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Comet assay analysis of multigenerational genomic instability (F0-F2) in Aedes aegypti exposed to gamma radiation in Sterile Insect Technique.

The use of irradiation in the Sterile Insect Technique (SIT) is a sustainable and environmentally friendly strategy for controlling Aedes aegypti populations by the release of sterile males. However, the potential toxic effects of radiation on mosquito genetic material, as well as the heritability of such damage, remain insufficiently understood. In this study, we evaluated gamma radiation-induced DNA damage (20, 30, 40, and 50 Gy) in male pupae (F0 generation) and assessed the persistence of these effects in subsequent generations (F1 and F2) using the comet assay in hemocytes. In the parental generation, a significant dose-response relationship was observed, with increasing radiation doses associated with higher damage index and damage frequency (p < 0.05). In the F1 generation, both larvae and adults exhibited significantly greater DNA damage than the control group, particularly at doses of 30 and 40 Gy, supporting the inheritance of radiation-induced genomic instability. In the F2 generation, genotoxic effects were attenuated, although residual damage remained detectable in adults, suggesting partial recovery of genomic stability, possibly influenced by DNA repair mechanisms and/or selective pressures. No viable offspring were obtained at 50 Gy, confirming the sterilizing efficacy of higher doses. Integration of comet assay results with micronucleus data and reproductive parameters reinforces the association between DNA damage, mutagenic effects, and reduced fertility. These findings indicate that radiation-induced genotoxic effects may persist beyond the irradiated generation but tend to decline across generations. Overall, this study provides insights into the balance between achieving sterility and preserving biological quality in SIT programs, contributing to optimizing radiation doses and enhancing the safety and efficacy of vector control strategies.

Comet assay

Control and management of insect populations by chemosterilants.

Chemosterilants, i.e., chemical compounds that interfere with the reproduction potential of sexually reproducing organisms, can be used in three new approaches to insect control. In the sterile-insect release technique, the principal problem is to develop compounds and methods for their application that would not result in introducing harmful residues into the environment. Because of the unusual and often unique circumstances connected with releasing large numbers of sterilized insects, the residue problem and its cost-benefit aspects must be examined individually for each intended control or eradication program. In the direct application technique, chemosterilants must meet the same efficiency and safety standards required from approved insecticides. Combined insecticidal and sterilizing activity is characteristic for some compounds now being investigated. In the genetic technique, chemosterilants may be used for inducing heritable changes in the insect's genome under laboratory conditions, and such procedures would not present any residue problems. Only the first two chemosterilant techniques are approaching practical application, and their safety aspects require detailed evaluation and assessment.

Chemosterilants

Abiotic conditions can modify the penetrance of transgene-based lethality systems for insect population control.

Modern genetic biocontrol techniques for insect pest management, when compared to chemical insecticide spraying, offer high species specificity and reduced environmental impact, and some of these methods require the environmental release of genetically modified (GM) insects. Because organisms exposed to different environments often show variability in phenotype and gene expression, it is likely that GM insects will also experience environmentally mediated variation, potentially compromising pest control efficiency. This study examines the impact of temperature and nutrition on the early embryonic Tet-off conditional lethality system in Drosophila melanogaster. By independently manipulating parental and offspring environments, we assessed how exposure to variable environments influenced the probability of larval hatching and the transcript abundance of the transgenic system. Our findings revealed that: (i) transgene performance distinctly responds to temperature and nutrition; (ii) thermal stress has a greater impact when embryos, rather than parents, are exposed; and (iii) extreme nutritional conditions can markedly reduce the penetrance of transgenic lethality. Although changes in transgene transcript abundance were observed across environments, these changes did not fully explain the phenotypic variation, suggesting that factors downstream of transcription probably drive variation in transgenic lethality.

Animals

Metabolism of radiolabeled insecticides in insects and related arthropods: a critical study of various techniques.

