Integrated spectroscopic and computational analyses unravel the molecular interaction of pesticide azinphos-methyl with bovine beta-lactoglobulin.

Organophosphorus are typically hazardous chemicals used in the pharmaceutical, agricultural, and other industries. They pose a serious risk to human life and can be fatal upon direct exposure. Hence, studying the interaction between such compounds with proteins is crucial for environmental, health, and food safety. In this study, we investigated the interaction mechanism between azinphos-methyl (AZM) and ß-lactoglobulin (BLG) at pH 7.4 using a combination of biophysical techniques. Intrinsic fluorescence investigations revealed that BLG fluorescence was quenched in the presence of increasing AZM concentrations. The quenching mechanism was identified as static, as evidenced by a decrease in the fluorescence quenching constant (1.25 × 104, 1.18 × 104, and 0.86 × 104 M-1) with an increase in temperatures. Thermodynamic calculations (ΔH > 0; ΔS > 0) affirmed the formation of a complex between AZM and BLG through hydrophobic interactions. The BLG's secondary structure was found to be increased due to AZM interaction. Ultraviolet -visible spectroscopy data showed alterations in BLG conformation in the presence of AZM. Molecular docking highlighted the significant role of hydrophobic interactions involving residues such as Val43, Ile56, Ile71, Val92, Phe105, and Met107 in the binding between BLG and AZM. A docking energy of -6.9 kcal mol-1, and binding affinity of 1.15 × 105 M-1 suggest spontaneous interaction between AZM and BLG with moderate to high affinity. These findings underscore the potential health risks associated with the entry of AZM into the food chain, emphasizing the need for further consideration of its impact on human health.

Cuticular proteins in codling moth (Cydia pomonella) respond to insecticide and temperature stress.

The insect cuticle consists of chitin and cuticular proteins (CPs), which stabilize the body shape and provide an effective physical barrier against the external environment. They are also potential target sites for developing environmentally friendly insect management through the utilization of physiology-based methods. The codling moth, Cydia pomonella, is a pest afflicting fruit orchards worldwide. This study used a comparative genomic approach, whole-genome resequencing, and transcriptome data to understand the role that CPs played in the environmental adaptation of the codling moth. A total of 182 putative CPs were identified in the codling moth genome, which were classified into 12 CP families. 119 CPR genes, including 54 RR-1, 60 RR-2, and 5 RR-3 genes were identified and accounted for 65.4% of the total CPs. Eight and seven gene clusters are formed in RR1 and RR2 subfamily and the ancestor-descendant relationship was explained. Five CPAP genes were highly expressed during the egg stage and exposed to high temperature, which indicated their potential role in aiding codling moth eggs in acclimating to varying external heat conditions. Moreover, six CPs belonging to the CPR and CPLCP families were identified in association with insecticide resistance by population resequencing. Their expression levels increased after exposure to insecticides, suggesting they might be involved in codling moth resistance to the insecticides azinphos-methyl or deltamethrin. Our results provide insight into the evolution of codling moth CPs and their association with high temperature adaptation and insecticide resistance, and provide an additional information required for further analysis of CPs in environmental adaptation.

Interactive effects of high temperature and pesticide exposure on oxidative status, apoptosis, and renin expression in kidney of goldfish: Molecular and cellular mechanisms of widespread kidney damage and renin attenuation.

