Soma et al. Malar J (2017) 16:357 DOI 10.1186/s12936-017-2012-8 Malaria Journal RESEARCH Open Access Does mosquito mass‑rearing produce an inferior mosquito? Dieudonné D. Soma1,2,3*, Hamidou Maïga1,2, Wadaka Mamai2,4, Nanwintoun S. Bimbile‑Somda1,2, Nelius Venter5, Adel B. Ali2, Hanano Yamada2, Abdoulaye Diabaté1, Florence Fournet6, Georges A. Ouédraogo3, Rosemary S. Lees7, Roch K. Dabiré1 and Jeremie R. L. Gilles2* Abstract Background: The success of the sterile insect technique depends, among other things, on continuous releases of sexually competitive sterile males within the target area. Several factors (including high rearing density and physi‑ cal manipulation, such as larvae and pupae separation) can infuence the quality of males produced in mass-rearing facilities. The diferent steps in mass production in the laboratory may modify the behaviour of mosquitoes, directly or through loss of natural characters as a result of adaptation to lab rearing, and lead to the competitiveness of sterile male being reduced. In the present study, the objective was to evaluate the efect of mass-rearing conditions on ster‑ ile male sexual competitiveness in semi-feld cages compared to routine small scale laboratory rearing methods. Methods: Anopheles arabiensis immature stages were reared both on a large scale using a rack and tray system developed by the FAO/IAEA (MRS), and on a small scale using standard laboratory rearing trays (SRS). Mosquito life his‑ tory traits such as pupation rate, emergence rate, adult size as well as the efect of irradiation on adult longevity were evaluated. Moreover, 5–6 day old mosquitoes were released into feld cages and left for two nights to mate and the mating competitiveness between sterile mass-reared males and fertile males reared on a small scale when compet‑ ing for small scale reared virgin females was investigated. Resulting fertility in a treatment ratio of 1:1:1 (100 irradiated males: 100 non-irradiated males: 100 virgin females) was compared to control cages with 0:100:100 (non-irradiated control) and 100:0:100 (irradiated control). Results: No signifcant diferences in life history parameters were observed between rearing methods. The competi‑ tiveness index of mass reared males (0.58) was similar to males reared on a small scale (0.59). A residual fertility rate of 20% was observed in the irradiated control (100:0:100), measured as the percentage of eggs collected from the cages which developed to adulthood. No signifcant diference was observed (t 0.2896, df 4, P 0.7865) between the rearing treatments (MRS and SRS) in the fertility rate, a measure of mating =competitiveness.= = Conclusions: The results showed that the FAO/IAEA mass-rearing process did not afect mosquito life history param‑ eters or the mating competitiveness of males. Keywords: Anopheles arabiensis, Sterile insect technique, Mass-rearing, Competitiveness Background malaria control strategies are mainly based on the use Malaria remains a serious threat to world health, caus- of insecticides, chemoprevention and case management ing an estimated 212 million malaria cases and 429,000 [1]. Vector control has been and continues to be an efec- deaths in 2015 [1]. With no vaccine available, current tive means of disease control [1, 2], though the current malaria control strategies are being undermined by the rapid spread of resistance to common insecticide classes *Correspondence: [email protected]; [email protected] in major malaria vectors [3–6] and of Plasmodium to 2 Insect Pest Control Laboratory, Joint FAO/IAEA Division of Nuclear available anti-malaria drugs [7]. Terefore, innovative Techniques in Food and Agriculture, Vienna, Austria Full list of author information is available at the end of the article © The Author(s) 2017. This article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/ publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated. Soma et al. Malar J (2017) 16:357 Page 2 of 8 and/or alternative strategies are needed for more efec- loaded with around 15,000 pupae where emerging adults tive vector control [8, 9]. has access to 5% sugar solution using a Whatman fl- Te sterile insect technique (SIT), involving release of ter paper (58 × 58 cm). Females were ofered defrosted males sterilized using gamma or X-rays, could be used in bovine blood in the Hemotek membrane feeder (Dis- the context of integrated vector control as it has already covery Workshops, UK) [19] for 2 h. Eggs were collected been by many control programmes of insect pests or from large cages and quantifed according to methods disease vectors (as reviewed by Lees et al. [10]). Te