Spatial resolution and optical sensitivity in the compound eyes of two common European wasps, Vespula germanica and Vespula vulgaris.

Vespula germanica and Vespula vulgaris are two common European wasps that have ecological and economic importance due to their artificial introduction to many different countries and environments. Their success has undoubtedly been aided by their capacity for visually guided hunting, foraging, learning, and using visual cues in the context of homing and navigation. However, the visual systems of V. germanica and V. vulgaris have not received any deep attention. We used electrophysiology, together with optical and anatomical techniques, to measure the spatial resolution and optical sensitivity of the compound eyes of both species. We found that both wasps have high anatomical spatial resolution with narrow interommatidial angles (Δφ between 1.0 and 1.5°) and a distinct acute zone in the fronto-ventral part of the eye. These narrow interommatidial angles are matched to photoreceptors having narrow angular sensitivities (acute zone acceptance angles Δρ below 1.3°), indicating eyes of high spatial resolution that are well suited to their ecological needs. Additionally, we found that both species possess an optical sensitivity that is typical of other day-flying hymenopterans.

Wax Protrusions on Anti-Adhesive Plant Surfaces and Their Interactions with Insect Adhesive Pads: A Mechanical Interpretation.

Insect attachment devices enhance adhesion to complex-geometry substrates by increasing the real contact area. In nature, insects mainly interact with plant surfaces that are often covered by 3D wax structures. Here, we describe, discuss, and give a mechanical interpretation of plant waxes and the possible fracture mechanisms of these wax structures during their interactions with the adhesive pads of insects. It is argued that these plant surface microstructures significantly influence insect adhesion through reducing the contact area and contaminating the insect pads.

Mark-Release-Recapture (MRR) of Sterile Male Aedes albopictus (Skuse) in Sri Lanka: Field Performance of Sterile Males and Estimation of the Wild Mosquito Population Density.

Dengue is an important mosquito-borne disease in Sri Lanka. The Sterile Insect Technique (SIT) is an environment-friendly and novel method that can suppress dengue vector mosquitoes in Sri Lanka. This study aimed to evaluate the field performance of sterile males and the density of wild male Aedes albopictus (Skuse) using a Mark-Release-Recapture (MRR) assay. Laboratory-colonized male pupae were exposed to 50 Gy gamma using a Co60 source. Sterile males (approx. 10,000) marked with fluorescent dust were released weekly for 4 consecutive weeks (January-February 2021) in a geographically isolated 30 ha site in Gampaha. Results show sterile males could disperse up to 543.8 m with a mean distance of 255.1 ± 44.6 m and survive up to 6 days with a mean life expectancy of 3.55 ± 2.32 days. A high field mating competitiveness of sterile males based on a Fried value of 0.47 ± 0.007 and significant induced sterility in the wild eggs in the second generation were found. The mean wild male mosquito population density was 163 males/ha. The data generated will be useful for designing future trials in Sri Lanka and other countries with similar situations.

Ultrastructure of the Spermiogenesis in Halyomorpha halys (Hemiptera: Pentatomidae): X-Irradiation and New Insights on the Centriolar Region Organization.

Halyomorpha halys (Heteroptera: Pentatomidae) is an insect pest native to Asia that has spread over the last two decades to most of the North America, parts of South America, Europe and North Africa. Its impact is significant as it can feed on more than 300 host plants, rendering affected fruits and vegetable crops unsellable or of lower quality. Various chemical and biological methods have been used to control this pest, with varying degrees of success. The sterile insect technique (SIT) is a pest control method involving the sterilization of insects via ionizing radiation and their subsequent mass release into the field. In the present contribution, the spermiogenesis of H. halys was studied from an ultrastructural point of view in both irradiated and non-irradiated adult males. In both cases, we observed ultrastructural characteristics typical of hemipteran sperm cells: bridges connecting the mitochondrial derivatives and the axonemal microtubules, the absence of accessory bodies, and the presence of two or three crystalline inclusions within the mitochondrial derivatives, an acrosome composed of tightly packed tubules, and an atypical, plaque-shaped microtubular organizing center (MTOC) in the centriolar region. Moreover, in the same region, we seldom observed the presence of two centrioles in the spermatids, one of which disappeared at a later stage of maturation. This feature is a novelty for insect spermiogenesis. The cysts of irradiated adults were not all uniformly affected by the radiation. However, irradiated cysts sometimes exhibited a general disorganization of sperm arrangement, incomplete divisions of sperm cells resulting in multiple copies of the same organelle within the same cell, failure to reabsorb the cytoplasm, and the lack of axonemes. Finally, rod-shaped viruses or virus-like particles were observed in vasa deferentia independently of irradiation.

