Hamidou Maïga

Assessment of Alternative Mating Strategies in Anopheles gambiae: Does Mating Occur Indoors?

Abstract Mating in Anopheles gambiae has been observed only in outdoor swarms. Here we evaluate whether mating also occurs indoors. Mark-release-recapture of virgin males and females in natural houses showed that mating occurred over a single day even when mosquitoes can leave the house through exit traps and without adaptation to laboratory conditions. In these experiments, insemination rate in the M molecular form of An. gambiae (and An. arabiensis) was higher than that of the S form (15 versus 6%). Under these conditions, smaller females of the M form mated more frequently than larger females of that form. Sampling mosquitoes throughout the day showed that both sexes enter houses around sunrise and leave around sunset, staying indoors together from dawn to dusk. In an area dominated by the M form, the daily rate of insemination in samples from exit traps was ≈5% higher than in those from entry traps, implying that mating occurred indoors. Importantly, frequency of cross mating between the molecular forms was as high as that between members of the same form, indicating that, indoors, assortative mating breaks down. Altogether, these results suggest that indoor mating is an alternative mating strategy of the M molecular form of An. gambiae. Because naturally occurring mating couples have not yet been observed indoors, this conclusion awaits validation.

Effect of three larval diets on larval development and male sexual performance of Anopheles gambiae s.s.

Population replacement/elimination strategies based on mass-release of sterile or otherwise genetically modified (male) mosquitoes are being considered in order to expand the malaria vector control arsenal on the way to eradication. A challenge in this context, is to produce male mosquitoes that will be able to compete and mate with wild females more efficiently than their wild counterparts, i.e. high fitness males. This study explored the effect of three larval food diets developed by the International Atomic Energy Agency on the overall fitness and mating performance of male Anopheles gambiae s.s. mosquitoes (Kisumu strain). Larval development (pupation and emergence rate, development time) was monitored, and adult wing length and energy reserves at emergence (i.e. lipids, sugars, glycogen and proteins) were measured. Male sexual performance was assessed through an insemination test whereby one male and 10 virgin females were maintained together in the same cage in order to record the number of inseminated females per 24h. Our results show that males reared on Diets 2 and 3 performed best during larval development. Males provided with treatment 2.2 had a shorter development time and performed best in insemination tests. However, these males had the lowest overall lifespan, suggesting a trade-off between longevity and sexual performances which needs to be taken into consideration when planning release. The results from this work were discussed in the context of sterile insect techniques or genetic control methods which is today one of the strategy in the overall mosquito control and elimination efforts.

Role of nutritional reserves and body size in Anopheles gambiae males mating success

A better knowledge of the different parameters that account for male mating success in the wild is critical to the development of genetic control strategies. In this study, we measured energy budgets (total sugar and glycogen) as the daily energetic investment in swarming males of An. gambiae s.s. M and S molecular forms from two different field locations, VK7 and Soumousso. We also looked at the difference between energetic reserves in mated males compared to unmated ones, and assessed wing length in both molecular forms to explore whether this phenotypic trait was involved in swarming behavior or mating success. The current study showed that the energetic cost of 25 min of swarming was around 50% of the males sugar (M form: 48.5%, S form: 56.2%) and glycogen (M form: 53.1%, S form: 59%) reserves. However, no difference in carbohydrate content was observed between mated and unmated males. Mated males were found to be bigger than unmated ones, while intermediate size of males is advantageous in mating system, both in M and S molecular forms and when collected in two different locations. Regardless of the collection location, no difference in wing size was observed in swarming males collected early or late during a particular swarm. The results are discussed in the context sexual selection in different ecological locations.

Variation in energy reserves and role of body size in the mating system of Anopheles gambiae

ABSTRACT: Anopheles gambiae mates in flight. Males gather at stationary places at sunset and compete for incoming females. Factors that account for male mating success are not known but are critical for the future of any genetic control strategy. The current study explored variations in nutritional reserves (sugars, glycogen, lipids, and proteins) in wild-caught swarming and resting males and evaluated the effect of body size and wing symmetry on male mating success. Our results showed that glycogen and sugar reserves are mobilized for flight. Males consume proportionally 5.9-fold as much energy derived from sugars in swarming activities than when they are at rest. Mated males were on average bigger than unmated ones (P<0.0001). A strong correlation between the left and right wings in both mated and unmated males was found and additional analysis on fluctuating asymmetry did not show any indication of mated males being more symmetrical than unmated ones. The distribution of wing size of mated males was focused around a central value, suggesting that intermediate size of males is advantageous in the An. gambiae mating system. The results are discussed in the context of sexual selection.

