Efficacy Of Sterile Insect Technique Research Articles

Sequence and expression analysis of the spermatogenesis-specific gene cognates, wampa and Prosα6T, in Drosophila suzukii.

The sterile insect technique (SIT) is a highly effective biologically-based method for the population suppression of highly invasive insect pests of medical and agricultural importance. The efficacy of SIT could be significantly enhanced, however, by improved methods of male sterilization that avoid the fitness costs of irradiation. An alternative sterilization method is possible by gene-editing that targets genes essential for sperm maturation and motility, rendering them nonfunctional, similar to the CRISPR-Cas9 targeting of β2-tubulin in the genetic model system, Drosophila melanogaster. However, since genetic strategies for sterility are susceptible to breakdown or resistance in mass-reared populations, alternative targets for sterility are important for redundancy or strain replacement. Here we have identified and characterized the sequence and transcriptional expression of two genes in a Florida strain of Drosophila suzukii, that are cognates of the D. melanogaster spermatocyte-specific genes wampa and Prosalpha6T. Wampa encodes a coiled-coil dynein subunit required for axonemal assembly, and the proteasome subunit gene, Prosalpha6T, is required for spermatid individualization and nuclear maturation. The reading frames of these genes differed from their NCBI database entries derived from a D. suzukii California strain by 44 and 8 nucleotide substitutions/polymorphisms, respectively, though all substitutions were synonymous resulting in identical peptide sequences. Expression of both genes is predominant in the male testis, and they share similar transcriptional profiles in adult males with β2-tubulin. Their amino acid sequences are highly conserved in dipteran species, including pest species subject to SIT control, supporting their potential use in targeted male sterilization strategies.

Sterile insect technique with accidental releases of sterile females. Impact on mosquito-borne diseases control when viruses are circulating

The sterile insect technique (SIT) is a technique to control some vectors of diseases by releasing sterile males. However, during these releases, sterilized females can be (accidentally) released and since only females are vectors of diseases, it is important to study their impact when arthropod viruses are circulating. To that aim, we develop and study an entomological–epidemiological model, considering either permanent or periodic releases. Qualitative analyses of the continuous and periodic models are conducted. We highlight a critical sterile males release rate, ΛMcrit, above which the control of wild population is always effective, using massive releases. Estimating the basic reproduction number of the epidemiological model, R02, we show that if it is above a certain threshold, R0,∗2, that depends on the basic offspring number, N, and the release rate of sterile females, the epidemiological risk can only be controlled using (very) massive releases. Otherwise, we can estimate the basic reproduction number of the SIT epidemiological model, R0,SIT2, that shapes the stability property of the (periodic) disease-free equilibrium. We show that it might be possible to take R0,SIT2 below 1 using non-massive, but large enough, releases. However, practically, it seems more efficient to consider massive releases, followed by small releases once the vector population is small enough.In addition to SIT, we also recommend mechanical control, i.e. the reduction of breeding sites, that greatly improves the efficacy of SIT, in terms of duration or size of the releases.Our results reveal that outside an epidemic period, the release of sterile females is not an issue, as long as the sterile males release rate is greater than ΛMcrit. Within an epidemic period, we show that sterile females releases do not really impact the SIT efficiency, as long as the release rate, ΛF, is lower than a critical value, ΛFcrit, that depends on the mosquito and epidemiological threshold parameters, N, and R02. To illustrate numerically our theoretical results, we consider Dengue parameters. We estimate all thresholds and also the effective reproduction number, Reff2, and highlight the importance of early permanent or periodic SIT control to prevent or mitigate the risk of a Dengue epidemic, with and without sterile females releases.

Suppression of female fertility in Aedes aegypti with a CRISPR-targeted male-sterile mutation

Aedes aegypti spread devastating viruses such as dengue, which causes disease among 100 to 400 million people annually. A potential approach to control mosquito disease vectors is the sterile insect technique (SIT). The strategy involves repeated release of large numbers of sterile males, which reduces insect populations because the sterile males mate and thereby suppress the fertility of females that would otherwise mate with fertile males. While SIT has been successful in suppressing certain agricultural pests, it has been less effective in depressing populations of Ae. aegypti This limitation is in part because of the fitness effects resulting from mutagenizing the mosquitoes nonspecifically. Here, we introduced and characterized the impact on female fertility of an Ae. aegypti mutation that disrupts a gene that is specifically expressed in testes. We used CRISPR/Cas9 to generate a null mutation in the Ae. aegypti β2-tubulin (B2t) gene, which eliminates male fertility. When we allowed wild-type females to first mate with B2t mutant males, most of the females did not produce progeny even after being subsequently exposed to wild-type males. We also introduced B2t mutant and wild-type males simultaneously with wild-type females and found that a larger number of B2t mutant males relative to the wild-type males was effective in significantly suppressing female fertility. These results raise the possibility of employing B2t sterile males to improve the efficacy of SIT in suppressing populations of Ae. aegypti through repeated releases and thereby reduce the transmission of viruses by these invasive mosquitoes.