The Use of Insect Pigment in Art Works.

In this compilation, the focus is on the Cochineal insect (Dactylopius coccus Costa, 1835 (Hemiptera: Dactylopiidae)), a creature native to South America that produces a potent natural red pigment known as "carmine". This pigment, utilized for obtaining the color red, has been an integral part of the art world for thousands of years. Indigenous cultures, in particular, have employed the dye extracted from this insect in the creation of textile dyes and paintings. Moreover, the Cochineal insect and its unique pigments have not only supported artistic expression but also captivated and inspired artists. During the Renaissance period, artists preferred the carmine pigment produced by the females of the Cochineal insect for obtaining bright and vivid red tones. This study delves into the history of the Cochineal insect, its role in art, and its perception in the modern world. Famous paintings created with dyes obtained from the Cochineal insect are discussed, exploring how pigments have found a place in the art world and how artists have utilized this extraordinary source to create distinctive works.

Ultrastructure and Spectral Characteristics of the Compound Eye of Asiophrida xanthospilota (Baly, 1881) (Coleoptera, Chrysomelidae).

In this study, the morphology and ultrastructure of the compound eye of Asi. xanthospilota were examined by using scanning electron microscopy (SEM), transmission electron microscopy (TEM), micro-computed tomography (µCT), and 3D reconstruction. Spectral sensitivity was investigated by electroretinogram (ERG) tests and phototropism experiments. The compound eye of Asi. xanthospilota is of the apposition type, consisting of 611.00 ± 17.53 ommatidia in males and 634.8 0 ± 24.73 ommatidia in females. Each ommatidium is composed of a subplano-convex cornea, an acone consisting of four cone cells, eight retinular cells along with the rhabdom, two primary pigment cells, and about 23 secondary pigment cells. The open type of rhabdom in Asi. xanthospilota consists of six peripheral rhabdomeres contributed by the six peripheral retinular cells (R1~R6) and two distally attached rhabdomeric segments generated solely by R7, while R8 do not contribute to the rhabdom. The orientation of microvilli indicates that Asi. xanthospilota is unlikely to be a polarization-sensitive species. ERG testing showed that both males and females reacted to stimuli from red, yellow, green, blue, and ultraviolet light. Both males and females exhibited strong responses to blue and green light but weak responses to red light. The phototropism experiments showed that both males and females exhibited positive phototaxis to all five lights, with blue light significantly stronger than the others.

The Food Source and Gut Bacteria Show Effects on the Invasion of Alien Pests-A Case of Bactrocera dorsalis (Hendel) (Diptera: Tephritidae).

How alien pests invade new areas has always been a hot topic in invasion biology. The spread of the Bactrocera dorsalis from southern to northern China involved changes in food sources. In this paper, in controlled conditions, we take Bactrocera dorsalis as an example to study how plant host transformation affects gut bacteria by feeding it its favorite host oranges in the south, its favorite host peaches and apples in the north, and feeding it cucumbers as a non-favorite host plant, thereby further affecting their fitness during invasion. The result showed that, after three generations of feeding on cucumbers, Bactrocera dorsalis took longer to develop as a larva while its longevity and fecundity decreased and pre-adult mortality increased. Feeding it cucumbers significantly reduced the overall diversity of gut microbiota of Bactrocera dorsalis. The relative abundance of Enterobacter necessary for survival decreased, while the Empedobacter and Enterococcus increased, resulting in decreased carbohydrate transport and metabolism and increased lipid transport and metabolism. Feeding Bactrocera dorsalis Empedobacter brevis and Enterococcus faecalis resulted in a 26% increase in pre-adult mortality and a 2-3 d increase in adult preoviposition period (APOP). Additionally, Enterococcus faecalis decreased the longevity of female and male adults by 17 and 12 d, respectively, and decreased fecundity by 11%. We inferred that the shifted plant hosts played an important role in posing serious harm to Bactrocera dorsalis invading from the south to the north. Therefore, after an invasion of Bactrocera dorsalis into northern China, it is difficult to colonize cucumbers for a long time, but there is still a risk of short-term harm. The findings of this study have established that the interactions between an insect's food source and gut bacteria may have an important effect on insect invasions.

Two-Sex Life Table Analysis for Optimizing Beauveria bassiana Application against Spodoptera exigua (Hübner) (Lepidoptera: Noctuidae).

