Sterile Insect Technique (SIT) field trial targeting the suppression of Aedes albopictus in Greece.

The sterile insect technique (SIT) involves releasing large numbers of sterile males to outcompete wild males in mating with females, leading to a decline in pest populations. In the current study, we conducted a suppression trial in Greece against the invasive dengue vector mosquito Aedes albopictus (Skuse) through the weekly release of sterile males for 22 weeks from June to September 2019. Our approach included the long-distance transport of sterile mosquitoes, and their release at a density of 2,547 ± 159 sterile males per hectare per week as part of an area-wide integrated pest management strategy (AW-IPM). The repeated releases of sterile males resulted in a gradual reduction in egg density, reaching 78% from mid-June to early September. This reduction remained between 70% and 78% for four weeks after the end of the releases. Additionally, in the SIT intervention area, the ovitrap index, representing the percentage of traps containing eggs, remained lower throughout the trial than in the control area. This trial represents a significant advance in the field of mosquito control, as it explores the viability and efficacy of producing and transporting sterile males from a distant facility to the release area. Our results provide valuable insights for future SIT programmes targeting Ae. Albopictus, and the methodology we employed can serve as a starting point for developing more refined and effective release protocols, including the transportation of sterile males over long distances from production units to intervention areas.

Title: Essai sur le terrain de la Technique de l'Insecte Stérile (TIS) ciblant la suppression d'Aedes albopictus en Grèce. Abstract: La technique de l'insecte stérile (TIS) consiste à libérer un grand nombre de mâles stériles pour supplanter les mâles sauvages lors de l'accouplement avec les femelles, entraînant ainsi un déclin des populations de nuisibles. Dans la présente étude, nous avons mené un essai de suppression en Grèce contre le moustique vecteur invasif de la dengue, Aedes albopictus (Skuse), par le biais de la libération hebdomadaire de mâles stériles pendant 22 semaines de juin à septembre 2019. Notre approche comprenait le transport sur de longues distances de moustiques stériles, et leur lâcher à une densité de 2 547 ± 159 mâles stériles par hectare et par semaine dans le cadre d'une stratégie de lutte intégrée contre les nuisibles à l'échelle de la zone (AW-IPM). Les lâchers répétés de mâles stériles ont entraîné une réduction progressive de la densité des œufs, atteignant 78 % de la mi-juin au début septembre. Cette réduction est restée entre 70 % et 78 % pendant quatre semaines après la fin des lâchers. De plus, dans la zone d'intervention de la TIS, l'indice d'oviposition, représentant le pourcentage de pièges contenant des œufs, est resté plus faible que dans la zone témoin tout au long de l'essai. Cet essai représente une avancée significative dans le domaine de la lutte contre les moustiques, car il explore la viabilité et l'efficacité de la production et du transport de mâles stériles depuis une installation éloignée vers la zone de lâcher. Nos résultats fournissent des informations précieuses pour les futurs programmes de TIS ciblant Ae. albopictus et la méthodologie que nous avons utilisée pourra servir de point de départ pour développer des protocoles de libération plus raffinés et plus efficaces, y compris le transport de mâles stériles sur de longues distances depuis les unités de production jusqu'aux zones d'intervention.

Effect of 2 sex-sorting time schedules on SIT facility management.

Improvements are needed in mosquito mass-rearing to effectively implement the sterile insect technique (SIT). However, managing this technique is challenging and resource intensive. SIT relies on mass rearing, sterilization, and release of adult males to reduce field populations. Maintaining an acceptable level of female presence, who can transmit viruses through biting, is crucial. Females are also essential for facility sustainability. Sex sorting plays a vital role in the production process, and our current mechanical sorting approach aims to obtain a high number of adult males with minimal female contamination within 24 h of pupation. Utilizing protandry helps control female contamination. While the 24-h sorting period achieves desired contamination levels, it may not yield enough females to sustain breeding lines, leading to increased labor costs that impact project sustainability. By delaying the sorting procedure to 48 h, we obtained sufficient females to sustain breeding lines, achieving a balance between male production and female contamination using the automatic version of the Fay-Morlan device as the sorting tool.

Aedes albopictus Sterile Male Production: Influence of Strains, Larval Diet and Mechanical Sexing Tools.

The sterile insect technique (SIT) is a biologically based method of pest control, which relies on the mass production, sterilization, and release of sterile males of the target species. Since females can transmit viruses, it is important to develop a mass rearing system to produce a large number of males with a low presence of females. We evaluated the effects of different strains, larval diets and sexing tools on male productivity and residual female presence for the application of SIT against Aedes albopictus. Strains coming from Italy, Germany, Greece, and Montenegro, with different levels of colonization, were reared with three larval diets: IAEA-BY, BLP-B and SLP-BY. Developed pupae were sexed using two different mechanical methods: sieve or Fay-Morlan separator. The results proved that adoption of the Fay-Morlan separator increased the productivity and limited the female presence. The IAEA-BY diet showed the lowest female contamination. Strains with a high number of breeding generations showed a decreased productivity and an increased female presence. Increased female presence was found only in extensively reared strains and only when the sorting operation was conducted with sieves. We hypothesize that extensive colonization may determine a size reduction which limits the sexing tool efficiency itself.