suc- described by Maïga et al. [20], and reared to adulthood cess of the SIT component depends, among other things, on either a mass-rearing scale (mass-rearing MRS) or a on continuous releases of sexually competitive sterile small rearing scale (SRS) for all experiments. male within the target area [11]. It is desirable that the released males be as sexually competitive as wild males Mass‑rearing scale [10]. Te diferent processes of mass production in the Immature stages were reared in the larval mass- laboratory may modify the behaviour of mosquitoes, by rearing unit, a tiltable steel rack holding 50 trays directly impacting their quality or over time through loss (100 × 60 × 3 cm) developed at the FAO/IAEA IPCL of natural characteristics during adaptation to lab rearing [21]. Each tray was flled with 4 L of deionized water the [12]. Tis can contribute to reducing the competitiveness day before adding the eggs to allow the water to reach of the sterile males, in addition to the impact of irradia- room temperature (28–30 °C). An aliquot of 4000 eggs tion that can afect competitiveness when too high doses was dispensed into a plastic ring foating on the surface of are used or handling methods are not optimized [13]. the water in each of the ffty rearing trays. Te IAEA lar- Te rearing history of the colony and a lack of genetic val diet (1%) was used to feed larvae [22], and pupae were diversity induced by the laboratory colonization might collected by tilting the rack and separating them from also alter male sexual vigour under feld conditions [14]. remaining larvae following the IAEA guidelines [23]. Terefore, studies in semi-feld cages are necessary to better evaluate the competitiveness of sterile mass-reared Small rearing scale males and to determine the minimum required release Aliquots of 750 eggs obtained from mass-rearing cages ratio of sterile male to wild male that could impact wild were hatched and reared in small plastic laboratory trays insect populations. (30 × 40 × 7 cm) flled with 1 L of deionized water [24]. Te semi-feld cage system provides a very valuable A 1% solution of the IAEA diet was used to feed larvae: measure of these parameters, as demonstrated in previ- 10 mL per tray for the frst 3 days, 20 mL on the 4th day ous studies in Anopheles coluzzii [15] and Anopheles ara- and 30 mL on each remaining day as described by Mamai biensis [16, 17]. However, all mosquitoes used in these et al. [24]. Pupae were removed on a daily basis using a previous experiments were reared using routine labo- pipette, counted and placed into small bowls contain- ratory equipment at a small scale and no studies have ing 50 mL of the same water treatment as they had been assessed the competitiveness of sterile males reared using reared as larvae to homogenize rearing conditions. Pupae the FAO/IAEA mass-rearing unit. collected from each rearing method were divided into Tis study aimed to assess the efect of mass-rearing two groups, one for irradiation and one for adult size and conditions on mosquito life history traits and sterile male longevity measurements. sexual competitiveness in semi-feld cages compared to routine small scale laboratory rearing methods. Te ef- Irradiation of pupae cacy of the sterilization process was evaluated in terms Pupae collected from both rearing conditions were sep- of residual fertility which was further assessed when full arated by sex under a stereomicroscope by observing sterilization was not reached by observing whether the the shape of their genitalia [25]. Individuals pupating larvae that hatched can reach adulthood. between 9:00 a.m. and 3:00 p.m. each day were collected for irradiation at 11:00 a.m. the following day, so that Methods 20–26 h old male pupae were irradiated with gamma Mosquito strain and rearing conditions rays generated by a Cobalt-60 Gammacell ® (Nordion Te Anopheles arabiensis Dongola strain, originat- 220) source at the IPCL (Seibersdorf, Austria) at a dose ing from the northern state of Sudan was used in all of 75 Gy. To avoid possible variability related to radia- experiments. Adults were reared at a temperature of tion exposure, pupae originating from MRS and SRS 27 ± 1 °C, 60 ± 10% relative humidity (RH) and main- were irradiated at the same time, 75 pupae per batch tained under a light regime (light: dark) of 12:12 h includ- with most of the rearing water removed. Te precise ing 1 h each of dusk and dawn. Adult mosquitoes were dose received by pupae in each treatment was measured ® mass-reared using large cages (200 × 100 × 20 cm) [18] with a dosimetry system using Gafchromic HD-810 flm Soma et al.