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Background Human African trypanosomiasis (HAT) is a neglected tropical disease caused by Trypanosoma brucei gambiense transmitted by tsetse flies in sub-Saharan West Africa. In southern Chad the most active and persistent focus is the Mandoul focus, with 98% of the reported human cases, and where African animal trypanosomosis (AAT) is also present. Recently, a control project to eliminate tsetse flies (Glossina fuscipes fuscipes) in this focus using the sterile insect technique (SIT) was initiated. However, the release of large numbers of sterile males of G. f. fuscipes might result in a potential temporary increase in transmission of trypanosomes since male tsetse flies are also able to transmit the parasite. The objective of this work was therefore to experimentally assess the vector competence of sterile males treated with isometamidium for Trypanosoma brucei brucei. Methods An experimental infection was set up in the laboratory, mimicking field conditions: the same tsetse species that is present in Mandoul was used. A T. b. brucei strain close to T. b. gambiense was used, and the ability of the sterile male tsetse flies fed on blood with and without a trypanocide to acquire and transmit trypanosomes was measured. Results Only 2% of the experimentally infected flies developed an immature infection (midgut) while none of the flies developed a metacyclic infection of T. b. brucei in the salivary glands. We did not observe any effect of the trypanocide used (isometamidium chloride at 100 mg/l) on the development of infection in the flies. Conclusions Our results indicate that sterile males of the tested strain of G. f. fuscipes were unable to cyclically transmit T. b. brucei and might even be refractory to the infection. The data of the research indicate that the risk of cyclical transmission of T. brucei by sterile male G. f. fuscipes of the strain colonized at IAEA for almost 40 years appears to be small. Graphical Abstract

International audience; Mosquito-borne diseases represent a major threat to humankind. Recently, the incidence of malaria has stopped decreasing while that of dengue is increasing exponentially. Alternative mosquito-control methods are urgently needed. The sterile insect technique (SIT) has seen significant developments recently and may play an important role. However, testing and implementing SIT for vector control is challenging, and a phased conditional approach (PCA) is recommended, that is, advancement to the next phase depends on completion of activities in the previous one. We herewith present a PCA to test the SIT against mosquitoes within an area-wide-integrated pest-management programme, taking into account the experience gained with plant and livestock pests and the recent developments of the technique against mosquitoes.

International audience; Spatio-temporally heterogeneous environments may lead to unexpected population dynamics. Knowledge is needed on local properties favouring population resilience at large scale. For pathogen vectors, such as tsetse flies transmitting human and animal African trypanosomosis, this is crucial to target management strategies. We developed a mechanistic spatio-temporal model of the age-structured population dynamics of tsetse flies, parametrized with field and laboratory data. It accounts for density- and temperature-dependence. The studied environment is heterogeneous, fragmented and dispersal is suitability-driven. We confirmed that temperature and adult mortality have a strong impact on tsetse populations. When homogeneously increasing adult mortality, control was less effective and induced faster population recovery in the coldest and temperature-stable locations, creating refuges. To optimally select locations to control, we assessed the potential impact of treating them and their contribution to the whole population. This heterogeneous control induced a similar population decrease, with more dispersed individuals. Control efficacy was no longer related to temperature. Dispersal was responsible for refuges at the interface between controlled and uncontrolled zones, where resurgence after control was very high. The early identification of refuges, which could jeopardize control efforts, is crucial. We recommend baseline data collection to characterize the ecosystem before implementing any measures.; hôtes-vecteurs-parasites-environnement dans les maladies tropicales négligées dues aux trypanosomatides', Cirad

Microsatellite loci still represent valuable resources for the study of the population biology of non-model organisms. Discovering or adapting new suitable microsatellite markers in species of interest still represents a useful task, especially so for non-model organisms as tsetse flies (genus Glossina), which remain a serious threat to the health of humans and animals in sub-Saharan Africa. In this paper, we present the development of new microsatellite loci for four species of Glossina: two from the Morsitans group, G. morsitans morsitans (Gmm) from Zimbabwe, G. pallidipes (Gpalli) from Tanzania; and the other two from the Palpalis group, G. fuscipes fuscipes (Gff) from Chad, and G. palpalis gambiensis (Gpg) from Guinea. We found frequent short allele dominance and null alleles. Stuttering could also be found and amended when possible. Cryptic species seemed to occur frequently in all taxa but Gff. This explains why it may be difficult finding ecumenical primers, which thus need adaptation according to each taxonomic and geographic context. Amplification problems occurred more often in published old markers, and Gmm and Gpg were the most affected (stronger heterozygote deficits). Trinucleotide markers displayed selection signature in some instances (Gmm). Combining old and new loci, for Gmm, eight loci can be safely used (with correction for null alleles); and five seem particularly promising; for Gpalli, only five to three loci worked well, depending on the clade, which means that the use of loci from other species (four morsitans loci seemed to work well), or other new primers will need to be used; for Gff, 14 loci behaved well, but with null alleles, seven of which worked very well; and for G. palpalis sl, only four loci, needing null allele and stuttering corrections seem to work well, and other loci from the literature are thus needed, including X-linked markers, five of which seem to work rather well (in females only), but new markers will probably be needed. Finally, the high proportion of X-linked markers (around 30%) was explained by the non-Y DNA quantity and chromosome structure of tsetse flies studied so far.

