Sterile Insect Research Articles

Probiotic consortium modulating the gut microbiota composition and function of sterile Mediterranean fruit flies

The sterile insect technique (SIT) remains a successful approach in managing pest insects. However, the long-term mass rearing and sterilizing radiation associated with SIT have been observed to induce physiological and ecological fitness decline in target insects. This decline may be attributed to various factors, including commensal microbiota dysbiosis, selection procedures, loss of heterozygosity, and other complex interactions.. There is evidence that the bacterial symbiont of insects may play critical roles in digestion, development, reproduction, and behavior. Probiotics are an increasingly common approach for restoring the intestinal microbiota structure and fitness parameters of sterile insects, particularly in the Vienna 8 genetic sexing strain (V8-GSS) of the Mediterranean fruit fly (medfly), Ceratitis capitata. Here, we explore the influence of the previously isolated bacterial strain, Lactococcus lactis, Enterobacter sp., and Klebsiella oxytoca, administration as probiotic consortia (LEK-PC) to the larvae and/or adult diet over the course of 20 rearing generations on fitness parameters. The experiment was carried out in four colonies: a control colony (C), one to which probiotics were not added, one to which probiotics were added to the larval medium (L+), one to which probiotics were added to the adult medium (A+), and one to which probiotics were added to both the larval and adult mediums (AL+). Emergence, flight ability, survival under stress conditions, and mating competitiveness, were all significantly improved by the LEK-PC treatment independently of the administration stage. The intestinal microbiota structure of various medfly V8-GSS colonies also underwent a significant shift, despite the fact that the core microbial community was unaffected by the LEK-PC administration stage, according to 16S metagenomics sequencing. Comparison of the metabolic function prediction and associated carbohydrate enzymes among colonies treated with “LEK-PC” showed an enrichment of metabolic functions related to carbohydrates, amino acids, cofactors, and vitamins metabolism, as well as, glycoside hydrolase enzymes in the AL+ colony compared to the control. This study enriches the knowledge regarding the benefits of probiotic treatment to modulate and restore the intestinal microbiota of C. capitata sterile males for a better effectiveness of the SIT.

MODELING THE IMPACT OF RAINFALL AND TEMPERATURE ON STERILE INSECT CONTROL STRATEGIES IN A TROPICAL ENVIRONMENT

The sterile insect technique (SIT) is a biological control technique that can be used either to eliminate or decay a wild mosquito population under a given threshold to reduce the nuisance or the epidemiological risk. In this work, we propose a model using a differential system that takes into account the variations of rainfall and temperature over time and study their impacts on sterile males’ releases strategies. Our model is as simple as possible to avoid complexity while being able to capture the temporal variations of an Aedes albopictus mosquito population in a domain treated by SIT, located in Réunion island. The main objective is to determine what period of the year is the most suitable to start a SIT control to minimize the duration of massive releases and the number of sterile males to release, either to reduce the mosquito nuisance, or to reduce the epidemiological risk. Since sterilization is not [Formula: see text] efficient, we also study the impact of different levels of residual fertility within the released sterile males population. Our study shows that rainfall plays a major role in the dynamics of the mosquito and the SIT control, that the best period to start a massive SIT treatment lasts from July to December, that residual fertility has to be as small as possible, at least for nuisance reduction. Indeed, when the main objective is to reduce the epidemiological risk, we show that residual fertility is not necessarily an issue. Increasing the size of the releases is not always interesting. We also highlight the importance of combining SIT with mechanical control, i.e., the removal of breeding sites, in particular when the initial mosquito population is large. Last but not least our study shows the usefulness of the modeling approach to derive various simulations to anticipate issues and demand in terms of sterile insects’ production.

