Increased Glutathione S-transferase Activity Research Articles

Role of Glutathione in Chalcone Derivative Induced Apoptosis of Brugia malayi and its Possible Therapeutic Implication.

Oxidative stress is an essential component of innate response against microbes. The oxidative impact has a very subtle connection with apoptosis. Our previous work indicated presumptive evidence of apoptosis by the chalcone derivatives against the human lymphatic filarial parasite. Evidence suggests the involvement of glutathione-S-transferase (GST) in the mechanism of action of chalcone drugs. In the present study, we explored the implications of redox status in apoptosis of the parasite by this drug. Treatment with the representative drug, 4t, significantly decreased GSH level and increased GST activity in the Brugia malayi microfilariae (Mf) in comparison to Mf without 4t treatment. Drug-induced loss of motility of the parasites was reversed by the treatment with GSH (41%) and NAC (19%). A significant fall in rGST activity was observed due to drug addition, which could be reversed by the addition of GSH co-substrate, but not with the re-addition of rGST, indicating a vital role of GSH. In silico study demonstrated a favorable drug-GST enzyme interaction. Oxidative stress was reflected by increased protein carbonylation and intracellular reactive oxygen species level, in the drug-treated parasite. Mitochondrial oxygen consumption was reduced by the drug, which was reversed on the addition of GSH. Mitochondrial dysfunction was confirmed by MTT and cytochrome c assay. Apoptosis was confirmed by the inhibition in PARP activity. We conclude that the depletion of GSH by chalcone with concomitant mitochondrial dysfunction revealed a novel rationale of apoptosis in the parasite. Such a mechanism might have wide therapeutic implications.

Target-Site Mutations and Glutathione S-Transferases Are Associated with Acequinocyl and Pyridaben Resistance in the Two-Spotted Spider Mite Tetranychus urticae (Acari: Tetranychidae).

Simple SummaryThe two-spotted spider mite Tetranychus urticae is a difficult-to-control pest due to its short life cycle and rapid resistance development. In this study, we characterized field strains collected in 2001 and 2003 that have been selected for acequinocyl resistance and pyridaben resistance, respectively. These strains displayed resistance ratios of 1798.6 and 5555.6, respectively, and were screened for cross-resistance against several currently used acaricides. The acequinocyl resistant strain exhibited pyridaben cross-resistance, but the pyridaben resistant strain showed no cross-resistance. The acequinocyl resistant strain exhibited point mutations in cytb (I256V and N321S) and PSST (H92R). In contrast, the pyridaben resistant strain exhibited the H92R but not the I256V and N321S point mutations. In addition, the increased GST metabolism and GST delta expression might be related to acequinocyl resistance in Tetranychus urticae. We hope that the data and patterns described here can now be exploited in the continued quest for rational resistance management strategies.The two-spotted spider mite Tetranychus urticae is a difficult-to-control pest due to its short life cycle and rapid resistance development. In this study, we characterized field strains collected in 2001 and 2003 that were selected for acequinocyl resistance (AR) and pyridaben resistance (PR), respectively. These strains displayed resistance ratios of 1798.6 (susceptible vs. AR) and 5555.6 (susceptible vs. PR), respectively, and were screened for cross-resistance against several currently used acaricides. The AR strain exhibited pyridaben cross-resistance, but the PR strain showed no cross-resistance. The AR strain exhibited point mutations in cytb (I256V, N321S) and PSST (H92R). In contrast, the PR strain exhibited the H92R but not the I256V and N321S point mutations. In some cases increased glutathione S-transferase (GST) activity has previously been linked to enhanced detoxification. The AR strain exhibited approximately 2.3-, 1.8-, and 2.2-fold increased GST activity against 1-chloro-2,4-dinitrobenzene (CDNB), 1,2-dichloro-4-nitrobenzene (DCNB), and 4-nitrobenzyl chloride (NBC), respectively. Among the five GST subclass genes (delta, omega, mu, zeta, and kappa), the relative expression of delta class GSTs in the AR strain were significantly higher than the PR and susceptible strain. These results suggest that the I256V and N321S mutations and the increased GST metabolism and GST delta overexpression might be related to acequinocyl resistance in T. urticae.

