Discrimination of 15 Amazonian Anopheline Mosquito Species by Polymerase Chain Reaction-Restriction Fragment Length Polymorphism.

Precise identification of anopheline species is paramount for incrimination of malaria vectors and implementation of a sustainable control program. Anopheline mosquitoes are routinely identified morphologically, a technique that is time-consuming, needs high level of expertise, and prone to misidentifications especially when considering Amazonian species. The aim of this study was therefore to develop a DNA-based identification technique to supplement traditional morphological identification methods for the discrimination of anopheline mosquitoes collected in French Guiana. The internal transcribed spacer 2 (ITS2) region of ribosomal DNA (rDNA) for anopheline species was amplified by polymerase chain reaction (PCR), and digested with AluI/MspI restriction enzymes. PCR-restriction fragments length polymorphism (RFLP) assay was compared to sequencing of the ITS2 region for validation. Fifteen Anopheles species have shown distinct PCR-RFLP profiles. A concordance of 100% was obtained when identification by PCR-RFLP was compared to sequencing of ITS2. A high throughput, fast, and cost-effective PCR-RFLP assay has been developed for unambiguous discrimination of fifteen anopheline mosquito species from French Guiana including primary and suspected secondary malaria vectors.

Spatial and Seasonal Dynamics of Anopheles Mosquitoes in Saint-Georges de l'Oyapock, French Guiana: Influence of Environmental Factors.

Little is known about the Anopheles fauna of Saint-Georges de l'Oyapock, a persistent malaria-endemic municipality in French Guiana. This study aimed to update the knowledge of local Anopheles diversity, and their ecology and role in malaria transmission. Sampling sessions were implemented between September 2013 and October 2014. Four species were identified from the 3,450 specimens collected: Anopheles darlingi Root, An. braziliensis, An. triannulatus s.l., and An. nuneztovari s.l. Anopheles darlingi was the predominant species. Its involvement in malaria transmission was suspected due to 1) its abundance, 2) the presence of a density peak during the malaria emergence period, and 3) a dynamic correlated with malaria cases observed two months later. Present and past studies show that the influence of environmental conditions on malaria vector dynamics is high, and may vary drastically according to the local context. This supports evidence that control strategies must be designed at fine scales.

Risk of daytime transmission of malaria in the French Guiana rain forest.

Between 2008 and 2014, there were 1070 malaria cases reported in French Guiana among members of the armed forces. Most of the malaria outbreaks investigated were multifactorial and followed missions conducted at illegal gold mining sites. For example, a malaria outbreak occurred in September 2013, three weeks after the deployment of 15 soldiers at Dagobert, which is such a site. The attack rate was 53%, with seven Plasmodium vivax infections and one coinfection with both Plasmodium vivax and Plasmodium falciparum. Two months later, an entomological investigation in the field caught 321 anopheles by the human landing catch method. Among them, 282 were Anopheles darlingi. One specimen was PCR-positive for P. vivax, for an infection rate of 0.4% (1/282). In 15.7% of these cases, the An. darlingi was caught during the day. The existence of daytime biting activity by An. darlingi in the Guianese forest might play a key role in malaria outbreaks among military personnel. This finding requires that the Army Health Service adapt its recommendations concerning malaria prevention in French Guiana.

Unravelling the relationships between Anopheles darlingi (Diptera: Culicidae) densities, environmental factors and malaria incidence: understanding the variable patterns of malarial transmission in French Guiana (South America).

Anopheles darlingi, one of the main malaria vectors in the Neotropics, is widely distributed in French Guiana, where malaria remains a major public-health problem. Elucidation of the relationships between the population dynamics of An. darlingi and local environmental factors would appear to be an essential factor in the epidemiology of human malaria in French Guiana and the design of effective vector-control strategies. In a recent investigation, longitudinal entomological surveys were carried out for 2-4 years in one village in each of three distinct endemic areas of French Guiana. Anopheles darlingi was always the anopheline mosquito that was most frequently caught on human bait, although its relative abundance (as a proportion of all the anophelines collected) and human biting rate (in bites/person-year) differed with the study site. Seasonality in the abundance of human-landing An. darlingi (with peaks at the end of the rainy season) was observed in only two of the three study sites. Just three An. darlingi were found positive for Plasmodium (either P. falciparum or P. vivax) circumsporozoite protein, giving entomological inoculation rates of 0·0-8·7 infectious bites/person-year. Curiously, no infected An. darlingi were collected in the village with the highest incidence of human malaria. Relationships between malaria incidence, An. darlingi densities, rainfall and water levels in the nearest rivers were found to be variable and apparently dependent on land-cover specificities that reflected the diversity and availability of habitats suitable for the development and reproduction of An. darlingi.