Modeling the impact of directional movement on mosquito population suppress

Due to mosquito habitat heterogeneity, its directional movement has been observed. In this paper, we develop a reaction–advection–diffusion model to explore mosquito populations suppression with sterile insect technique in spatially heterogeneous environments. By employing theoretical analysis, we show that the dynamic behavior of mosquitoes is closely related to their directional movement when the sterile mosquito amount is small. Numerical simulations further verify the theoretical analysis.

Evaluation of Peregrinus maidis transformer-2 as a target for CRISPR-based control.

The corn planthopper, Peregrinus maidis, is an economically important pest of corn and sorghum. Here we report the initial steps towards developing a CRISPR-based control method, precision guided sterile insect technique (pgSIT), for this hemipteran pest. Specifically, we evaluated the potential of transformer-2 (tra-2) as a target for sterilizing insects. First, we identified tra-2 transcripts within our P. maidis transcriptome database and performed RNA interference (RNAi) to confirm functional conservation. RNAi-mediated knockdown of Pmtra-2 in nymphs transformed females into pseudomales with deformed ovipositors resembling male claspers. While males showed no overt difference in appearance, they were indeed sterile. Importantly, the results were similar to those observed in another planthopper, Nilaparvata lugens. We also used CRISPR/Cas9 genome editing to assess the impact of tra-2 knockout in injectees. CRISPR-mediated knockout of Pmtra-2 had lethal effects on embryos, and hence not many injectees reached adulthood. However, mosaic knockout of Pmtra-2 did impact female and male fertility, which supports the use of tra-2 as a target for pgSIT in this hemipteran species.

Radiation dose fractionation and its potential hormetic effects on male Glossina palpalis gambiensis (Diptera: Glossinidae): a comparative study of reproductive and flight quality parameters.

One of the most critical factors for implementing the sterile insect technique for the management of tsetse is the production of large quantities of highly competitive sterile males in the field. Several factors may influence the biological quality of sterile males, but optimizing the irradiation protocols to limit unwanted somatic cell damage could improve male performance. This study evaluated the effect of fractionation of gamma radiation doses on the fertility and flight quality of male Glossina palpalis gambiensis. Induced sterility was assessed by mating irradiated males with virgin fertile females. Flight quality was assessed using a standard protocol. The male flies were irradiated as pupae on day 23-27 post larviposition with 110Gy, either in a single dose or in fractionations of 10+100Gy and 50+60Gyseparated by 1-, 2- and 3-day intervals or 55+55Gyseparated by 4-, 8-, and 24-hour intervals. All treatments induced more than 90% sterility in females mated with irradiated males, as compared with untreated males. No significant differences were found in emergence rate or flight propensity between fractionated and single radiation doses, nor between the types of fractionations. Overall, the 50(D0)+60(D1) Gy dose showed slightly higher induced sterility, flight propensity, and survival of males under feeding regime. Dose fractionation resulted in only small improvements with respect to flight propensity and survival, and this should be traded off with the required increase in labor that dose fractionation entails, especially in larger control programs.

Modelling mosquito population suppression based on competition system with strong and weak Allee effect.

Mosquito-borne diseases are threatening half of the world's population. To prevent the spread of malaria, dengue fever, or other mosquito-borne diseases, a new disease control strategy is to reduce or eradicate the wild mosquito population by releasing sterile mosquitoes. To study the effects of sterile insect technique on mosquito populations, we developed a mathematical model of constant release of sterile Aedes aegypti mosquitoes with strong and weak Allee effect and considered interspecific competition with Anopheles mosquitoes. We calculated multiple release thresholds and investigated the dynamical behavior of this model. In order to get closer to reality, an impulsive differential equation model was also introduced to study mosquito suppression dynamics under the strategy of releasing $ c $ sterile male mosquitoes at each interval time $ T $. Finally, the relationship between the releasing amount or the waiting period and the number of days required to suppress mosquitoes was illustrated by numerical simulations.

Sterile Insect Technique: Successful Suppression of an Aedes aegypti Field Population in Cuba

Sterile Insect Technique: Successful Suppression of an Aedes aegypti Field Population in Cuba

Animal trypanosomosis eliminated in a major livestock production region in Senegal following the eradication of a tsetse population.

