Application Of Sterile Insect Technique Research Articles

Enterobacter sp. AA26 as a Protein Source in the Larval Diet of Drosophila suzukii.

Simple SummaryDrosophila suzukii has caused considerable damages to a variety of soft fruit crops. The sterile insect technique (SIT) is one of the most promising candidates that has recently attracted significant research efforts. A SIT package requires highly productive and cost-efficient mass rearing of the insects to produce a sufficient amount of males that will be sterilized and consequently released in the target area. Operational costs of mass-rearing facilities can be remarkably high, mainly due to the larval diet used for rearing. Gut symbiotic bacteria have been shown to enhance the productivity and development of fruit flies when used as supplements or protein source of their larval diet. In this study, we evaluated whether Enterobacter sp. AA26 could replace inactive brewer’s yeast as a protein source in D. suzukii larval diet and effects on the biological quality of the flies are discussed. The Spotted-Wing Drosophila fly, Drosophila suzukii, is an invasive pest species infesting major agricultural soft fruits. Drosophila suzukii management is currently based on insecticide applications that bear major concerns regarding their efficiency, safety and environmental sustainability. The sterile insect technique (SIT) is an efficient and friendly to the environment pest control method that has been suggested for the D. suzukii population control. Successful SIT applications require mass-rearing of the strain to produce competitive and of high biological quality males that will be sterilized and consequently released in the wild. Recent studies have suggested that insect gut symbionts can be used as a protein source for Ceratitis capitata larval diet and replace the expensive brewer’s yeast. In this study, we exploited Enterobacter sp. AA26 as partial and full replacement of inactive brewer’s yeast in the D. suzukii larval diet and assessed several fitness parameters. Enterobacter sp. AA26 dry biomass proved to be an inadequate nutritional source in the absence of brewer’s yeast and resulted in significant decrease in pupal weight, survival under food and water starvation, fecundity, and adult recovery.

Transport from production facility to release locations caused a decline in quality of sterile Queensland fruit fly received for SIT application

AbstractFruit flies are among the most destructive horticultural pests. For Queensland fruit fly (Qfly), Bactrocera tryoni (Froggatt) (Diptera: Tephritidae), the sterile insect technique (SIT) is the main control method used in southeastern Australia. SIT involves the mass rearing of Qfly, sterilisation of pupae by irradiation, transport to release centres, and release of sterile Qfly to disperse into the environment. Mating of sterile and wild flies results in no offspring; thus, the wild population rapidly decreases. Previously, research has mainly focused on improvements within the production facility. However, transport from production facility to release sites is also an essential component in all SIT programs. Here, we examined historical records from the Victorian SIT program between 2002 and 2013 in three seasons (spring, summer, and autumn). We compared quality control (QC) data of irradiated fly pupae at the mass‐rearing facility in New South Wales (limited transport) to those of pupae transported to a release centre in Victoria (extensive transport). After extensive transport, pupal weight (−0.3 mg, or −3%) and flight ability (an overall 26% decline) were lower across all seasons, but particularly in spring. Adult emergence was lower across all seasons (an overall 23% decline), but particularly in summer. All three QC parameters were more variable after extensive transport. We discuss potential causes of the observed decreased pupal quality and avenues of future research to mitigate transport losses.

Sex separation ofAedesspp. mosquitoes for sterile insect technique application: a review

AbstractSeparation of the sexes is necessary for the application of the sterile insect technique (SIT) in mosquitoes due to the hematophagous habits and disease vector activity of the females. In this review we analyze the history, current status, and future perspectives for the development of genetic sexing strains (GSS) ofAedesmosquitoes (Diptera: Culicidae). Various genetic control methods for mosquitoes are reviewed, as are their need for sex‐separation methods. We focus on areas of opportunity where GSS developed with classical genetic methods can be used. Regulatory restrictions and social acceptance of various control methods are analyzed. We conclude that the development of GSS by classical methods represents the most viable option for separation of the sexes and the application of large‐scale SIT programs within an area‐wide integrated vector management (AW‐IVM) approach.