One of many noteworthy consequences of increasing societal reliance on pesticides is their predominance in aquatic environments. These pernicious chemicals interact with high temperatures from global climate change, heat waves, and natural variations to create unstable environments that negatively impact organisms' health. To understand these conditions, we examined the dose-dependent effects of environmentally relevant pesticide mixtures (metolachlor, linuron, isoproturon, tebuconazole, aclonifen, atrazine, pendimethalin, and azinphos-methyl) combined with elevated temperatures (22 control vs. 32°C for 4-week exposure) on renin, dinitrophenyl protein (DNP, an indicator of reactive oxygen species, ROS), 3-nitrotyrosine protein (NTP, an indicator of reactive nitrogen species, RNS), superoxidase dismutase (SOD, an antioxidant), and catalase (CAT, an antioxidant) expressions in the kidneys of goldfish (Carassius auratus). Histopathological analysis showed widespread damage to kidney tissues in high temperature and pesticide co-exposure groups, including rupture of the epithelial layer, hemorrhaging, and degeneration of tubular epithelium. Quantitative real-time polymerase chain reaction (qRT-PCR) and immunohistochemical analyses demonstrated significant declines in renin receptor-like mRNA and protein expressions in kidney tissues under combined exposure to high temperature and pesticides compared with controls; conversely, expression of DNP, NTP, SOD, and CAT increased in kidney tissues under the same conditions. Apoptotic cells were also increased in co-exposure groups as assessed by in situ terminal deoxynucleotidyl transferase dUTP nick labeling (TUNEL) assay. The enhanced apoptosis in kidneys of heat and pesticides co-exposed fish was associated with increased caspase-3 (a protease enzyme) mRNA levels. Our results demonstrated that high temperature and pesticides induced oxidative/nitrative stress (i.e., ROS/RNS), damaged tissues, increased cellular apoptosis, and suppressed renin expression in kidneys of goldfish.

Experimental and Established Oximes as Pretreatment before Acute Exposure to Azinphos-Methyl.

Poisoning with organophosphorus compounds (OPCs) represents an ongoing threat to civilians and rescue personal. We have previously shown that oximes, when administered prophylactically before exposure to the OPC paraoxon, are able to protect from its toxic effects. In the present study, we have assessed to what degree experimental (K-27; K-48; K-53; K-74; K-75) or established oximes (pralidoxime, obidoxime), when given as pretreatment at an equitoxic dosage of 25% of LD01, are able to reduce mortality induced by the OPC azinphos-methyl. Their efficacy was compared with that of pyridostigmine, the only FDA-approved substance for such prophylaxis. Efficacy was quantified in rats by Cox analysis, calculating the relative risk of death (RR), with RR=1 for the reference group given only azinphos-methyl, but no prophylaxis. All tested compounds significantly (p ≤ 0.05) reduced azinphos-methyl-induced mortality. In addition, the efficacy of all tested experimental and established oximes except K-53 was significantly superior to the FDA-approved compound pyridostigmine. Best protection was observed for the oximes K-48 (RR = 0.20), K-27 (RR = 0.23), and obidoxime (RR = 0.21), which were significantly more efficacious than pralidoxime and pyridostigmine. The second-best group of prophylactic compounds consisted of K-74 (RR = 0.26), K-75 (RR = 0.35) and pralidoxime (RR = 0.37), which were significantly more efficacious than pyridostigmine. Pretreatment with K-53 (RR = 0.37) and pyridostigmine (RR = 0.52) was the least efficacious. Our present data, together with previous results on other OPCs, indicate that the experimental oximes K-27 and K-48 are very promising pretreatment compounds. When penetration into the brain is undesirable, obidoxime is the most efficacious prophylactic agent already approved for clinical use.

Differential effects of azinphos-methyl and chlorpyrifos on polyamine oxidative metabolism during the embryonic development of Rhinella arenarum and its relation to oxidative stress.