African animal trypanosomosis (AAT) was one of the main disease-related constraints to the development of intensive livestock production systems in the Niayes region of Senegal, a 30km wide strip of land along the coast between Dakar and Saint-Louis. To overcome this constraint, the Government of Senegal initiated an area-wide integrated pest management programme combining chemical control tactics with the sterile insect technique to eradicate a population of the tsetse fly Glossina palpalis gambiensis Vanderplank, 1949 (Diptera, Glossinidae) in this area. The project was implemented following a phased conditional approach, and the target area was divided into three blocks treated sequentially. This study aims to assess the temporal dynamics of the prevalence of Trypanosoma spp. during the implementation of this programme. Between 2009 and 2022, 4,359 blood samples were collected from cattle and screened for trypanosomes using both the buffy coat and ELISA techniques, and PCR tests since 2020. The seroprevalence decreased from 18.9% (95%CI: 11.2-26.5) in 2009 to 0% in 2017-2022 in block 1, and from 92.9% (95%CI: 88.2-97) in 2010 to 0% in 2021 in block 2. The parasitological and serological data confirm the entomological monitoring results, i.e., that there is a high probability that the population of G. p. gambiensis has been eradicated from the Niayes and that the transmission of AAT has been interrupted in the treated area. These results indicate the effectiveness of the adopted approach and show that AAT can be sustainably removed through the creation of a zone free of G. p. gambiensis.

Diversity and Species Composition of Midgut Symbiotic Bacteria in Culex quinquefasciatus Mosquitoes in Gampaha District, Sri Lanka.

Mosquitoes, notorious for their deadly impact as disease vectors, also hold economic value owing to their roles in disease transmission. The present study focuses on the importance of understanding mosquito gut microbiota for implementing innovative vector control strategies, thereby mitigating disease transmission. The study was conducted in the Gampaha Medical Office of Health (MOH) area of Sri Lanka with the focus of elucidating the microbial diversity within the midgut of Culex quinquefasciatus, a crucial step to support ongoing paratransgenesis efforts. Sampling was performed by utilizing standard mosquito sampling techniques and their midgut homogenates were plated on Plate Count Agar to isolate bacteria, which were then identified through biochemical tests. Subsequently, the most abundant bacterial families were subjected to DNA extraction, PCR amplification, and gene sequencing for species identification. The study revealed the presence of four bacterial families (Staphylococcaceae, Streptococcaceae, Neisseriaceae, and Moraxellaceae) in adult mosquitoes, while larvae harbored an additional family, Micrococcaceae. Interestingly, the relative distribution of midgut bacteria varied significantly among field-caught larval and adult strains from different study areas (chi-square = 1.673; P < 0.05), indicating similar bacterial flora across mosquito life stages and geographical locations. Of particular interest is the identification of Lysinibacillus sphaericus, a bacterium with potential for paratransgenesis applications. Given the high mosquito density in the study area, leveraging paratransgenesis for Cx. quinquefasciatus control is recommended. Furthermore, insights into gut microbes could inform the integration of gut microflora from modified strains into existing Sterile Insect Technique (SIT) and Incompatible Insect Technique (IIT) approaches in Sri Lanka.

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.

Feedback stabilization and observer design for sterile insect technique models.

This paper focuses on the feedback global stabilization and observer construction for a sterile insect technique model. The sterile insect technique (SIT) is one of the most ecological methods for controlling insect pests responsible for worldwide crop destruction and disease transmission. In this work, we construct a feedback law that globally asymptotically stabilizes an SIT model at extinction equilibrium. Since the application of this type of control requires the measurement of different states of the target insect population, and, in practice, some states are more difficult or more expensive to measure than others, it is important to know how to construct a state estimator, which from a few well-chosen measured states, estimates the other ones, as the one we build in the second part of our work. In the last part of our work, we show that we can apply the feedback control with estimated states to stabilize the full system.