The Effect of Radiation on the Gut Bacteriome of Aedes albopictus.

The sterile insect technique (SIT) has been developed as a component of area-wide integrated pest management approaches to control the populations of Aedes albopictus, a mosquito vector capable of transmission of dengue, Zika and chikungunya viruses. One of the key factors for the success of SIT is the requirement of high biological quality sterile males, which upon their release would be able to compete with wild males for matings with wild females in the field. In insects, gut bacteriome have played a catalytic role during evolution significantly affecting several aspects of their biology and ecology. Given the importance of gut-associated bacterial species for the overall ecological fitness and biological quality of their hosts, it is of interest to understand the effects of radiation on the gut-associated bacteriome of Ae. albopictus. In this study, the effect of radiation on the composition and density levels of the gut-associated bacterial species at the pupal stage as well as at 1- and 4-day-old males and females was studied using 16S rRNA gene-based next generation sequencing (NGS) and quantitative PCR (qPCR) approaches. Age, diet, sex, and radiation were shown to affect the gut-associated bacterial communities, with age having the highest impact triggering significant changes on bacterial diversity and clustering among pupae, 1- and 4-day-old adult samples. qPCR analysis revealed that the relative density levels of Aeromonas are higher in male samples compared to all other samples and that the irradiation triggers an increase in the density levels of both Aeromonas and Elizabethkingia in the mosquito gut at specific stages. Our results suggest that Aeromonas could potentially be used as probiotics to enhance protandry and sex separation in support of SIT applications against Ae. albopictus, while the functional role of Elizabethkingia in respect to oxidative stress and damage in irradiated mosquitoes needs further investigation.

A Novel Genetic Sexing Strain of Anastrepha ludens for Cost-Effective Sterile Insect Technique Applications: Improved Genetic Stability and Rearing Efficiency.

Simple SummaryTephritid flies, including the Mexican fruit fly Anastrepha ludens, are key agricultural pests responsible for billions of dollars of damage each year due to the female flies which lay eggs and develop maggots in the fruits of hundreds of species of economically-important plants. Integrated pest management practices such as the sterile insect technique were developed which rely on the mass rearing and release of millions of sterile males of the same pest species in order to suppress the pest reproductive capacity. The presence of females in early bisexual strains often caused high rearing and release costs plus damage to crops by oviposition. Therefore, using classical genetic tools, genetic sexing strains were developed for some fruit flies in order to produce male-only colonies which enhanced the technique efficacy, cost-efficiency and reduced undesirable side-effects such as fruit damage by the mass reared females. In this paper the process for the development and characterization of a new genetic sexing strain (GSS) for A. ludens is documented, using the low-dose irradiation and line selection techniques. The new GUA10 GSS, when compared with its predecessor Tapachula-7, promises to further increase the sterile insect technique performance given its higher quality, yield and genetic stability.Anastrepha ludens (Loew) is one of the most destructive insect pests damaging several fruits of economic importance. The sterile insect technique (SIT) is used under an area-wide integrated pest management approach, to suppress these pest populations. Mass rearing facilities were initially established to produce sterile males of bi-sexual strains in support of SIT. The first genetic sexing strain (GSS) for A. ludens, Tapachula-7, based on pupal color dimorphism, was a key development since the release of males-only significantly increases the SIT efficiency. In this study, we document the development of a novel pupal color-based GSS. Twelve radiation-induced translocation lines were assessed as potential GSS in terms of recombination rates and rearing efficiency at a small scale. The best one, GUA10, was cytogenetically characterized: it was shown to carry a single translocation between the Y chromosome and chromosome 2, which is known to carry the black pupae marker. This GSS was further evaluated at medium and large scales regarding its genetic stability, productivity and quality versus Tapachula-7. GUA10 presented better genetic stability, fecundity, fertility, production efficiency, flying ability, and male mating, clear indicators that GUA10 GSS can significantly improve the efficacy and cost-effectiveness of SIT applications against this pest species.