Field Performance Assessment of Irradiated Aedes albopictus Males Through Mark-Release-Recapture Trials With Multiple Release Points.

Mark-release-recapture (MRR) trials have been conducted in Northern Italy to evaluate the capacity of radio-substerilized Aedes albopictus males to survive, disperse, and engage in mating in the field. Two MRR sessions with the human landing collection method (HLC) were conducted with the simultaneous release of irradiated males marked with four different pigment colors. The survival and dispersal rates seem to be influenced more by environmental factors such as barriers, shading, and vegetation rather than weather parameters. In this study, we confirmed a positive linear relationship between the sterile adult male's daily survival rate and the relative humidity previously reported in similar experimental conditions and a different dispersal capacity of the released A. albopictus males in low- (NDVI index <0.4) and high (NDVI index >0.4)-vegetated areas. Consistent with previous studies, A. albopictus males have their maximal dispersion in the first days after release, while in the following days the males become more stationary. The similar field performances obtained with marked and unmarked radio-sterilized and untreated A. albopictus males on similar environments confirm the negligible effects of irradiation and marking procedures on the quality of the males released. The similar sterile to wild (S/W) male ratio measured in high- and low-vegetation areas in the release sites indicates a similar distribution pattern for the wild and the released sterile males. According to the MRR data collected, the Lincoln index estimated different A. albopictus mean population densities in the study areas equal to 7,000 and 3,000 male/ha, respectively.

The Possible Role of Microorganisms in Mosquito Mass Rearing.

In Europe, one of the most significant mosquitoes of public health importance is Aedes albopictus (Skuse), an allochthonous species of Asian origin. One of the most promising control methods against Aedes albopictus is the sterile insect technique (SIT), which consists of mass rearing the target species, separation of males from females, and male exposure to sterilizing ionizing radiation. Once released in the environment, the sterile males are expected to search for wild females to mate with. If mating occurs, no offspring is produced. The quality of sterile males is a crucial aspect in SIT programs in order to optimize effectiveness and limit production costs. The integration of probiotic microorganisms in larval and adult mosquito diets could enhance the quality parameters of the released sterile males. In this review, we attempt to give the most representative picture of the present knowledge on the relationships between gut microbiota of mosquitoes and the natural or artificial larval diet. Furthermore, the possible use of probiotic microorganisms for mosquito larvae rearing is explored. Based on the limited amount of data found in the literature, we hypothesize that a better understanding of the interaction between mosquitoes and their microbiota may bring significant improvements in mosquito mass rearing for SIT purposes.

Reduction in Egg Fertility of Aedes albopictus Mosquitoes in Greece Following Releases of Imported Sterile Males.

Aedes albopictus, an invasive mosquito species, is currently well established in many European countries, posing high risks to human health. A preliminary trial using repetitive releases of irradiated sterile male mosquitoes was designed, implemented and evaluated for the first time in Greece. The main scope of this trial was to investigate the efficacy of sterile insect technique (SIT) on wild population egg hatch rate in Greece using mass-produced sterile male mosquitoes from another country (Italy). The study was conducted in Vravrona area, close to Athens International Airport (Attica Region). The number of eggs laid in ovitraps was similar in all intervention and control plots. However, a significant reduction in egg hatch rate was recorded in the SIT plot in comparison with both control plots starting two weeks after the first release. This trial validates the logistics (transportation, releases handling and monitoring) as a major step towards implementing efficient, environmentally safe control approaches as an additional tool against the invasive Aedes species in Greece and more widely in Europe.

Field Competitiveness of Aedes albopictus (Diptera: Culicidae) Irradiated Males in Pilot Sterile Insect Technique Trials in Northern Italy.

Vector-borne diseases account for 17% of infectious diseases, leading to more than one million deaths each year. Mosquitoes are responsible for 90% of the casualties and alternative control methods to insecticides are urgently needed, especially against Aedes vectors. Aedes albopictus is a particularly important species, causing major public health problems because it is a vector of several arboviruses and has a strong invasive behavior. Various genetic control methods have been proposed to be integrated into the management strategies of Aedes species, among which the sterile insect technique (SIT), which proved efficient against various insect pests and vectors. However, the ability of released irradiated sterile male mosquitoes to compete with their wild counterparts and induce sterility in wild females, which is critical to the success of this strategy, remained poorly defined. Here, we assessed the field competitiveness of Ae. albopictus irradiated male using data from eight release trials implemented in Northern Italy for 3 yr. Sterile males were capable of inducing a good level of sterility in the wild female population, however, with high variability in time and space. The field competitiveness of the released males was strongly negatively correlated with the ratio of sterile to wild males. This should be taken into consideration when designing future programs to suppress field populations of Aedes mosquitoes.