The organophosphorus pesticides azinphos-methyl (AZM) and chlorpyrifos (CPF) exert their toxic action by inhibition of acetylcholinesterase, but non-target processes such as polyamine metabolism can also be affected. Our objective was to evaluate the effects of different concentrations of AZM (0.5-, 2- and 9 mg L-1) and CPF (0.5- and 1 mg L-1) on polyamine oxidative metabolism along Rhinella arenarum embryonic development and to explore its relationship to oxidative stress. Free and conjugated polyamines were measured by HPLC. The activity of spermine oxidase (SMOX), N1-acetylpolyamine oxidase (PAOX) and diamine oxidase (DAO) were measured through kinetic spectrofluorometry. Free putrescine and spermine were significantly increased in open mouth embryos exposed to AZM. Free polyamine levels were not affected by CPF exposure. In embryos exposed to AZM, DAO was increased in tail bud stage and SMOX was increased in open mouth stage, while embryos exposed to CPF showed an increase of PAOX activity in tail bud stage and a decrease of DAO and SMOX activity in open mouth stage. Polyamine levels and oxidative degradation enzymes respond differently if R. arenarum embryos are exposed to AZM or CPF, despite that both insecticides belong to the same chemical family. The early increase of DAO and PAOX would play a protective role to guarantee the normal progression of embryonic development. The increased production of reactive species might contribute to an oxidative stress situation generated by exposure to the insecticides and to the alteration of the antioxidant defense system. In tail bud stage embryos, PAOX and SMOX were positively correlated to acetylcholinesterase activity and reduced glutathione levels (GSH), and negatively correlated to the antioxidant enzymes catalase (CAT) and glutathione S-transferase (GST). In complete operculum embryos, a negative correlation between antioxidant parameters and polyamine levels and polyamine oxidative metabolism was observed, except for SMOX, which showed a low positive correlation with CAT and GSH and a negative correlation to PAOX and DAO. We suggest the use of DAO and PAOX as biomarkers of exposure to AZM and CPF, respectively, as they respond earlier than the classical biomarker acetylcholinesterase.

Ultrasound-assisted dispersive liquid-liquid microextraction followed by ion mobility spectrometry for the simultaneous determination of bendiocarb and azinphos-ethyl in water, soil, food and beverage samples.

A sensitive and fast ultrasound-assisted dispersive liquid-liquid microextraction procedure combined with ion mobility spectrometry has been developed for the simultaneous extraction and determination of bendiocarb and azinphos-ethyl. Experimental parameters affecting the analytical performance of the method were optimized: type and volume of extraction solvent (chloroform, 150 µL), pH (9.0), type and volume of buffer (ammonium buffer pH = 9.0, 4.5 mL) and extraction time (3.0 min). Under optimum conditions, the linearity was found to be in the range of 2-40 and 6-100 ng/mL and the limits of detection (LOD) were 1.04 and 1.31 ng/mL for bendiocarb and azinphos-ethyl, respectively. The method was successfully validated for the analysis of bendiocarb and azinphos-ethyl in different samples such as waters, soil, food and beverage samples.

Environmental concentrations of azinphos-methyl cause different toxic effects without affecting the main target (cholinesterases) in the freshwater gastropod Biomphalaria straminea.

Organophosphate insecticides (OPs) are commonly used in Argentina and around the world for pest control in food crops. They exert their toxicity through the inhibition of the enzyme acetylcholinesterase. In the present study, we aimed to evaluate biochemical and reproductive effects in Biomphalaria straminea, a freshwater gastropod naturally distributed in Argentina, of subchronic exposures to environmental azinphos-methyl concentrations (20 and 200 µg L-1). For biochemical parameters, adult organisms were exposed for 14 days and the activity of cholinesterases (ChEs), carboxylesterases (CEs), glutathione S-transferase (GST), catalase (CAT), superoxide dismutase (SOD), the production of reactive oxygen species (ROS), the total antioxidant capacity (TAC), glycogen and proteins were determined. For reproductive parameters, the egg masses of B. straminea were exposed to azinphos-methyl for one month, and the hatching time and success as well as the offspring survival were registered. We found different toxic effects elicited by the insecticide on the studied biomarkers. CEs activity was significantly inhibited while CAT and GST activities, ROS production and TAC were significantly increased, with respect to the solvent control group. ChE and SOD activities and protein and glycogen contents were not altered by azinphos-methyl. The hatching time and success were not statistically different from control. Nevertheless, the offspring survival was severely affected by the insecticide. Our results show that the primary target of the insecticide (ChE) was not inhibited but CEs, GST, CAT, ROS, TAC and offspring survival were sensitive biomarkers and valuable endpoints for subchronic toxicity assessments in this species.