Pupation Substrate Type and Volume Affect Pupation, Quality Parameters and Production Costs of a Reproductive Colony of Ceratitis capitata (Diptera: Tephritidae) VIENNA 8 Genetic Sexing Strain.

Simple SummaryThe Mediterranean fruit fly, Ceratitis capitata, is a pest of fruit and vegetables in many parts of the world. The Sterile Insect Technique (SIT) is used as an environmentally friendly method for the control and/or eradication of this pest. A critical step in the production of sterile insects in biofactories is when the larvae complete their development and begin metamorphosis to become pupae. Artificial substrates are used to facilitate pupation, but little research has been devoted to this critical moment in the life cycle of the insect. We tested seven pupation substrates and five substrate volumes for a reproductive colony of C. capitata VIENNA 8 genetic sexing strain for SIT application in Mexico. One type of cellulose and sawdust at relatively low ratios of substrate:larvae enhanced pupation and were more cost-effective to produce one million pupae than other substrates including fine wheat bran currently used in a SIT mass-rearing facility in Mexico. Our study contributes to improving rearing processes involved in SIT-based pest control.Adequate pupation substrates and substrate volume are critical factors in the mass-rearing of insects for Sterile Insect Technique (SIT) applications. To identify an ideal pupation substrate for a reproductive colony of Ceratitis capitata (Wiedemann) VIENNA 8 genetic sexing strain, we first examined pupation in cellulose from recycled paper (cellulose I), sawdust, fine wheat bran, vermiculite and coconut fiber using a volume of 2.5–12.5 mL of substrate for each 5 mL volume of fly larvae. We found a positive relationship between substrate volume and pupation, with cellulose I generating the highest proportions of pupation and coconut fiber the lowest. Higher proportions of female flies (white pupae) pupated in sawdust. The proportion of female fliers increased as substrate volume rose in sawdust and coconut fiber, whereas it decreased in vermiculite and cellulose. In a second experiment, we tested three types of cellulose differing in physicochemical characteristics (celluloses I, II and III), sawdust, and fine wheat bran using a substrate:larvae ratio of 1:1. The three types of cellulose produced the highest pupation levels. The highest proportions of female fliers were observed in sawdust, and cellulose types III and II. Cellulose III and sawdust at relatively low volumes were more cost-effective to produce one million pupae than other substrates, including fine wheat bran used in a mass-rearing facility in Mexico.

Lessons from Drosophila: Engineering Genetic Sexing Strains with Temperature-Sensitive Lethality for Sterile Insect Technique Applications.

Simple SummaryThe sterile insect technique is a pest control strategy used to suppress or eliminate regional populations of insects that pose significant threats to agriculture or human health. The process involves mass-rearing, sterilization and release of male insects who fail to produce viable offspring when they mate with wild females, which leads to a population decline. Females are essential for colony propagation in rearing facilities and their selective removal prior to sterile releases remains an ongoing challenge. Developing genetic sexing strains with conditional temperature sensitive lethal mutations offers one strategy to eliminate female embryos through heat treatment, while males carry a wild type allele translocated to the Y-chromosome (or sex determination locus) to maintain their fitness. Here we review point mutations in Drosophila melanogaster genes that cause temperature sensitive phenotypes with the potential or ability to cause embryonic lethality. Re-engineering these known temperature sensitive mutations in other insects using CRISPR/Cas9 technology presents new opportunities to engineer genetic sexing strains for the sterile insect technique.A major obstacle of sterile insect technique (SIT) programs is the availability of robust sex-separation systems for conditional removal of females. Sterilized male-only releases improve SIT efficiency and cost-effectiveness for agricultural pests, whereas it is critical to remove female disease-vector pests prior to release as they maintain the capacity to transmit disease. Some of the most successful Genetic Sexing Strains (GSS) reared and released for SIT control were developed for Mediterranean fruit fly (Medfly), Ceratitis capitata, and carry a temperature sensitive lethal (tsl) mutation that eliminates female but not male embryos when heat treated. The Medfly tsl mutation was generated by random mutagenesis and the genetic mechanism causing this valuable heat sensitive phenotype remains unknown. Conditional temperature sensitive lethal mutations have also been developed using random mutagenesis in the insect model, Drosophila melanogaster, and were used for some of the founding genetic research published in the fields of neuro- and developmental biology. Here we review mutations in select D. melanogaster genes shibire, Notch, RNA polymerase II 215kDa, pale, transformer-2, Dsor1 and CK2α that cause temperature sensitive phenotypes. Precise introduction of orthologous point mutations in pest insect species with CRISPR/Cas9 genome editing technology holds potential to establish GSSs with embryonic lethality to improve and advance SIT pest control.

