Ecotoxicological Effects of Titanium Aluminum Carbide Composites on Biochemical and Metabolic Parameters of Galleria mellonella.

Metal composites have been extensively used in various fields such as automotive industry, medicine and pharmacy. However, the high exposure of these chemicals may have an adverse effect on the living organisms. In this study, the effect of titanium aluminum carbide (Ti3AlC2) on the model organism Galleria mellonella was investigated. The change in the metabolic enzymes such as alanine transferase, aspartate transferase, gamma-glutamyl transferase, lactate dehydrogenase, amylase, creatine kinase, alkaline phosphatase in the hemolymph of G. mellonella which was exposed to Ti3AlC2 was determined. The contents of the bilirubin, albumin, uric acid and the total protein were also measured after the Ti3AlC2 exposure on the model organism. The results of our study clearly indicate that Ti3AlC2 has adverse effects on the model organism G. mellonella.

Oxidative and genotoxic effect of piperazine on Galleria mellonella (Lepidoptera: Pyralidae) hemolymph.

Recently, there are many studies suggesting antibacterial, antifungal, and anthelmintic agents as alternative chemicals to insecticides. In this study, the oxidative and genotoxic effect of Piperazine, a clinically important hexahydropyrazine anthelmintic, on Galleria mellonella L. hemolymph tissue by adding artificial diet were investigated. Galleria mellonella larvae were reared until 7th larval stage in artificial diet containing 0.001, 0.01, 0.1, and 1 g piperazine per 100 g of diet. Using hemolymph collected from 7th-instar larvae, the amount of lipid peroxidation final product malondialdehyde (MDA), protein oxidation product protein carbonyl (PCO), and detoxification enzymes glutathione S-transferase (GST) and cytochrome P450 monooxygenase (cyt P450) activity, comet assay were measured. According to the results obtained, when the piperazine high concentrations tested with the control group were compared, statistically significant differences were found in MDA, PCO content, cyt P450, GST activity, and comet assay in the hemolymph of the insect. While MDA content was 0.01 ± 0.0021 nmol/mg protein in the control group, this amount increased approximately 2-fold at the highest concentration (0.0231 ± 0.0050 nmol/mg protein). On the other hand, when the control group and the highest piperazine concentration were compared in the GST and cyt P450 activity, it was determined that there was a statistically significant increase. We obtained similar results in comet assay and micronucleus formation data. This study showed that the tested piperazine concentrations caused significant changes in the detoxification capacity, oxidative stress, and genotoxic markers in the insect's hemolymph tissue.

Effects of Midazolam on Antioxidant Levels, Biochemical and Metabolic Parameters in Eurygaster integriceps Puton (Hemiptera: Scutelleridae) Eggs Parasitized by Trissolcus semistriatus Nees (Hymenoptera: Scelionidae).

Eurygaster integriceps Puton (Hemiptera: Scutelleridae) is among the most important insect pests of wheat (Triticum sativum L.) and barley (Hordeum vulgare L.) grown in the Middle East. Biological and chemical methods are insufficient to control E. integriceps populations below economic thresholds. In this study, we investigated the effects of midazolam, a clinical drug, on selected metabolic enzyme activity, antioxidant levels, and biochemical parameters in E. integriceps eggs parasitized by Trissolcus semistriatus Nees (Hymenoptera: Scelionidae). Increasing concentrations of midazolam caused cell damage in the parasitized eggs due to its oxidative effects. Transferase enzymes, such as, aspartate transferase, alanine transferase, and gamma glutamyl transferase activities were altered following exposure. Metabolic enzymes, such as, creatine kinase, alkaline phosphatase, amylase, and lactate dehydrogenase also were adversely affected. Levels of the non-enzymatic antioxidants uric acid, bilirubin, and albumin also were altered.

Effects of Copper Oxide Nanoparticles on Immune and Metabolic Parameters of Galleria mellonella L.

In this study, the effects of dietary CuO nanoparticles (NPs) on metabolic enzyme activity, biochemical parameters, and total (THC) and differential hemocyte counts (DHC) were determined in Galleria mellonella larvae. Using concentrations of 10, 100, 1000 mg/L and the LC10 and LC30 levels of CuO NPs, we determined that the NPs negatively impacted metabolic enzyme activity and biochemical parameters in larval hemolymph. Compared with the control, the greatest increase in THC was observed in larvae fed on diets with 100 mg L-1 of CuO NPs. Plasmatocytes and granulocytes were among the most numerous hemocytes in all treatments. These results suggest that dietary CuO NPs effects the metabolic metabolism and immune system of G. mellonella and provide indirect information regarding the toxic effects of CuO NPs in mammalian immune system given similarities between mammalian blood cells and insect hemocytes.