The production of a large number of mosquitoes of high biological qualities and reliable sex sorting before release are key challenges when applying the sterile insect technique as part of an area-wide integrated pest management approach. There is a need to fully evaluate the production capacity of the equipment developed in order to plan and maintain a daily production level for large-scale operational release activities. This study aimed to evaluate the potential use of the FAO/IAEA larval rearing unit for Aedes aegypti and the subsequent female contamination rate after sex sorting with a Fay-Morlan glass separator. Trays from each rack were tilted and their contents sorted either for each individual tray or after mixing the content of all trays from the rack. The pupal production and the female contamination rate were estimated with respect to day of collection, position of the tray, type of pupae collection, and sorting operator. Results showed significant daily variability of pupal production and female contamination rate, with a high male pupal production level achieved on the second day of collection and estimated female contamination of male pupae reached around 1%. Neither tray position nor type of pupae collection affected the pupal production and female contamination rate. However, the operator had a significant effect on the female contamination rate. These results highlight the need to optimize pupal production at early days of collection and to develop a more effective and automated method of sex separation.; La production d’un grand nombre de moustiques de haute qualité biologique et le tri des sexes avant les lâchers sont des défis clés lors de l’application de la technique des insectes stériles, dans le cadre d’une approche de lutte intégrée contre les ravageurs à l’échelle d’une zone. Il est nécessaire d’évaluer pleinement la capacité de production des équipements développés afin de planifier et de maintenir un niveau de production quotidien pour les activités de libération opérationnelle à grande échelle. Cette étude visait à évaluer l’utilisation potentielle de l’unité d’élevage larvaire FAO/AIEA pour Aedes aegypti et le taux de contamination par des femelles après le tri sexuel avec un séparateur en verre Fay–Morlan. Les plateaux de chaque rack ont été inclinés et leur contenu trié soit pour chaque plateau, soit après avoir mélangé le contenu de tous les plateaux du rack. La production de pupes et le taux de contamination par des femelles ont été estimés en fonction du jour de collecte, de la position du bac, du type de collecte des pupes et de l’opérateur du tri. Les résultats ont montré une variabilité quotidienne significative de la production de pupes et du taux de contamination par des femelles, avec un niveau élevé de production de pupes mâles atteint le deuxième jour de collecte et la contamination estimée des pupes mâles par des femelles a atteint environ 1 %. Ni la position du plateau ni le type de collecte des pupes n’ont affecté la production de pupes et le taux de contamination par des femelles. Cependant, l’opérateur avait un effet significatif sur le taux de contamination par les femelles. Ces résultats mettent en évidence la nécessité d’optimiser la production des pupes dès les premiers jours de la collecte et de développer une méthode de séparation des sexes plus efficace et automatisée.

Background Tsetse flies (Diptera: Glossinidae) and tabanids (Diptera: Tabanidae) are haematophagous insects of medical and veterinary importance due to their respective role in the biological and mechanical transmission of trypanosomes. Few studies on the distribution and relative abundance of both families have been conducted in Mozambique since the country’s independence. Despite Nicoadala, Mozambique, being a multiple trypanocidal drug resistance hotspot no information regarding the distribution, seasonality or infection rates of fly-vectors are available. This is, however, crucial to understanding the epidemiology of trypanosomosis and to refine vector management. Methods For 365 days, 55 traps (20 NGU traps, 20 horizontal traps and 15 Epsilon traps) were deployed in three grazing areas of Nicoadala District: Namitangurine (25 traps); Zalala (15 traps); and Botao (15 traps). Flies were collected weekly and preserved in 70% ethanol. Identification using morphological keys was followed by molecular confirmation using cytochrome c oxidase subunit 1 gene. Trap efficiency, species distribution and seasonal abundance were also assessed. To determine trypanosome infection rates, DNA was extracted from the captured flies, and submitted to 18S PCR-RFLP screening for the detection of Trypanosoma. Results In total, 4379 tabanids (of 10 species) and 24 tsetse flies (of 3 species), were caught. NGU traps were more effective in capturing both the Tabanidae and Glossinidae. Higher abundance and species diversity were observed in Namitangurine followed by Zalala and Botao. Tabanid abundance was approximately double during the rainy season compared to the dry season. Trypanosoma congolense and T. theileri were detected in the flies with overall infection rates of 75% for tsetse flies and 13% for tabanids. Atylotus agrestis had the highest infection rate of the tabanid species. The only pathogenic trypanosome detected was T. congolense. Conclusions Despite the low numbers of tsetse flies captured, it can be assumed that they are still the cyclical vectors of trypanosomosis in the area. However, the high numbers of tabanids captured, associated to their demonstrated capacity of transmitting trypanosomes mechanically, suggest an important role in the epidemiology of trypanosomosis in the Nicoadala district. These results on the composition of tsetse and tabanid populations as well as the observed infection rates, should be considered when defining strategies to control the disease.

International audience; Glossina palpalis gambiensis is a riverine tsetse species endemic in West Africa and thriving in riparian vegetation of the savannah areas from Burkina Faso and Mali to Guinea and Senegal. It is a major vector of human and animal trypanosomosis (sleeping sickness and nagana, respectively) in that region. G. p. gambiensis is an opportunistic species, feeding on a wide range of hosts from reptiles to pigs and cattle, with humans as one of its preferred hosts. Like most tsetse species, it has a narrow range of acceptable temperature and humidity, a low reproduction rate, and is thus very sensitive to climate change but can adapt to human modification of its environment and survive in polluted and densely populated areas. Its presence in the Niayes area of Senegal, where rainfall is below 500 mm a year, and in the Parc de Hahn of Dakar reveals an extraordinary plasticity. In the Niayes area it is presently targeted by an eradication program, including a sterile insect technique component. Trends Trends in in Parasitology Parasitology TRANSMISSION FACTS: G. p. gambiensis can transmit all Trypanosoma spp., particularly Trypanosoma brucei gambiense in humans and Trypanosoma vivax in cattle. It picks up the bloodstream form of trypanosomes from a host and injects the metacyclic form into the skin of another host after an extrinsic cycle of 10-30 days depending on the parasite species. It is a day-biter, with a peak of activity conditioned by temperature. Its distribution, density, lifespan and infection rate are also temperature-dependent. Its learning capability increases the hunting efficiency of older flies, that is, the host selected for the first bloodmeal can influence host selection for the second meal.