Genetic analysis of medfly populations in an area of sterile insect technique applications

The sterile insect technique (SIT) is widely used in integrated pest management programs for the control of the Mediterranean fruit fly (medfly), Ceratitis capitata. The genetic interactions between the released individuals from the genetic sexing strains (GSS), used for SIT applications worldwide, and wild individuals have not been studied. Under the hypothesis that a number of Vienna GSS individuals released to the field might not be completely sterile and may produce viable offspring, we have analyzed medfly Spanish field populations to evaluate the presence of Vienna strain genetic markers. To this goal, we have used contrasted nuclear and mitochondrial genetic markers, and two novel sets of nuclear polymorphisms with the potential to be markers to discriminate between Vienna and wild individuals. Nuclear Vienna markers located on the 5th chromosome of Vienna males have been found in 2.2% (19 from 875) of the Spanish wild medfly females captured at the area where SIT is applied. In addition, a female-inherited mitochondrial Vienna marker has been found in two from the 19 females showing nuclear Vienna markers. The detection of several of these markers in single individuals represents evidence of the introgression of Vienna strain into natural populations. However, alternative explanations as their presence at low frequency in wild populations in the studied areas cannot be fully discarded. The undesired release of non-fully sterile irradiated GSS individuals into the field and their interactions with wild flies, and the potential environmental implications should be taken into account in the application of the SIT.

Spatio-temporal distribution of sexual calling behaviour in domesticated, sterile and wild Queensland fruit fly males under field cage conditions.

The sterile insect technique (SIT) is used in Australia to contain and eradicate outbreaks of Bactrocera tryoni (Froggatt) (Diptera: Tephritidae) in fruit-fly-free regions, and to manage populations in some endemic regions. To assess the likely impact of SIT on wild B. tryoni populations it is important to assess the colocation and synchrony of male calling between sterile and wild flies. We observed the location and timing of calling behaviours of marked mass-reared (fertile and sterile) and wild B. tryoni males in walk-in field cages. We found that wild males called further from the canopy centre than mass-reared (fertile or sterile) males. Mass-reared (fertile or sterile) males called earlier in the evening than wild males and, consequently, mass-reared males called when temperature and light intensity were higher than when wild males called. Male calling is a prerequisite to mating among dacine fruit flies. Therefore, our observations of spatio-temporal divergence in male calling behaviour may lead to assortative mating between mass-reared and wild B. tryoni in SIT applications. The importance of these spatio-temporal differences warrants further inquiry, with particular focus on how environmental conditions modify calling behaviour and avenues to ameliorate differences between sterile and wild flies. © 2021 Society of Chemical Industry.

Manipulation of Gut Symbionts for Improving the Sterile Insect Technique: Quality Parameters of Bactrocera dorsalis (Diptera: Tephritidae) Genetic Sexing Strain Males After Feeding on Bacteria-Enriched Diets.