Sterile Insect Technique in an Integrated Vector Management Program against Tiger Mosquito Aedes albopictus in the Valencia Region (Spain): Operating Procedures and Quality Control Parameters.

Aedes albopictus and Aedes aegypti are the main vectors of arboviral diseases such as dengue, Zika and chikungunya viruses. About a third of the world population is currently at risk of contracting Aedes-borne epidemics. In recent years, A. albopictus has drastically increased its distribution in many countries. In the absence of efficient mosquito vector control methods, the sterile insect technique (SIT) is presented as a very promising and environment-friendly control tool. The Agriculture Department of the Valencian Region is promoting an ongoing pilot project to evaluate the efficacy of an integrated vector management program (IVM) based on the use of the SIT as the main method of control. The laboratory studies for evaluating the entomological efficacy of SIT through the phased conditional testing process recommended by World Health Organization and the International Atomic Energy Agency (WHO-IAEA) are addressed. This study describes the routine operating procedures and quality control parameters for the medium-scale rearing of sterile male A. albopictus. More than 15 million sterile males have been produced and released in an area of 80 ha between 2018 and 2020. Of the initial L1 larvae, we recovered 17.2% of male pupae after sex sorting to be sterilized and released on the field, while the rest of the pupae remained available to maintain the rearing colony. The residual percentage of females after sex sorting was on average 0.17%. The obtained values in terms of production and quality control as well as the proposed rearing methodology can be useful for designing a medium-scale mosquito-rearing pipeline.

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

Simple SummaryAedes albopictus, also known as the Asian tiger mosquito, is an invasive mosquito well established in Europe, posing high risks of transmission of dengue and chikungunya viruses that are detrimental to human health. The newly invaded areas that experienced no intense mosquito annoyance in the past are now suffering because of the Asian tiger mosquito. Due to the large number of diverse breeding sites and the poor community participation in mosquito habitat elimination programs, traditional control efforts, such as intense chemical control efforts in public areas, have rather low efficacy. The sterile insect technique (SIT) is a method of insect control with successful applications against several agricultural insect pests and it has been proposed as a control method against invasive mosquito species. Weekly release of 2280 to 2995 sterile male mosquitoes/hectare resulted in increased levels of egg sterility of Ae. albopictus at a single release site of 5 hectares. This first SIT pilot trial against Ae. albopictus in Greece showed encouraging results that justify the continuation with larger scale trials.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.

Deconstructing the eradication of new world screwworm in North America: retrospective analysis and climate warming effects.

Before its eradication from North America, the subtropical‐tropical new world screwworm fly Cochliomyia hominivorax (Coquerel) invaded southwestern temperate areas of the U.S.A., where it caused myiasis in wildlife and livestock. Outbreaks of the fly occurred during years when adult migrants were carried northward on North American monsoon winds from the northern areas of Mexico and south Texas. We deconstruct, retrospectively, the biology and the effect of weather on the eradication of the fly in North America. Screwworm was found to be an ideal candidate for eradication using the sterile insect technique (SIT) because females mate only once, whereas males are polygynous, and, although it has a high reproductive potential, field population growth rates are low in tropical areas. In northern areas, eradication was enhanced by cool‐cold weather, whereas eradication in tropical Mexico and Central America is explained by the SIT. Despite low average efficacy of SIT releases (approximately 1.7%), the added pressure of massive SIT releases reduced intrinsically low fly populations, leading to mate‐limited extinction. Non‐autochthonous cases of myiasis occur in North America and, if the fly reestablishes, climate warming by 2045–2055 will expand the area of favourability and increase the frequency and severity of outbreaks.

Exploitation of the Medfly Gut Microbiota for the Enhancement of Sterile Insect Technique: Use of Enterobacter sp. in Larval Diet-Based Probiotic Applications.