Comparative Probabilistic Assessment of Occupational Pesticide Exposures Based on Regulatory Assessments.

Implementation of probabilistic analyses in exposure assessment can provide valuable insight into the risks of those at the extremes of population distributions, including more vulnerable or sensitive subgroups. Incorporation of these analyses into current regulatory methods for occupational pesticide exposure is enabled by the exposure data sets and associated data currently used in the risk assessment approach of the Environmental Protection Agency (EPA). Monte Carlo simulations were performed on exposure measurements from the Agricultural Handler Exposure Database and the Pesticide Handler Exposure Database along with data from the Exposure Factors Handbook and other sources to calculate exposure rates for three different neurotoxic compounds (azinphos methyl, acetamiprid, emamectin benzoate) across four pesticide-handling scenarios. Probabilistic estimates of doses were compared with the no observable effect levels used in the EPA occupational risk assessments. Some percentage of workers were predicted to exceed the level of concern for all three compounds: 54% for azinphos methyl, 5% for acetamiprid, and 20% for emamectin benzoate. This finding has implications for pesticide risk assessment and offers an alternative procedure that may be more protective of those at the extremes of exposure than the current approach.

Effects of azinphos-methyl on enzymatic activity and cellular immune response in the hemolymph of the freshwater snail Chilina gibbosa.

The use of a battery of biomarkers, especially those more closely related to species integrity, is desired for more complete ecotoxicological assessments of the effects of pesticide contamination on aquatic organisms. The phosphorodithioate azinphos-methyl has been intensively used in agriculture worldwide and have been found in the habitat of Chilina gibbosa, a freshwater snail endemic to South America. This snail has been proposed as a good model organism for ecotoxicity bioassays on the basis of studies focused mainly on enzymatic responses in whole tissue homogenates. Our aim was to evaluate the effect of an acute 48 h exposure to an environmental concentration of azinphos-methyl on C. gibbosa hemolymph enzymatic activity and cellular immune response. Our results show that cholinesterase activity was strongly inhibited (94%) in hemolymph of exposed snails. Carboxylesterase activity measured with p-nitrophenyl butyrate and glutathione S-transferase activity were augmented 47% and 89% respectively after exposure. No differences were found for hemolymph carboxylesterase activity measured with p-nitrophenyl acetate. These results differ from those reported for whole tissue homogenates and reveal that tissue-specific responses of enzymatic biomarkers exist in this species. Regarding immune cell response, hemocytes were identified for the first time for C. gibbosa. Their viability and phagocytic activity decreased after azinphos-methyl exposure although total number of circulating cells did not differ between treatments. We conclude that concentrations of azinphos-methyl that can be found in the environment can compromise both hemolymph cholinesterase activity and the immune system of C. gibbosa. Furthermore, we propose that carboxylesterase and glutathione S-transferase activities measured in hemolymph and hemocyte viability and phagocytic activity could be incorporated as sensitive biomarkers to evaluate the effects of pesticide exposure on this and related species.

Human Oral Buccal Microbiomes Are Associated with Farmworker Status and Azinphos-Methyl Agricultural Pesticide Exposure.