Ecotoxicological effects of dietary titanium dioxide nanoparticles on metabolic and biochemical parameters of model organism Galleria mellonella (Lepidoptera: Pyralidae).

Nanoparticles (NPs) are now being used in many industrial activities, such as mining, paint and glass industries. The frequent industrial use of NPs contributes to environmental pollution and may cause cellular and oxidative damage in native organisms. In this study, the toxic effects of titanium dioxide nanoparticles (TiO2 NPs) were investigated using Galleria mellonella larvae as a model insect species. Alterations in cell damage indicators, such as alanine transferase, aspartate transferase, lactate dehydrogenase, non-enzymatic antioxidants and biochemical parameters, were determined in the hemolymph of G. mellonella larvae exposed to TiO2 NPs at different concentrations (5, 50, 250 and 1250 µg/mL) in their diets. TiO2 NPs caused concentration-dependent cellular damage in the hemolymph of G. mellonella larvae and increased the levels of the non-enzymatic antioxidants uric acid and bilirubin. In addition, total protein in hemolymph significantly decreased at the highest concentration (1250 µg/mL) of TiO2 NPs. Level of the urea increased at the highest concentration (1250 µg/mL) of TiO2 NPs, whereas the amount of glucose was not affected. These findings demonstrated that TiO2 NPs caused concentration-dependent toxic effects on G. mellonella larvae.

Drosophila melanogaster Response to Feeding with Neomycin-Based Medium Expressed in Fluctuating Asymmetry.

The fruit fly Drosophila melanogaster is a model species used for a wide range of studies. Contamination of Drosophila cultures with bacterial infection is common and is readily eradicated by antibiotics. Neomycin antibiotics can cause stress to D. melanogaster's larvae and imagoes, which may affect the interpretation of the results of research using culture from neomycin-based medium. In the present study, fluctuating asymmetry (FA), one of the important bioindicators of stress, was measured. Larvae and imagoes of a wild-type D. melanogaster strain were exposed to various concentrations of neomycin. The size of anal papillae and selected wing veins were measured using scanning electron and light microscopy, respectively. Next, the FA was checked. The values obtained for larval anal papillae appeared to be concentration-dependant; the FA indices increased with the concentration of neomycin. The wing FA presented a large but variable correlation, depending on the measured vein. However, the mean length of veins was the highest for the control group, with neomycin-exposed groups showing lower values. The research showed that neomycin may cause sublethal stress in D. melanogaster, which manifests in increased FA indices. This suggests that neomycin can cause physiological and developmental stress in insects, which should be taken into account when interpreting the results of studies using these model organisms.

The Use of Dietary Antifungal Agent Terbinafine in Artificial Diet and Its Effects on Some Biological and Biochemical Parameters of the Model Organism Galleria mellonella (Lepidoptera: Pyralidae).

Diet quality widely affects the survival, development, fecundity, longevity, and hatchability of insects. We used the greater wax moth Galleria mellonella (Linnaeus) to determine the effects of the antifungal, antibiotic terbinafine on some of its' biological parameters. The effects of terbinafine on malondialdehyde (MDA) and protein carbonyl (PCO) contents and the activity of the detoxification enzyme, glutathione S-transferase (GST), in the midgut of seventh-instar larvae of G. mellonella were also investigated. The insects were reared on an artificial diet containing terbinafine at concentrations of 0.001, 0.01, 0.1, and 1 g. The survival rates at all development stages of G. mellonella were significantly decreased at all terbinafine concentrations. The females from a control diet produced 82.9 ± 18.1 eggs; however, this number was significantly reduced to 51.4 ± 9.6 in females given a 0.1 g terbinafine diet. The highest concentration of terbinafine (1 g) completely inhibited egg laying. Terbinafine significantly increased MDA content and GST activity in the midgut tissue of seventh-instar larvae in a dose-dependent manner. Relative to controls, these low dietary concentrations of terbinafine significantly increased midgut PCO content; a 0.1 g terbinafine concentration raised PCO content from 155.19 ± 21.8 to 737.17 ± 36.4 nmol/mg protein. This study shows concentration-dependent effects on the biological traits of the greater wax moth G. mellonella, including the oxidative status and detoxification capacity of the midgut. Low terbinafine concentrations may be possible for use as an antifungal agent in insect-rearing diets.