Keeping a close eye on the searching behaviour of the mass-reared parasitoid Dichasmimorpha longicaudata: addressing a standardised test

ABSTRACT Tephritid fruit flies are responsible for many losses in the fruit market due to direct damage to fruits and their quarantine restrictions. Augmentative parasitoid releases are an effective method of biological control that is complementary to the sterile insect technique (SIT) currently used for the management of many tephritid species. However, mass-produced parasitoids must exhibit high-quality attributes that are crucial for the reduction of pest populations. Females must be able to disperse and locate host larvae following kairomones. Nevertheless, there is currently no standardised, fast-processing method to assess the quality of searching parasitoids. Since parasitoid effectiveness is linked to flight ability, we developed a test with a larval-odour device that aimed to measure the quality of female parasitoids via their ability to locate host larvae by their odour. As an odour source, we applied a used or clean artificial host diet on a 3 × 2 cm piece of filter paper. We diluted the diets in ethanol at 1/1, 1/2, 1/3, and 1/4 (% mass/vol) to obtain an odour gradient. Female wasps landed and attempted to oviposit more frequently on devices treated with used diet than on those treated with clean diet. We suggest that this test can be implemented as a simple and fast method to evaluate the searching behaviour of mass-reared parasitoids of fruit fly larvae.

Side effects of X-ray irradiation on flight ability of Cydia pomonella moth.

The sterile insect technique (SIT) has proven to be an effective approach in managing the population of major invasive pests. Our previous studies showed that irradiation of Cydia pomonella males at a dosage of 366 Gy X-rays resulted in complete sterility. However, the mating competitiveness of sterilized males is significantly compromised, which can be attributed to a decline in their ability to fly. In this study, we examined the flight patterns of both male and female adults of C. pomonella. The results revealed significant variations in the average flight speed of both genders at different stages of maturity, with females displaying longer flight duration and covering greater distances. Effect of irradiation on the flight performance of 3-day-old male moths was further evaluated, as they demonstrated the longest flight distance. The findings indicated a significant decrease in flight distance, duration, and average speed, due to wing deformities caused by irradiation, which also limited the dispersal distance of moths in orchards, as indicated by the mark-and-recapture assay. Reverse-transcription quantitative polymerase chain reaction analysis revealed a down-regulation of flight-related genes such as Flightin, myosin heavy chain, and Distal-less following radiation exposure. These findings demonstrate that X-ray irradiation at a radiation dose of 366 Gy has a detrimental effect on the flight ability of male C. pomonella adults. These insights not only contribute to a better understanding of how radiation sterilization diminishes the mating competitiveness of male moths, but also aid in the development and improvement of SIT practices for the effective control of C. pomonella. © 2023 Society of Chemical Industry.

Comparative transcriptome analysis reveals the molecular mechanism of sterility induced by irradiation of Plutella xylostella (Linnaeus)

Plutella xylostella (Linnaeus) is an important pest of cruciferous plants, which is harmful all over the world, causing serious economic losses, and its drug resistance is increasing rapidly. The sterile insect technique (SIT) is a green control method and does not cause resistance. In this study, transcriptomics and bioinformatics were used to explore the effects of irradiation on the reproductive function of Plutella xylostella, and the response mechanism of sterility under irradiation was initially revealed. We identified 3342 (1682 up-regulated, 1660 down-regulated), 1963 (1042 up-regulated, 921 down-regulated) and 1531 (721 up-regulated, 810 down-regulated) differentially expressed genes (DEGs) in the 200 Gy vs CK (Control Check), 400 Gy vs CK and 400 Gy vs 200 Gy groups, respectively. GO and KEGG analyses were performed for DEGs in each group. The results showed that 200 Gy activated the downstream phosphorylation pathway and inhibited the cytochrome p450 immune response mechanism. 400 Gy promoted protein decomposition and absorption pathways, autophagy pathways, etc. Down-regulated genes were concentrated in the transformation process of energy metabolizing substances such as ATP, phosphorylation signaling pathway, and insulin, while up-regulated genes were concentrated in biological regulation and metabolic processes. Eight genes in the phosphorylation pathway were selected for qRT-PCR verification, and the results showed that the phosphorylation of different dose groups was regulated in different ways. 400 Gy used positive feedback regulation, while the phosphorylation of F1 used negative feedback regulation.

