Increased Glutathione S-transferase Activity Research Articles

Biochemical and histopathological responses in Nile tilapia exposed to a commercial insecticide mixture containing dinotefuran and lambda-cyhalothrin

The indiscriminate use of pesticides has led to an increased risk of environmental contamination and pest resistance worldwide, favoring the development of less hazardous formulations. The commercial insecticide ZEUS® (Ihara, Brazil) combining dinotefuran and lambda-cyhalothrin was recently formulated in order to meet the environmental sustainability and food security. However, little is known about the potential toxic effects of ZEUS® to aquatic species. Thus, we report, for the first time, the biochemical and histological responses in tilapia (Oreochromis niloticus) following 96 h exposure to 0.01 mg/L, 0.05 mg/L and 0.1 mg/L ZEUS®. Different biochemical endpoints, including acetylcholinesterase (AChE), gamma-glutamyltransferase (GGT) and alkaline phosphatase (ALP), were assessed as potential biomarkers of insecticide effects. Glutathione S-transferase (GST) was evaluated as a marker of phase II biotransformation, and histopathological changes were measured to indicate gill alterations following ZEUS® exposure. After 96 h exposure, ZEUS® treatment increased GST activity in the liver of fish exposed to the highest concentration, while the intermediate dose increased both renal GGT and hepatic ALP activities. These findings reflect the importance of the liver and kidneys in the detoxification of ZEUS® and highlight the need to understand further toxicity effects. Likewise, the histopathological analysis of gills provided evidence that ZEUS® caused moderate damages. Despite biomarkers alterations reported for O. niloticus following ZEUS® exposure, by comparing our findings with data on toxicity of individual compounds, the commercial ZEUS® mixture seems to present similar or even lower adverse effects on freshwater fish.

Pyrethroid insecticide and milkweed cardenolide interactions on detoxification enzyme activity and expression in monarch caterpillars

Declines of the monarch butterfly population have prompted large-scale plantings of milkweed to restore the population. In North America, there are >73 species of milkweed to choose from for these nationwide plantings. However, it is unclear how different milkweed species affect monarch caterpillar physiology, particularly detoxification enzyme activity and gene expression, given the highly variable cardenolide composition across milkweed species. Here, we investigate the effects of a high cardenolide, tropical milkweed species and a low cardenolide, swamp milkweed species on pyrethroid sensitivity as well as detoxification enzyme activity and expression in monarch caterpillars. Caterpillars fed on each species through the fifth-instar stage and were topically treated with bifenthrin after reaching this final-instar stage. Esterase, glutathione S-transferase, and cytochrome P450 monooxygenase activities were quantified as well as the expression of selected esterase, glutathione S-transferase, ABC transporter, and cytochrome P450 monooxygenase transcripts. There were no significant differences in survival 24 h after treatment with bifenthrin. However, bifenthrin significantly increased glutathione S-transferase activity in caterpillars feeding on tropical milkweed and significantly decreased esterase activity in caterpillars feeding on tropical and swamp milkweed. Significant differential expression of ABC transporter, glutathione S-transferase, and esterase genes was observed for caterpillars feeding on tropical and swamp milkweed and not receiving bifenthrin treatment. Furthermore, significant differential expression of glutathione S-transferase and esterase genes was observed for bifenthrin-treated and -untreated caterpillars feeding on tropical milkweed relative to swamp milkweed. These results suggest that feeding on different milkweed species can affect detoxification and development mechanisms with which monarch caterpillars rely on to cope with their environment.

Biofilm forming rhizobacteria affect the physiological and biochemical responses of wheat to drought

Plant growth promoting rhizobacteria (PGPR) can attenuate the adverse effects of water deficit on plant growth. Since drought stress tolerance of bacteria has earlier been associated to biofilm formation, we aimed to investigate the role of bacterial biofilm formation in their PGPR activity upon drought stress. To this end, a biofilm-forming bacterial collection was isolated from the rhizospheres of native arid grassland plants, and characterized by their drought tolerance and evaluated on their plant growth promoting properties. Most bacterial strains formed biofilm in vitro. Most isolates were drought tolerant, produced auxins, showed 1-aminocyclopropane-1-carboxylic acid (ACC) deaminase activity and solubilized mineral phosphate and potassium, but at considerably different levels. Greenhouse experiments with the most promising isolates, B1, B2 and B3, under three levels of water deficit and two wheat varieties led to an increased relative water content and increased harvest index at both moderate and severe water deficit. However, the bacteria did not affect these plant parameters upon regular watering. In addition, decreased hydrogen peroxide levels and increased glutathione S-transferase activity occurred under water deficit. Based on these results, we conclude that by improving root traits and antioxidant defensive system of wheat, arid grassland rhizospheric biofilm forming bacilli may promote plant growth under water scarcity.