Swarming behaviour in natural populations of Anopheles gambiae and An. coluzzii: review of 4 years survey in rural areas of sympatry, Burkina Faso (West Africa)

The swarming behaviour of natural populations of Anopheles gambiae and An. coluzzii (formerly known as An. gambiae S and M forms, respectively) were investigated through longitudinal surveys conducted between July 2006 and October 2009 in two rural areas of south-western Burkina Faso where these forms are sympatric. In both sites, the majority of swarms were recorded above visual markers localised among houses. In Soumousso, a wooded area of savannah, 108 pairs caught in copula from 205 swarms were sampled; in VK7, a rice growing area, 491 couples from 250 swarms were sampled. If segregated swarms were the norm in both sites, many visual markers were shared by the two forms of An. gambiae. Furthermore, mixed swarms were collected annually in frequencies varying from one site to another, though no mixed inseminations were recorded, corroborating the low hybrid rate previously reported in the field. The occurrence of inter-specific mate-recognition mechanisms, which allow individuals to avoid hybridisation, is discussed.

Targeting male mosquito swarms to control malaria vector density

Malaria control programs are being jeopardized by the spread of insecticide resistance in mosquito vector populations. It has been estimated that the spread of resistance could lead to an additional 120000 deaths per year, and interfere with the prospects for sustained control or the feasibility of achieving malaria elimination. Another complication for the development of resistance management strategies is that, in addition to insecticide resistance, mosquito behavior evolves in a manner that diminishes the impact of LLINs and IRS. Mosquitoes may circumvent LLIN and IRS control through preferential feeding and resting outside human houses and/or being active earlier in the evening before people go to sleep. Recent developments in our understanding of mosquito swarming suggest that new tools targeting mosquito swarms can be designed to cut down the high reproductive rate of malaria vectors. Targeting swarms of major malaria vectors may provide an effective control method to counteract behavioral resistance developed by mosquitoes. Here, we evaluated the impact of systematic spraying of swarms of Anopheles gambiae s.l. using a mixed carbamate and pyrethroid aerosol. The impact of this intervention on vector density, female insemination rates and the age structure of males was measured. We showed that the resulting mass killing of swarming males and some mate-seeking females resulted in a dramatic 80% decrease in population size compared to a control population. A significant decrease in female insemination rate and a significant shift in the age structure of the male population towards younger males incapable of mating were observed. This paradigm-shift study therefore demonstrates that targeting primarily males rather than females, can have a drastic impact on mosquito population.

Mating competitiveness of sterile male Anopheles coluzzii in large cages

BackgroundUnderstanding the factors that account for male mating competitiveness is critical to the development of the sterile insect technique (SIT). Here, the effects of partial sterilization with 90 Gy of radiation on sexual competitiveness of Anopheles coluzzii allowed to mate in different ratios of sterile to untreated males have been assessed. Moreover, competitiveness was compared between males allowed one versus two days of contact with females.MethodsSterile and untreated males four to six days of age were released in large cages (~1.75 sq m) with females of similar age at the following ratios of sterile males: untreated males: untreated virgin females: 100:100:100, 300:100:100, 500:100:100 (three replicates of each) and left for two days. Competitiveness was determined by assessing the egg hatch rate and the insemination rate, determined by dissecting recaptured females. An additional experiment was conducted with a ratio of 500:100:100 and a mating period of either one or two days. Two controls of 0:100:100 (untreated control) and 100:0:100 (sterile control) were used in each experiment.ResultsWhen males and females consort for two days with different ratios, a significant difference in insemination rate was observed between ratio treatments. The competitiveness index (C) of sterile males compared to controls was 0.53. The number of days of exposure to mates significantly increased the insemination rate, as did the increased number of males present in the untreated: sterile male ratio treatments, but the number of days of exposure did not have any effect on the hatch rate.DiscussionThe comparability of the hatch rates between experiments suggest that An. coluzzii mating competitiveness experiments in large cages could be run for one instead of two days, shortening the required length of the experiment. Sterilized males were half as competitive as untreated males, but an effective release ratio of at least five sterile for one untreated male has the potential to impact the fertility of a wild female population. However, further trials in field conditions with wild males and females should be undertaken to estimate the ratio of sterile males to wild males required to produce an effect on wild populations.