In a longitudinal agricultural community cohort sampling of 65 adult farmworkers and 52 adult nonfarmworkers, we investigated agricultural pesticide exposure-associated changes in the oral buccal microbiota. We found a seasonally persistent association between the detected blood concentration of the insecticide azinphos-methyl and the taxonomic composition of the buccal swab oral microbiome. Blood and buccal samples were collected concurrently from individual subjects in two seasons, spring/summer 2005 and winter 2006. Mass spectrometry quantified blood concentrations of the organophosphate insecticide azinphos-methyl. Buccal oral microbiome samples were 16S rRNA gene DNA sequenced, assigned to the bacterial taxonomy, and analyzed after "centered-log-ratio" transformation to handle the compositional nature of the proportional abundances of bacteria per sample. Nonparametric analysis of the transformed microbiome data for individuals with and without azinphos-methyl blood detection showed significant perturbations in seven common bacterial taxa (>0.5% of sample mean read depth), including significant reductions in members of the common oral bacterial genus Streptococcus Diversity in centered-log-ratio composition between individuals' microbiomes was also investigated using principal-component analysis (PCA) to reveal two primary PCA clusters of microbiome types. The spring/summer "exposed" microbiome cluster with significantly less bacterial diversity was enriched for farmworkers and contained 27 of the 30 individuals who also had azinphos-methyl agricultural pesticide exposure detected in the blood. IMPORTANCE: In this study, we show in human subjects that organophosphate pesticide exposure is associated with large-scale significant alterations of the oral buccal microbiota composition, with extinctions of whole taxa suggested in some individuals. The persistence of this association from the spring/summer to the winter also suggests that long-lasting effects on the commensal microbiota have occurred. The important health-related outcomes of these agricultural community individuals' pesticide-associated microbiome perturbations are not understood at this time. Future investigations should index medical and dental records for common and chronic diseases that may be interactively caused by this association between pesticide exposure and microbiome alteration.

Characterization and in vitro sensitivity of cholinesterases of gilthead seabream (Sparus aurata) to organophosphate pesticides.

The characterization of cholinesterase activity in brain and muscle of gilthead seabream was carried out using four specific substrates and three selective inhibitors. In addition, K m and V max were calculated from the Michaelis-Menten equation for ASCh and BSCh substrates. Finally, the in vitro sensitivity of brain and muscle cholinesterases to three organophosphates (OPs) was also investigated by estimating inhibition kinetics. The results indicate that AChE is the enzyme present in the brain, whereas in muscle, a typical AChE form is present along with an atypical form of BChE. Very low ChE activity was found in plasma with all substrates used. The inhibitory potency of the studied OPs on brain and muscle AChEs based on bimolecular inhibition constants (k i ) was: omethoate < dichlorvos < azinphosmethyl-oxon. Furthermore, muscle BChE was found to be several orders of magnitude (from 2 to 4) more sensitive than brain and muscle AChE inhibition by dichlorvos and omethoate.

Reversible cholinesterase inhibitors as pre-treatment for exposure to organophosphates: assessment using azinphos-methyl.

Pre-treatment with reversible acetylcholinesterase (AChE) inhibitors before organophosphorous compound (OPC) exposure can reduce OPC-induced mortality. However, pyridostigmine, the only substance employed for such prophylaxis, is merely efficacious against a limited number of OPCs. In search of more efficacious and broad-range alternatives, we have compared in vivo the ability of five reversible AChE inhibitors (pyridostigmine, physostigmine, ranitidine, tacrine and K-27) to reduce mortality induced by the OPC azinphos-methyl. Protection was quantified using Cox analysis by determining the relative risk (RR) of death in rats that were administered these AChE inhibitors in equitoxic dosage (25% of LD01) 30 min before azinphos-methyl exposure. Azinphos-methyl-induced mortality was significantly reduced by all five tested compounds as compared with the reference group that was only exposed to azinphos-methyl without prior pre-treatment (RR = 1). The most efficacious prophylactic agents were K-27 (RR = 0.15) and physostigmine (RR = 0.21), being significantly more efficacious than ranitidine (RR = 0.62) and pyridostigmine (RR = 0.37). Pre-treatment with tacrine (RR = 0.29) was significantly more efficacious than pre-treatment with ranitidine, but the difference between tacrine and pyridostigmine was not significant. Our results indicate that prophylactic administration of the oxime K-27 may be a promising alternative in cases of imminent OPC exposure.

Comparative assessment of in vitro and in vivo toxicity of azinphos methyl and its commercial formulation.