Background The sterile insect technique (SIT) is a vector control strategy relying on the mass release of sterile males into wild vector populations. Current sex separation techniques are not fully efficient and could lead to the release of a small proportion of females. It is therefore important to evaluate the effect of irradiation on the ability of released females to transmit pathogens. This study aimed to assess the effect of irradiation on the survival and competence of Anopheles arabiensis females for Plasmodium falciparum in laboratory conditions. Methods Pupae were irradiated at 95 Gy of gamma-rays, and emerging females were challenged with one of 14 natural isolates of P. falciparum. Seven days post-blood meal (dpbm), irradiated and unirradiated-control females were dissected to assess the presence of oocysts, using 8 parasite isolates. On 14 dpbm, sporozoite dissemination in the head/thorax was also examined, using 10 parasites isolates including 4 in common with the 7 dpbm dissection (oocyst data). The survivorship of irradiated and unirradiated-control mosquitoes was monitored. Results Overall, irradiation reduced the proportion of mosquitoes infected with the oocyst stages by 17% but this effect was highly inconsistent among parasite isolates. Secondly, there was no significant effect of irradiation on the number of developing oocysts. Thirdly, there was no significant difference in both the sporozoite infection rate and load between the irradiated and unirradiated-control mosquitoes. Fourthly, irradiation had varying effects on female survival with either a negative effect or no effect. Conclusions The effect of irradiation on mosquito competence strongly varied among parasite isolates. Because of such isolate variability and, the fact that different parasite isolates were used to collect oocyst and sporozoite data, the irradiation-mediated reduction of oocyst prevalence was not confirmed for the sporozoite stages. Our data indicate that irradiated female An. arabiensis could contribute to malaria transmission, and highlight the need for perfect sexing tools, which would prevent the release of females as part of SIT programmes.

International audience; Tsetse flies (Diptera: Glossinidae) are the main vectors of animal and human trypanosomoses in Africa. The Sterile Insect Technique (SIT) has proven effective in controlling tsetse flies when applied to isolated populations but necessitates the production of large numbers of sterile males. A new approach, called boosted SIT, combining SIT with the contamination of wild females by sterile males coated with biocides has been proposed for large-scale control of vector populations. The aim of the study was to evaluate this new approach using pyriproxyfen on the riverine species Glossina palpalis gambiensis (Vanderplank, 1949) in the laboratory. The contamination dose and persistence of pyriproxyfen on sterile males, the impact of pyriproxyfen on male survival, and the dynamics of pyriproxyfen transfer from a sterile male to a female during mating, as well as the impact of pyriproxyfen on pupal production and adult emergence, were evaluated in the laboratory. For this purpose, a method of treatment by impregnating sterile males with a powder containing 40% pyriproxyfen has been developed. The results showed that the pyriproxyfen has no impact on the survival of sterile males. pyriproxyfen persisted on sterile males for up to 10 days at a dose of 100 ng per fly. In addition, the horizontal transfer of pyriproxyfen from a treated sterile male to a female during mating could be measured with an average of 50 ng of pyriproxyfen transferred. After contacts without mating, the average quantity transferred was more than 10 ng. Finally, the pyriproxyfen powder was very effective on G. p. gambiensis leading to 0% emergence of the pupae produced by contaminated females. These promising results must be confirmed in the field. A large-scale assessment of this boosted pyriproxyfen-based SIT approach will be carried out against tsetse flies in Senegal (West Africa). On the African continent, tsetse flies (Diptera: Glossinidae) are the main vectors of the parasites responsible for Human African Trypanosomosis (HAT), or sleeping sickness, and Animal African Trypanosomosis (AAT), also called Nagana 1. These diseases caused by Trypanosomatidae belong to the group of neglected tropical diseases (NTDs), occurring in developing areas 2. Sub-Saharan African countries suffer from a significant impact of NTDs on public health and economic development 3. Glossina palpalis gambiensis (Vanderplank, 1949) is one of the main vectors of the HAT parasite in West Africa and is responsible for the persistence of many residual outbreaks of this disease in forest and savannah areas 4. There is still no vaccine against HAT and curative treatments are difficult to access for the most exposed and vulnerable populations 5. Glossina palpalis gambiensis is also involved in the transmission of AAT, which affects livestock and reduces animal production thus limiting the availability of food resources 6,7. Despite the millions of doses of trypanocides administered, nearly 3 million cattle die each year in Africa from AAT. Annual direct and indirect agricultural losses attributed to this disease are estimated at

International audience; The Sterile Insect Technique (SIT), used to control insect populations, consists of releasing males sterilized by ionizing radiations. Wild females that mate with these males can no longer produce viable offspring, which may drives the population decline. Although this method has proved its efficiency, its effect may be more limited for fast-reproducing large-population species, such Aedes albopictus. A novel approach, named ”boosted SIT” has been designed to strengthen the SIT technique: It consists of coating sterile males with a biocide that will be transferred to the mated females, which will then contaminate the oviposition sites. This study is aimed at exploring demographic effects of both techniques (SIT and boosted SIT) through their inclusion in a weather-driven abundance model of the Aedes albopictus population dynamics in the geographical context of La Reunion Island. Sensitivity analysis showed that the date to start the release, as well as the quantity of sterile males released and their competitiveness, are of key importance for both control methods. According to our results, boosted SIT allows 1) Increasing the effectiveness of the SIT when the sterile males released are of medium quality in terms of competitiveness, and 2) extending the optimal window to start the control period.

International audience; In genetic control programmes, including the sterile insect technique (SIT), it is crucial to release insects of the highest quality with maximum survival. It is likely that male mosquitoes will follow the trend of other insects in SIT programmes and be stored, transported and eventually released under chilled conditions. The aim of our study was to investigate the impact of different chilling temperatures on male Aedes aegypti and Ae. albopictus survival by exposing them to a range of temperatures for different durations. Ae. aegypti were found to be less sensitive to the impact of chilling, with only 6 degrees C causing a marginal decrease in survival in comparison to non-chilled controls. Conversely, Ae. albopictus displayed a significantly reduced survival at all chilling temperatures even when exposed for a short time. In both species, longer exposure to low temperatures reduced survival. Our results uncovered that Ae. albopictus are more sensitive to chilling, regardless of the temperature, when compared to Ae. aegypti. Such results indicate differences in thermal tolerances between species and the necessity of conducting experiments on a species by species basis when determining temperature limits for any insect destined for release as part of a genetic control programme.