Reusing larval rearing water and its effect on development and quality of Anopheles arabiensis mosquitoes

BackgroundThere is growing interest in applying the sterile insect technique (SIT) against mosquitoes. Mass production of mosquitoes for large-scale releases demands a huge amount of water. Yet, many arid and/or seasonally arid countries face the difficulties of acute water shortage, deterioration of water quality and environmental constraints. The re-use of water to rear successive generations of larvae is attractive as a way to reduce water usage and running costs, and help to make this control method viable.MethodsTo determine whether dirty larval water was a suitable rearing medium for Anopheles arabiensis, in place of the ‘clean’ dechlorinated water routinely used, a series of three experiments was carried out to evaluate the effect of dirty water or mixed clean and dirty water on several parameters of insect quality. Batches of 100 fresh eggs were distributed in dirty water or added to clean water to test the effect of dirty water on egg hatching, whereas first-instar larvae were used to determine the effect on immature development time, pupation, adult emergence, body size, and longevity. Moreover, to assess the effect of dirty water on larval mortality, pupation rate, adult emergence, and longevity, L4 larvae collected after the tilting or larvae/pupae separation events were returned either to the dirty water or added to clean water.ResultsResults indicated that reusing dirty water or using a 50:50 mix of clean and dirty water did not affect egg hatching. Moreover, no difference was found in time to pupation, larval mortality or sex ratio when first-instar larvae were added to clean water, dirty water, or a 75:25, 50:50 or 25:75 mix of clean and dirty water and reared until emergence. When late-instar larvae were put back into their own rearing water, there was no effect on pupation rate, emergence rate or female longevity, though male longevity was reduced. When reared from first-instar larvae, however, dirty water decreased pupation rate, emergence rate, body size, and adult longevity.ConclusionsRe-used larval-rearing water has no impact on egg hatching, development time or mortality of the immature stages of An. arabiensis. However, dirty water is not suitable for the production of high quality adult mosquitoes. Recycling processes to improve water quality and increase insect quality will be investigated, since it may have important implications for the implementation of the SIT in areas where clean water is a scarce or costly resource.

Enhancements to the mass-rearing cage for the malaria vector, Anopheles arabiensis for improved adult longevity and egg production

Innovations in mosquito mass‐rearing techniques are essential in the quest to develop SIT (sterile insect technique) methods to fight mosquito vectors of disease. This study reports modifications to the Food and Agriculture Organisation/International Atomic Energy Agency (FAO/IAEA) mass‐rearing cage (MRC) for mosquitoes to support the behaviour of adult Anopheles arabiensis Patton (Diptera: Culicidae) and to maximize egg production. The effects of an improved sugar‐feeding device, and the addition of resting sites and a black cloth shroud to create an artificial horizon (visual contrast of light vs. dark at the edge) were assessed for their effect on adult longevity and egg production. Egg production of adults resulting from larvae reared in individual free‐standing trays vs. those reared in the same trays in the FAO/IAEA larval rearing rack was also compared. Finally, the effect of blood feeding and frequency of egg collection from the MRC on average egg production per batch was investigated. Overall, the modifications to the MRC enhanced adult longevity, and the improved cage prototype allowed the collection of more eggs overall from a cohort of adults than was possible using the original and previous cage prototypes. These stepwise improvements are important for the development of economical and logistically efficient mass‐rearing systems for the malaria vector An. arabiensis.

Cost-effective larval diet mixtures for mass rearing of Anopheles arabiensis Patton (Diptera: Culicidae)

BackgroundLarval nutrition, particularly diet quality, is a key driver in providing sufficient numbers of high quality mosquitoes for biological control strategies such as the sterile insect technique. The diet currently available to mass rear Anopheles arabiensis, referred here to as the “IAEA diet”, is facing high costs and difficulties concerning the availability of the bovine liver powder component. To promote more affordable and sustainable mosquito production, the present study aimed to find alternative diet mixtures. Eight cheaper diet mixtures comprised of varying proportions of tuna meal (TM), bovine liver powder (BLP), brewer’s yeast (BY), and chickpea (CP) were developed and evaluated through a step by step assessment on An. arabiensis larvae and adult life history traits, in comparison to the IAEA diet which served as a basis and standard.ResultsFour mixtures were found to be effective regarding larval survival to pupation and to emergence, egg productivity, adult body size and longevity. These results suggest that these different diet mixtures have a similar nutritional value that support the optimal development of An. arabiensis larvae and enhance adult biological quality and production efficiency, and thus could be used for mass rearing.ConclusionsOur study demonstrated that four different diet mixtures, 40 to 92% cheaper than the IAEA diet, can result in a positive assessment of the mosquitoes’ life history traits, indicating that this mosquito species can be effectively mass reared with a significant reduction in costs. The mixture comprised of TM + BY + CP is the preferred choice as it does not include BLP and thus reduces the cost by 92% compared to the IAEA diet.