Larval Rearing and Nutrition of the Polyphagous Tephritid Pest Anastrepha ludens on Artificial Diets with Calcium Alginate, Agar, or Carrageenan as Gelling Agents at Various Concentrations and across Extreme Larval Density Conditions.

Research on larval rearing and nutrition of tephritid flies on artificial diets is key for the sterile insect technique. Here, we examined the effects of the type of gel (calcium alginate, agar, or carrageenan), at varying percentages in artificial diets for the polyphagous pest Anastrepha ludens, on the physicochemical and nutritional traits of the diets, and the effects of the type of gel, the gel content and the larval density (larvae/g of diet) used in production, quality parameters for mass-reared tephritids, diet removal (an indirect estimation of diet consumption), and nutritional traits of flies. Regardless of the gel content, calcium alginate diets were firmer and more resistant to penetration than the agar and carrageenan diets. The larval recovery, pupation, pupal weight, and flight ability of A. ludens were lower in calcium alginate diets than in agar and carrageenan diets. Diet removal was higher in calcium alginate diets; however, low levels of ammonium and high levels of uric acid in excretions from larvae on these diets suggest an alteration in protein metabolism. The firmness and penetration resistance characteristics of calcium alginate diets may have limited movement and feeding of larvae, but this could be overcome by the collective feeding of large groups of larvae. Our findings provide insights into the mechanism governing gel-diet rearing systems for A. ludens.

Accelerating sexual maturation of male Anastrepha ludens (Diptera: Tephritidae) fruit flies by adding two juvenile hormone analogues.

Improving the mating competitiveness and survival of sterile males are direct means to increase the effectiveness of the sterile insect technique (SIT). Some insecticide growth regulators, such as the juvenile hormone analogue (JHA) methoprene, have been used to improve the mating competitiveness of male tephritid flies by reducing their sexual maturation period. However, the application of methoprene reduces fly resistance to stress and decreases survival. Here, we compared the effects of methoprene and pyriproxyfen (PPF), another JHA, in Anastrepha ludens males. PPF is an insect growth regulator that exhibits higher negative effects on the larval molting process than methoprene or natural juvenile hormone. Both compounds were administered at two doses (0.05% and 0.10%) via the male diet immediately after emergence. Our results show that both PPF and methoprene reduced male sexual maturation. However, PPF-treated males exhibited a shorter maturation period and obtained more matings at a given age than methoprene-treated males. No significant differences were observed between the two PPF doses tested (0.05% and 0.10%). Male survival was equally reduced by the two compounds. Our results demonstrate that PPF accelerated sexual development without reducing the mating propensity of sterile male flies and can be used as a suitable alternative for methoprene. © 2023 Society of Chemical Industry.

β-tubulin functions in spermatogenesis in Lygus hesperus Knight

Lygus hesperus Knight is an important insect pest of crops across western North America, with field management heavily reliant on the use of chemical insecticides. Because of the evolution of resistance to these insecticides, effective and environmentally benign pest management strategies are needed. Traditional sterile insect technique (SIT) has been successfully employed to manage or eradicate some insect pests but involves introducing irradiated insects with random mutations into field populations. New genetically-driven SIT techniques are a safer alternative, causing fixed mutations that manipulate individual genes in target pests to produce sterile individuals for release. Here, we identified seven β-tubulin coding genes from L. hesperus and show that Lhβtub2 is critical in male sperm production and fertility. Lhβtub2 is expressed primarily in the male testes and targeting of this gene by RNA interference or gene editing leads to male sterility.