Toxicity and Sublethal Effects of Fluxametamide on the Key Biological Parameters and Life History Traits of Diamondback Moth Plutella xylostella (L.)

Fluxametamide, a novel isoxazoline insecticide, is newly registered for the control of various lepidopteran, coleopteran and thysanopteran insect pests on lethal and sublethal levels. In the present study, the toxicity and sublethal effects of fluxametamide on diamondback moth Plutella xylostella (L.), an invasive lepidopteran foliage feeder of cruciferous vegetables, were assessed to explore its bio-ecological impact on pest populations. The toxicity of fluxametamide to the third instar larvae of P. xylostella was 0.18 mg L−1 (LC50) at 72 h bioassay. After treatment with LC10 and LC30 concentrations of fluxametamide, the fourth instar larval duration, the rate of deformed pupa and adults, and the adult pre-oviposition period were significantly increased, whereas the pupation rate and pupal weight were significantly decreased in the F0 generation. In the F1 generation, sublethal effects of fluxametamide were indicated by a reduced fecundity, rate of pupation and adult emergence, pupal weight, and adult longevity, however a significant increase in eggs and total larval duration, deformed adults rate, and total longevity and pre-oviposition period was observed in the offspring. The intrinsic rate of increase (r), finite rate of increase (λ) and net reproductive rate (R0) of sublethal treatments were significantly lower than the control. The relative fitness of F1 was 0.68 and 0.64 in LC10 and LC30 treatments, respectively. The LC30 fluxametamide treatment exhibited increased glutathione S-transferase activities (elevated 1.433-fold) in P. xylostella. Our results suggest that in addition to its high lethal toxicity, the sublethal concentrations of fluxametamide might suppress the reproduction, development and survival of the P. xylostella population and its progeny, which can help to optimize integrated pest management program.

Molecular Cloning of a TCHQD Class Glutathione S-Transferase and GST Function in Response to GABA Induction of Melon Seedlings under Root Hypoxic Stress

Glutathione-S-transferase (GST), a versatile enzyme that occurs widely in plants, plays a key role in plant resistance to environmental stresses. Previous results have demonstrated that GST proteins are involved in alleviating root hypoxic injury caused by gamma-aminobutyric acid (GABA); however, the induction mechanism of the GST gene in the melon under root hypoxic stress and its functional mechanisms remain unclear. In this study, based on gene cloning and bioinformatics analysis, GST gene expression and activity and glutathione (GSH) content were assessed under root hypoxic and normoxic conditions with or without GABA. The results showed that the CmGST locus includes an 804 bp gene sequence that encodes 267 amino acids. The sequence was highly similar to those of other plant TCHQD GSTs, and the highest value (94%) corresponded to Cucumis sativus. Real-time PCR results showed that the CmGST gene was induced by root hypoxic stress and GABA, and this induction was accompanied by increased GST activity and GSH content. Root hypoxic stress significantly upregulated CmGST expression in melon roots (0.5–6 d), stems, and leaves (0.5–4 d), and GST activity and GSH content were also significantly increased. Exogenous GABA treatment upregulated CmGST gene expression, GST activity, and GSH content, particularly under root hypoxic conditions. As a result, CmGST expression in GABA-treated roots and leaves at 0.5–4 d and stems at 0.5–6 d was significantly higher than that under root hypoxic stress alone. This study provides evidence that the TCHQD CmGST may play a vital role in how GABA increases melon hypoxia tolerance by upregulating gene expression and improving metabolism.

Chemoprotection mediated by açaí berry (Euterpe oleracea) in white shrimp Litopenaeus vannamei exposed to the cyanotoxin saxitoxin analyzed by in vivo assays and docking modeling.