Single and repeated low-dose UVB radiation exposures affect the visual system

The visual system is an important biological indicator of effects induced by ultraviolet (UV) radiation. However, research has extensively investigated the effects of high-dose UV radiation in a single exposure, thus, the differential of this work was to investigate the effects of UVB radiation in low doses in single and repeated exposure. Therefore, we investigated the effects of repeated exposure to environmental UVB doses (0.09 J/cm2) on the retina and optic lobes of the crab Neohelice granulata. We evaluated the reactive oxygen species (ROS) concentration, antioxidant capacity against peroxyl radicals (ACAP) levels, catalase (CAT) and glutathione S-transferase (GST) activities and lipoperoxidation (LPO) levels and performed histological analysis. The crabs were exposed to UVB radiation for 1 or 60 days, while the control group was exposed to visible light. In the retina region, increases in ROS concentration and CAT and GST activities after the single exposure were observed. After 60 days of exposure, we observed an increase in ACAP levels. In the optic lobes, we observed an increase in GST activity and a decrease in LPO levels after the single exposure. However, we observed an increase in ROS concentration after 60 days of exposure. Moreover, after 60 days of exposure, infiltrating hemocytes in the retina and disorganization in neuron cell bodies of the external medulla were observed. In this sense, single and repeated exposure to low doses of UVB radiation induced changes in oxidative status and inflammatory process in the visual system of the crab Neohelice granulata.

Chlorothalonil causes redox state change leading to oxidative stress generation in Danio rerio

A diverse range of chemicals are used in agriculture to increase food production on a large scale, and among them is the use of pesticides such as chlorothalonil, a broad-spectrum fungicide used in the control of foliar fungal diseases. This study aimed to elucidate the effects of chlorothalonil on biochemical biomarkers of oxidative stress in tissues of the fish Danio rerio. To achieve this, animals were exposed for 4 and 7 days, to nominal concentrations of chlorothalonil at 0 μg/L (DMSO, 0.001%), 0.1 μg/L and 10 μg/L, and after the exposure period, the tissues (gills and liver) were removed for biochemical analysis. Antioxidant capacity against peroxyl radicals (ACAP) and enzyme activities, such as superoxide dismutase (SOD), catalase (CAT), glutathione S-transferase (GST) and glutamate cysteine ligase (GCL), were evaluated in both tissues. In addition, the concentration of reactive oxygen species (ROS), reduced glutathione (GSH) and lipid peroxidation (LPO) levels were also analysed. A significant increase in ROS concentration, ACAP levels, GST and GCL activities and a significant reduction of LPO levels in gills exposed to the highest concentration were observed after 4 days. However, there was a significant reduction of ACAP and CAT activity, as well as a significant increase of GST activity and LPO levels in gills exposed to the lower concentration after 7 days. The liver was less affected, presenting a significant reduction in CAT activity and LPO levels after 4 days. However, a significant increase in SOD activity and LPO levels occurred after 7 days. These results indicate that chlorothalonil, after 4 days, caused activation of the antioxidant defence system in gills of animals exposed to the highest concentration. However, after 7 days, the lowest concentration of this compound caused oxidative stress in this same organ. Also, the results show that gills were more affected than the liver, probably because gills can be involved in chlorothalonil metabolisation. Therefore, it is possible that the liver could be exposed to lower chlorothalonil concentrations or less toxic metabolites due to the metabolism taking place in the gills.

Curcumin protects remote organs against injury that is caused by intestinal ischemia and reperfusion

In addition to several local pathophysiological consequences, intestinal injury that is caused by ischemia and reperfusion can result in the development of lesions in remote organs. Curcumin has therapeutic potential because of its antiinflammatory and antioxidant effects. The present study evaluated the effects of curcumin on oxidative and inflammatory parameters in the liver and kidneys in rats that were subjected to intestinal ischemia and reperfusion. The rats were subjected to 45 min. of ischemia followed by 7 days of reperfusion and treated daily with 60 mg kg-1 curcumin. The liver and kidneys were collected, weighed, and biochemically analyzed. Intestinal ischemia and reperfusion increased levels of lipid hydroperoxide (LOOH), decreased levels of reduced glutathione (GSH), and increased the enzymatic activity of superoxide dismutase (SOD), glutathione S-transferase (GST), and myeloperoxidase (MPO) in the liver. Intestinal ischemia and reperfusion decreased kidney weight and increased GST activity in the kidneys. Curcumin prevented these changes in the liver and kidneys. Intestinal ischemia and reperfusion mainly affected the liver, promoting inflammation and oxidative stress. The kidneys underwent repair much earlier than the liver, in which they did not present alterations of MPO or main parameters of oxidative stress after 7 days of reperfusion. Treatment with curcumin had beneficial effects, ameliorating or even preventing injury that was caused by intestinal ischemia and reperfusion in the liver and kidneys in rats

Toxicity evaluation of the active ingredient acetamiprid and a commercial formulation (Assail® 70) on the non-target gastropod Biomphalaria straminea (Mollusca: Planorbidae).

