J.R.L. Gilles

An Inexpensive and Effective Larval Diet for Anopheles arabiensis (Diptera: Culicidae): Eat Like a Horse, a Bird, or a Fish?

ABSTRACT A successful sterile insect technique program depends upon mass production of goodquality sterile insects for release into a target area. Specifically, to control Anopheles arabiensis Patton (Diptera: Culicidae) in a pilot area in northern Sudan, 1 million An. arabiensis sterile males per day are needed. To reach this production objective, mosquito mass rearing is indispensable and larval diet, a key parameter for the production of healthy male mosquitoes, needs to be cost-effective. The Koi Floating Blend fish food, previously used at the Food and Agriculture Organization/International Atomic Energy Agency Insect Pest Control Laboratory for the routine rearing of the An. arabiensis colony, is no longer available. The aim of this study was to find a cheap and effective substitute for the discontinued diet. Several candidate powdered diets that are commercially available were tested to determine the best diet. By using mixture experiment principles and response surface methodology, the combination of two components, bovine liver powder and tuna meal, showed the best results in terms of larval survival, developmental, rate and adult size. The addition of a vitamin mixture further improved the diet. These positive production results coupled with the relatively low cost of our blend demonstrated the possibility of its use for mass rearing purpose.

Density-Dependent Effects in Experimental Larval Populations of Anopheles arabiensis (Diptera: Culicidae) Can be Negative, Neutral, or Overcompensatory Depending on Density and Diet Levels

ABSTRACT Anopheles arabiensis Patton (Diptera: Culicidae) larvae were reared from hatching to the adult stage in the laboratory under a range of diet and larval concentrations using a factorial design. The range circumscribed most of the larval densities and diet concentrations that would allow larval growth and survival using the particular diet formulation and water volume we tested. We determined how these variables affected three outcomes, as follows: larval development rate, survival, and wing length. As has been reported previously, negative density dependence of survival as a function of increased larval density was the prevalent effect on all outcomes when diet was limiting. When diet was not limiting, density dependence was not observed, and three cases of overcompensatory survival were seen. We discuss these results in the context of diet and larval densities for mass rearing and the effect of larval competition on control strategies.

Efficiency of three diets for larval development in mass rearing Aedes albopictus (Diptera: Culicidae).

ABSTRACT A fundamental step in establishing a mass production system is the development of a larval diet that promotes high adult performance at a reasonable cost. To identify a suitable larval diet for Aedes albopictus (Skuse), three diets were compared: a standard laboratory diet used at the Centro Agricoltura Ambiente, Italy (CAA) and two diets developed specifically for mosquito mass rearing at the FAO/IAEA Laboratory, Austria. The two IAEA diets, without affecting survival to the pupal stage, resulted in a shorter time to pupation and to emergence when compared with the CAA diet. At 24 h from pupation onset, 50 and 90% of the male pupae produced on the CAA and IAEA diets, respectively, had formed and could be collected. The diet received during the larval stage affected the longevity of adult males with access to water only, with best results observed when using the CAA larval diet. However, similar longevity among diet treatments was observed when males were supplied with sucrose solution. No differences were observed in the effects of larval diet on adult male size or female fecundity and fertility. Considering these results, along with the relative costs of the three diets, the IAEA 2 diet is found to be the preferred choice for mass rearing of Aedes albopictus, particularly if a sugar meal can be given to adult males before release, to ensure their teneral reserves are sufficient for survival, dispersal, and mating in the field.

A new larval tray and rack system for improved mosquito mass rearing.

ABSTRACT The requirement for efficient mosquito mass rearing technology has been one of the major obstacles preventing the large scale application of the Sterile Insect Technique against mosquitoes. At the Food and Agriculture Organization/International Atomic Energy Agency (FAO/IAEA) Insect Pest Control Laboratories we developed a larval rearing unit based on the use of a stainless steel rack that operates 50 thermoformed ABS plastic trays and is expected to be able to successfully rear 140,000–175,000 Anopheles arabiensis (Patton) adult mosquitoes per rack. The mechanized rearing unit is simple to handle, maintains minimal water temperature variation and negligible water evaporation and allows normal larval development. The mosquito mass-rearing tray was designed to provide a large surface area of shallow water that would closely mimic natural breeding sites. The trays stack into a dedicated rack structure and filling and draining were easily performed. The close stacking of the trays in the rack and the possibility to tightly line up several racks makes this rearing unit a valid solution for maximal use of the space thus reducing construction, heating, and cooling costs. The low amount of labor required to operate the system also reduces labor costs that represent one of the main expenditures in any mass rearing facility operation. Preliminary experiments performed on Aedes albopictus (Skuse) also confirm the possibility of successfully extending the use of this technology to other mosquito species. Our larval rearing unit could enhance any mosquito control strategy in which large-scale releases of mosquitoes are needed to suppress or replace natural populations.