Saxitoxin (STX) is a neurotoxic cyanotoxin that also generate reactive oxygen species, leading to a situation of oxidative stress and altered metabolism. The Amazonian fruit açaí Euterpe oleracea possesses a high concentration of antioxidant molecules, a fact that prompted us to evaluate its chemoprotection activity against STX toxicity (obtained from samples of Trichodesmium sp. collected in the environment) in the shrimp Litopenaeus vannamei. For 30 days, shrimps were maintained in 16 aquaria containing 10 shrimps (15% salinity, pH 8.0, 24°C, 12C/12D photoperiod) and fed twice daily with a diet supplemented with lyophilized açaí pulp (10%), in addition to the control diet. After, shrimps (7.21±0.04g) were exposed to the toxin added to the feed for 96h. Four treatments were defined: CTR (control diet), T (lyophilized powder ofTrichodesmium sp. 0.8μg/g), A (10% of açaí) and the combination T+A. HPLC analysis showed predominance of gonyautoxin-1 concentrations (GTX-1) and gonyautoxin-4 concentrations (GTX-4). The results of molecular docking simulations indicated that all variants of STX, including GTX-1, can be a substrate of isoform mu of the glutathione-S-transferase (GST) enzyme since these molecules obtained similar values of estimated Free Energy of Binding (FEB), as well as similar final positions on the binding site. GSH levels were reduced in muscle tissues of shrimp in the T, A, and T+A treatments. Increased GST activity was observed in shrimp hepatopancreas of the T treatment and the gills of the A and T+A treatments. A decrease of protein sulfhydryl groups (P-SH) was observed in gills of shrimps from T+A treatment. A reduction in malondialdehyde (MDA) levels was registered in the hepatopancreas of the T+A treatment in respect to the Control, T, and A treatments. The use of açaí supplements in L. vannamei feed was able to partially mitigate the toxic effects caused by Trichodesmium sp. extracts, and points to mu GST isoform as a key enzyme for saxitoxin detoxification in L. vannamei, an issue that deserves further investigation.

The effects of mepiquat chloride (DPC) on the soluble protein content and the activities of protective enzymes in cotton in response to aphid feeding and on the activities of detoxifying enzymes in aphids

BackgroundMepiquat chloride (DPC) enhances the resistance of cotton plants, and it is widely used as a growth regulator. DPC can stimulate photosynthesis, stabilize the structure of cotton leaves, and affect population reproduction and energy substances in Aphis gossypii Glover (cotton aphids), but interactions between DPC and cotton aphids remain unclear. In this study, we analyzed the physiological responses of cotton to DPC, and the toxicity of DPC toward cotton aphids, before and after feeding, to explore the DPC-induced defense mechanism against cotton aphids.ResultsMeasurements of protective enzyme activity in cotton showed that the soluble protein contents, peroxidase (POD) activity, and catalase (CAT) activity in cotton treated with different concentrations of DPC were higher than in the control. Superoxide dismutase (SOD) activity was higher than that of the control when the concentration of DPC was < 0.1 g/L. Under aphid feeding stress, POD activity in cotton treated with a low insect population density was significantly lower than in the controls, but the reverse was true for cotton treated with a high insect population density, and SOD activity was positively correlated with population density. The activities of detoxification enzymes in field and laboratory experiments showed that DPC promoted the specific activity of glutathione S-transferase (GST) in cotton aphids, while the specific activities of carboxylesterase (CarE) and acetylcholinesterase (AchE) were decreased.ConclusionsDPC enhanced the aphid resistance in cotton by increasing the soluble protein content and the activity of protective enzymes. It also had a toxic effect on cotton aphids by increasing GST activity (the main DPC target). DPC increased the soluble protein content and protective enzymes activity in cotton under aphid stress, and thereby enhanced tolerance to cotton aphids. It conclude that DPC interferes with cotton aphids through indirect (DPC induced cotton defense responses) and direct (DPC toxicity to cotton aphids) ways, which plays a positive role in interfering with cotton aphids.