Neonicotinoids emerged as an environmentally safe alternative to previous generations of insecticides becoming one of the most widely applied in modern agriculture. Nevertheless, they have been reported to affect several non-target organisms. Most toxicity studies focus on the effects on pollinators or terrestrial invertebrates and evaluate either the active ingredient or the commercial formulation. In the present study, we aimed to assess the long-term effects of the active ingredient acetamiprid and a broadly used commercial formulation (Assail® 70) on the non-target freshwater gastropod Biomphalaria straminea using a battery of biomarkers. A 14 day-exposure of adult organisms to both active ingredient and commercial formulation increased carboxylesterase activity and glutathione content, inhibited superoxide dismutase activity and decreased reactive oxygen species levels. The commercial formulation additionally increased glutathione S-transferase activity and inhibited catalase activity. The results indicate a greater toxicity of the commercial formulation than that of the active ingredient alone. Cholinesterase activity, development and offspring survival of B. straminea were not impaired. We conclude that the toxicity of acetamiprid on this gastropod species is mainly related to effects on detoxification and oxidative metabolism responses. This study provides novel information about the adverse effects of the active ingredient and a commercial formulation of a widely used neonicotinoid on a non-target aquatic species.

Modification of the Bacterial Cell Wall—Is the Bioavailability Important in Creosote Biodegradation?

Creosote oil, widely used as a wood preservative, is a complex mixture of different polycyclic aromatic compounds. The soil contamination result in the presence of a specific microcosm. The presented study focuses on the most active strains involved in bioremediation of long-term creosote-contaminated soil. In three soil samples from different boreholes, two Sphingomonas maltophilia (S. maltophilia) and one Paenibacillus ulginis (P. ulginis) strain were isolated. The conducted experiments showed the differences and similarities between the bacteria strains capable of degrading creosote from the same contaminated area. Both S. maltophilia strains exhibit higher biodegradation efficiency (over 50% after 28 days) and greater increase in glutathione S-transferase activity than P. ulginis ODW 5.9. However, S. maltophilia ODW 3.7 and P. ulginis ODW 5.9 were different from the third of the tested strains. The growth of the former two on creosote resulted in an increase in cell adhesion to Congo red and in the total membrane permeability. Nevertheless, all three strains have shown a decrease in the permeability of the inner cell membrane. That suggests the complex relationship between the cell surface modifications and bioavailability of the creosote to microorganisms. The conducted research allowed us to broaden the current knowledge about the creosote bioremediation and the properties of microorganisms involved in the process.

Side-effects of a neonicotinoid insecticide (actara®) on a non-target larvivorous fish Gambusia affinis: Growth and biomarker responses

Actara® 25 WG belongs to neonicotinoids, which is the most widely used insecticide class for controlling various insect pests in the world. The present study was aimed to assess the potential hazards of this insecticide on the mosquitofish Gambusia affinis following a chronic exposure. The female adults were collected during the reproduction period and exposed to various concentrations (10, 20 and 40 mg.L−1) during 28 days. The effect of actara was investigated on morphometric and selected biomarker measurements. Results showed that the compound was found to affect significantly the condition factor at days 21 and 28 of exposure with the highest concentration. However, the treatment had no effect on hepato-somatic and gonado-somatic indices. Biochemical analyses revealed an increase in glutathione S-transferase activity and carbonylated protein content, and a decrease in acetylcholinesterase activity for the three tested concentrations while the vitellin level was reduced only for the highest concentration. Thus, this product can pose side-effects on this non-target freshwater fish.

Enzyme response of activated sludge to a mixture of emerging contaminants in continuous exposure.