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Background The sterile insect technique (SIT) requires mass-rearing of the target species, irradiation to induce sexual sterility and transportation from the mass-rearing facility to the target site. Those treatments require several steps that may affect the biological quality of sterile males. This study has been carried out to evaluate the relative impact of the chilling, irradiation and transport on emergence rate, flight ability and survival of sterile male tsetse flies Glossina palpalis gambiensis. Results Chilling, irradiation and transport all affected the quality control parameters studied. The emergence rate was significantly reduced by long chilling periods and transport, i.e. from 92% at the source insectary to 78% upon arrival in Dakar. Flight ability was affected by all three parameters with 31% operational flies lossed between the source and arrival insectaries. Only survival under stress was not affected by any of the treatments. Conclusion The chilling period and transport were the main treatments which impacted significantly the quality of sterile male pupae. Therefore, the delivery of sterile males was divided over two shipments per week in order to reduce the chilling time and improve the quality of the sterile males. Quality of the male pupae may further be improved by reducing the transport time and vibration during transport. Keys words: Tsetse flies, mass-rearing conditions, sterile insect technique, quality

A spatially and temporally heterogeneous environment may lead to unexpected population dynamics. Knowledge still is needed on which of the local environment properties favour population maintenance at larger scale. For pathogen vectors, such as tsetse flies transmitting human and animal African trypanosomosis, such a knowledge is crucial to design relevant management strategies. We developed an original mechanistic spatio-temporal model of tsetse fly population dynamics, accounting for combined effects of spatial complexity, density-dependence, and temperature on the age-structured population, and parametrized with field and laboratory data. We confirmed the strong impact of temperature and adult mortality on tsetse populations. We showed that the coldest cells with the smallest variations in temperature acted as refuges when adult mortality was homogeneously increased, control being less effective in such refuges. In contrast, targeting the cells contributed the most to population management, i.e. those of highest carrying capacity and the most impacted by increased mortality, resulted in a decline in population size with a similar efficacy, but resulted in more dispersed individuals, control efficacy being no longer related to temperature. Population resurgence after control was slow, but could be very high locally in refuges, with highly contrasted situations after a heterogeneous control, refuges being located at the interface between controlled and uncontrolled zones. Our results highlighted the importance of baseline data collection to characterize the targeted ecosystem before any control measure is implemented.

Este artículo se encuentra disponible en la página web de la revista en la siguiente URL: https://parasitesandvectors.biomedcentral.com/articles/10.1186/s13071-018-3221-x Background: Several mosquito population suppression strategies based on the rearing and release of sterile males have provided promising results. However, the lack of an efficient male selection method has hampered the expansion of these approaches into large-scale operational programmes. Currently, most of these programmes targeting Aedes mosquitoes rely on sorting methods based on the sexual size dimorphism (SSD) at the pupal stage. The currently available sorting methods have not been developed based on biometric analysis, and there is therefore potential for improvement. We applied an automated pupal size estimator developed by Grupo Tragsa with laboratory samples of Anopheles arabiensis, Aedes albopictus, Ae. polynesiensis, and three strains of Ae. aegypti. The frequency distribution of the pupal size was analyzed. We propose a general model for the analysis of the frequency distribution of mosquito pupae in the context of SSD-sorting methods, which is based on a Gaussian mixture distribution functions, thus making possible the analysis of performance (% males recovery) and purity (% males on the sorted sample). Results: For the three Aedes species, the distribution of the pupae size can be modeled by a mixture of two Gaussian distribution functions and the proposed model fitted the experimental data. For a given population, each size threshold is linked to a specific outcome of male recovery. Two dimensionless parameters that measure the suitability for SSD-based sorting of a specific batch of pupae are provided. The optimal sorting results are predicted for the highest values of SSD and lowest values of intra-batch variance. Rearing conditions have a strong influence in the performance of the SSD-sorting methods and non-standard rearing can lead to increase pupae size heterogeneity. Conclusions: Sex sorting of pupae based on size dimorphism can be achieved with a high performance (% males recovery) and a reasonably high purity (% males on the sorted sample) for the different Aedes species and strains. The purity and performance of a sex sorting operation in the tested Aedes species are linked parameters whose relation can be modeled. The conclusions of this analysis are applicable to all the existing SSD-sorting methods. The efficiency of the SSDsorting methods can be improved by reducing the heterogeneity of pupae size within rearing containers. The heterogeneity between batches does not strongly affect the quality of the sex sorting, as long as a specific separation threshold is not pre-set before the sorting process. For new developments, we recommend using adaptive and precise threshold selection methods applied individually to each batch or to a mix of batches. Adaptive and precise thresholds will allow the sex-sorting of mixed batches in operational conditions maintaining the target purity at the cost of a reduction in performance. We also recommend a strategy whereby an acceptable level of purity is pre-selected and remains constant across the different batches of pupae while the performance varies from batch to batch to fit with the desired purity.