Extract of Tagetes erecta L., a medicinal plant rich in lutein, promotes gastric healing and reduces ulcer recurrence in rodents

Ethnopharmacological relevanceTagetes erecta L. (Asteraceae), popularly known as Aztec Marigold, is used in South America to treat several ailments. Despite reports that T. erecta flowers are used in folk medicine to treat gastrointestinal diseases, there is no study regarding its gastric healing effects. Aim of the studyThe effect of dry extract of T. erecta L. (DETe) in gastric healing and gastric ulcer recurrence was evaluated, contributing to the validation of the antiulcer potential of this medicinal plant. MethodsRats were treated orally with vehicle (1 ml/kg), omeprazole (20 mg/kg), or DETe (3, 30 or 300 mg/kg) for 7 days, twice a day. The lesion area was evaluated, and the levels of reduced glutathione (GSH) and lipoperoxides (LOOH) and the activity of the superoxide dismutase (SOD), catalase (CAT), glutathione S-transferase (GST), and myeloperoxidase (MPO) were measured. The ulcer recurrence was evaluated in mice and induced by interleukin (IL)-1β (1 μg/kg, i.p). The recurred area, gastric wall thickness, GSH and cytokines levels, MPO and N-acetylglucosaminidase (NAG) activities were measured. ResultsDETe accelerated the healing of gastric ulcers only at 300 mg/kg, reducing the ulcerated area by 66%. In parallel, DETe reduced LOOH levels, SOD, CAT and MPO activities, while increasing GST activity and mucin amount. In the recurrence model, DETe reduced the lesion area by 94%, and in parallel decreased the gastric wall thickness and TNF levels, while increasing IL-10 amount. ConclusionsCorroborating the popular use of T. erecta, DETe favors the antioxidant system and reduce gastric inflammation, accelerating the gastric healing process and reducing the ulcer recurrence.

The role of nanoplastics on the toxicity of the herbicide phenmedipham, using Danio rerio embryos as model organisms

Once in the aquatic ecosystems, nanoplastics (NPls) can interact with other contaminants acting as vectors of transport and altering their toxicological effects towards organisms. Thus, the present study aims to investigate how polystyrene NPls (44 nm) interact with the herbicide phenmedipham (PHE) and affect its toxicity to zebrafish embryos. Single exposures to 0, 0.015, 0.15, 1.5, 15 and 150 mg/L NPls and 0.02, 0.2, 2 and 20 mg/L PHE were performed. Embryos were also exposed to the binominal combinations: 0.015 mg/L NPls + 2 mg/L PHE, 0.015 mg/L NPls + 20 mg/L PHE, 1.5 mg/L NPls + 2 mg/L PHE and 1.5 mg/L NPls + 20 mg/L PHE. Due to the low solubility of PHE in water, a solvent control was performed (0.01% acetone). PHE was quantified. Mortality, heartbeat and hatching rate, malformations appearance, locomotor behavior and biomarkers related to oxidative stress, neurotransmission and energy budgets were analyzed. During 96 h, NPls and PHE single and combined exposures did not affect embryos development. After 120 h, NPls induced hyperactivity and PHE induced hypoactivity. After 96 h, NPls increased catalase activity and PHE increased glutathione S-transferases activity. On the combination 0.015 mg/L NPls + 20 mg/L PHE, hyperactivity behavior was found, similar to 0.015 mg/L NPls, and cholinesterase activity was inhibited. Additionally, the combination 1.5 mg/L NPls + 20 mg/L PHE increased both catalase and glutathione S-transferases activities. The combination NPls with PHE affected more biochemical endpoints than the single exposures, showing the higher effect of the binominal combinations. Dissimilar interactions effects – no interaction, synergism and antagonism – between NPls and PHE were found. The current study shows that the effects of NPls on bioavailability and toxicity of other contaminants (e.g. PHE) cannot be ignored during the assessment of NPls environmental behavior and risks.

Effect of Heavy Metals on the Activity of Catalase and Glutathione-S-Transferase in Nile Tilapia Fish (Oreochromis niloticus)

This study evaluated the response of antioxidant enzymes, such as Catalase (CAT) and Glutathione-S-transferase (GST) in freshwater Nile tilapia fish Oreochromis niloticus (O. niloticus) exposed to heavy metals (HMs) including copper (Cu), lead (Pb) and cadmium (Cd). Fish were expose to various concentrations of Cu2+, Pb2+ (0, 0.02, 0.05, 0.2 mg/l) and Cd2+ (0, 0.005, 0.01, 0.05 mg/l) for 15, 30, 45 and 60 days. The results indicated that enzyme activity was different according to the exposure time, concentration and type of heavy metals. CAT activity increased significantly beginning at day 45 of HMs exposure. After 60 days of exposure, CAT activity was steady with Cu and Pb but inhibited by Cd. While, GST induction was earlier observed from day 15 of HMs exposure. The increase of GST activity was found with the increase of exposure time in the treatment with Cu and Cd but not with Pb. Interestingly, GST activity was inhibited by Pb at longer exposure (45 and 60 days). Among tested metals, Cu showed the weaker effect on the activity of CAT and GST in comparison with Pb and Cd suggesting that these enzymes were less sensitive to Cu than other tested metals.