The relevant information about the impacts caused by presence of emerging pollutants in mixtures on the ecological environment, especially on the more vulnerable compartments such as activated sludge (AS) is relatively limited. This study investigated the effect of ibuprofen (IBU) and triclosan (TCS), alone and in combination to the performance and enzymatic activity of AS bacterial community. The assays were carried out in a pilot AS reactor operating for two-weeks under continuous dosage of pollutants. The microbial activity was tracked by measuring oxygen uptake rate, esterase activity, oxidative stress and antioxidant enzyme activities. It was found that IBU and TCS had no acute toxic effects on reactor biomass concentration. TCS led to significant decrease of COD removal efficiency, which dropped from 90% to 35%. Continuous exposure to IBU, TCS and their mixtures increased the activities of glutathione s-transferase (GST) and esterase as a response to oxidative damage. A high increase in GST activity was associated with non-reversible toxic damage while peaks of esterase activity combined with moderate GST increase were attributed to an adaptive response.

Molecular characterization, expression and enzyme activity of three glutathione S-transferase genes from Eriocheir sinensis under pesticide stresses

Glutathione S-transferases (GSTs) are a multifunctional protein superfamily that can catalyze the detoxification processes in an organism. In the present study, we determined the structure and function of GSTs in Chinese mitten crab (Eriocheir sinensis) by gene cloning, expression, and enzyme activity in order to investigate the metabolic detoxification of GSTs in the hepatopancreas and muscles under three pesticide (trichlorfon, β-cypermethrin and avermectin) stresses. Multiple sequence alignment analysis showed that all the three Es-GST genes possessed N-terminal, and C-terminal domain as well as G-binding sites, while Es-GST2 and Es-GST3 contained Mu-type GST-specific Mu-loop structures. Phylogenetic tree analysis revealed that the three Es-GSTs belonged to the Mu-type GST of crustaceans. The quantitative real-time PCR revealed that the three Es-GSTS were expressed in 9 tissues of Eriocheir sinensis, with highest expression in hepatopancreas and muscle. The expression of the three Es-GSTS significantly increased in the hepatopancreas and muscle under the three pesticide stresses compared to the control group, and a steady increase in GST activity was observed. The study showed that the three Es-GSTs belong to the Mu-type GST of the crustaceans and might play an important role in the metabolic detoxification in Eriocheir sinensis.

Simultaneous exposure to vinylcyclohexene and methylmercury in Drosophila melanogaster: biochemical and molecular analyses

BackgroundExposure to vinylcyclohexene (VCH) and methylmercury (MeHg+) can induce oxidative stress and gene modulation. Several studies have been evaluating the effects of VCH and MeHg+, but little is known about interactive effects between them. This work aimed to assess the exposure and co-exposure effects of MeHg+ and VCH on oxidative stress and gene modulation in Drosophila melanogaster.MethodsReactive species production, glutathione S-transferase (GST) and acetylcholinesterase (AChE) activities were evaluated after exposure and co-exposure to VCH (1 mM) and MeHg+ (0.2 mM) for one or three days in the head and body (thorax and abdomen) of flies. The expression of genes related to redox state and inflammatory response was evaluated after exposure and co-exposure to VCH and MeHg+ for three days.ResultsSurvival decreased only in flies co-exposed to VCH and MeHg+ for three days. All treatments increased total reactive species production after one day of exposure. However, no significant changes were observed in the head after three days of exposure. One day of exposure to VCH caused an increase in the head GST activity, whereas MeHg+ induced an increase after three days of exposure. Regarding the body, all treatments increased GST activity after one day of exposure, but only the flies exposed to MeHg+ presented an increase in GST activity after three days of exposure. Treatments did not alter AChE activity in the head. As for gene expression, there was a significant increase in the Relish transcription factor gene in the flies’ body, but Nrf2, Keap1, Jafrac1, TrxR1, and NF-κβ were not altered.ConclusionThe results suggest that exposure to VCH and MeHg+ induce oxidative stress and activation of an inflammatory response in fruit flies.

An imazethapyr-based herbicide formulation induces genotoxic, biochemical, and individual organizational effects in Leptodactylus latinasus tadpoles (Anura: Leptodactylidae).

