual bird blood samples emanating from 241 flocks were analysed, and the association between avian influenza seroprevalence and these landcape variables was quantified using logistic regression models. No evidence of the presence of H5 or H7 avian influenza subtypes was found, suggesting that only low pathogenic avian influenza (LPAI) circulated. Three predominant land cover classes were identified around the poultry farms: grassland savannah, rice paddy fields and wetlands. A significant negative relationship was found between LPAI seroprevalence and distance to the closest body of water. We also found that LPAI seroprevalence was higher in farms characterised by predominant wetlands or rice landscapes than in those surrounded by dry savannah. Results from this study suggest that if highly pathogenic avian influenza H5N1 virus were introduced in Madagascar, the environmental conditions that prevail in Lake Alaotra region may allow the virus to spread and persist.

Description and analysis of the cattle trade network in the Madagascar highlands: potential role in the diffusion of Rift Valley fever virus.

In 2008-2009 a Rift Valley Fever (RVF) outbreak occurred in the Anjozorobe area, a temperate and mountainous area of the Madagascar highlands. The results of a serosurvey conducted in 2009 suggested recurrent circulation of RVF virus (RVFV) in this area and potential involvement of the cattle trade in RVFV circulation. The objective of this study was to describe the cattle trade network of the area and analyse the link between network structure and RVFV circulation. Five hundred and sixteen animals that tested negative in 2009 were sampled again in 2010. The 2009-2010 cattle-level seroconversion rate was estimated at 7% (95% CI: 5-10%). Trade data from 386 breeders of 48 villages were collected and analysed using social network analysis methodology, nodes being villages and ties being any movements of cattle connecting villages. The specific practice of cattle barter, known as kapsile, that involves frequent contacts between cattle of two breeders, was observed in addition to usual trade. Trade data were analysed using a logistic model, the occurrence of seroconversion at the village level being the outcome variable and the network centrality measures being the predictors. A negative association was observed between the occurrence of seroconversion in the village and introduction of cattle by trade (p=0.03), as well as the distance to the nearest water point (p=0.002). Conversely, the practice of kapsile, was a seroconversion risk factor (p=0.007). The kapsile practice may be the support for inter-village RVFV circulation whereas the trade network is probably rather implicated in the introduction of RVFV to the area from other parts of Madagascar. The negative association of the distance to the nearest water point suggests that after RVFV introduction, a substantial part of transmission may be due to vectors.

An unexpected recurrent transmission of Rift Valley fever virus in cattle in a temperate and mountainous area of Madagascar.

Rift Valley fever is an acute, zoonotic viral disease of domestic ruminants, caused by a phlebovirus (Bunyaviridae family). A large outbreak occurred in Madagascar in 2008-2009. The goal of the present study was to evaluate the point prevalence of antibodies against Rift Valley Fever Virus (RVFV) in cattle in the Anjozorobe district, located in the wet and temperate highland region of Madagascar and yet heavily affected by the disease, and analyse environmental and trade factors potentially linked to RVFV transmission. A serological study was performed in 2009 in 894 bovines. For each bovine, the following variables were recorded: age, location of the night pen, minimum distance from the pen to the nearest water point and the forest, nearest water point type, and herd replacement practices. The serological data were analyzed using a generalized linear mixed model. The overall anti-RVFV IgG seroprevalence rate was 28% [CI95% 25-31]. Age was statistically linked to prevalence (p = 10(-4)), being consistent with a recurrent RVFV circulation. Distance from the night pen to the nearest water point was a protective factor (p = 5.10(-3)), which would be compatible with a substantial part of the virus transmission being carried out by nocturnal mosquito vectors. However, water point type did not influence the risk of infection: several mosquito species are probably involved. Cattle belonging to owners who purchase animals to renew the herd were significantly more likely to have seroconverted than others (p = 0.04): cattle trade may contribute to the introduction of the virus in this area. The minimum distance of the night pen to the forest was not linked to the prevalence. This is the first evidence of a recurrent transmission of RVFV in such an ecosystem that associates a wet, temperate climate, high altitude, paddy fields, and vicinity to a dense rain forest. Persistence mechanisms need to be further investigated.