The toxic effects of Gusathion (GUS), which is a commercial organophosphate (OP) pesticide, and also its active ingredient, azinphos methyl (AzM), are evaluated comparatively with in vitro and in vivo studies. Initially, the 96-h LC50 values of AzM and GUS were estimated for two different life stages of Xenopus laevis, embryos, and tadpoles. The actual AzM concentrations in exposure media were monitored by high-performance liquid chromatography. Also, the sub-lethal effects of these compounds to tadpoles were determined 24 h later at exposure concentrations of 0.1 and 1 mg/L using selected biomarker enzymes such as acetylcholinesterase (AChE), carboxylesterase (CaE), glutathione S-transferase (GST), glutathione reductase, lactate dehydrogenase, and aspartate aminotrasferase. Differences in AChE inhibition capacities of AzM and GUS were evaluated under in vitro conditions between frogs and fish in the second part of this study. The AChE activities in a pure electrical eel AChE solution and in brain homogenates of adult Cyprinus carpio, Pelophylax ridibundus, and X. laevis were assayed after in vitro exposure to 0.05, 0.5, 5, and 50 mg/L concentrations of AzM and GUS. According to in vivo studies AChE, CaE and GST are important biomarkers of the effect of OP exposure while CaE may be more effective in short-term, low-concentration exposures. The results of in vitro studies showed that amphibian brain AChEs were relatively more resistant to OP exposure than fish AChEs. The resistance may be the cause of the lower toxicity/lethality of OP compounds to amphibians than to fish.

Resistance in cholinesterase activity after an acute and subchronic exposure to azinphos-methyl in the freshwater gastropod Biomphalaria straminea.

Organophosphorous and carbamates insecticides are ones of the most popular classes of pesticides used in agriculture. Its success relies on their high acute toxicity and rapid environmental degradation. These insecticides inhibit cholinesterase and cause severe effects on aquatic non-target species, particularly in invertebrates. Since the properties of cholinesterases may differ between species, it is necessary to characterize them before their use as biomarkers. Also organophosphorous and carbamates inhibit carboxylesterases and the use of both enzymes for biomonitoring is suggested. Azinphos-methyl is an organophosphorous insecticide used in several parts of the word. In Argentina, it is the most applied insecticide in fruit production in the north Patagonian region. It was detected with the highest frequency in superficial and groundwater of the region. This work aims to evaluate the sensitivity of B. straminea cholinesterases and carboxylesterases to the OP azinphos-methyl including estimations of 48 h NOEC and IC50 of the pesticide and subchronic effects at environmentally relevant concentrations. These will allow us to evaluate the possibility of using cholinesterase and carboxylesterase of B. straminea as sensitive biomarkers. Previously a partial characterization of these enzymes will be performed. As in most invertebrates, acetylthiocholine was the preferred hydrolyzed substrate of B. straminea ChE, followed by propionylthiocholine and being butyrylthiocholine hydrolysis very low. Cholinesterase activity of B. straminea was significantly inhibited by the selective cholinesterases inhibitor (eserine) and by the selective inhibitor of mammalian acethylcholinesterase (BW284c51). In contrast, iso-OMPA, a specific inhibitor of butyrylcholinesterase, did not inhibit cholinesterase activity. These results suggest that cholinesterase activity in total soft tissue of B. straminea corresponds to acethylcholinesterase. Carboxylesterases activity was one order of magnitude higher than cholinesterase. A greater efficiency (Vmax/Km) was obtained using acetylthiocholine and p-nitrophenyl butyrate. Acute exposure to azinphos-methyl did not cause inhibition of cholinesterase activity until 10 mg L(-1) used. Carboxylesterases towards p-nitrophenyl butyrate was inhibited by azinphos-methyl being the IC502.20±0.75 mg L(-1) of azinphos-methyl. Subchronic exposure to environmental concentrations of azinphos-methyl (0.02 and 0.2 mg L(-1)) produced a decrease in survival, protein content and carboxylesterases activity despite no inhibition of cholinesterase activity was observed. B. straminea cholinesterase is not a sensible biomarker. On the contrary, carboxylesterases activity was inhibited by azinphos-methyl. Carboxylesterases could be protecting cholinesterase activity and therefore, protecting the organism from neurotoxicity. This work confirms the advantages of measuring cholinesterases and carboxylesterases jointly in aquatic biomonitoring of pesticide contamination. This becomes relevant in order to find more sensitive biomarkers and new strategies to protect non-target aquatic organisms from pesticide contamination.