Genotoxic, biochemical, and individual organizational effects on Leptodactylus latinasus tadpoles were evaluated after exposure to an imazethapyr (IMZT)-based commercial herbicide formulation, Pivot® H (10.59% IMZT). A determination of the value of the lethal concentration (LC50) was determined as a toxicological endpoint. Alterations in animal behavior and morphological abnormalities as well as cholinesterase (ChE), catalase (CAT), and glutathione S-transferase (GST) activities were employed as individual sublethal endpoints. Micronuclei frequencies (MNs), binucleated cells (BNs), blebbed nuclei (BLs), lobed nuclei (LBs), notched nuclei (NTs), erythroplastids (EPs), and evaluation of DNA strand breaks were employed as genotoxic endpoints. All biomarkers were evaluated after 48 and 96 h of exposure to concentrations of IMZT within 0.07-4.89 mg/L. LC5096h values of 1.01 and 0.29 mg/L IMZT were obtained for Gosner stages 25 and 36, respectively. Irregular swimming, diamond body shape, and decreased frequency of keratodonts were detected at both sampling times. Results showed that IMZT increased GST activity and MN frequency at 48 and 96 h of exposure. Other nuclear abnormalities were also observed in the circulating erythrocytes of tadpoles, i.e., NT and BL values after 48 h, and LN, BL, and EP values after 96 h. Finally, results showed that IMZT within 0.07-0.22 mg/L increased the genetic damage index in tadpoles exposed for both exposure times (48 and 96 h). This study is the first to report the sublethal biochemical effects of IMZT in anurans and is also the first report using L. latinasus tadpoles as a bioindicator for ecotoxicological studies.

Cigarette smoking and antioxidant defences in packed red blood cells prior to storage.

Red blood cells from smoking donors can have more lesions from oxidative stress, decreasing the benefits of blood transfusion. We aimed to explore the effect of cigarette smoking on the oxidative status of packed red blood cells (PRBCs) prior to storage. We compared serum vitamin C, plasmatic malondialdehyde (MDA), and non-protein thiol groups (GSH) levels in PRBCs, as well glutathione peroxidase (GPx) and glutathione s-transferase (GST) activity in PRBCs from smoking (n=36) and non-smoking (n=36) donors. We also correlated urinary cotinine levels with these parameters. Cigarette smoking was associated with decreased serum levels of vitamin C and GPx, and increased GST activity in PRBCs. We found negative correlations between cotinine, GPx activity and vitamin C levels, and a positive correlation between cotinine and GST activity. Cigarette smoking changed antioxidant defences of PRBCs prior to storage and these parameters are correlated with cotinine levels. Increased RBC antioxidants such as GST may reflect an exposure to oxidants during erythropoiesis. Because of the inability of mature RBCs to resynthesise antioxidants, PRBCs from smokers may have higher risk of storage lesions than those from non-smoker donors.

Alternative NADH dehydrogenase extends lifespan and increases resistance to xenobiotics in Drosophila

Mitochondrial alternative NADH dehydrogenase (aNDH) was found to extend lifespan when expressed in the fruit fly. We have found that fruit flies expressing aNDH from Ciona intestinalis (NDX) had 17–71% lifespan prolongation on media with different protein-tocarbohydrate ratios except NDX-expressing males that had 19% shorter lifespan than controls on a high protein diet. NDX-expressing flies were more resistant to organic xenobiotics, 2,4-dichlorophenoxyacetic acid and alloxan, and inorganic toxicant potassium iodate, and partially to sodium molybdate treatments. On the other hand, NDX-expressing flies were more sensitive to catechol and sodium chromate. Enzymatic analysis showed that NDX-expressing males had higher glucose 6-phosphate dehydrogenase activity, whilst both sexes showed increased glutathione S-transferase activity.

The impact of polar fraction of the fine particulate matter on redox responses in different rat tissues.