Oxidative Effects of Gemifloxacin on Some Biological Traits of Drosophila melanogaster (Diptera: Drosophilidae).

In recent times, several studies have been undertaken on the artificial mass-rearing of insects in which clinical antibacterial antibiotics, including gemifloxacin, which is a DNA gyrase and topoisomerases inhibitor, are amended into the diet to control microbial contamination and preserve diet. The findings of these studies have suggested the possibility that these antibiotics influence the biological traits of insects in relation to their oxidative effects. This study investigated the effects of gemifloxacin on Drosophila melanogaster (Meigen) survival rates, development times, and male-female adult longevity. And we also determined the effects of gemifloxacin on lipid peroxidation product, malondialdehyde, protein carbonyl levels, and glutathione S-transferase activity of fruit fly eggs. First instars were fed on artificial diets containing 150, 300, 600, and 900 mg/liter concentrations of gemifloxacin until adult emergence. Our results indicate that sublethal effects of gemifloxacin are likely to significantly impair adult fitness and life-history parameters in D. melanogaster, probably because of its oxidative effects.

The Effect of Neomycin on Survival and Development of Pimpla turionellae L. (Hymenoptera: Ichneumonidae) Reared on a Natural Host.

Understanding the effects of diet on metabolic events is crucial for biological control programs of parasitoid insects. As bioindicators of long-term physiological stress: survivorship of fifth instar larvae, pupation, adult survival, and developmental time for stages of endoparasitoid Pimpla turionellae L. (Hymenoptera: Ichneumonidae) were investigated by rearing the parasitoid on the host, Galleria mellonella L. (Lepidoptera: Pyralidae) pupae were treated with neomycin. First instar larvae of G. mellonella were reared on artificial diets containing 0.005, 0.01, or 0.5 g neomycin (g/100 g of diet) until seventh instar larvae; the pupae from these larvae were used as a host for rearing P. turionellae. In the control group, the pupae from larvae reared on artificial diets without neomycin were used as a host. Survivorship of fifth instar, pupal, and adult stages of P. turionellae L. reared on G. mellonella pupae as a host fed with different concentrations of neomycin were significantly decreased in comparison to the control group. Approximately eighty percent of P. turionellae L. pupae were produced from control host pupae, while other neomycin concentrations significantly decreased the pupation of the parasitoid. Pimpla turionellae L. larvae reared on control host pupae reached fifth instar in about 9.6 ± 0.61 d, while the larvae reared from a host pupae exposed to the highest antibiotic concentration completed their development to the fifth instar in about 7.4 d. These results showed that neomycin, and possibly its metabolites, contaminated P. turionellae L. larvae from a host and affected larval stages of the parasitoid.

Inhibition of eicosanoid signaling leads to increased lipid peroxidation in a host/parasitoid system.

We posed the hypothesis that inhibition of eicosanoid biosynthesis leads to increased lipid peroxidation in insects. Here we report that rearing the greater wax moth, Galleria mellonella, on media supplemented with selected inhibitors of eicosanoid biosynthesis throughout the larval, pupal and adult life led to major alterations in selected oxidative and antioxidative parameters of wax moth and its ectoparasitoid, Bracon hebetor. The highest dietary dexamethasone (Dex), esculetin (Esc) and phenidone (Phe) led to increased malondialdehyde (MDA) levels and to elevated catalase (CAT) and glutathione-S-transferase (GST) activities in all developmental stages of host larvae. Dietary Phe resulted in increased MDA levels, and CAT activity in G. mellonella adults by about 4-fold and about 2-fold, respectively. The Phe effect on GST activity in all stages of the wax moth was expressed in a dose-dependent manner, increased to 140nmol/mg protein/min in larvae. MDA levels were increased by over 30-fold in adult wasps reared on Dex- and Esc-treated hosts. CAT and GST activities were increased in adult parasitoids reared on Esc-and Phe-treated hosts. GST activity of Dex-treated parasitoid larvae increased from about 4 to over 30nmol/mg protein/min. Dietary Phe led to increased GST activity, by about 25-fold, in adult wasps. These data indicate that chronic inhibition of eicosanoid biosynthesis leads to increased oxidative stress, strongly supporting our hypothesis. The significance of this work lies in understanding the roles of eicosanoids in insect biology. Aside from other well-known eicosanoids actions, we propose that eicosanoids mediate reductions in oxidative stress.

Ingestion of the anti-bacterial agent, gemifloxacin mesylate, leads to increased gst activity and peroxidation products in hemolymph of Galleria mellonella l. (lepidoptera: pyralidae).