Swarming is a key part of the natural system of reproduction of anopheline mosquito populations, and a better understanding of swarming and mating systems in a targeted species in its natural habitat would contribute to better design control strategies with a greater chance of success. Our study investigated the monthly occurrence of swarming and the mating frequency (within swarms) of Anopheles arabiensis in Dioulassoba, Burkina Faso and their relationship with local environmental factors. Mosquitoes collected from swarms were described in terms of body size, recent sugar meal intake, and female repletion, insemination, and Plasmodium falciparum infection status. Swarms of An. arabiensis were found in each month of the year. Both start and end times of swarming varied significantly between months, correlating with the time of sunset. Swarming mostly started after or coincided with sunset from late July to early October but occurred before sunset from late October to early July. Swarming duration, the number of mosquitoes and mating pairs per swarm, and time to first mating were significantly different between months in an inverse relationship with the monthly rainfall. The number of mating pairs was strongly and positively correlated with swarm size. Almost all the females caught in copula were inseminated but a very few were blood fed; no P. falciparum infection was observed. Males caught in copula and in solo were similar in body size and in the proportion which had taken a recent sugar meal. Our investigations showed that An. arabiensis reproductive activities are most frequent during the dry season, suggesting either the species' preference for dry climatic conditions or a lack of available breeding sites during the rainy season due to the seasonal flooding in this area. Targeting interventions to kill mosquitoes in swarms or to achieve an over-flooding ratio of sterile males during the rainy season would increase their efficiency in reducing the population density of this vector.

The production of a large number of mosquitoes of high biological qualities and reliable sex sorting before release are key challenges when applying the sterile insect technique as part of an area-wide integrated pest management approach. There is a need to fully evaluate the production capacity of the equipment developed in order to plan and maintain a daily production level for large-scale operational release activities. This study aimed to evaluate the potential use of the FAO/IAEA larval rearing unit for Aedes aegypti and the subsequent female contamination rate after sex sorting with a Fay-Morlan glass separator. Trays from each rack were tilted and their contents sorted either for each individual tray or after mixing the content of all trays from the rack. The pupal production and the female contamination rate were estimated with respect to day of collection, position of the tray, type of pupae collection, and sorting operator. Results showed significant daily variability of pupal production and female contamination rate, with a high male pupal production level achieved on the second day of collection and estimated female contamination of male pupae reached around 1%. Neither tray position nor type of pupae collection affected the pupal production and female contamination rate. However, the operator had a significant effect on the female contamination rate. These results highlight the need to optimize pupal production at early days of collection and to develop a more effective and automated method of sex separation.Développement larvaire et production de pupes d’Aedes aegypti dans le système d’élevage de masse de la FAO/AIEA et facteurs influençant l’efficacité de la séparation des sexes.La production d’un grand nombre de moustiques de haute qualité biologique et le tri des sexes avant les lâchers sont des défis clés lors de l’application de la technique des insectes stériles, dans le cadre d’une approche de lutte intégrée contre les ravageurs à l’échelle d’une zone. Il est nécessaire d’évaluer pleinement la capacité de production des équipements développés afin de planifier et de maintenir un niveau de production quotidien pour les activités de libération opérationnelle à grande échelle. Cette étude visait à évaluer l’utilisation potentielle de l’unité d’élevage larvaire FAO/AIEA pour Aedes aegypti et le taux de contamination par des femelles après le tri sexuel avec un séparateur en verre Fay–Morlan. Les plateaux de chaque rack ont été inclinés et leur contenu trié soit pour chaque plateau, soit après avoir mélangé le contenu de tous les plateaux du rack. La production de pupes et le taux de contamination par des femelles ont été estimés en fonction du jour de collecte, de la position du bac, du type de collecte des pupes et de l’opérateur du tri. Les résultats ont montré une variabilité quotidienne significative de la production de pupes et du taux de contamination par des femelles, avec un niveau élevé de production de pupes mâles atteint le deuxième jour de collecte et la contamination estimée des pupes mâles par des femelles a atteint environ 1 %. Ni la position du plateau ni le type de collecte des pupes n’ont affecté la production de pupes et le taux de contamination par des femelles. Cependant, l’opérateur avait un effet significatif sur le taux de contamination par les femelles. Ces résultats mettent en évidence la nécessité d’optimiser la production des pupes dès les premiers jours de la collecte et de développer une méthode de séparation des sexes plus efficace et automatisée.

International audience; Glossina palipides salivary gland hypertrophy virus (GpSGHV) infects tsetse flies, which are vectors for African trypanosomosis. This virus represents a major challenge in insect mass rearing and has hampered the implementation of the sterile insect technique programs in the Member States of the International Atomic Energy Agency. GpSGHV virions consist of long rod-shaped particles over 9000Å in length, but little is known about their detailed structural organization. We show by cryo electron microscopy and cryo electron tomography that the GpSGHV virion has a unique, non-icosahedral helical structure. Its envelope exhibits regularly spaced spikes that protrude from the lipid bilayer and are arranged on a four-start helix. This study provides a detailed insight into the 3D architecture of GpSGHV, which will help to understand the viral life cycle and possibly allow the design of antiviral strategies in the context of tsetse fly infections.

Este artículo se encuentra disponible en la página web de la revista en la siguiente URL: https://www.nature.com/articles/s41598-018-34469-6 Vector-borne diseases are responsible for more than one million deaths per year. Alternative methods of mosquito control to insecticides such as genetic control techniques are thus urgently needed. In genetic techniques involving the release of sterile insects, it is critical to release insects of high quality. Sterile males must be able to disperse, survive and compete with wild males in order to inseminate wild females. There is currently no standardized, fast-processing method to assess mosquito male quality. Since male competitiveness is linked to their ability to fly, we developed a flight test device that aimed to measure the quality of sterile male mosquitoes via their capacity to escape a series of flight tubes within two hours and compared it to two other reference methods (survival rate and mating propensity). This comparison was achieved in three different stress treatment settings usually encountered when applying the sterile insect technique, i.e. irradiation, chilling and compaction. In all treatments, survival and insemination rates could be predicted by the results of a flight test, with over 80% of the inertia predicted. This novel tool could become a standardised quality control method to evaluate cumulative stress throughout the processes related to genetic control of mosquitoes.