One environmentally friendly method used to manage Bactrocera dorsalis (Hendel), a key agricultural pest of substantial economic importance, is the sterile insect technique (SIT). Nevertheless, several deficiencies related to this strategy impair the success of the SIT, including the inferior performance of released sterile males compared with wild males, which could be partly solved by the utilization of gut symbionts as probiotic dietary components. In this study, a culture-dependent method was used to isolate and characterize gut-associated bacterial species in adult B. dorsalis genetic sexing strain (GSS) males. In addition, three bacterial isolates from the Enterobacteriaceae family, namely, Enterobacter sp., Morganella morganii, and Moellerella wisconsensis, were used as supplements in larval and adult diets to assess their effects on the life-history traits of irradiated males. Consistent with many previous studies, Enterobacter spp. was shown to be beneficial, with some quality control indices, such as adult size, pupal weight, survival rate under stress and nutritionally rich conditions, and mating competitiveness, being significantly increased, while slight nonsignificant increases in emergence rate and flight ability were observed. Conversely, the M. morganii and M. wisconsensis strains both had negative effects on irradiated male fitness and mating competitiveness. Our results, in combination with those of earlier studies, can contribute to improving the effectiveness of SIT application by enhancing the different aspects of augmentative rearing and biological traits of pests under laboratory rearing conditions.

Identification and characterization of four Drosophila suzukii cellularization genes and their promoters

BackgroundThe spotted-wing Drosophila (Drosophila suzukii) is a widespread invasive pest that causes severe economic damage to fruit crops. The early development of D. suzukii is similar to that of other Drosophilids, but the roles of individual genes must be confirmed experimentally. Cellularization genes coordinate the onset of cell division as soon as the invagination of membranes starts around the nuclei in the syncytial blastoderm. The promoters of these genes have been used in genetic pest-control systems to express transgenes that confer embryonic lethality. Such systems could be helpful in sterile insect technique applications to ensure that sterility (bi-sex embryonic lethality) or sexing (female-specific embryonic lethality) can be achieved during mass rearing. The activity of cellularization gene promoters during embryogenesis controls the timing and dose of the lethal gene product.ResultsHere, we report the isolation of the D. suzukii cellularization genes nullo, serendipity-α, bottleneck and slow-as-molasses from a laboratory strain. Conserved motifs were identified by comparing the encoded proteins with orthologs from other Drosophilids. Expression profiling confirmed that all four are zygotic genes that are strongly expressed at the early blastoderm stage. The 5′ flanking regions from these cellularization genes were isolated, incorporated into piggyBac vectors and compared in vitro for the promoter activities. The Dsnullo promoter showed the highest activity in the cell culture assays using D. melanogaster S2 cells.ConclusionsThe similarities in the gene coding and 5′ flanking sequence as well as in the expression pattern of the four cellularization genes between D. melanogaster and D. suzukii, suggest that conserved functions may be involved in both species. The high expression level at the early blastoderm stage of the four cellularization genes were confirmed, thus their promoters can be considered in embryonic lethality systems. While the Dsnullo promoter could be a suitable candidate, all reported promoters here are subject to further in vivo analyses before constructing potential pest control systems.

Development and characterization of a pupal-colour based genetic sexing strain of Anastrepha fraterculus sp. 1 (Diptera: Tephritidae)

BackgroundArea-wide integrated pest management programs (AW-IPM) incorporating sterile insect technique (SIT) have been successful in suppressing populations of different fruit fly species during the last six decades. In addition, the development of genetic sexing strains (GSS) for different fruit fly species has allowed for sterile male-only releases and has significantly improved the efficacy and cost effectiveness of the SIT applications. The South American Fruit Fly Anastrepha fraterculus (Diptera: Tephritidae) is a major agricultural pest attacking several fruit commodities. This impedes international trade and has a significant negative impact on the local economies. Given the importance of sterile male-only releases, the development of a GSS for A. fraterculus would facilitate the implementation of an efficient and cost-effective SIT operational program against this insect pest species.ResultsFor potential use in a GSS, three new morphological markers (mutants) were isolated in a laboratory strain of A. fraterculus sp. 1, including the black pupae (bp) gene located on chromosome VI. The black pupa phenotype was used as a selectable marker to develop genetic sexing strains by linking the wild type allele (bp+) to the Y-chromosome -via irradiation to induce a reciprocal Y-autosome translocation. Four GSS were established and one of them, namely GSS-89, showed the best genetic stability and the highest fertility. This strain was selected for further characterization and cytogenetic analysis.ConclusionsWe herein report the development of the first genetic sexing strain of a major agricultural pest, A. fraterculus sp. 1, using as a selectable marker the black pupae genetic locus.