Aedes albopictus (Skuse) males in laboratory and semi-field cages: Release ratios and mating competitiveness

To control the container-breeding mosquito and major vector of dengue and chikungunya Aedes albopictus, the sterile insect technique (SIT) is proposed as a component of integrated vector management programs in endemic areas. For the technique to be successful, released males, sterilized with 35 Gy of ionizing radiation during the pupal stage, must be able to compete for mating opportunities with wild counterparts and successfully copulate with wild females to induce sterility in the population. Any reduction in competitiveness can be compensated for by increasing the ratio of released sterile to wild males, a ratio which must be optimized for effectiveness and efficiency. Fruit fly SIT programs use field enclosures to test the competitiveness of sterile males to monitor the quality of the colony and adjust release ratios. This is laborious and time consuming, and for mosquito programs it would be advantageous if similarly useful results could be obtained by smaller scale laboratory tests, conducted on a more regular basis. In the present study we compared the competitiveness, as measured by hatching rate of resulting egg batches, of irradiated males measured in small and large laboratory cages and semi-field enclosures in a greenhouse setting, when competing in a 1:1, 3:1, and 5:1 ratio with fertile males. The sterile males were found to be equally competitive when compared to unirradiated counterparts, and a 5:1 ratio was sufficient to reduce, but not eliminate, the fertility of the female populations, irrespective of cage size. Variability in hatch rate in eggs laid by individual females and so-called indeterminate matings, when we could not be certain whether a female had mated a fertile or a sterile male, could be investigated by closer investigation of mating status and the frequency of multiple matings in Ae. albopictus. The laboratory results are encouraging for the effectiveness of the SIT using irradiated males of this species, and we support further assessment in the field.

X-Ray-Induced Sterility in Aedes albopictus (Diptera: Culicidae) and Male Longevity Following Irradiation

ABSTRACT The mosquito Aedes albopictus (Skuse, 1895) is a potent vector of several arboviral diseases, most notably chikungunya and dengue fever. In the context of the sterile insect technique (SIT), the sterilization of the male mosquitoes before their release can be achieved by gamma-ray irradiation. As gamma-ray irradiators are becoming increasingly problematic to purchase and transport, the suitability of an X-ray irradiator as an alternative for the sterilization of Ae. albopictus males was studied. The sterilization of up to 200,000 pupae at one time can be achieved with relative ease, and the sterility results obtained were comparable with those achieved by gamma irradiation, where 99% sterility is induced with a dose of 40 Gy. A significant reduction of longevity was observed in the latter stages of the males’ life after irradiation treatments, especially at doses >40 Gy, which is consistent with the negative effects on longevity induced by similar radiation doses using gamma rays. Females irradiated at 40 Gy were not only 100% sterile, but also failed to oviposit entirely, i.e., all of the females laid 0 eggs. Overall, it was found that the X-ray irradiator is generally suitable for the sterilization process for sterile insect technique programs, as it showed a high processing capacity, practicality, high effectiveness, and reproducibility.

Mass Production Cage for Aedes albopictus (Diptera: Culicidae)

ABSTRACT Mass production is an important component of any pest or vector control program that requires the release of large number of insects. As part of efforts to develop an area-wide program involving the sterile insect technique (SIT) for the control of mosquitoes, the Insect Pest Control Laboratory of the Food and Agriculture Organization—International Atomic Energy Agency (FAO—IAEA Joint Division) has developed a mass production cage (Aedes MPC) for brood stock colonies in a mass production system for Aedes albopictus (Skuse, 1895). A preliminary experiment using Plexiglas cages was carried out to estimate the impact of cage volume on egg productivity. Transparent Plexiglas cages of different dimensions but loaded with the same adult density were tested. Egg productivity (number of eggs laid per adult female) and adult survival were recorded and analyzed. According to the results, the optimal volume of 100 liters has been chosen to develop the Aedes MPC. The numbers of adults introduced into the Aedes MPC did not affect the egg production and adult survival in comparison with the Plexiglas cage experiment data, confirming the possible use of Aedes MPC for mass-rearing procedures. Finally, the modification of Aedes MPC and creation of a new prototype model of MPC (Anopheles MPC) to effectively contain Anopheles arabiensis (Patton, 1905) adults is discussed with major changes pioneered to oviposition devices and systems for automatic collection of the eggs.