The relationship between homocysteine, oxidative stress, and cognition in mild cognitive impairment

AbstractBackgroundHomocysteine (Hcy) generates reactive oxygen species and has been associated with cardiovascular disease and impaired cognition. While the relationship between Hcy and cognition has been extensively studied in other disorders, it has not been well characterized in mild vascular cognitive impairment (mVCI). Due to Hcy’s effect on vasculature and increased oxidative stress burden, individuals with elevated Hcy may be at greater risk of developing vascular dementia (VaD). This study investigated the relationship between Hcy, oxidative stress, and cognition.MethodParticipants were mVCI patients enrolled in a cardiac rehabilitation program. Cognition was assessed via National Institute of Neurological Disorders and Stroke and the Canadian Stroke Network (NINDS‐CSN) recommended battery, and blood was analyzed for Hcy and glutathione S‐transferase (GST), which modulates reactive oxygen species. Statistical analyses were performed using linear regression models in SPSS 26.ResultThere are 26 participants (73% male, 69% Caucasian, mean (SD) age=68.1 (7.2), years of education=15.7 (3.3)) with cardiovascular risk factors (62% hypertensive, 65% dyslipidemia, 31% diabetic, BMI=29.8 (5.1)) enrolled in the study to date. Mean plasma Hcy concentration was 13.4 μmol/L, with 9 participants classified as elevated (≥15 μmol/L). In preliminary analyses, plasma Hcy (stand. β=0.09, p=0.65) did not significantly predict executive function (F(2,23)=0.77, p=0.47) or verbal memory (F(2,23) =1.62, p=0.22). Hcy (stand. β = 0.684, p=0.013) significantly predicted GST activity (F(2,11) = 4.542, p=0.013, adj. R2 = 0.353), with a medium effect size.ConclusionFindings suggest that increased Hcy may be associated with compensatory increased GST activity in mVCI. A larger sample size will allow further analysis of cognition with additional covariates in this at‐risk population with increased oxidative stress.

Evaluation of antioxidant response and Na+-K+-ATPase activity in zebrafish exposed to polyethylene microplastics: Shedding light on a physiological adaptation

Although the toxicity of microplastics has already been demonstrated in different animal models, our knowledge about the response of freshwater fish to this pollutant is still limited. Thus, we aimed to evaluate the impact of exposure of zebrafish adults (Danio rerio) to environmentally relevant concentrations of polyethylene microplastics (PE-MPs) (5 and 50 µg/L) and at different times of exposure (10 and 20 days). Initially, scanning electron microscope image illustrated size and format of the particle and FTIR analysis confirmed the presence of PE-MPs in the gastrointestinal tract of fish (at both concentrations tested). Subsequently, an alteration of oxidative and antioxidant responses was evaluated in the liver and brain. The results showed that catalase (CAT) activity, in liver, was significantly decreased, as was glutathione S-transferases (GSTs) activity (on the 10th experimental day). However, after 20 days of exposure, we observed a concentration-dependent increase in GST activity in liver of the animals exposed to PE-MPs. Furthermore, the lipid peroxidation (LPO) level was significantly increased by exposure to MPs, especially in the brain, after 20 days of exposure. The increase in Na+-K+-ATPase activity in the animals' gills was correlated with the increased production of malondialdehyde (MDA), which suggests the existence of a compensatory mechanism in which the high activity of this enzyme would be necessary to regulate the loss of ions caused by the increase in the processes of LPO, which has never been previously demonstrated. Thus, our study sheds light on a new physiological adaptation to deal with the oxidative effects of PE-MPs, in addition to supporting the future use of the assessment of Na+/K+-ATPase activity as a biomarker of the toxicity of these pollutants.