Evaluation of four methods for collecting malaria vectors in French Guiana.

In French Guiana, malaria transmission is mainly due to Anopheles darlingi Root, but other species also are involved. Investigation and surveillance must be carried out on all the species to unravel malaria transmission patterns. In this study, we aimed to compare the ability of Mosquito Magnet- and CDC-Light Trap-based methods and human landing collection for evaluating abundance of Anopheles species. Human landing collections exhibited the best results, followed by the Mosquito Magnet and the CDC-Light Trap methods. Studies have to be done to further investigate these methods and other traps have to be tested.

Aedes aegypti in French Guiana: susceptibility to a dengue virus.

Twenty-seven samples of Aedes aegypti (F1 generation) from French Guiana were tested for their susceptibility to dengue serotype 2 virus. Very high infection rates were observed by indirect fluorescent antibody (IFA) test. Ae. aegypti samples were pooled according to two groups: the first group (N=10) represented mosquitoes from the urbanized area of Cayenne and surroundings, and the second group (N=17) corresponded to mosquitoes collected in the countryside. Infection rates were found to be similar in these two cases. These findings are discussed in relation with the history of Ae. aegypti in this part of the world.

Some epidemiological and entomological aspects of the dengue situation and the vector aedes aegypti (L) in French Guiana

General epidemiological and entomological aspect studies of the dengue fever (DF) situation and the mosquito vector, Aedes aegypti, in French Guiana were studied for two years, between 1st October 1993 and 30th September 1995. The methodology included epidemiological data gathered from patients recently infected by a dengue virus, and entomological investigations with sampling of all stages (eggs, larvae, pupae and adults) of Ae.aegypti in the suspected dengue-infected places. DF cases were reported from all parts of the country, mostly from the main city of Cayenne and its surroundings. Likewise, Ae.aegypti was found in all investigated inhabited areas, and infected vectors were collected in four locations. Sampling of immature Ae.aegypti showed that the small outdoor containers were the most important breeding-sites in French Guiana. The duration of development of Ae. aegypti was significantly different according to the type of breeding-site. The vertical transmission of Df viruses under naturual conditions in French Guiana was demonstrated and 2 DEN-2 and 2 DEN-4 strains were isolated. We conclude that DF viruses are endemic in French Guiana, all inhabitied areas of the country are susceptible to epidemic DF and the reponsible vector, Ae.aegypti, can also act as a reservoir for the dengue virus (AU)

[Aedes aegypti in French Guiana. Some aspects of history, general ecology and vertical transmission of the dengue virus]. / Aedes Aegypti en Guyane Franaise. Quelques aspects de i'histoire, de l'écologie générale et de la transmission verticale des virus de la dengue.

Aedes aegypti is suspected to be present in the country since the late 18th century, and was responsible of urban yellow fever epidemics in the last century. This mosquito was identified for the first time in French Guiana in 1902. More recently, in 1940, an eradication campaign started and Aedes aegypti was eradicated between 1950 and 1963, date of the reinfestation. During the past 30 years, some dengue outbreaks occurred every 2 to 6 years, and the first dengue haemorrhagic fever epidemic spread over the country in 1992. Actually, Ae. aegypti is distributed almost all inhabited areas of French Guiana: in the towns, villages, smaller human settlements, and was also found in a wild area. The most frequent Ae. aegypti breeding-sources are the outside discarded small containers, other less frequent breeding-sites are the outside flower pots and the outside big containers. The type of breeding-source significantly influences the duration of larval and pupal development. In French Guiana, Ae. aegypti is the only vector of dengue. The vertical transmission of dengue viruses under field conditions was demonstrated. Dengue is thus endemic in the country and has almost the same distribution as Ae. aegypti, with most probably the same possibilities of extension. Ae. aegypti can be considered not only as vector and an amplificator of dengue in French Guiana, but also as a reservoir, even if occasional.