Gemifloxacin mesylate (GEM) is a synthetic, fourth-generation fluoroquinolone antibacterial antibiotic that has a broad spectrum of activity against bacteria. GEM inhibits DNA synthesis by inhibiting DNA gyrase and topoisomerase IV activities. Recent research into insect nutrition and mass-rearing programs, in which antibiotics are incorporated into the culture media to maintain diet quality, raised a question of whether clinical antibiotics influence the health or biological performance of the insects that ingest these compounds. Because some antibiotics are pro-oxidant compounds, we addressed the question with experiments designed to assess the effects of GEM (mesylate salt) on oxidative stress indicators, using Galleria mellonella larvae. The insects were reared from first-instar larvae to adulthood on artificial diets amended with GEM at 0.001, 0.01, 0.1, or 1.0%. Feeding on the 1% diets led to significantly increased hemolymph contents of the lipid peroxidation product, malondialdehyde and protein oxidation products, protein carbonyl. All GEM concentrations led to increased hemolymph glutathione S-transferase activity. We inferred that although it was not directly lethal to G. mellonella larvae, dietary exposure to GEM exerts measurable oxidative damage, possibly on insects generally. Long-term, multigenerational effects remain unknown.

Effects of Antiviral Agent, Acyclovir, on the Biological Fitness of Galleria mellonella (Lepidoptera: Pyralidae) Adults.

The effects of a synthetic purine nucleoside analog, antiviral agent, acyclovir (ACV), on adult longevity, fecundity, and hatchability of a serious honeycomb pest, greater wax moth Galleria mellonella L. were investigated by adding 0.01, 0.1, 1.0, and 3.0% ACV into artificial and natural diets. Control larvae were reared on diet without ACV. The artificial diet containing the lowest level of ACV, 0.01%, raised egg production from a number of 12.9 ± 0.6 to 163.2 ± 1.3. The hatching rate of these eggs was increased from 49.2 ± 2.4% to 68.2 ± 3.2%. Higher concentrations of ACV in natural food significantly increased both egg production and egg hatching rate. Female reared on old dark combs as natural diet exposed to 1.0% of ACV produced 167.5 ± 5.8 eggs with 93.2 ± 6.8% hatched. This study emphasizes the importance of determining the dietary impact of an antimicrobial agent as a food additive to a particular species of insect before its using for dietary antimicrobial purpose.

Solanum tuberosum and Lycopersicon esculentum Leaf Extracts and Single Metabolites Affect Development and Reproduction of Drosophila melanogaster.

Glycoalkaloids are secondary metabolites commonly found in Solanaceae plants. They have anti-bacterial, anti-fungal and insecticidal activities. In the present study we examine the effects of potato and tomato leaf extracts and their main components, the glycoalkaloids α-solanine, α-chaconine and α-tomatine, on development and reproduction of Drosophila melanogaster wild-type flies at different stages. Parental generation was exposed to five different concentrations of tested substances. The effects were examined also on the next, non-exposed generation. In the first (exposed) generation, addition of each extract reduced the number of organisms reaching the pupal and imaginal stages. Parent insects exposed to extracts and metabolites individually applied showed faster development. However, the effect was weaker in case of single metabolites than in case of exposure to extracts. An increase of developmental rate was also observed in the next, non-exposed generation. The imagoes of both generations exposed to extracts and pure metabolites showed some anomalies in body size and malformations, such as deformed wings and abdomens, smaller black abdominal zone. Our results further support the current idea that Solanaceae can be an impressive source of molecules, which could efficaciously be used in crop protection, as natural extract or in formulation of single pure metabolites in sustainable agriculture.

A Review of Bioinsecticidal Activity of Solanaceae Alkaloids.

Only a small percentage of insect species are pests. However, pest species cause significant losses in agricultural and forest crops, and many are vectors of diseases. Currently, many scientists are focused on developing new tools to control insect populations, including secondary plant metabolites, e.g., alkaloids, glycoalkaloids, terpenoids, organic acids and alcohols, which show promise for use in plant protection. These compounds can affect insects at all levels of biological organization, but their action generally disturbs cellular and physiological processes, e.g., by altering redox balance, hormonal regulation, neuronal signalization or reproduction in exposed individuals. Secondary plant metabolites cause toxic effects that can be observed at both lethal and sublethal levels, but the most important effect is repellence. Plants from the Solanaceae family, which contains numerous economically and ecologically important species, produce various substances that affect insects belonging to most orders, particularly herbivorous insects and other pests. Many compounds possess insecticidal properties, but they are also classified as molluscides, acaricides, nematocides, fungicides and bactericides. In this paper, we present data on the sublethal and lethal toxicity caused by pure metabolites and crude extracts obtained from Solanaceae plants. Pure substances as well as water and/or alcohol extracts cause lethal and sublethal effects in insects, which is important from the economical point of view. We discuss the results of our study and their relevance to plant protection and management.