Glyphosate and glufosinate ammonium, herbicides commonly used on genetically modified crops, and their interaction with microplastics: Ecotoxicity in anuran tadpoles

The effects of glyphosate (GLY)-based and glufosinate ammonium (GA)-based herbicides (GBH and GABH, respectively) and polyethylene microplastic particles (PEMPs) on Scinax squalirostris tadpoles were assessed. Tadpoles were exposed to nominal concentrations of both herbicides (from 1.56 to 100 mg L−1) and PEMPs (60 mg L−1), either alone or in combination, and toxicity evaluated at 48 h. Acetylcholinesterase (AChE), carboxylesterase (CbE), and glutathione-S-transferase (GST) activities were analyzed at the three lowest concentrations (1.56, 3.12 and 6.25 mg L−1, survival rates >85%) of both herbicides alone and with PEMPs. Additionally, the thermochemistry of the interactions between the herbicides and polyethylene (PE) was analyzed by Density Functional Theory (DFT). The median-lethal concentration (LC50) was 43.53 mg L−1 for GBH, 38.56 mg L−1 for GBH + PEMPs, 7.69 for GABH, and 6.25 mg L−1 for GABH+PEMPs. The PEMP treatment increased GST but decreased CbE activity, whereas GBH and GABH treatments increased GST but decreased AChE activity. In general, the mixture of herbicides with PEMPs increased the effect observed in the individual treatments: the highest concentration of GBH + PEMPs increased GST activity, whereas GABH+PEMP treatments decreased both AChE and CbE activities. DFT analysis revealed spontaneous interactions between the herbicides and PE, leading to the formation of bonds at the herbicide-PE interface, significantly stronger for GA than for GLY. The experimental and theoretical findings of our study indicate that these interactions may lead to an increase in toxicity when pollutants are together, meaning potential environmental risk of these combinations, especially in the case of GA.

Functional divergence of three glutathione transferases in two biotypes of the English grain aphid, Sitobion avenae

AbstractThe English grain aphid, Sitobion avenae (Fabricius) (Hemiptera: Aphididae), is a significant pest of cereal crops, but molecular factors and mechanisms that underpin its ability to develop differential biotypes on variable host plants are still not well understood. In this study, we investigated the interactions between two plant secondary metabolites (i.e., gramine and gallic acid) and three glutathione S‐transferases (GSTs) of S. avenae. Using artificial diets complemented or not with one of these two plant compounds, we found that gramine had relatively stronger negative effects on fitness of S. avenae biotype 3 (adapted to barley), and gallic acid on that of biotype 4 (adapted to wheat). Gramine significantly induced overexpression of SaveGST1 and SaveGST2 in biotype 4, but not in biotype 3. Gramine also reduced SaveGST3 expression in biotype 4, but not in biotype 3, suggesting biotype‐specific effect of GSTs’ regulation. In the treatments with gallic acid, the overexpression of SaveGST1, but not SaveGST2 or SaveGST3, was significantly induced in both biotypes, suggesting a critical role of SaveGST1 in detoxification of phenolic compounds such as gallic acid. The total constitutive GST activity was much higher in biotype 4 than in biotype 3. Significant increase in GST activity was obtained by the addition of both secondary metabolites in biotype 4, but not in biotype 3, showing significantly higher expression plasticity of GSTs in biotype 4. Thus, both constitutive and induced expression of GSTs could affect the adaptability of S. avenae on plants with variable secondary compounds, and thus contribute to the divergence of biotypes in this aphid species.

Delta class glutathione S-transferase (TuGSTd01) from the two-spotted spider mite Tetranychus urticae is inhibited by abamectin

GSTs (Glutathione S-transferases) are known to catalyze the nucleophilic attack of the sulfhydryl group of reduced glutathione (GSH) on electrophilic centers of xenobiotic compounds, including insecticides and acaricides. Genome analyses of the polyphagous spider mite herbivore Tetranychus urticae (two-spotted spider mite) revealed the presence of a set of 32 genes that code for secreted proteins belonging to the GST family of enzymes. To better understand the role of these proteins in T. urticae, we have functionally characterized TuGSTd01. Moreover, we have modeled the structure of the enzyme in apo form, as well as in the form with bound inhibitor. We demonstrated that this protein is a glutathione S-transferase that can conjugate glutathione to 1-chloro-2,4-dinitrobenzene (CDNB). We have tested TuGSTd01 activity with a range of potential substrates such as cinnamic acid, cumene hydroperoxide, and allyl isothiocyanate; however, the enzyme was unable to process these compounds. Using mutagenesis, we showed that putative active site variants S11A, E66A, S67A, and R68A mutants, which were residues predicted to interact directly with GSH, have no measurable activity, and these residues are required for the enzymatic activity of TuGSTd01. There are several reports that associate some T. urticae acaricide resistance with increased activity of GSTs . However, we found that TuGSTd01 is not able to detoxify abamectin; in fact, the acaricide inhibits the enzyme with Ki = 101 μM. Therefore, we suggest that the increased GST activity observed in abamectin resistant T. urticae field populations is a part of the compensatory feedback loop. In this case, the increased production of GSTs and relatively high concentration of GSH in cells allow GSTs to maintain physiological functions despite the presence of the acaricide.