Particulate matter (PM) contains different chemical substances that have been associated with health effects and an increased risk of mortality due to their toxicity. In this study, fine particulate matter (PM2.5) samples were collected in a region with rural characteristics (Seropédica (Se)) and another with some industries (Duque de Caxias (DC)) (Brazil, RJ). Rats were exposed to PM2.5 extracts daily for 25 days at different dilutions: 10×, 5×, and a concentrated solution (CS). Biochemical analyses were investigated for total antioxidant capacity (ACAP), lipid peroxidation (LPO) levels, reduced glutathione (GSH) concentration, activity of glutamate cysteine ligase (GCL), and activity of glutathione S-transferase (GST). The liver showed a significant increase in GCL (DC-5×, DC-CS and Se-CS) and GST activities (DC-CS and Se-CS) in both regions when compared to the control group. In the renal cortex, GCL activity decreased in most of the tested groups while GST activity increased only in the 5× groups of both regions (DC and Se). In the renal medulla, GCL activity decreased for Se-10× and DC-CS but increased for Se-5×, and GST activity increased in the Se-10×, DC-5×, and DC-CS groups. Lung GCL increased in all groups for both regions. Moreover, this organ also showed an increase in GST activity when higher metal concentrations were present (5× and CS). TBARS levels were increased for all tissues in most tested concentrations. These data indicate that soluble compounds (e.g., metals) from PM2.5 sampled in areas with different pollution indexes can change the redox status and cause damage to different tissues.

Quercetin Mediated Salt Tolerance in Tomato through the Enhancement of Plant Antioxidant Defense and Glyoxalase Systems.

Quercetin (Qu) is a strong antioxidant among the phenolic compounds having physiological and biochemical roles in plants. Hence, we have studied the Qu evolved protection against salinity in tomato (Solanum lycopersicum L.). Salinity caused ionic toxicity by increasing Na+ content in seedlings along with nutritional starvation of K+, Ca2+, and Mg2+. While osmotic stress was detected by higher free proline (Pro) content and lower leaf relative water content (LRWC) in salt-stressed seedlings. Salt toxicity also induced higher H2O2 generation, malondialdehyde (MDA) content and lipoxygenase (LOX) activity as a sign of oxidative stress. Tomato seedlings suffered from methylglyoxal (MG) toxicity, degradation of chlorophyll, along with lower biomass accumulation and growth due to salt exposure. However, Qu application under salinity resulted in lower Na+/K+ due to reduced Na+ content, higher LRWC, increased Pro, and reduction of H2O2 and MDA content, and LOX activity, which indicated alleviation of ionic, osmotic, and oxidative stress respectively. Quercetin caused oxidative stress, lessening through the strengthening of both enzymatic and non-enzymatic antioxidants. In addition, Qu increased glutathione S-transferase activity in salt-invaded seedlings, which might be stimulated reactive oxygen species (ROS) scavenging along with higher GSH content. As a result, toxic MG was detoxified in Qu supplemented salt-stressed seedlings by increasing both Gly I and Gly II activities. Moreover, Qu insisted on better plant growth and photosynthetic pigments synthesis in saline or without saline media. Therefore, exogenous applied Qu may become an important actor to minimize salt-induced toxicity in crops.

Behavioural and biochemical effects of one-week exposure to aflatoxin B1 and aspartame in male Wistar rats

Food products are susceptible to contamination by mycotoxins, and aflatoxin B1 (AFB1) stands as the most toxic among them. AFB1 intoxication results in distinct signs, including widespread systemic toxicity. Aspartame (ASP) is an artificial sweetener used as a sugar substitute in many products, and compelling evidence indicates ASP can be toxic. Interestingly, mechanisms underlying ASP and AFB1 toxicity involve oxidative stress. In this context, concomitant use of ASP and AFB1 in a meal may predispose to currently unidentified behavioural and biochemical changes. Therefore, we evaluated the effect of AFB1 (250 μg/kg, intragastrically (i.g.)) and/or ASP (75 mg/kg, i.g.) exposure for 7 days on behavioural and biochemical markers of oxidative stress in male Wistar rats. AFB1 and/or ASP increased hepatic glutathione S-transferase (GST) activity when compared to controls. In the kidneys, increased GST activity was detected in AFB1 and AFB1+ASP groups. In addition, AFB1 and or ASP elicited behavioural changes in the open field, marble burying and splash tests, however no additive effects were detected. Altogether, present data suggest AFB1 and ASP predispose to anxiety- and obsessive-compulsive-like symptoms, as well as to enzymatic defence system imbalance in liver and kidney of Wistar rats.