Using Moderate Transgene Expression to Improve the Genetic Sexing System of the Australian Sheep Blow Fly Lucilia cuprina.

Simple SummaryPopulations of pest insects can be suppressed through repeated mass releases of sterilized insects. This is particularly effective if only sterile males are released. We previously developed several genetically modified strains of the Australian sheep blowfly that produce only males when raised on diet that lacked tetracycline. A disadvantage of the some of the engineered strains was that females would lay few eggs unless fed a diet with a low dose of tetracycline. In this study we show that effective male-only strains can be made by combining driver/effector lines that have moderate transgene expression/activity. Furthermore, the strain does not require tetracycline in the adult diet for female fertility. This “moderate expression/activity” strategy could be more generally applied to other pests that can be genetically modified.The sterile insect technique (SIT) is a promising strategy to control the Australian sheep blow fly Lucilia cuprina, a major pest of sheep. We have previously developed a transgenic embryonic sexing system (TESS) for this pest to facilitate the potential SIT application. TESS carry two transgenes, a tetracycline transactivator (tTA) driver and a tTA-activated pro-apoptotic effector. TESS females die at the embryonic stage unless tetracycline is supplied in the diet. However, undesired female sterility was observed in some TESS strains without tetracycline due to expression of tTA in ovaries. Here we investigate if TESS that combine transgenes with relatively low/moderate expression/activity improves the fertility of TESS females. tTA driver lines were evaluated for tTA expression by quantitative real time PCR and/or by crossing with a tTA-activated RFPex effector line. Fertility and lethality tests showed that a TESS strain containing a driver line with moderate tTA expression and an effector line showing moderate pro-apoptotic activity could recover the fertility of parental females and eliminated all female offspring at the embryonic stage. Consequently, such a strain could be further evaluated for an SIT program for L. cuprina, and such a “moderate strategy” could be considered for the TESS development in other pest species.

Improving the Phenotypic Properties of the Ceratitis capitata (Diptera: Tephritidae) Temperature-Sensitive Lethal Genetic Sexing Strain in Support of Sterile Insect Technique Applications.

The genetic sexing strain (GSS) of the Mediterranean fruit fly (Ceratitis capitata (Wiedemann)) Vienna 8D53− is based on a male-linked translocation system and uses two selectable markers for male-only production, the white pupae (wp) and the temperature sensitivity lethal (tsl) genes. In this GSS, males emerge from brown pupae and are resistant to high temperatures while females emerge from white pupae, are sensitive to high temperatures. However, double homozygous females (wp tsl/wp tsl) exhibit a slower development rate compared to heterozygous males (wp+tsl+/wp tsl) during the larval stage, which was attributed to the pleiotropic effects of the tsl gene. We present the first evidence that this slower development is due to a different gene, here namely slow development (sd), which is closely linked to the tsl gene. Taking advantage of recombination phenomena between the two loci, we report the isolation of a novel temperature sensitivity lethal strain using the wp mutation as a morphological marker, which showed faster development (wp tsl FD) during the larval stage and increased in its temperature sensitivity compared with the normal tsl strain. Moreover, the introgression of this novel wp tsl FD combined trait into the Vienna 8D53− GSS, resulted in a novel Vienna 8D53− FD GSS, where females showed differences in the thermal sensibility, larval development speed, and productivity profiles. The modification of these traits and their impact on the mass rearing of the GSS for sterile insect technique applications are discussed.

Egg Sterilisation of Irradiated Nezara viridula (Hemiptera: Pentatomidae).