Spodoptera exigua (Hübner) (Lepidoptera: Noctuidae) is a highly dispersive, polyphagous insect pest that severely defoliates crops. Excessive reliance on synthetic insecticides leads to ecological pollution and resistance development, urging scientists to probe eco-friendly biopesticides. Here, we explore the virulence of an entomopathogenic fungus, Beauveria bassiana, against S. exigua, resulting in 88% larval mortality. Using an age-stage, two-sex life table, we evaluated the lethal and sublethal effects of B. bassiana on the demographic parameters of S. exigua, including survival, development, and reproduction. Sublethal (LC20) and lethal concentrations (LC50) of B. bassiana impacted the parental generation (F0), with these effects further influencing the demographic parameters of the first filial generation (F1). The infected F1 offsprings showed a reduced intrinsic rate of increase (r), mean generation time (T), and net reproduction rate (R0). Larval developmental duration varied significantly between the control (10.98 d) and treated groups (LC20: 10.42; LC50: 9.37 d). Adults in the treated groups had significantly reduced lifespans (M: 8.22; F: 7.32 d) than the control (M: 10.00; F: 8.22 d). Reduced fecundity was observed in the B. bassiana-infected groups (LC20: 313.45; LC50: 223.92 eggs/female) compared to the control (359.55 eggs/female). A biochemical assay revealed elevated levels of detoxification enzymes (esterases, glutathione S-transferases, and acetylcholinesterase) in the F0 generation after B. bassiana infection. However, the enzymatic activity remained non-significant in the F1 generation likely due to the lack of direct fungal exposure. Our findings highlight the enduring effects of B. bassiana on the biological parameters and population dynamics of S. exigua, stressing its use in eco-friendly management programs.

Properties of Chitin and Its Regenerated Hydrogels from the Insect Zophobas morio Fed Citrus Biomass or Polystyrene.

The potential of insects as a recycling tool has recently attracted attention. In this study, chitin was extracted with 1 M HCl for 24 h at 20 °C, followed by 1 M NaOH for 5 h at 90 °C, and bleached with 2.5% v/v NaOCl for 2 h at 20 °C from Zophobas morio (ZM) insects fed citrus waste biomass (OP) or polystyrene foam (PS). The highest survival rate was found in the OP group. The properties of the resulting chitin material are reported, as well as the preparation of hydrogels using a DMAc/LiCl solvent. All chitins obtained were α-chitin. The degrees of deacetylation, crystallinity, molecular weight, and solubility in DMAc/LiCl were similar between the PS and biomass feeds, and they showed similar viscosities in the DMAc/LiCl solution. All hydrogels obtained had similar properties and viscoelastic behavior, indicating that the resultant chitins and their hydrogels from ZM were similar between those fed with citrus biomass and those fed with PS.

Water Stress and Black Cutworm Feeding Modulate Plant Response in Maize Colonized by Metarhizium robertsii.

Plants face many environmental challenges and have evolved different strategies to defend against stress. One strategy is the establishment of mutualistic associations with endophytic microorganisms which contribute to plant defense and promote plant growth. The fungal entomopathogen Metarhizium robertsii is also an endophyte that can provide plant-protective and growth-promoting benefits to the host plant. We conducted a greenhouse experiment in which we imposed stress from deficit and excess soil moisture and feeding by larval black cutworm (BCW), Agrotis ipsilon, to maize plants that were either inoculated or not inoculated with M. robertsii (Mr). We evaluated plant growth and defense indicators to determine the effects of the interaction between Mr, maize, BCW feeding, and water stress. There was a significant effect of water treatment, but no effect of Mr treatment, on plant chlorophyl, height, and dry biomass. There was no effect of water or Mr treatment on damage caused by BCW feeding. There was a significant effect of water treatment, but not Mr treatment, on the expression of bx7 and rip2 genes and on foliar content of abscisic acid (ABA), 2,4-dihydroxy-7-methoxy-1,4-benzoxazin-3-one (DIMBOA), and gibberellin 19 (GA19), whereas GA53 was modulated by Mr treatment. Foliar content of GA19 and cis-Zeatin (cZ) was modulated by BCW feeding. In a redundancy analysis, plant phenology, plant nutrient content, and foliar DIMBOA and ABA content were most closely associated with water treatments. This study contributes toward understanding the sophisticated stress response signaling and endophytic mutualisms in crops.