International audience; Major efforts are currently underway to develop novel, complementary methods to combat mosquito-borne diseases. Mosquito genetic control strategies (GCSs) have become an increasingly important area of research on account of their species-specificity, track record in targeting agricultural insect pests, and their environmentally non-polluting nature. A number of programs targeting Aedes and Anopheles mosquitoes, vectors of human arboviruses and malaria respectively, are currently being developed or deployed in many parts of the world. Operationally implementing these technologies on a large scale however, beyond proof-of-concept pilot programs, is hampered by the absence of adequate sex separation methods. Sex separation eliminates females in the laboratory from male mosquitoes prior to release. Despite the need for sex separation for the control of mosquitoes, there have been limited efforts in recent years in developing systems that are fit-for-purpose. In this special issue of Parasites and Vectors we report on the progress of the global Coordinated Research Program on Exploring genetic, molecular, mechanical and behavioural methods for sex separation in mosquitoes that is led by the Insect Pest Control Subprogramme of the Joint FAO/IAEA Division of Nuclear Techniques in Food and Agriculture with the specific aim of building efficient sex separation systems for mosquito species. In an effort to overcome current barriers we briefly highlight what we believe are the three main reasons why progress has been so slow in developing appropriate sex separation systems: the availability of methods that are not scalable, the difficulty of building the ideal genetic systems and, finally, the lack of research efforts in this area.

The sterile insect technique (SIT) is an environmentally safe and proven technology to suppress wild populations. To further advance its utility, a novel CRISPR-based technology termed precision guided SIT (pgSIT) is described. PgSIT mechanistically relies on a dominant genetic technology that enables simultaneous sexing and sterilization, facilitating the release of eggs into the environment ensuring only sterile adult males emerge. Importantly, for field applications, the release of eggs will eliminate burdens of manually sexing and sterilizing males, thereby reducing overall effort and increasing scalability. Here, to demonstrate efficacy, we systematically engineer multiple pgSIT systems in Drosophila which consistently give rise to 100% sterile males. Importantly, we demonstrate that pgSIT-generated sterile males are fit and competitive. Using mathematical models, we predict pgSIT will induce substantially greater population suppression than can be achieved by currently-available self-limiting suppression technologies. Taken together, pgSIT offers to potentially transform our ability to control insect agricultural pests and disease vectors.

Background The Government of Senegal has embarked several years ago on a project that aims to eradicate Glossina palpalis gambiensis from the Niayes area. The removal of the animal trypanosomosis would allow the development more efficient livestock production systems. The project was implemented using an area-wide integrated pest management strategy including a sterile insect technique (SIT) component. The released sterile male flies originated from a colony from Burkina Faso. Methodology/Principal Findings Monitoring the efficacy of the sterile male releases requires the discrimination between wild and sterile male G. p. gambiensis that are sampled in monitoring traps. Before being released, sterile male flies were marked with a fluorescent dye powder. The marking was however not infallible with some sterile flies only slightly marked or some wild flies contaminated with a few dye particles in the monitoring traps. Trapped flies can also be damaged due to predation by ants, making it difficult to discriminate between wild and sterile males using a fluorescence camera and / or a fluorescence microscope. We developed a molecular technique based on the determination of cytochrome oxidase haplotypes of G. p. gambiensis to discriminate between wild and sterile males. DNA was isolated from the head of flies and a portion of the 5’ end of the mitochondrial gene cytochrome oxidase I was amplified to be finally sequenced. Our results indicated that all the sterile males from the Burkina Faso colony displayed the same haplotype and systematically differed from wild male flies trapped in Senegal and Burkina Faso. This allowed 100% discrimination between sterile and wild male G. p. gambiensis. Conclusions/Significance This tool might be useful for other tsetse control campaigns with a SIT component in the framework of the Pan-African Tsetse and Trypanosomosis Eradication Campaign (PATTEC) and, more generally, for other vector or insect pest control programs., Author Summary The Government of Senegal has embarked since several years on a project that aims to create a tsetse-free area in the Niayes. The project was implemented using an area-wide integrated pest management (AW-IPM) strategy where the sterile flies used for the sterile insect technique (SIT) component were derived from a colony originating from Burkina Faso. Monitoring the efficacy of the sterile male releases requires the discrimination between wild and sterile males that are sampled in monitoring traps. Before being released, sterile males were marked with a fluorescent dye powder. The marking was however not infallible with some sterile flies only slightly marked or some wild flies contaminated with a few dye particles in the monitoring traps, making it difficult to discriminate between wild and sterile males using a UV camera. We developed a molecular technique based on the cytochrome oxidase gene that efficiently discriminates between wild and sterile males. This tool might be useful for other tsetse control campaigns with a SIT component or for other vector or insect pest control programs.

Alternative vector control methods to manage the Zika virus outbreak: more haste, less speed

Issues of male fertility must be addressed to support the development of a sterile insect technique (SIT) programme for the control of Aedes albopictus Skuse (Diptera: Culicidae) populations on Reunion Island in the Indian Ocean. The mating ability of a local strain of Ae. albopictus was tested using several batches of females and different cage sizes under laboratory conditions. Individual males were able to inseminate up to 14 females at an average of 9.5 females per male when exposed to 20 females over 7 days. Males filled between three and 27 spermathecal capsules at an average of 15.5 capsules per male. The average number of females inseminated per male was 5.3 when two virgin females were introduced to one male and replaced every day for 12 days, and 8.6 when 10 virgin females were introduced to one male and replaced every day for 14 days. A continuous decrease in the number of both inseminated females and filled spermathecal capsules was observed over time, until no mating occurred after 14 days. The high number of females inseminated by one male and the duration of male activity may have strong implications for SIT control of mosquitoes.

International audience; : 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 1.5 and 4.5m above the ground at the 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.

International audience; 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.

