Christophe Paupy

Aedes albopictus, an arbovirus vector: from the darkness to the light.

The Asian tiger mosquito, Aedes albopictus (Skuse, 1894), is an invasive species that can be found on all continents. The species, originally considered a secondary vector of viruses such as Dengue viruses, has recently been suggested to play a role in the transmission of Chikungunya virus in several countries bordering the Indian Ocean, Central Africa and Europe. Here we review the current geographic range and the relevant biological traits of A. albopictus in order to explain its rapid spread. We examine and discuss recent changes in its role as a vector, particularly in the transmission of arboviruses, and its importance in the current and future emergence of pathogens. Finally, we report conventional and innovative ways to control A. albopictus.

Zika Virus in Gabon (Central Africa) – 2007: A New Threat from Aedes albopictus?

Background Chikungunya and dengue viruses emerged in Gabon in 2007, with large outbreaks primarily affecting the capital Libreville and several northern towns. Both viruses subsequently spread to the south-east of the country, with new outbreaks occurring in 2010. The mosquito species Aedes albopictus, that was known as a secondary vector for both viruses, recently invaded the country and was the primary vector involved in the Gabonese outbreaks. We conducted a retrospective study of human sera and mosquitoes collected in Gabon from 2007 to 2010, in order to identify other circulating arboviruses. Methodology/Principal Findings Sample collections, including 4312 sera from patients presenting with painful febrile disease, and 4665 mosquitoes belonging to 9 species, split into 247 pools (including 137 pools of Aedes albopictus), were screened with molecular biology methods. Five human sera and two Aedes albopictus pools, all sampled in an urban setting during the 2007 outbreak, were positive for the flavivirus Zika (ZIKV). The ratio of Aedes albopictus pools positive for ZIKV was similar to that positive for dengue virus during the concomitant dengue outbreak suggesting similar mosquito infection rates and, presumably, underlying a human ZIKV outbreak. ZIKV sequences from the envelope and NS3 genes were amplified from a human serum sample. Phylogenetic analysis placed the Gabonese ZIKV at a basal position in the African lineage, pointing to ancestral genetic diversification and spread. Conclusions/Significance We provide the first direct evidence of human ZIKV infections in Gabon, and its first occurrence in the Asian tiger mosquito, Aedes albopictus. These data reveal an unusual natural life cycle for this virus, occurring in an urban environment, and potentially representing a new emerging threat due to this novel association with a highly invasive vector whose geographic range is still expanding across the globe.

Comparative role of Aedes albopictus and Aedes aegypti in the emergence of dengue and Chikungunya in Central Africa

Since its discovery in Nigeria in 1991, Aedes albopictus has invaded much of Central Africa, a region where Ae. aegypti also occurs. To assess the relationship between the invasion by Ae. albopictus and the recent emergence of dengue virus (DENV) and chikungunya virus (CHIKV), we undertook vector competence experiments on populations collected from Cameroon and conducted field investigations during concurrent epidemics of DENV and CHIKV in Gabon. Overall, infection and dissemination rates were not significantly different between Ae. albopictus and Ae. aegypti when exposed to titers of 10(8.1) mosquito infectious dose 50/mL and 10(7.5) plaque forming units/mL of DENV type 2 and CHIKV, respectively. Field investigations showed that Ae. albopictus readily bit man, was abundant, and outnumbered Ae. aegypti to a large extent in Gabon, particularly in suburban environments. Nevertheless, Ae. aegypti was predominant in the more urbanized central parts of Libreville. In this city, CHIKV and DENV were detected only in Ae. albopictus. These data strongly suggest that Ae. albopictus acted as the major vector of both viruses in Libreville in 2007, impacting on the epidemiology of DENV and CHIKV in this area.

Worldwide patterns of genetic differentiation imply multiple ‘domestications’ of Aedes aegypti, a major vector of human diseases

Understanding the processes by which species colonize and adapt to human habitats is particularly important in the case of disease-vectoring arthropods. The mosquito species Aedes aegypti, a major vector of dengue and yellow fever viruses, probably originated as a wild, zoophilic species in sub-Saharan Africa, where some populations still breed in tree holes in forested habitats. Many populations of the species, however, have evolved to thrive in human habitats and to bite humans. This includes some populations within Africa as well as almost all those outside Africa. It is not clear whether all domestic populations are genetically related and represent a single ‘domestication’ event, or whether association with human habitats has developed multiple times independently within the species. To test the hypotheses above, we screened 24 worldwide population samples of Ae. aegypti at 12 polymorphic microsatellite loci. We identified two distinct genetic clusters: one included all domestic populations outside of Africa and the other included both domestic and forest populations within Africa. This suggests that human association in Africa occurred independently from that in domestic populations across the rest of the world. Additionally, measures of genetic diversity support Ae. aegypti in Africa as the ancestral form of the species. Individuals from domestic populations outside Africa can reliably be assigned back to their population of origin, which will help determine the origins of new introductions of Ae. aegypti.