The metabolism of radiolabeled insecticides in insects and acarina is studied largely by coupling radiotracer techniques with analytical methods, such as TLC, paper and column chromatography, gel-permeation chromatography, and enzymatic assays. These techniques in various combinations yield both the identification and quantification of the metabolites. Other analytical methods such as gas chromatography or IR spectrometry may also be used to obtain additional support for identification of metabolites. In the absence of authentic chromatographic standards, however, NMR and mass spectrometry are necessary in the identification of the unknown compound. The quantity of the radiolabeled insecticide administered should be within the toxicological range of the insect. Therefore, the dosage-mortality response of the insect using unlabeled material should be determined. A dose should be selected that keeps insect mortality to a minimum in order to avoid complications in the computation of the balance data. The radiolabeled insecticide is usually applied topically to the insect. Alternately, the material may be administered by dipping in a solution containing the radiolabeled compound or by exposure to filter paper impregnated with radiolabeled material. Administration of the radiolabeled material by the oral route presents several problems. Sterile rearing conditions are mandatory to avoid contamination of treated diet with microorganisms. Some knowledge of the insect's feeding rhythm is desirable so that the labeled diet is given at peak feeding time. Synthetic diets should be adjusted to pH 7.0. These precautions minimize degradation of the insecticide in the diet prior to consumption by the insect. Precise doses of radiolabeled materials may be administered by injection. The technique is mainly useful in metabolism studies of intermediate materials resulting from the biotransformation of the parent compound.

Animals

Spiroplasma associated with flowers of the tulip tree (Liriodendron tulipifera L.).

Spiroplasmas were isolated and cultivated from nonsurface-sterilized petals and bracts excised from flowers of Liriodendron tulipifera L. (tulip tree) in Anne Arundel, Charles, and Prince George's Counties in Maryland, and East Lyme County, Connecticut. All isolates grew at 30 and at 37 degrees C. Morphology of colonies on solid agar (1%) medium containing serum differed among isolates: some isolates formed highly diffuse, barely visible colonies; others formed distinct colonies with granular centers surrounded by minute surface or submerged 'satellite' colonies. Cellular morphology and motility of organisms in broth and in agar culture were typical of the spiroplasmas pathogenic in plants and insects. In viscous media containing methylcellulose, spiroplasmas freely suspended in the medium swam, and organisms in contact with glass slide or coverslip "crawled" across the solid surface. Frowth inhibition, metabolic inhibition, and organism deformation tests failed to reveal a serologic relationship between spiroplasma strain 23-6 from tulip tree flowers and spiroplasma strain AS 576 from honey bee.

Agar

[The queen technic for ant control. I. The effect of tepa on laboratory colonies of the pharaoh ant].

The present paper discussed at the example of Tepa [Tris (1-aziridinyl) phosphine oxide] in which manner the control of pharaoh's ant in the sense of the "queen technique" with chemosterilants is possible; which points of view must be considered and which condition an acceptable substance should be fulfilled. The application of Tepa is ralized according to various techniques (e.g. dipping, baiting, tarsal contact), in the course of which the baiting technique is preferred. The influence of different concentrations in various baits on the brood, workers and queens are described. In baits with 1% of the substance a permanent sterilization could be achieved. In combination with other population depressing factors (larval and worker mortality) resulted in eradication of the colonies. The action extended also to the larvae of the sexuals, so that the queenless colonies failed in the production of new queens and males. Histological investigations showed in the females at higher concentrations distinct pathological alterations (pycnosis, vacuolizations, proliferation in the follicles epithelium); whereas the spermatogenesis in the males is decreased but not entirely suppressed. At lower concentrations only the fertility but not the fecundity was influenced. A handing-over of recessive lethal mutations to the progeny does not take place. The practical points of view for the use of chemosterilants in the control of pharaoh's ant are discussed. Whereas the low biological stability of Tepa does not exclude an introduction in the practice, the high mutagenic activity prevents an application in the field.

Animals