International audience; The enormous burden placed on populations worldwide by mosquito-borne diseases, most notably malaria and dengue, is currently being tackled by the use of insecticides sprayed in residences or applied to bednets, and in the case of dengue vectors through reduction of larval breeding sites or larviciding with insecticides thereof. However, these methods are under threat from, amongst other issues, the development of insecticide resistance and the practical difficulty of maintaining long-term community-wide efforts. The sterile insect technique (SIT), whose success hinges on having a good understanding of the biology and behaviour of the male mosquito, is an additional weapon in the limited arsenal against mosquito vectors. The successful production and release of sterile males, which is the mechanism of population suppression by SIT, relies on the release of mass-reared sterile males able to confer sterility in the target population by mating with wild females. A five year Joint FAO/IAEA Coordinated Research Project brought together researchers from around the world to investigate the pre-mating conditions of male mosquitoes (physiology and behaviour, resource acquisition and allocation, and dispersal), the mosquito mating systems and the contribution of molecular or chemical approaches to the understanding of male mosquito mating behaviour. A summary of the existing knowledge and the main novel findings of this group is reviewed here, and further presented in the reviews and research articles that form this Acta Tropica special issue.

International audience; Male insects are expected to optimize their reproductive strategy according to the availability of sperm or other ejaculatory materials, and to the availability and reproductive status of females. Here, we investigated the reproductive strategy and sperm management of male and virgin female Aedes albopictus, a mosquito vector of chikungunya and dengue viruses. The dynamics of semen transfer to the female bursa inseminalis and spermathecae were observed. Double-mating experiments were conducted to study the effect of time lapsed or an oviposition event between two copulations on the likelihood of a female double-insemination and the use of sperm for egg fertilization; untreated fertile males and radio-sterilised males were used for this purpose. Multiple inseminations and therefore the possibility of sperm competition were limited to matings closely spaced in time. When two males consecutively mated the same female within a 40 min interval, in ca. 15% of the cases did both males sire progeny. When the intervals between the copulations were longer, all progeny over several gonotrophic cycles were offspring of the first male. The mating behavior of males was examined during a rapid sequence of copulations. Male Ae. albopictus were parceling sperm allocation over several matings; however they would also attempt to copulate with females irrespective of the available sperm supply or accessory gland secretion material. During each mating, they transferred large quantities of sperm that was not stored for egg fertilization, and they attempted to copulate with mated females with a low probability of transferring their genes to the next generation. The outcomes of this study provided in addition some essential insights with respect to the sterile insect technique (SIT) as a vector control method.

The complete genome of Sibine fusca densovirus was cloned and sequenced. The genome contained 5,012 nucleotides (nt), including inverted terminal repeats (ITRs) of 230 nt with terminal hairpins of 161 nt. Its DNA sequence and monosense organization with 3 open reading frames (ORFs) is typical of the genus Iteravirus in the subfamily Densovirinae of the Parvoviridae .

Reunion Island suffers from high densities of the chikungunya and dengue vector Aedes Reunion Island suffers from high densities of the chikungunya and dengue vector Aedes albopictus. The sterile insect technique (SIT) offers a promising strategy for mosquito-borne diseases prevention and control. For such a strategy to be effective, sterile males need to be competitive enough to fulfil their intended function by reducing wild mosquito populations in natura. We studied the effect of irradiation on sexual maturation and mating success of males, and compared the sexual competitiveness of sterile versus wild males in the presence of wild females in semi-field conditions. For all untreated or sterile males, sexual maturation was completed within 13 to 20 h post-emergence and some males were able to inseminate females when 15 h old. In the absence of competition, untreated and sterile males were able to inseminate the same number of virgin females during 48 h, in small laboratory cages: an average of 93% of females was inseminated no matter the treatment, the age of males, and the sex ratio. Daily mating success of single sterile males followed the same pattern as for untreated ones, although they inseminated significantly fewer females after the ninth day. The competitiveness index of sterile males in semi-field conditions was only 0.14 when they were released at 1-day old, but improved to 0.53 when the release occurred after a 5-day period in laboratory conditions. In SIT simulation experiments, a 5:1 sterile to wild male ratio allowed a two-fold reduction of the wild population's fertility. This suggests that sterile males could be sufficiently competitive to mate with wild females within the framework of an SIT component as part of an AW-IPM programme for suppressing a wild population of Ae. albopictus in Reunion Island. It will be of interest to minimise the prerelease period in controlled conditions to ensure a good competitiveness without increasing mass rearing costs.

The riverine tsetse species Glossina palpalis gambiensis Vanderplank 1949 (Diptera: Glossinidae) inhabits riparian forests along river systems in West Africa. The government of Senegal has embarked on a project to eliminate this tsetse species, and African animal trypanosomoses, from the Niayes area using an area-wide integrated pest management approach.A stratified entomological sampling strategy was therefore developed using spatial analytical tools, remote sensing and mathematical modeling. A preliminary phyto-sociological census identified eight types of suitable habitat, which could be discriminated from LandSat 7ETM+ satellite images and denominated wet areas. At the end of March 2009, 683 unbaited Vavoua traps had been deployed, and the observed infested area in the Niayes was 525 km2. In the remaining area, a mathematical model was used to assess the risk that flies were present despite a sequence of zero catches. The analysis showed that this risk was above 0.05 in 19% of this area that will be considered as infested during the control operations. The remote sensing analysis that identified the wet areas allowed a restriction of the area to be surveyed to 4% of the total surface area (7,150 km2), whereas the mathematical model provided an efficient method to improve the accuracy and the robustness of the sampling protocol. The study thus allowed defining the limits of the target population for the eradication campaign. This entomological sampling procedure might be used for other vector or pest control scenarios. (Texte intégral)