The Mediterranean fruit fly (medfly), Ceratitis capitata, is a pest of worldwide substantial economic importance, as well as a Tephritidae model for sterile insect technique (SIT) applications. The latter is partially due to the development and utilization of genetic sexing strains (GSS) for this species, such as the Vienna 8 strain, which is currently used in mass rearing facilities worldwide. Improving the performance of such a strain both in mass rearing facilities and in the field could significantly enhance the efficacy of SIT and reduce operational costs. Recent studies have suggested that the manipulation of gut symbionts can have a significant positive effect on the overall fitness of insect strains. We used culture-based approaches to isolate and characterize gut-associated bacterial species of the Vienna 8 strain under mass rearing conditions. We also exploited one of the isolated bacterial species, Enterobacter sp., as dietary supplement (probiotic) to the larval diet, and we assessed its effects on fitness parameters under the standard operating procedures used in SIT operational programs. Probiotic application of Enterobacter sp. resulted in improvement of both pupal and adult productivity, as well as reduced rearing duration, particularly for males, without affecting pupal weight, sex ratio, male mating competitiveness, flight ability and longevity under starvation.

The Effect of Long-Distance Transportation on the Fitness of Irradiated False Codling Moth (Lepidoptera: Tortricidae) for Use in a Sterile Insect Release Program.

The effect of cold immobilization and long-distance transport of irradiated Thaumatotibia leucotreta (Meyrick) on the flight ability of male (♂) and female (♀) moths, the longevity of male and female moths, and the realized fecundity of mating pairs CIM (chilled irradiated moths) ♀ × CIM♂, CIM♀ × NIP (nonirradiated pupae) ♂, NIP♀ × CIM♂, and NIP♀ × NIP♂ was examined to improve application of the sterile insect technique (SIT). Adult moths treated with 150 Gy of gamma radiation were immobilized with cold temperature between 4 and 6°C inside a polyurethane cooler box and transported for 12 h by road from Citrusdal, Western Cape Province, to Addo, Eastern Cape Province. Nonirradiated moths were transported as pupae inside a cardboard tray and removed by hand after which male and female pupae were separated and placed inside containers for eclosion. Male and female moths were individually placed inside petri dishes to determine longevity or paired with irradiated and nonirradiated counterparts to evaluate realized fecundity before incubation in 100% darkness at 25°C and 75% relative humidity. Flight tests were conducted indoors at 25°C by release of individual moths per hand. A significant decrease in flight ability and longevity of irradiated false codling moth was found after handling, cold immobilization, and transport, although critically, realized fecundity was not affected. Because of the impact of long-distance transport on quality of the released insects as well as the efficacy of SIT, comprehensive protocols for this critical step in the process need to be developed for a pestiferous insect with phytosanitary status such as false codling moth.

Engineered Female-Specific Lethality for Control of Pest Lepidoptera

The sterile insect technique (SIT) is a pest control strategy involving the mass release of radiation-sterilized insects, which reduce the target population through nonviable matings. In Lepidoptera, SIT could be more broadly applicable if the deleterious effects of sterilization by irradiation could be avoided. Moreover, male-only release can improve the efficacy of SIT. Adequate methods of male-only production in Lepidoptera are currently lacking, in contrast to some Diptera. We describe a synthetic genetic system that allows male-only moth production for SIT and also replaces radiation sterilization with inherited female-specific lethality. We sequenced and characterized the doublesex (dsx) gene from the pink bollworm (Pectinophora gossypiella). Sex-alternate splicing from dsx was used to develop a conditional lethal genetic sexing system in two pest moths: the diamondback moth (Plutella xylostella) and pink bollworm. This system shows promise for enhancing existing pink bollworm SIT, as well as broadening SIT-type control to diamondback moth and other Lepidoptera.

Wolbachia Symbiont Infections Induce Strong Cytoplasmic Incompatibility in the Tsetse Fly Glossina morsitans