Reduced Fitness in Adults From Larval, Galleria mellonella (Lepidoptera: Pyralidae) Reared on Media Amended With the Antihelmintic, Mebendazole.

Benzimidazole antihelmintics, including mebendazole, have a broad antiparasitic spectrum. These drugs play a major role in the treatments of parasites of intestines or other organs of vertebrates, humans, and other animals.The impact of mebendazole on the biology of the greater wax moth, Galleria mellonella (L.), was assessed by observation of several developmental parameters as follows: survivorship, developmental time, and adult longevity. Sublethal toxicity was measured through reproductive parameters such as fecundity and hatchability.The larvae were reared on artificial diet from first-instar larvae to the adult stage in the laboratory. The diets contained mebendazole at different concentrations of 0.005, 0.05, 0.5, or 1.0%. Control diet did not containme bendazole and produced seventh-instar larvae in 96.6±1.67% of cases, whereas the addition of mebendazole into diet at 1.0% significantly decreased survivorship of seventh-instar larvae to 79.9±4.08%. The diet with the highest concentration of mebendazole decreased survivorship in the adult stage from 79.9±2.35 to 56.6±4.73%, and shortened the developmental time for adult emergence from 36.7±0.48 to 34.1±0.63 d. All mebendazole concentrations shortened adult longevity and significantly decreased fecundity and hatch ability of G. mellonella. The highest dietary concentration of this antihelmintic significantly decreased the egg number to 28.6±2.89 and hatching rate to 51.7±1.85%. The present study demonstrates that mebendazole exhibits significant adverse effects on greater wax moth, leading to deteriorated life table parameters and decreased adult fitness.

Potato leaf extract and its component, α-solanine, exert similar impacts on development and oxidative stress in Galleria mellonella L.

Plants synthesize a broad range of secondary metabolites that act as natural defenses against plant pathogens and herbivores. Among these, potato plants produce glycoalkaloids (GAs). In this study, we analyzed the effects of the dried extract of fresh potato leaves (EPL) on the biological parameters of the lepidopteran, Galleria mellonella (L.) and compared its activity to one of the main EPL components, the GA α-solanine. Wax moth larvae were reared from first instar on a diet supplemented with three concentrations of EPL or α-solanine. Both EPL and α-solanine affected survivorship, fecundity, and fertility of G. mellonella to approximately the same extent. We evaluated the effect of EPL and α-solanine on oxidative stress in midgut and fat body by measuring malondialdehyde (MDA) and protein carbonyl (PCO) contents, both biomarkers of oxidative damage. We evaluated glutathione S-transferase (GST) activity, a detoxifying enzyme acting in prevention of oxidative damage. EPL and α-solanine altered MDA and PCO concentrations and GST activity in fat body and midgut. We infer that the influence of EPL on G. mellonella is not enhanced by synergistic effects of the totality of potato leaf components compared to α-solanine alone.

The influence of dietary α-solanine on the waxmoth Galleria mellonella L.

Plant allelochemicals are nonnutritional chemicals that interfere with the biology of herbivores. We posed the hypothesis that ingestion of a glycoalkaloid allelochemical, α-solanine, impairs biological parameters of greater wax moths Galleria mellonella. To test this idea, we reared wax moths on artificial diets with 0.015, 0.15, or 1.5 mg/100 g diet of α-solanine. Addition of α-solanine to the diet affected survival of seventh-instar larvae, pupae, and adults; and female fecundity and fertility. The diet containing the highest α-solanine concentration led to decreased survivorship, fecundity, and fertility. The diets supplemented with α-solanine led to increased malondialdehyde and protein carbonyl contents in midgut and fat body and the effect was dose-dependent. Dietary α-solanine led to increased midgut glutathione S-transferase activity and to decreased fat body glutathione S-transferase activitiy. We infer from these findings that α-solanine influences life history parameters and antioxidative enzyme activities in the midgut and fat body of G. mellonella.

Effect of boric acid on antioxidant enzyme activity, lipid peroxidation, and ultrastructure of midgut and fat body of Galleria mellonella.