E.F. Knipling, on the occasion of receiving the World Food Prize in 1992, stated that "major advances in coping with most of the major arthropod pest problems will require a change in the tactics and the strategies to manage those pests. They must change for the currently used limited scale, reactive, broad-spectrum measures to preventive approaches that are target specific and applied on an area-wide basis". Since then, practices to manage these pests have hardly changed and pesticide use is still on the rise every year to the detriment of the environment, human health and the ecosystem. The need for more target specific, sustainable, environmentally-friendly and cost-effective control tactics and pest management strategies remains to date more pertinent than ever. Integrated pest management (IPM) aims at the careful integration of several control tactics which are the most opportune for the particular ecosystem, while safeguarding human health and the environment. It has remained the dominant paradigm of pest control over the last half century. IPM can be applied on a field-by-field basis (localised IPM), or against an entire pest population within a delimited geographic area (area-wide (AW-IPM). AW-IPM seeks to treat all habitats, also those that are of no economic interest to the farmer, so that none produces migrants to re-establish significant infestations in areas of concern. This is in contrast to the field-by-field approach that focuses narrowly on defending the valued entity (crop, livestock, people, building, etc.). In most cases, AW-IPM leads to more effective and sustainable management of the insect pest. The sterile insect technique (SIT) is a species-specific autocidal control tactic that interferes with the reproductive potential of the target insect pest. It is a very powerful tool that is most effective when insect populations are low and when applied on an area-wide basis. The SIT within an AW-IPM context can be applied for the suppression, eradication, containment or prevention of insect pest populations. There are numerous AW-IPM programmes where the SIT has been used successfully, especially against dipteran and lepidopteran pests, and examples will be given to illustrate this. On economic, environmental and biological grounds, the case for SIT is compelling.

Background Separating males and females at the early adult stage did not ensure the virginity of females of Anopheles arabiensis (Dongola laboratory strain), whereas two years earlier this method had been successful. In most mosquito species, newly emerged males and females are not able to mate successfully. For anopheline species, a period of 24 h post-emergence is generally required for the completion of sexual maturation, which in males includes a 180° rotation of the genitalia. In this study, the possibility of an unusually shortened sexual maturity period in the laboratory-reared colony was investigated. Methods The effect of two different sex-separation methods on the virginity of females was tested: females separated as pupae or less than 16 h post-emergence were mated with males subjected to various doses of radiation. T-tests were performed to compare the two sex-separation methods. The rate of genitalia rotation was compared for laboratory-reared and wild males collected as pupae in Dongola, Sudan, and analysed by Z-tests. Spermatheca dissections were performed on females mated with laboratory-reared males to determine their insemination status. Results When the sex-separation was performed when adults were less than 16 h post-emergence, expected sterility was never reached for females mated with radio-sterilized males. Expected sterility was accomplished only when sexes were separated at the pupal stage. Observation of genitalia rotation showed that some males from the laboratory strain Dongola were able to successfully mate only 11 h after emergence and 42% of the males had already completed rotation. A small proportion of the same age females were inseminated. Wild males showed a much slower genitalia rotation rate. At 17 h post-emergence, 96% of the laboratory-reared males had completed genitalia rotation whereas none of the wild males had. Conclusion This colony has been cultured in the laboratory for over one hundred generations, and now has accelerated sexual maturation when compared with the wild strain. This outcome demonstrates the kinds of selection that can be expected during insect colonization and maintenance, particularly when generations are non-overlapping and similar-age males must compete for mates.

The sterile insect technique is a promising environmentally friendly method for mosquito control. This technique involves releasing laboratory-produced sterile males into a target field site, and its effectiveness may be affected by the extent of adult mosquito predation. Sterile males undergo several treatments. Therefore, it is vital to understand which treatments are essential in minimizing risks to predation once released. The present study investigates the predation propensity of four mantis species (Phyllocrania paradoxa, Hymenopus coronatus, Blepharopsis mendica, Deroplatys desiccata) and two gecko species (Phelsuma standingi, P. laticauda) on adult Aedes aegypti, Ae. albopictus and Anopheles arabiensis mosquitoes in a laboratory setting. First, any inherent predation preferences regarding mosquito species and sex were evaluated. Subsequently, the effects of chilling, marking, and irradiation, on predation rates were assessed. The selected predators effectively preyed on all mosquito species regardless of the treatment. Predation propensity varied over days for the same individuals and between predator individuals. Overall, there was no impact of laboratory treatments of sterile males on the relative risk of predation by the test predators, unless purposely exposed to double the required sterilizing irradiation dose. Further investigations on standardized predation trials may lead to additional quality control tools for irradiated mosquitoes.

Genetic control methods of mosquito vectors of malaria, dengue, yellow fever, and Zika are becoming increasingly popular due to the limitations of other techniques such as the use of insecticides. The sterile insect technique is an effective genetic control method to manage insect populations. However, it is crucial to release sterile mosquitoes by air to ensure homogeneous coverage, especially in large areas. Here, we report a fully automated adult mosquito release system operated from an uncrewed aerial vehicle or drone. Our system, developed and tested in Brazil, enabled a homogeneous dispersal of sterile male Aedes aegypti while maintaining their quality, leading to a homogeneous sterile-to-wild male ratio due to their aggregation in the same sites. Our results indicate that the released sterile males were able to compete with the wild males in mating with the wild females; thus, the sterile males were able to induce sterility in the native female population. The use of drones to implement the sterile insect technique will lead to improvements in areal coverage and savings in operational costs due to the requirement of fewer release sites and field staff.; Genetic control methods of mosquito vectors of malaria, dengue, yellow fever, and Zika are becoming increasingly popular due to the limitations of other techniques such as the use of insecticides. The sterile insect technique is an effective genetic control method to manage insect populations. However, it is crucial to release sterile mosquitoes by air to ensure homogeneous coverage, especially in large areas. Here, we report a fully automated adult mosquito release system operated from an uncrewed aerial vehicle or drone. Our system, developed and tested in Brazil, enabled a homogeneous dispersal of sterile male Aedes aegypti while maintaining their quality, leading to a homogeneous sterile-to-wild male ratio due to their aggregation in the same sites. Our results indicate that the released sterile males were able to compete with the wild males in mating with the wild females; thus, the sterile males were able to induce sterility in the native female population. The use of drones to implement the sterile insect technique will lead to improvements in areal coverage and savings in operational costs due to the requirement of fewer release sites and field staff.