Evaluation of the effects of two anaesthetic protocols on oxidative status and DNA damage in red-eared sliders (Trachemys scripta elegans) undergoing endoscopic coeliotomy.

The aim of this study was to assess how red-eared sliders (Trachemys scripta elegans) respond to anaesthesia itself and coelioscopy. For that purpose, the turtles were anaesthetised with ketamine-medetomidine or propofol, and the activities of superoxide dismutase (SOD), catalase (CAT) and glutathione S-transferase (GST) and the level of malondialdehyde (MDA) were determined by spectrophotometry. The possible genotoxic effects of the anaesthetic agents were estimated by comet assay. A total of 24 turtles were included in this study. The animals were divided into four groups according to the anaesthetic protocol and according to whether endoscopy would be performed. Significantly decreased activities of CAT were found only in the propofol group and in turtles undergoing coelioscopy. Both anaesthetic protocols induced significantly increased MDA levels, while no differences were observed after the intervention. A significant increase in GST activity was detected in turtles after both anaesthetic protocols, but after coelioscopy significant changes in GST activity were found only in the propofol group. However, no differences in SOD activity and no DNA damages were detected in either group. These findings suggest that ketamine-medetomidine may be more suitable anaesthetic agents in red-eared sliders than propofol.

Effects of Temperature and Glyphosate on Fatty Acid Composition, Antioxidant Capacity, and Lipid Peroxidation in the Gastropod Lymneae sp.

Little is known about the potential effects of glyphosate on freshwater gastropods and possible interactions between glyphosate and other stressors. A two-way factorial experiment was conducted to investigate the effects of temperature (20 °C/25 °C) and glyphosate (0 µg/L/200 µg/L) on Lymnaea sp. After 21 days, antioxidant capacity (superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx) and glutathione-S-transferase (GST)), malondialdehyde content (MDA), and fatty acid (FA) composition of Lymnaea sp. tissue were measured. Temperature had an effect on SOD activity and GPx activity. In contrast, an increase in GST activity was observed in glyphosate-exposed snails, highlighting the role of GST in the glyphosate detoxification process. Differences in temperature and glyphosate did not affect lipid peroxidation (MDA); however, we observed a trend suggesting the presence of higher MDA content in glyphosate-exposed snails at 20 °C. The FA groups were generally not strongly affected by the treatments, except for omega−9 (n-9) that was markedly lower at the higher temperature. Changes were also observed in individual FA as a response to glyphosate and/or temperature. For example, a significant decrease in 18:1n9 was observed at 25 °C. Our results showed that antioxidant capacity and FA profiles were mainly affected by temperature, while glyphosate seemed to have a lesser impact.

ACC deaminase-producing Ensifer adhaerens KS23 enhances proximate nutrient of Pisum sativum L. cultivated in high altitude.

A phytohormone producing, N2-fixing and 1-aminocyclopropane-1-carboxylate (ACC) deaminase synthesizing bacterium Ensifer adhaerens KS23 effectively increased the yield and nutritional contents of Pisum sativum var. Arkel. The isolate KS23 showed positive ACC deaminase activity with 174.2 (nmol of α-ketobutyrate/g-1 biomass½ h-1) a 9.7-fold increase in glutathione S-transferase activity. The proximate analysis exhibited an increased yield of protein (21.45%), carbohydrate (38.90%), sulphur (29.94%) starch (27.52%), total ash (35.57%), fat content (27.5%), nitrogen (24.06%) and hydrogen (17.91%) in treated seeds of P. sativum as compared to untreated crop seeds in field trials at Srikot village, Srinagar-246,174 (Garhwal) India. The most desirable essential and non-essential amino-acids content was also enhanced simultaneously by E. adhaerens KS23 as compared to non-treated crop seeds. This study revealed the enhancement of various nutritional contents resulting in quality improvement and an increase in growth productivity of pea. This study provides an encouraging result that may benefit the marginal income of farmers belonging mainly to hilly regions who are dependent on traditional methods of farming and thus improving their economy.