Assessing the sterility and quality of gamma-irradiated pepper weevils, Anthonomus eugenii (Coleoptera: Curculionidae), toward the development of the sterile insect technique.

The pepper weevil (PW), Anthonomus eugenii, is an economically significant pest of cultivated Capsicum spp. pepper crops in North America where it remains a challenge to manage because of its cryptic immature life stages. The sterile insect technique (SIT) is a genetic pest management tactic that relies on the release of insects that have been sterilized with ionizing radiation to lower the population reproductive rate. Toward developing an effective PW-SIT program, this study has, for the first time, investigated the effects of gamma irradiation on the sterility and survival of this species. Among the array of doses tested, we found that pupal PW males and females irradiated at 110 Gy produced no adult offspring. Furthermore, females mated with a male irradiated at 110 Gy had high egg sterility (97.3%), and irradiated females nearly completely failed to lay eggs (97.5%). Individuals irradiated at this dose had a shortened lifespan (lethal time to 50% mortality values of 12 and 11 days for males and females, respectively) and quantitatively reduced spontaneous flight activity. The eclosion rate of PW pupae was not significantly reduced by any radiation treatment. This study suggests that PWs irradiated at a gamma radiation dose of 110 Gy as pupae could feasibly be used in a PW-SIT program, because both males and females were 100% sterile at this dose. These findings will inform the development of a SIT program that could considerably improve the sustainability and effectiveness of PW management in greenhouse and field pepper crops worldwide. © 2023 His Majesty the King in Right of Canada and The Authors. Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry. Reproduced with the permission of the Minister of Agriculture and Agri Food Canada.

Dengue a global threat: An insight into management and control

Abstract Background : Dengue, whose severe forms have the potential to be fatal, is the most prevalent and rapidly proliferating vector-borne viral disease worldwide. It is evolving as a significant public health and economic dilemma for both rural and urban communities. Objectives : Given the urgent need for control measures, the purpose of this study is to evaluate both traditional and contemporary preventive strategies. Results : Traditional dengue control measures, while somewhat effective, tend to provide only temporary relief. In contrast, biotechnological innovations such as paratransgenesis, sterile insect technology, and genetically modified vectors have increased the effectiveness of conventional methods. In addition, the novel discovery of vaccines and immunotherapies offers a promising avenue for the management of dengue. Discussion &amp; Conclusion: Integration of time-tested techniques with the most recent biotechnological interventions emerges as a crucial dengue-prevention strategy. While these developments offer cause for optimism, additional research is required to determine the long-term implications and viability of these techniques. Implications : The findings have significant implications for theory, policymaking, and medical practice, highlighting the need for a combined approach to dengue management.

Biotechnology and Solutions: Insect-Pest-Resistance Management for Improvement and Development of Bt Cotton (Gossypium hirsutum L.)

Cotton (Gossypium spp. L.) is a major origin of natural fiber, and is projected at 117 million bales worldwide for 2021/22. A variety of biotic and abiotic stresses have considerable negative impacts on cotton. The significantly decreased applications of chemical insecticidal sprays in the agro-ecosystem have greatly affected the biodiversity and dynamics of primary and secondary insects. Various control measures were taken around the globe to increase production costs. Temperature, drought, and salinity, and biotic stresses such as bacteria, viruses, fungi, nematodes, insects, and mites cause substantial losses to cotton crops. Here, we summarize a number of biotic and abiotic stresses upsetting Bt cotton crop with present and future biotechnology solution strategies that include a refuge strategy, multi-gene pyramiding, the release of sterile insects, seed mixing, RNAi, CRISPR/Cas9, biotic signaling, and the use of bioagents. Surveillance of insect resistance, monitoring of grower compliance, and implementation of remedial actions can lead to the sustainable use of cotton across the globe.