The use of the bacterium Wolbachia is an attractive alternative method to control vector populations. In mosquitoes, as in members of the Culex pipiens complex, Wolbachia induces a form of embryonic lethality called cytoplasmic incompatibility, a sperm-egg incompatibility occurring when infected males mate either with uninfected females or with females infected with incompatible Wolbachia strain(s). Here we explore the feasibility of the Incompatible Insect Technique (IIT), a species-specific control approach in which field females are sterilized by inundative releases of incompatible males. We show that the Wolbachia wPip(Is) strain, naturally infecting Cx. p. pipiens mosquitoes from Turkey, is a good candidate to control Cx. p. quinquefasciatus populations on four islands of the south-western Indian Ocean (La Réunion, Mauritius, Grande Glorieuse and Mayotte). The wPip(Is) strain was introduced into the nuclear background of Cx. p. quinquefasciatus mosquitoes from La Réunion, leading to the LR[wPip(Is)] line. Total embryonic lethality was observed in crosses between LR[wPip(Is)] males and all tested field females from the four islands. Interestingly, most crosses involving LR[wPip(Is)] females and field males were also incompatible, which is expected to reduce the impact of any accidental release of LR[wPip(Is)] females. Cage experiments demonstrate that LR[wPip(Is)] males are equally competitive with La Réunion males resulting in demographic crash when LR[wPip(Is)] males were introduced into La Réunion laboratory cages. These results, together with the geographic isolation of the four south-western Indian Ocean islands and their limited land area, support the feasibility of an IIT program using LR[wPip(Is)] males and stimulate the implementation of field tests for a Cx. p. quinquefasciatus control strategy on these islands.

Protecting vegetables with a screen in peri-urban areas of tropical countries could reduce or even prevent often indiscriminate insecticide applications by small-scale farmers. The advantages of such an approach are protection of human health by reducing insecticide sprays, reducing environmental pollution from insecticide residues and increasing effectiveness of crop protection. Tunnel screens are well adapted to farmers cultivating intensively on small plots. Two trials were conducted to test the ability of screened tunnels to protect Brassica oleracea crops. The first was carried out onstation and the second in partnership with three farmers in Cotonou, Benin, West Africa. Tunnel screens impregnated with deltamethrin were found to be particularly well adapted to protect young plants in seedling nurseries against infestations by the aphid Lipaphis erysimi (Kaltenbach). The number of diamondback moth Plutella xylostella (Linnaeus) and borer Hellula undalis (Fabricius) on cabbages protected with the tunnel screen was significantly lower than that of plots conventionally treated with insecticides. The tunnel screen was not efficient against the armyworm Spodoptera littoralis (Boisduval) which laid eggs on the screen. After planting out, the use of a temporary screen from 1700 to 0900 h gave better control against pests than the use of a permanent screen possibly due to the impact of natural enemies during the day. The field trials showed that the protection of cabbage with a tunnel screen could be an economically viable method. The costs of pesticides are on average US$ 45 per 100 m 2 for one crop cycle compared with US$ 24 per 100 m 2 for tunnel screen material (assuming that this material can be used for 10 consecutive crop cycles). In addition, there are environmental benefits from a reduction of pesticide use. Farmers will have to cope with the initial investment for the screen material, which is, however, very cost-effective and locally available. Tunnel screens for vegetable protection can be easily combined with other integrated pest-management techniques.

The knock-down, mortality and 'irritancy' effects of three synthetic repellents (DEET, IR3535 and KBR 3023) on Aedes aegypti (L) (Diptera: Culicidae) were evaluated in the laboratory in the absence of animal bait. Filter paper tests were carried out to assess the knock-down effect (KDt(50) and KDt(95)) and mortality (LC(50) and LC(95)) induced by each repellent. 'Irritancy' tests were carried out to compare the flight response (time to first take-off, or FT) to increasing concentrations of repellents (2-7%) and at five distances from the treated surface (0-40 mm). DEET had an insecticidal effect (KDt(50) = 9.7 min at 7%; CL(50)= 1165 mg/m(2)), whereas IR3535 and KBR 3023 did not. Relative to an untreated control, IR3535 was an irritant (relative irritancy or RI > 1) at doses of 5% and 7% (RI = 17.7 and 9.9, respectively), whereas DEET was an irritant at lower concentrations (RI = 12.3 at 2% DEET). KBR 3023 was the weakest irritant over the same range of concentrations (RI(max) = 3.6 at 6%). DEET was more of an irritant (RI(20) = 9.4) than IR3535 (RI(20) = 2.9) over a range of distances (0-20 mm), and KBR 3023 was not an irritant unless mosquitoes made contact with the treated surface. All three repellents had a significant effect on mosquitoes, but DEET exhibited a more complex mode of action than the others due to its insecticidal properties. The repellents do not behave as a single class of compounds with a common mode of action, but most probably affect different physiological systems in insects. The physiological and molecular mechanisms of repellents, especially DEET, should be investigated to ensure a better use of these molecules for skin applications and/or for treating materials against mosquitoes.

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.