Pyriproxyfen induces intracellular calcium overload and alters antioxidant defenses in Danio rerio testis that may influence ongoing spermatogenesis.

We investigated the invitro effects of pyriproxyfen on ionic balance in the testis of the zebrafish by measuring 45Ca2+ influx. Invivo pyriproxyfen treatment was carried out to study oxidative stress, and conduct morphological analysis of the testis and liver. Whole testes were incubated invitro with/without pyriproxyfen (10-12, 10-9 or 10-6M; 30min) and 45Ca2+ influx determined. To study pyriproxyfen's mechanism of action, inhibitors/activators of ionic channels or pumps/exchangers, protein kinase inhibitors or a calcium chelator were added 15min before the addition of 45Ca2+ and pyriproxyfen. We evaluated the invivo effects of 7 day exposure to waterborne pyriproxyfen (10-9M) on reactive oxygen species (ROS) formation, lipid peroxidation, and reduced glutathione content (GSH), glutathione S-transferase (GST), superoxide dismutase (SOD), catalase (CAT) and γ-glutamyltransferase (GGT) activity. Morphological analyses of the testis and liver were carried out after invivo exposure of D.rerio to pyriproxyfen. Pyriproxyfen increased 45Ca2+ influx by opening the voltage-dependent T-type channels (T-type VDCC), inhibiting sarco/endoplasmic reticulum 45Ca2+-ATPase (SERCA) and the NCX exchanger (forward mode) and by mobilizing calcium from stores. The involvement of potassium channels and protein kinase C (PKC) was also demonstrated in pyriproxyfen-induced intracellular calcium elevation. Invivo pyriproxyfen treatment of D.rerio increased lipid peroxidation, decreased GSH content and increased GST activity in testes, in addition to increasing the number and size of spermatogonia cysts and inducing hepatocyte basophilia and dilation of blood vessels in the liver. The toxicity of pyriproxyfen is mediated by calcium overload, increased lipid peroxidation, and a diminished antioxidant capacity in the testis, due to GSH depletion, and altered spermatogenesis. The development of high basophilia in the liver suggests that pyriproxyfen may have estrogenic activity, possibly acting as an endocrine-disruptor. These findings indicate that these alterations may contribute to pyriproxyfen toxicity and spermatogenesis disruption.

Herbicide safener increases weed-management tools for control of annual grasses in wheat

AbstractAnnual grass weeds reduce profits of wheat farmers in the Pacific Northwest. The very-long-chain fatty acid elongase (VLCFA)-inhibiting herbicides S-metolachlor and dimethenamid-P could expand options for control of annual grasses but are not registered in wheat, because of crop injury. We evaluated a safener, fluxofenim, applied to wheat seed for protection of 19 soft white winter wheat varieties from S-metolachlor, dimethenamid-P, and pyroxasulfone herbicides; investigated the response of six varieties (UI Sparrow, LWW 15-72223, UI Magic CL+, Brundage 96, UI Castle CL+, and UI Palouse CL+) to incremental doses of fluxofenim; established the fluxofenim dose required to optimally protect the varieties from VLCFA-inhibiting herbicides; and assessed the impact of fluxofenim dose on glutathione S-transferase (GST) activity in three wheat varieties (UI Sparrow, Brundage 96, and UI Castle CL+). Fluxofenim increased the biomass of four varieties treated with S-metolachlor or dimethenamid-P herbicides and one variety treated with pyroxasulfone. Three varieties showed tolerance to the herbicides regardless of the fluxofenim treatment. Estimated fluxofenim doses resulting in 10% biomass reduction of wheat ranged from 0.55 to 1.23 g ai kg−1 seed. Fluxofenim doses resulting in 90% increased biomass after treatment with S-metolachlor, dimethenamid-P, and pyroxasulfone ranged from 0.07 to 0.55, 0.09 to 0.73, and 0.30 to 1.03 g ai kg−1 seed, respectively. Fluxofenim at 0.36 g ai kg−1 seed increased GST activity in UI Castle CL+, UI Sparrow, and Brundage 96 by 58%, 30%, and 38%, respectively. These results suggest fluxofenim would not damage wheat seedlings up to three times the rate labeled for sorghum, and fluxofenim protects soft white winter wheat varieties from S-metolachlor, dimethenamid-P, or pyroxasulfone injury at the herbicide rates evaluated.