Occurrence of natural Anopheles arabiensis swarms in an urban area of Bobo-Dioulasso city, Burkina Faso, West Africa

The swarming behaviour of natural populations of Anopheles arabiensis was investigated by conducting transect surveys on 10 consecutive days, around dusk, from March to April and from September to October 2012 in Dioulassoba, a district of Bobo-Dioulasso city in Burkina Faso (West Africa). Swarms were observed outside, around identified larval breeding sites on the banks of the Houet River, as well as in the open-air courtyards found at the centre of many homes in the region. Swarms were found to occur in open sunlit spaces, mostly located above physical or visual cues somehow visually distinct from the surrounding area. Overall 67 and 78 swarms were observed, respectively, during the dry season (March-April) and the rainy season (September-October) of 2012, between 1.5m and 4.5m above the ground at their centre. 964 mosquitoes were collected and analysed from dry season swarms, of which most were male, and all were An. arabiensis, as were the few resting mosquitoes collected indoors. Larvae collected from breeding sites found on the banks of the Houet River mostly consisted of An. arabiensis and only a minority of Anopheles coluzzii (formerly identified as An. gambiae M form). Of 1694 mosquitoes analysed from 78 swarms in the rainy season collections, a few An. gambiae (formerly known as An. gambiae S form) males were identified, and the remainders were An. arabiensis. The majority of larvae collected during the wet season from the same breeding sites were identified as An. arabiensis followed by An. coluzzii and An. gambiae. The same pattern of species composition was observed in resting mosquitoes, though the proportion of An. arabiensis was less overwhelming. These data support the conclusion that An. arabiensis is the most prevalent species in this area, though the difference in species composition when using different population sampling techniques is noteworthy. Further studies are required for more detailed investigations of male dispersal, feeding behaviour and mating patterns in this urban setting.

Different Blood and Sugar Feeding Regimes Affect the Productivity of Anopheles arabiensis Colonies (Diptera: Culicidae)

ABSTRACT The success of the sterile insect technique for the management of mosquito populations depends on the release of large numbers of competitive sterile male insects. Sustainable mosquito production can only be obtained when proper mass-rearing equipment and adequate methods are available, including those to feed blood to the female mosquitoes. The blood feeding apparatus Hemotek consists of a small aluminum plate to which a collagen membrane is fixed and filled with blood kept warm by an electric heating element. A larger aluminum plate was developed to feed a larger number of female mosquitoes with blood that is kept at a constant temperature. The effect of different blood feeding regimes (feeding frequency and time the blood is kept in the Hemotek) and sugar deprivation before blood feeding on egg production of female Anopheles arabiensis Patton was tested. Egg production was higher when blood was offered to the mosquitoes every day as compared with every 2 or 4 d. Sugar deprivation for 7 h before blood feeding enhanced egg production by 50% compared with female mosquitoes that had continuous access to sugar. Neither male nor female survival was impaired. Finally, we showed that the same blood could be kept warm and used over several hours to feed mosquitoes in multiple cages without any impact on egg production or hatch rate. Being able to use the same blood over extended periods would save considerable time, handling, and funds.

Anopheles arabiensis Sperm Production After Genetic Manipulation, Dieldrin Treatment, and Irradiation

ABSTRACT The use of the sterile insect technique relies on the release of sterilized mass-reared male insects which, before field releases, endure several unnatural treatments. In the case of Anopheles arabiensis (Patton) sterile insect technique program in Sudan, the genetic background of the original strain was first changed to create a genetic sexing strain that is based on a dieldrin-resistant mutation. Secondly, the eggs of the genetic sexing strain require treatment with dieldrin to allow complete elimination of female Ll larvae to enable the release of males only. Finally, male mosquitoes receive an irradiation dose of 70 Gy as pupae for sterilization. The effects of these treatments on sperm production were tested separately and in combination. Irradiation alone significantly decreased the initial sperm number and prevented new sperm production. However, the dieldrin treatment, aimed at eliminating females, appears to have an unexpected radioprotectant effect.