OxyR is required for oxidative stress resistance of the entomopathogenic bacterium Xenorhabdus nematophila and has a minor role during the bacterial interaction with its hosts.

Xenorhabdus nematophila is a Gram-negative bacterium, mutualistically associated with the soil nematode Steinernema carpocapsae, and this nemato-bacterial complex is parasitic for a broad spectrum of insects. The transcriptional regulator OxyR is widely conserved in bacteria and activates the transcription of a set of genes that influence cellular defence against oxidative stress. It is also involved in the virulence of several bacterial pathogens. The aim of this study was to identify the X. nematophila OxyR regulon and investigate its role in the bacterial life cycle. An oxyR mutant was constructed in X. nematophila and phenotypically characterized in vitro and in vivo after reassociation with its nematode partner. OxyR plays a major role during the X. nematophila resistance to oxidative stress in vitro. Transcriptome analysis allowed the identification of 59 genes differentially regulated in the oxyR mutant compared to the parental strain. In vivo, the oxyR mutant was able to reassociate with the nematode as efficiently as the control strain. These nemato-bacterial complexes harbouring the oxyR mutant symbiont were able to rapidly kill the insect larvae in less than 48 h after infestation, suggesting that factors other than OxyR could also allow X. nematophila to cope with oxidative stress encountered during this phase of infection in insect. The significantly increased number of offspring of the nemato-bacterial complex when reassociated with the X. nematophila oxyR mutant compared to the control strain revealed a potential role of OxyR during this symbiotic stage of the bacterial life cycle.

Bioactive Bioxanthracene and Cyclodepsipeptides from the Entomopathogenic Fungus Blackwellomyces roseostromatus BCC56290.

In the course of our ongoing research targeting the identification of potential biocontrol agents from entomopathogenic fungi (EPF), we explored a solid-state rice fungal extract of Blackwellomyces roseostromatus BCC56290 derived from infected lepidopteran larvae. Chemical and biological prospections afforded four unprecedentedly reported natural products differentiated into a dimeric naphthopyran bioxanthracene ES-242 derivative (1) and three cyclodepsipeptides (2-4) in addition to two known cyclodepsipeptides, cardinalisamides B (5) and C (6). Chemical structures of the isolated compounds were elucidated through comprehensive 1D/2D NMR and HR-ESI-MS data together with comparisons to the reported literature. The absolute configuration of the isolated cyclodepsipeptides was determined using Marfey's method. All isolated compounds were assessed for their antimicrobial, cytotoxic, and nematicidal activities with some compounds revealing significant activities.

A Literature Review on the Role of the Invasive Aedes albopictus in the Transmission of Avian Malaria Parasites.

The Asian tiger mosquito (Aedes albopictus) is an invasive mosquito species with a global distribution. This species has populations established in most continents, being considered one of the 100 most dangerous invasive species. Invasions of mosquitoes such as Ae. albopictus could facilitate local transmission of pathogens, impacting the epidemiology of some mosquito-borne diseases. Aedes albopictus is a vector of several pathogens affecting humans, including viruses such as dengue virus, Zika virus and Chikungunya virus, as well as parasites such as Dirofilaria. However, information about its competence for the transmission of parasites affecting wildlife, such as avian malaria parasites, is limited. In this literature review, we aim to explore the current knowledge about the relationships between Ae. albopictus and avian Plasmodium to understand the role of this mosquito species in avian malaria transmission. The prevalence of avian Plasmodium in field-collected Ae. albopictus is generally low, although studies have been conducted in a small proportion of the affected countries. In addition, the competence of Ae. albopictus for the transmission of avian malaria parasites has been only proved for certain Plasmodium morphospecies under laboratory conditions. Therefore, Ae. albopictus may play a minor role in avian Plasmodium transmission in the wild, likely due to its mammal-biased blood-feeding pattern and its reduced competence for the development of different avian Plasmodium. However, further studies considering other avian Plasmodium species and lineages circulating under natural conditions should be carried out to properly assess the vectorial role of Ae. albopictus for the Plasmodium species naturally circulating in its distribution range.

Disclosing the virome of Aedes, Anopheles and Culex female mosquitoes, Alto Pantanal of Mato Grosso, Brazil, 2019.