Geographic distribution and developmental sites of Aedes albopictus (Diptera: Culicidae) during a Chikungunya epidemic event.

Aedes albopictus is generally considered to have a low vectorial capacity because of its lack of host specificity. Nevertheless, it has been the sole vector of the Chikungunya virus in recent explosive epidemics on the islands of La Réunion and Mauritius. We report on investigations of the seasonal prevalence, container preferences, and geographic distribution of the species on La Réunion. Ae. albopictus showed strong ecological plasticity. In the warm wet season, small disposable containers were the principal urban breeding site, with 1939 positive containers in 750 houses. In the dry winter season, the species remained abundant throughout the island up to 800 m and was present to a maximum altitude of 1200 m. Natural containers were clearly important in this season, although productive sources were hard to find. The preferred natural developmental sites were bamboo stumps and rock holes, over 357 developmental sites observed in peri-urban and gully areas. Generalized logistic models indicated that the optimum sites contained clear water with high organic content and were situated in sites with moderate shade. Our data will provide input into the models of the epidemiology of the disease and design of vector control programs.

Recent Introduction and Rapid Dissemination of Chikungunya Virus and Dengue Virus Serotype 2 Associated With Human and Mosquito Coinfections in Gabon, Central Africa

BACKGROUND Chikungunya virus (CHIKV) and Dengue virus serotype 2 (DENV-2) were recently introduced in central Africa, along with Aedes albopictus. Simultaneous outbreaks of CHIKV and DENV-2 have subsequently occurred, in Cameroon in 2006 and Gabon in 2007. METHODS To study the spread of the 2 viruses, we conducted active surveillance of acute febrile syndromes throughout Gabon between 2007 and 2010. Diagnostic methods included quantitative real-time reverse-transcription polymerase chain reaction, and molecular characterization was based on partial envelope gene sequences. RESULTS Between 2007 and 2010, 4287 acutely febrile patients were investigated for CHIKV and DENV-2 infections, of whom 1567 were CHIKV-positive, 376 DENV-2-positive, and 37 coinfected. We diagnosed 153 CHIKV and 11 DENV-2 cases in 2008, and 5 CHIKV and 9 DENV-2 cases in 2009. In 2010, CHIKV and DENV-2 caused a second large simultaneous outbreak. Among 2826 acutely febrile patients examined during this outbreak, 1112 were CHIKV-positive, 288 DENV-2-positive, and 28 coinfected. Mosquitoes were collected near the homes of coinfected patients, and 1 Aedes albopictus specimen was found to be positive for both CHIKV and DENV-2. CONCLUSIONS These findings show the rapid dissemination of CHIKV and DENV-2 within a nonimmune population in a tropical African country, probably facilitated by the spread of Aedes albopictus. This has resulted in major simultaneous outbreaks with numerous coinfections in both human and mosquito.

Chikungunya virus, Cameroon, 2006.

We report the isolation of chikungunya virus from a patient during an outbreak of a denguelike syndrome in Cameroon in 2006. The virus was phylogenetically grouped in the Democratic Republic of the Congo cluster, indicating a continuous circulation of a genetically similar chikungunya virus population during 6 years in Central Africa.

Insecticide susceptibility of Aedes aegypti and Aedes albopictus in Central Africa

BackgroundAedes aegypti (Linnaeus, 1762) and Aedes albopictus (Skuse, 1894) are the main vectors of dengue (DENV) and chikungunya (CHIKV) viruses worldwide. As there is still no vaccine or specific treatment for DENV and CHIKV, vector control remains the cornerstone of prevention and outbreak control. Unfortunately, vector control programs are facing operational challenges with mosquitoes becoming resistant to commonly used insecticides in several areas through the world. Throughout Central Africa no recent data are available susceptible/resistant status of either vector species since the introduction/arrival of Ae. albopictus in this area. We therefore studied the level of resistance of these two major vectors to insecticides commonly used in Africa for mosquito control.ResultsAedes aegypti and Ae. albopictus were sampled in six urban localities of Cameroon (Garoua, Bertoua, Yaoundé, Bafia, Buea) and Gabon (Libreville). Larval bioassays, carried out to determine the lethal concentrations (LC50 and LC95) and resistance ratios (RR50 and RR95) suggested that both vector species were susceptible to Bti (Bacillus thuringiensis var israeliensis) and temephos. Bioassays were also performed on adults using WHO diagnostic test kits to assess phenotypic resistance to deltamethrin, DDT, fenitrothion and propoxur. These experiments showed that one population of Ae. aegypti (Libreville) and two populations of Ae. albopictus (Buea and Yaoundé) were resistant to DDT (mortality 36% to 71%). Resistance to deltamethrin was also suspected in Ae. albopictus from Yaoundé (83% mortality). All other field mosquito populations were susceptible to deltamethrin, DDT, fenitrothion and propoxur. No increase in the knockdown times (Kdt50 and Kdt95) was noted in the Yaoundé resistant population compared to other Ae. albopictus populations, suggesting the possible involvement of metabolic resistance to deltamethrin and DDT.ConclusionIn view of the recent increase in dengue and chikungunya outbreaks in Central Africa, these unique comparative data on the insecticide susceptibility of Ae. aegypti and Ae. albopictus could help public health services to design more effective vector control measures.