Simple SummaryCertain stink bugs are emerging as a serious threat to food production globally, as they invade new areas and feed on a wide range of crops. The control of these pests relies primarily on potentially harmful pesticides, but the increasing threat posed by stink bug pests has sparked investigation into environmentally-friendly methods that do not exert serious impacts on other species. One method receiving attention is the Sterile Insect Technique (SIT), which involves sterilising large numbers of a pest, through radiation exposure, and releasing them into the wild pest population where mating results in unfertilised eggs that do not hatch. In support of recent studies on SIT for the brown marmorated stink bug, we aimed to ascertain the feasibility of the method for another stink bug pest, the green vegetable bug. We exposed the insects to increasing levels of radiation and allowed mating to occur. Virtually all of the resulting eggs were sterile and did not hatch at the higher radiation doses tested. These results could be used to inform the potential development of SIT against stink bug pests, and in some circumstances, could form the basis of potential eradication programmes against new invasions.Nezara viridula Linnaeus (Hemiptera: Pentatomidae) is a polyphagous pest of a wide range of economically important crops. Because the control of this species and other pentatomids relies primarily on insecticide application, investigation into the Sterile Insect Technique (SIT) is warranted. We aimed to investigate the irradiation biology of N. viridula for the potential application of SIT against this pest. Male and female N. viridula were gamma-irradiated at doses between 4 and 28 Gy and mated with both irradiated and nonirradiated conspecifics. Sterility of the resulting eggs followed a dose-response in each case. Irradiated males crossed with untreated females showed higher F1 egg sterility than crosses where the female was irradiated. The greatest F1 egg sterility was observed when both parents were irradiated. There was no obvious dose-response for the longevity of irradiated males, and for the fecundity of nonirradiated females mated with irradiated males. The fecundity of irradiated females appeared to decrease with irradiation dose. These results can be applied to a potential future application of SIT against N. viridula, but predominantly supports the ongoing development of SIT for Halyomorpha halys Stål (Hemiptera: Pentatomidae) and hemipteran pests in general.

Manipulation of insect gut microbiota towards the improvement of Bactrocera oleae artificial rearing

AbstractBactrocera oleae (Rossi) (Diptera: Tephritidae) is the main pest of olive trees (Olea europaea L.), causing major damages in olive crops. Improvement of mass rearing is a prerequisite for the successful development of large‐scale sterile insect technique (SIT) applications. This can be achieved through the enrichment of artificial diets with gut bacteria isolates. We assessed the efficiency of three gut bacteria previously isolated from Ceratitis capitata (Wiedemann), and four isolated from B. oleae, as larval diet additives in both live and inactivated/dead forms. Our results showed that dead Enterobacter sp. AA26 increased pupal weight, whereas both live and dead cells increased pupal and adult production and reduced immature developmental time, indicating that its bacterial cells serve as a direct nutrient source. Live Providencia sp. AA31 improved pupal and adult production, enhanced male survival under stress conditions, and delayed immature development. Dead Providencia sp. AA31, however, did not affect production rates, indicating that live bacteria can colonize the insect gut and biosynthesize nutrients essential for larval development. Live and dead Bacillus sp. 139 increased pupal weight, accelerated immature development, and increased adult survival under stress. Moreover, live Bacillus sp. 139 improved adult production, indicating that Bacillus cells are a direct source of nutrients. Dead Serratia sp. 49 increased pupal and adult production and decreased male survival under stress conditions whereas live cells decreased insect production, indicating that the live strain is entomopathogenic, but its dead cells can be utilized as nutrient source. Klebsiella oxytoca, Enterobacter sp. 23, and Providencia sp. 22 decreased pupal and subsequent adult production and were harmful for B. oleae. Our findings indicate that dead Enterobacter sp. AA26 is the most promising bacterial isolate for the improvement of B. oleae mass rearing in support of future SIT or related population suppression programs.

RNA Interference Is Enhanced by Knockdown of double-stranded RNases in the Yellow Fever Mosquito Aedes Aegypti.

RNA interference (RNAi) techniques are being developed for a range of pest insect control technologies, including the sterile insect technique (SIT) and double-stranded RNA (dsRNA)-based insecticides. In SIT applications, where >99% of the released males should be sterile to meet industry standards, the efficiency of RNAi will need to be improved for many insect species if this technology is to be adopted. Endogenous dsRNases can impede dsRNA delivery in some insects, and, here, we investigated whether dsRNases in the midgut could limit RNAi efficacy in the mosquito Aedes aegypti. Ten putative dsRNases were identified in the Ae. aegypti genome, with two highly expressed in the midguts of larvae. Using an ex vivo assay, we observed that dsRNA was rapidly degraded within the mosquito larva’s gut. Double-stranded RNA targeting these two dsRNases, when fed to the larvae, effectively reduced gut dsRNase activity. When these dsRNase-specific dsRNAs were co-delivered with dsRNA targeting a cyan fluorescent protein (CFP) reporter gene, greater knockdown of CFP fluorescence was observed. These results suggest that inhibiting dsRNase activity could enable the implementation of RNAi-based mosquito control methods.