Tsetse flies are vectors of the protozoan parasite African trypanosomes, which cause sleeping sickness disease in humans and nagana in livestock. Although there are no effective vaccines and efficacious drugs against this parasite, vector reduction methods have been successful in curbing the disease, especially for nagana. Potential vector control methods that do not involve use of chemicals is a genetic modification approach where flies engineered to be parasite resistant are allowed to replace their susceptible natural counterparts, and Sterile Insect technique (SIT) where males sterilized by chemical means are released to suppress female fecundity. The success of genetic modification approaches requires identification of strong drive systems to spread the desirable traits and the efficacy of SIT can be enhanced by identification of natural mating incompatibility. One such drive mechanism results from the cytoplasmic incompatibility (CI) phenomenon induced by the symbiont Wolbachia. CI can also be used to induce natural mating incompatibility between release males and natural populations. Although Wolbachia infections have been reported in tsetse, it has been a challenge to understand their functional biology as attempts to cure tsetse of Wolbachia infections by antibiotic treatment damages the obligate mutualistic symbiont (Wigglesworthia), without which the flies are sterile. Here, we developed aposymbiotic (symbiont-free) and fertile tsetse lines by dietary provisioning of tetracycline supplemented blood meals with yeast extract, which rescues Wigglesworthia-induced sterility. Our results reveal that Wolbachia infections confer strong CI during embryogenesis in Wolbachia-free (GmmApo) females when mated with Wolbachia-infected (GmmWt) males. These results are the first demonstration of the biological significance of Wolbachia infections in tsetse. Furthermore, when incorporated into a mathematical model, our results confirm that Wolbachia can be used successfully as a gene driver. This lays the foundation for new disease control methods including a population replacement approach with parasite resistant flies. Alternatively, the availability of males that are reproductively incompatible with natural populations can enhance the efficacy of the ongoing sterile insect technique (SIT) applications by eliminating the need for chemical irradiation.

Bacterially enriched diet improves sexual performance of sterile male Mediterranean fruit flies

The Mediterranean fruit fly, Ceratitis capitata (Wiedemann) (Diptera: Tephritidae), (medfly) is a polyphagous and cosmopolitan agricultural pest, targeted in many areas for control by the Sterile Insect Technique (SIT). Our objective in the present study was to test the hypothesis that a bacterially enriched diet provided to sterile males will improve their sexual performance in competitive settings that emulate natural condi- tions. Specifically we determined how feeding on diets enriched with Klebsiella oxytoca affected the ability of sterile males to compete for wild females against wild males, their ability to inhibit female receptivity, and their survival. We found that enriching the sterile male diet with K.oxytoca significantly improved mating competitiveness in the labora- tory and in field cages. In addition, bacterially enriched sterile males inhibited female receptivity more efficiently than sugar fed males and survived longer duration of starvation. We conclude that inoculating mass reared sterile flies with bacteria prior to their release is a valid approach to improve the efficacy of SIT.

Sex ratio and the role of mild relative humidity in mating behaviour of red date palm weevil Rhynchophorus ferrugineus Oliv. (Coleoptera: Curculionidae) gamma‐irradiated adults

AbstractThe influence of gamma radiation on the mating behaviour of the red date palm weevil (RDPW), Rhynchophorus ferrugineus (Olivier), and the efficacy of sterile insect technique (SIT) under different levels of relative humidity (RH) were studied. No adverse effects of gamma radiation were observed on the mating behaviour parameters of the RDPW, such as mate recognition time, mating duration, mating frequency within a 30‐min period and duration between consecutive matings. However, the weevils were sexually stimulated during aggregation. RH significantly affected egg laying as well as egg hatching. Significantly lower egg laying and hatching were recorded at 25% RH than at higher humidity levels, suggesting that low humidity conditions are better for successful SIT manipulations in the field. Lifespan of irradiated RDPW males was significantly shorter than that of un‐irradiated controls, irrespective of RH level.

PeriodGene ofBactrocera cucurbitae(Diptera: Tephritidae) Among Strains with Different Mating Times and Sterile Insect Technique

AbstractThe efficacy of sterile insect technique (SIT) depends on successful mating of released males with wild females. If the time of mating in a day of mass-reared and released males differs from those of wild females, the efficiency of SIT decreases. Therefore, understanding the molecular mechanisms controlling mating time of the target pests is particularly important for SIT. The period (per) gene, which has been considered as a key clock gene controlling the mating time of the melon fly, Bactrocera cucurbitae (Coquillett) (Diptera: Tephritidae), was cloned from two strains having different times of mating during the day. DNA sites varied in the 5′ and 3′ untranslated regions and at synonymous sites, although protein sequences were identical. We also provide phylogenetic relationships among PER protein sequences of dipteran species including several tephritid pest species. The functional domains of PER in the melon fly are very similar to those in other tephritid species. A luciferase reporter assay showed that the melon fly PER can functionally complement that of Drosophila melanogaster (Meigen). The results implicate that the major genetic cause of the difference in circadian periods, and thus in reproductive isolation, is probably one or more other clock gene(s). Thus, the series of studies may provide a novel factor concerning genetic quality control of mass-reared insect pests for SIT, which depends on successful mating of released males and wild females.