The Effect of Niclosamide on Certain Biological and Biochemical Properties of Drosophila melanogaster

ABSTRACT Objective: Laboratory research on management strategies such as biological and chemical control usually demands rearing high numbers of high-quality insects. For this reason, there is a need for detailed information about which antimicrobial agents should be added to their diets in order to obtain high quality insects. Materials and methods: Niclosamide, which is an anthelmintic derivative of salisilanilid, was added in amounts of 100, 200, 400 and 800 mg/L into artificial diets of Drosophila melanogaster (Meigen). The effects on survival rate, developmental time and adult longevity of the insects were examined. Furthermore, the effect of this anthelmintic antibiotic on important oxidative stress indicators; lipid peroxidation product, malondialdehyde (MDA) and protein oxidation products; protein carbonyl (PCO) contents and a detoxification enzyme, glutathione S-transferase (GST) activity in 3rd instar larvae, pupae and adult stage of the insect were also investigated. Results: Compared with those insects on the control diet, the tested concentrations of niclosamide significantly decreased survival rate in all developmental stages of the insect. The control diet produced 94.0 ± 1.0 % of 3rd stage larvae whereas in the highest concentration (800 mg/L) this ratio decreased to 14.00 % ± 1.73. While 42.08 ± 0.50 % of the adults survived in the control diet, longevity decreased to 2.30 ± 0.15 days in the highest concentration. All concentrations of niclosamide resulted in increased MDA contents in the last larval stage of D. melanogaster and concentrations of 100, 200 and 800 mg/L niclosamide increased pupal MDA content in comparison to the control group. At low concentrations of niclosamide, PCO decreased in the last stage of larvae. The diet with 400 mg/L niclosamide concentration significiantly increased GST activity in pupal stage. Conclusion: The results of this work indicated that the negative effects of niclosamide on biological characteristics of D. melanogaster are due to an increase in the oxidative stress and crippled detoxification capacity of the insect.

Effects of atrazine on life parameters, oxidative stress, and ecdysteroid biosynthetic pathway in the marine copepod Tigriopus japonicus

Atrazine is a widely used pesticide which acts as an endocrine disruptor in various organisms. The aim of this study was to investigate adverse effects of atrazine on life parameters, oxidative stress, and ecdysteroid biosynthetic pathway in the marine copepod Tigriopus japonicus. In T. japonicus, no mortality was shown in response to atrazine up to 20 mg/L in acute toxicity assessment. In nauplii, retardation in the growth and prolonged molting and metamorphosis resulted under chronic exposure of atrazine at 20 mg/L. In addition, body sizes of T. japonicus nauplii were significantly decreased (P < 0.01 in length and P < 0.001 in width) in response to 20 mg/L of atrazine. Furthermore, atrazine induced oxidative stress by the generation of reactive oxygen species at all concentrations compared to the control in the nauplii. Also, significant increase in glutathione-S transferase activity was observed in adult T. japonicus at low concentration of atrazine. To understand effects of atrazine on ecdysteroid biosynthetic pathway-involved genes (e.g., neverland, CYP307E1, CYP306A1, CYP302A1, CYP3022A1 [CYP315A1], CYP314A1, and CYP18D1) were examined with mRNA expressions of ecdysone receptor (EcR) and ultraspiracle (USP) in response to 20 mg/L atrazine in nauplii and adults. In the nauplii, these genes were significantly downregulated (P < 0.05) in response to atrazine, compared to the control but not in the adult T. japonicus. These results suggest that atrazine can interfere in vivo life parameters by oxidative stress-induced retrogression and ecdysteroid biosynthetic pathway in this species.

Toxicity of oxalic acid and impact on some antioxidant enzymes on in vitro-reared honeybee larvae.

Nowadays, Varroa destructor is considered as a serious pest of honeybees (Apis mellifera) and its resistance to acaricides has been reported in Europe since the early 1990s. That is why new methods of treatment for Varroa mites are still in focus of many scientists. In our study, we determined the lethal concentration LC50 (72 h) of 2.425% oxalic acid solution following single spray exposure of honeybee larvae under laboratory conditions (Guideline OECD 237 2013). Potential sublethal effects of oxalic acid were monitored through the determination of the activity of antioxidant enzymes. Activation of primary antioxidant enzymes was observed at 1.75% of oxalic acid; 3.5% of oxalic acid brought on a statistically significant increase of glutathione S-transferase activity. This change was accompanied by an increase in thiobarbituric acid reactive substances, products of lipid peroxidation. Our results indicate that oxalic acid may be harmful to bee brood when present during application.