Genetic structure of the tiger mosquito, Aedes albopictus, in Cameroon (Central Africa).

Background Aedes albopictus (Skuse, 1884) (Diptera: Culicidae), a mosquito native to Asia, has recently invaded all five continents. In Central Africa it was first reported in the early 2000s, and has since been implicated in the emergence of arboviruses such as dengue and chikungunya in this region. Recent genetic studies of invasive species have shown that multiple introductions are a key factor for successful expansion in new areas. As a result, phenotypic characters such as vector competence and insecticide susceptibility may vary within invasive pest species, potentially affecting vector efficiency and pest management. Here we assessed the genetic variability and population genetics of Ae. albopictus isolates in Cameroon (Central Africa), thereby deducing their likely geographic origin. Methods and Results Mosquitoes were sampled in 2007 in 12 localities in southern Cameroon and analyzed for polymorphism at six microsatellite loci and in two mitochondrial DNA regions (ND5 and COI). All the microsatellite markers were successfully amplified and were polymorphic, showing moderate genetic structureamong geographic populations (FST = 0.068, P<0.0001). Analysis of mtDNA sequences revealed four haplotypes each for the COI and ND5 genes, with a dominant haplotype shared by all Cameroonian samples. The weak genetic variation estimated from the mtDNA genes is consistent with the recent arrival of Ae. albopictus in Cameroon. Phylogeographic analysis based on COI polymorphism indicated that Ae. albopictus populations from Cameroon are related to tropical rather than temperate or subtropical outgroups. Conclusion The moderate genetic diversity observed among Cameroonian Ae. albopictus isolates is in keeping with recent introduction and spread in this country. The genetic structure of natural populations points to multiple introductions from tropical regions.

The invaders: Phylogeography of dengue and chikungunya viruses Aedes vectors, on the South West islands of the Indian Ocean

Aedes albopictus and Aedes aegypti are the two main worldwide arbovirus vectors that have experienced invasion phases. Aedes aegypti is a pantropical species that spread centuries ago whereas Ae. albopictus started the main wave of invasion in the 1980s. Both species have been at various times on the different islands in Southwestern Indian Ocean (SWIO). This area provides an opportunity to examine the extent to which mosquitoes colonization patterns are influenced by different introductory events likely linked with human settlement and migration between the islands. To explore this hypothesis, we propose a CO1-based phylogeny using a large sampling of fresh Ae. albopictus, Ae. aegypti, and Ae. mascarensis, and 50-year-old dry specimens originating from different Indian Ocean islands. Our data allow us to hypothesize the existence of at least two waves of invasion for Ae. albopictus in the islands of SWIO. The first one most likely occurred several centuries ago with establishments in Madagascar, Mauritius and Reunion Island. The other one that appears to currently still on-going, reached almost all the islands of SWIO during the 1990s or later. The low genetic diversity found between the ancient invasive strain and the contemporary one, indicates with great certainty that Ae. albopictus is not indigenous to the islands of SWIO. Recently, in Madagascar, an invasive lineage of Ae. albopictus has expanded all over the island while Ae. aegypti populations have declined in urban areas. Three clusters of Aedes aegypti have been observed, two fitting with the wild form Ae. aegypti formosus and the other one fitting with the more domestic form Ae. ae. aegypti. Sequence of Ae. mascarensis, endemic to Mauritius suggest that this species might belong to Ae. aegypti species and on this basis we propose to classify it as a sub species or form of Ae. aegypti species. Given the increase of human population flux on these islands, the occurrence of these vectors and their ability to spread quickly are of high importance of arbovirus transmission and the epidemicity of the associated diseases in these islands.