Multiple Factors Determine the Structure of Bacterial Communities Associated With Aedes albopictus Under Artificial Rearing Conditions.

Insect symbionts are major manipulators of host’s behavior. Their effect on parameters such as fecundity, male mating competitiveness, and biological quality in general, can have a major influence on the effectiveness of the sterile insect technique (SIT). SIT is currently being developed and applied against human disease vectors, including Ae. albopictus, as an environment-friendly method of population suppression, therefore there is a renewed interest on both the characterization of gut microbiota and their exploitation in artificial rearing. In the present study, bacterial communities of eggs, larvae, and adults (both males and females) of artificially reared Ae. albopictus, were characterized using both culture-dependent and culture-independent approaches. Mosquito-associated bacteria corresponding to thirteen and five bacteria genera were isolated from the larval food and the sugar solution (adult food), respectively. The symbiont community of the females was affected by the provision of a blood meal. Pseudomonas and Enterobacter were either introduced or enhanced with the blood meal, whereas Serratia were relatively stable during the adult stage of females. Maintenance of these taxa in female guts is probably related with blood digestion. Gut-associated microbiota of males and females were different, starting early after emergence and continuing in older stages. Our results indicate that eggs contained bacteria from more than fifteen genera including Bacillus, Chryseobacterium, and Escherichia–Shigella, which were also main components of gut microbiota of female adults before and after blood feeding, indicating potential transmission among generations. Our results provided a thorough study of the egg- and gut-associated bacteria of artificially reared Ae. albopictus, which can be important for further studies using probiotic bacteria to improve the effectiveness of mosquito artificial rearing and SIT applications.

Efeito do cobalto-60 em fêmeas R.(B.) microplus na postura e na incubação de ovos

This study aimed to evaluate the effects of Co60 on the posture of engorged females of tick R. (B) microplus in the pre-laying phase and on the hatchability of their eggs when submitted to doses from 0 to 400 Gy where it was found that ionizing radiation affected approximately 70% the egg laying weight of these females, that the egg hatching rate of the Witness was 68.2% higher than the lowest ionizing dose, the 5 Gy Treatment, that the higher the radiation dose, the lower the index. of hatching and treatments that received doses higher than 20 Gy, there were no hatching of the postures, thus establishing a milestone to study the application of sterile insect technique in this specimen.

Aedes aegyptilines for combined sterile insect technique and incompatible insect technique applications: the importance of host genomic background

AbstractAedes aegyptiL. (Diptera: Culicidae), being the primary vector of pathogenic arboviruses, is a target for the development of novel genetic approaches to complement current conventional vector control strategies such as the combined sterile insect and incompatible insect technique (SIT/IIT). A transinfected line ofAe. aegypticarrying thewAlbBWolbachiastrain (WB2) was introgressed into two genomic backgrounds, Brazil and Mexico, producing two newAe. aegyptistrains (WB2‐BRA and WB2‐MEX). These strains were evaluated with respect to several life‐history traits such as fecundity, fertility, longevity, pupa size, pupation curve, and male mating competitiveness, as well as their response to irradiation. Our results show that the impact ofWolbachiainfection depends on the genomic background and that the Brazilian one had no significant effect, whereas the Mexican one negatively affected fertility, longevity, and pupal size. Interestingly,Wolbachia‐infectedAe. aegyptilines required a lower irradiation dose to achieve complete female sterility than the uninfected ones. The present findings are discussed given the potential use ofWolbachia‐infectedAe. aegyptilines in combined SIT/IIT population suppression programs.