The spread of resistance to pyrethroids in the major Afrotropical malaria vectors Anopheles gambiae s.s. necessitates the development of new strategies to control resistant mosquito populations. To test the effi cacy of nets treated with repellent and insecticide against susceptible and insecticideresistant An. gambiae mosquito populations, we impregnated mosquito bed nets with an insect repellent mixed with a low dose of organophosphorous insecticide and tested them in a rice-growing area near Bobo-Dioulasso, Burkina Faso. During the fi rst 2 weeks posttreatment, the mixture was as effective as deltamethrin alone and was more effective at killing An. gambiae that carried knockdown resistance (kdr) or insensitive acetylcholinesterase resistance (Ace1 R ) genes. The mixture seemed to not kill more susceptible genotypes for the kdr or Ace1 R alleles. Mixing repellents and organophosphates on bed nets could be used to control insecticideresistant malaria vectors if residual activity of the mixture is extended and safety is verifi ed. P yrethroids are the only class of insecticides that are recommended by the World Health Organization (WHO) and the Centers for Disease Control and Prevention for net impregnation to control malaria transmission (1,2). Unfortunately, malaria vector resistance to pyrethroids is becoming widespread across Africa. Pyrethroid resistance mechanisms can be divided into 2 groups: metabolic (alterations in the expression levels of activities of detoxifi cation enzymes) and target site (nonsilent point mutations within structural receptor genes, e.g., knockdown resistance [kdr] mutations) (3). Whether the spread of resistance genes will pose a serious threat to vector control programs that are based uniquely on pyrethroid use is uncertain. Some studies have shown that kdr resistance does not decrease the level of protection conferred by insecticide-treated nets (ITNs) (4) and that ITNs do not induce kdr selection (5). Conversely, more recent studies have reported a fi tness advantage for kdr-resistant phenotypes (6) and decreased effi cacy of ITNs in an area of pyrethroid resistance in Benin (7). The need for alternative insecticidal molecules is becoming increasingly clear; however, fewer novel active ingredients are available and the timeframe needed to satisfy the regulatory requirements of public health formulations is exceedingly long (8). Thus, the only option for managing insecticide resistance in malaria vectors is optimal use of existing compounds. Two such tactics have already been explored: 1) the alternating use of different classes of insecticides by rotation of active ingredients and mosaic treatments (9,10), and 2) the use of insecticide mixtures (10,11).We tested the ability of existing ingredients, a mixture of insect repellents and nonpyrethroid insecticides, to achieve vector death and excito-repellency (irritancy when mosquito contacts net and repellent activity in air) (12). The rationale behind this concept is that nonpyrethroid compounds can mimic the original features of...

The diversity and geographical distribution of fleas parasitizing small mammals have been poorly investigated on Indian Ocean islands with the exception of Madagascar where endemic plague has stimulated extensive research on these arthropod vectors. In the context of an emerging flea-borne murine typhus outbreak that occurred recently in Reunion Island, we explored fleas' diversity, distribution and host specificity on Reunion Island. Small mammal hosts belonging to five introduced species were trapped from November 2012 to November 2013 along two altitudinal transects, one on the windward eastern and one on the leeward western sides of the island. A total of 960 animals were trapped, and 286 fleas were morphologically and molecularly identified. Four species were reported: (i) two cosmopolitan Xenopsylla species which appeared by far as the prominent species, X. cheopis and X. brasiliensis; (ii) fewer fleas belonging to Echidnophaga gallinacea and Leptopsylla segnis. Rattus rattus was found to be the most abundant host species in our sample, and also the most parasitized host, predominantly by X. cheopis. A marked decrease in flea abundance was observed during the cool-dry season, which indicates seasonal fluctuation in infestation. Importantly, our data reveal that flea abundance was strongly biased on the island, with 81% of all collected fleas coming from the western dry side and no Xenopsylla flea collected on almost four hundred rodents trapped along the windward humid eastern side. The possible consequences of this sharp spatio-temporal pattern are discussed in terms of flea-borne disease risks in Reunion Island, particularly with regard to plague and the currently emerging murine typhus outbreak.

BackgroundChemicals are used on bed nets in order to prevent infected bites and to kill aggressive malaria vectors. Because pyrethroid resistance has become widespread in the main malaria vectors, research for alternative active ingredients becomes urgent. Mixing a repellent and a non-pyrethroid insecticide seemed to be a promising tool as mixtures in the laboratory showed the same features as pyrethroids.Methodology/Principal FindingsWe present here the results of two trials run against free-flying Anopheles gambiae populations comparing the effects of two insect repellents (either DEET or KBR 3023, also known as icaridin) and an organophosphate insecticide at low-doses (pirimiphos-methyl, PM) used alone and in combination on bed nets. We showed that mixtures of PM and the repellents induced higher exophily, blood feeding inhibition and mortality among wild susceptible and resistant malaria vectors than compounds used alone. Nevertheless the synergistic interactions are only involved in the high mortality induced by the two mixtures.ConclusionThese field trials argue in favour of the strategy of mixing repellent and organophosphate on bed nets to better control resistant malaria vectors.

Background: Arthropod borne virus infections are the cause of severe emerging diseases. Among the diseases due to arboviruses, dengue (DEN) and Rift Valley fever (RVF) are in the top ten in the list of diseases responsible of severe human cases worldwide. Understanding the effects of viral infection on gene expression in competent vectors is a challenge for the development of early diagnostic tools and may enable researchers and policy makers to better anticipate outbreaks in the next future. Methods: In this study, alterations in gene expression across the entire Aedes aegypti genome during infection with DENV and RVFV were investigated in vitro at two time points of infection, the early phase (24 h) and the late phase (6 days) of infection using the RNA sequencing approach Results: A total of 10 upregulated genes that share a similar expression profile during infection with both viruses at early and late phases of infection were identified. Family B and D clip-domain serine proteases (CLIP) were clearly overrepresented as well as C-type lectins and transferrin. Conclusions: Our data highlight the presence of 10 viral genes upregulated in Ae. aegypti during infection. They may also be targeted in the case of the development of broad-spectrum anti-viral diagnostic tools focusing the mosquito vectors rather than the mammalian hosts as they may predict the emergence of outbreaks.