Boric acid is widely used as an insecticide, acaricide, herbicide, and fungicide and also during various industrial processings. Hence, numerous populations are subjects to this toxic compound. Its action on animals is still not fully known and understood. We examined the effect of boric acid on larvae of greater wax moth (Galleria mellonella). The chemical appeared to be toxic for larvae, usually in a concentration-dependent manner. Exposed groups revealed increased lipid peroxidation and altered activity of catalase, superoxide dismutase, glutathione S-transferase, and glutathione peroxidase. We also observed changes of ultrastructure, which were in tune with biochemical assays. We suggest that boric acid has a broad mode of action, which may affect exposed larvae, and even if sublethal, they may lead to disturbances within exposed populations.

The influence of chronic eicosanoid biosynthesis inhibition on life history of the greater waxmoth, Galleria mellonella and its ectoparasitoid, Bracon hebetor.

Eicosanoids are oxygenated metabolites of three C20 polyunsaturated fatty acids, mainly arachidonic acid (AA; 20:4n-6), but also 20:3n-6 and 20:5n-3. Aside from their importance in biomedicine, eicosanoids act in invertebrate biology. Prostaglandins (PGs) influence salt and water transport physiology in insect rectal epithelia and in Malpighian tubules. PGs also influence a few insect behaviors, including releasing oviposition behavior and behavioral fever. Eicosanoids act in ovarian development and in insect immunity. Because eicosanoids act in several areas of insect biology, we posed the hypothesis that chronic inhibition of eicosanoid biosynthesis, in the absence of microbial challenge, can influence insect life table parameters, including developmental time, survival, adult longevity and parasitoid fecundity. Here we report that inhibiting eicosanoid biosynthesis throughout the larval life exerted minor influences on some life table parameters of the greater wax moth, Galleria mellonella and its ectoparasitoid, Bracon hebetor, however, the inhibitors strongly reduced the production and hatchability of the parasitoids' eggs. The significance of the work relates to the potentials of understanding and targeting eicosanoid systems as a platform for developing new technologies of insect pest management. As seen here, the impact of targeting eicosanoid systems is seen in crucial moments of insect life histories, such as reproduction or immune challenge rather than in overall larval development.

Eicosanoids mediate hemolymph oxidative and antioxidative response in larvae of Galleria mellonella L.

Antioxidant enzymes play a major role in the defense against pro-oxidative effects of xenobiotics and pro-oxidant plant allelochemicals in insects. We posed the hypothesis that eicosanoids also mediate antioxidant enzymatic defense reactions to pro-oxidant challenge. To test this idea, we reared first-instar larvae of Galleria mellonella (L.) with the lypoxygenase inhibitor, esculetin (0.001%), the phospholipase A(2) inhibitor, dexamethasone (0.001%) and the dual inhibitor of cyclooxygenase and lipoxygenase, phenidone (0.1%) to seventh-instars. Newly ecdysed seventh-instars were then fed on artificial diet containing 0.05% xanthotoxin (XA) for 2 days. Treating seventh-instar larvae of G. mellonella with XA induced lipid peroxidation and protein carbonylation as evident from the increased content of malondialdehyde (MDA) and protein carbonyls respectively, and antioxidative enzymatic response in a dose-dependent manner. High dietary XA concentrations (0.005 and 0.1%) were associated with increasing MDA and carbonyl content (by 3-fold) and antioxidant enzyme activities, superoxide dismutase (SOD) (by 3-fold) and catalase (CAT) (by 4-fold), and glutathione-dependent enzymes, glutathione S-transferase (GST) (by 15-fold) and glutathione peroxidase (GPx) (by 7-fold). Relative to control, eicosanoid biosynthesis inhibitors (EBIs) esculetin, dexamethasone and phenidone also resulted in impaired MDA content and antioxidant enzyme activities. However, carbonyl content did not differ between control- and EBIs-feeding larvae. Finally, MDA and carbonyl content, and antioxidant enzymes SOD, GST and GPx activities exhibited an incremental increase while CAT activity was decreased in the experimental larvae that had been reared on media amended with esculetin, dexamethasone and phenidone and then challenged with our standard XA challenge dose. Two of the markers indicated that significantly higher levels of oxidative stress were produced in the hemolymph tissue of larvae fed diets supplemented with EBIs and then challenged with XA. This oxidative stress was associated with elicited antioxidative responses by increasing SOD, GST and GPx and decreasing CAT activities in hemolymph. We infer from these findings that eicosanoids mediate insect antioxidant enzymatic responses to dietary pro-oxidants.