Using Illumina NextSeq sequencing and bioinformatics, we identified and characterized thirty-three viral sequences of unsegmented and multipartite viral families in Aedes spp., Culex sp. and Anopheles darlingi female mosquito pools from Porto São Luiz and Pirizal, Alto Pantanal. Seventeen sequences belong to unsegmented viral families, twelve represent putative novel insect-specific viruses (ISVs) within families Chuviridae (3/33; partial genomes) and coding-complete sequences of Xinmoviridae (1/33), Rhabdoviridae (2/33) and Metaviridae (6/33); and five coding-complete sequences of already-known ISVs. Notably, two putative novel rhabdoviruses, Corixo rhabdovirus 1 and 2, were phylogenetically related to Coxipo dielmovirus, but separated from other Alpharhabdovirinae genera, sharing Anopheles spp. as host. Regarding multipartite families, sixteen segments of different putative novel viruses were identified (13 coding-complete segments) within Durnavirales (4/33), Elliovirales (1/33), Hareavirales (3/33) and Reovirales (8/33) orders. Overall, this study describes twenty-eight (28/33) putative novel ISVs and five (5/33) already described viruses using metagenomics approach.

PdPLR1 effectively enhances resistance of Populus deltoides 'shalinyang' to Anoplophora glabripennis by positive regulation of lignan synthesis.

Anoplophora glabripennis (ALB) is one of the most devastating wood boring insects of poplars. Populus deltoides 'Shalinyang (PdS), a new poplar variety, shows strong resistance to ALB infestation. However, the molecular mechanism of insect resistance in PdS is unclear. Here, we found that lignan content was much higher in PdS phloem after ALB infestation than in healthy trees, and that adding lignan to artificial diet significantly reduced: larval weight; digestive enzyme activity (cellulase [CL], polygalacturonase [PG]); detoxification enzyme activity (carboxylesterase [CarE], glutathione S-transferase [GSH-ST]); and defense enzyme activity (Catalase [CAT]). We further identified the lignan biosynthesis-related PdPLR1 gene (Pinoresinol-lariciresinol reductase, PLR) based on transcriptome analysis, and it was significantly up-regulated in the PdS phloem attacked by ALB. Overexpression of PdPLR1 in Arabidopsis increased th lignan content. In contrast, silencing PdPLR1 in PdS significantly decreased expression levels of PdPLR1 and lignan content by 82.45% and 56.85%. However, silencing PdPLR1 increased the number of adults ovipositions and eggs hatching. The activity of CL, PG, CarE, GSH-ST and CAT and the biomass of larvae fed on phloem of PdS with silenced PdPLR1 were significantly higher than in the control. Taken together, up regulation of PdPLR1 enhanced PdS resistance to ALB by regulating lignan synthesis. Our findings provide in-depth insights into the molecular mechanisms of PdS-ALB interactions, which lay the foundation for understanding of defense in poplars to pest infection.

Sustainable scenarios in a plants-rhizobacteria-plant consumers system are in risk when biotic or abiotic factors change.

The ecological relationship among plants, rhizobacteria and plant consumers has attracted the attention of researchers due to its implications in field crops. It is known that, the rhizosphere is occupied not only by rhizobacteria which grant benefits to the plants but also by bacteria which are detrimental for them. In this work, we construct and analyze a plants-rhizobacteria-plant consumers system. In the modeling process, it is assumed that there is a conditioned interaction between plants and bacteria in the rhizosfera such that there is a mutualistic relationship at low densities of rhizobacteria and the relationship is parasitic or competitive at higher densities of them. Benefits granted by rhizobacteria include mechanisms that increase the plant growth and defense mechanisms against plant consumers. From the analysis of the model and its simplified version, we show that scenarios of coexistence of all populations can occur for a wide range of values of the parameters which describe biotic or abiotic factors; however, these scenarios are in risk since scenarios of exclusion of species can occur simultaneously due to the presence of bistability phenomena. The results obtained can be useful for the decision makers to design interventions strategies on field crops when plant growth-promoting rhizobacteria are used.

A highly resolved network reveals the role of terrestrial herbivory in structuring aboveground food webs.

Comparative studies suggest remarkable similarities among food webs across habitats, including systematic changes in their structure with diversity and complexity (scale-dependence). However, historic aboveground terrestrial food webs (ATFWs) have coarsely grouped plants and insects such that these webs are generally small, and herbivory is disproportionately under-represented compared to vertebrate predator-prey interactions. Furthermore, terrestrial herbivory is thought to be structured by unique processes compared to size-structured feeding in other systems. Here, we present the richest ATFW to date, including approximately 580 000 feeding links among approximately 3800 taxonomic species, sourced from approximately 27 000 expert-vetted interaction records annotated as feeding upon one of six different resource types: leaves, flowers, seeds, wood, prey and carrion. By comparison to historical ATFWs and null ecological hypotheses, we show that our temperate forest web displays a potentially unique structure characterized by two properties: (i) a large fraction of carnivory interactions dominated by a small number of hyper-generalist, opportunistic bird and bat predators; and (ii) a smaller fraction of herbivory interactions dominated by a hyper-rich community of insects with variably sized but highly specific diets. We attribute our findings to the large-scale, even resolution of vertebrate, insect and plant guilds in our food web.This article is part of the theme issue 'Connected interactions: enriching food web research by spatial and social interactions'.

Plasticity for the win: Flexible transcriptional response to host plant switches in the comma butterfly (Polygonia c-album).

Generalist plant-feeding insects are characterised by a broad host repertoire that can comprise several families or even different orders of plants. The genetic and physiological mechanisms underlying the use of such a wide host range are still not fully understood. Earlier studies indicate that the consumption of different host plants is associated with host-specific gene expression profiles. It remained, however, unclear if and how larvae can alter these profiles in the case of a changing host environment. Using the polyphagous comma butterfly (Polygonia c-album) we show that larvae can adjust their transcriptional profiles in response to a new host plant. The switch to some of the host plants, however, resulted in a larger transcriptional response and, thus, seems to be more challenging. At a physiological level, no correspondence for these patterns could be found in larval performance. This suggests that a high transcriptional but also phenotypic flexibility are essential for the use of a broad and diverse host range. We furthermore propose that host switch tests in the laboratory followed by transcriptomic investigations can be a valuable tool to examine not only plasticity in host use but also subtle and/or transient trade-offs in the evolution of host plant repertoires.

Comparison of volatile compound profiles derived from various livestock protein alternatives including edible-insect, and plant-based proteins.

In this study, the distinctive chemical fingerprints that contribute to the flavor characteristics of various protein materials, such as insects, plant-based protein, and livestock, were investigated. In edible-insects (Tenebrio molitor and Protaetia brevitarsis), aldehydes and cyclic volatile compounds were the predominant volatile components and had distinct flavor characteristics such as cheesy, sharp, green, floral, and sweet. In contrast, the relatively high levels of pyrazines and furans in plant-based protein materials, such as textured vegetable and pea protein. They included unique flavor properties characterized by sweet, fatty, grassy, creamy, and roasted. The primary volatile chemical group detected in livestock protein materials, such as a pork and a beef, was ketones. The pork sample showed specific flavors, such as alcoholic, green, and fruity, while a beef presented distinctive flavor, including creamy, fruity, and alcoholic. Based on the results, this research provided the understanding of the flavor aspects of diverse protein materials.

Crofton weed derived isomers of ageraphorone as potent antifeedant against Plutella xylostella (L.).

Global agricultural production is significantly hampered by insect pests, and the demand for natural pragmatic pesticides with environmental concern remains unfulfilled. Ageratina adenophora (Spreng.) also known as Crofton weed, is an invasive perennial herbaceous plant that is known to possess multiple bioactive compounds. In our study, two isomers of ageraphorone metabolites i.e, 10 Hα-9-oxo-ageraphorone (10HA) and 10 Hß-9-oxo-ageraphorone (10HB), were identified from Crofton weed, exhibiting potent antifeedant and larvicidal activities against Plutella xylostella. For antifeedant activity, the median effective concentration (EC50) values for 10HA and 10HB in the choice method were 2279 mg/L and 3233 mg/L, respectively, and for the no choice method, EC50 values were 1721 mg/L and 2394 mg/L, respectively. For larvicidal activity, lethal concentration (LC50) values for 10HA and 10HB were 2421 mg/L and 4109 mg/L at 48 h and 2101 mg/L and 3550 mg/L at 72 h. Furthermore, both in- vivo and in-vitro studies revealed that the isomers 10HA and 10HB exhibited potent detoxifying enzymes inhibition activity such as carboxylesterase and glutathione S-transferases. Molecular docking and MD simulation analysis provide insight into the possible interaction between isomers of ageraphorone metabolites and Carboxylic Ester Hydrolase protein (Gene: pxCCE016b) of P. xylostella, which led to a finding that CarEH protein plays a significant role in the detoxification of the two compounds in P. xylostella. Finally, our findings show that the primary enzymes undergoing inhibition by isomers of ageraphorone metabolites, causing toxicity in insects, are Carboxylesterase and glutathione S-transferase.