Acute toxicity and biochemical effects of azinphos methyl in the amphipod Hyalella curvispina.

We evaluated the acute toxicity and biochemical effects of the organophosphorus pesticide azinphos methyl (AM) in the amphipod Hyalella curvispina that inhabits ponds and irrigation channels of an intensive fruit-producing region in Rio Negro and Neuquén valley, North Patagonia, Argentina. The analysis by nonlinear regression of data from the 96 h-acute toxicity tests indicated the coexistence of two subpopulations of H. curvispina with different susceptibilities to AM. The 96 h-LC50 for the resistant subpopulation (166 ± 56 µg/L) was 216-fold higher than the 96h-LC50 value for the susceptible one (0.77 ± 1.33 µg/L).The two subpopulations could not be distinguished based on the biochemical measurements in control amphipods. Cholinesterase activity was significantly inhibited in AM-exposed amphipods in a concentration-dependent manner. The IC50 value obtained after 96 h of exposure (2.18 ± 1.95 µg/L) was significantly lower than the 48 h-IC50 value (29.6 ± 17.4 µg/L). Carboxylesterase activity was significantly inhibited after 48 h of exposure to 12.5 and 62.5 µg/L AM (inhibition, 51%). This enzyme was thus able to protect cholinesterase from inhibition at 12.5 µg/L AM. Reduced glutathione and catalase showed a significant increase after 24 h of exposure as an adaptive response to AM, whereas glutathione S-transferase activity was not significantly modified. The analysis of species sensitivity distribution showed that both subpopulations of H. curvispina were more tolerant to AM than most amphipod species, and that the susceptible subpopulation was more sensitive to AM than the other local aquatic species analyzed. The maximum concentration of AM in drainage water within the fruit-producing area reported by other studies would affect most of the amphipod species (99%) and also a 44% of local aquatic ones. The results obtained in this study point out the usefulness of including amphipods like H. curvispina in ecotoxicity studies and monitoring programs to perform pesticide risk assessments.

Simulated solar light phototransformation of organophosphorus azinphos methyl at the surface of clays and goethite.

The photochemical behavior of the pesticide azinphos methyl at the surface of clays (kaolinite, bentonite) and goethite was studied using Suntest setup (λ > 300 nm). The quantum yield on the clays was found to be roughly three times lower than that in aqueous solution. However, the photochemical efficiency was much higher at the surface of goethite owing to its photocatalytic activity through the hydroxyl radical production. The added humic substances on kaolonite show an inhibition of azinphos methyl degradation while the incorporation of iron(III) aquacomplexes leads to an important increase of the disappearance together with the formation of iron(II). Hydroxyl radical species were found to be formed either by excitation of goethite or clays. The goethite support acts as a more efficient catalyst for the formation of these reactive oxygen species. The photodecomposition reactions observed were (i) hydrolysis process leading to the formation of benzotriazone and the oxidation of the P = S bond giving rise to the formation of the oxon derivative, and (ii) homolytic cleavage of the N-C and C-S bonds of the organophosphorus bridge leading to the formation of dimers that appear to be specific to the irradiation at the surface of solid supports since they were not observed when the irradiation was performed in aqueous media: a statement that is related to the presence of aggregates at the surface of solid supports.

Breaking the take home pesticide exposure pathway for agricultural families: workplace predictors of residential contamination.

BACKGROUND: Pesticides used in agriculture can be taken into worker homes and pose a potential risk for children and other family members. This study focused on identification of potential intervention points at the workplace. METHODS: Workers (N = 46) recruited from two tree fruit orchards in Washington State were administered a 63-item pesticide safety questionnaire. Dust was collected from commuter vehicles and worker homes and analyzed for four organophosphorus (OP) pesticides (azinphosmethyl, phosmet, chlorpyrifos, malathion). RESULTS: Geometric mean azinphosmethyl concentrations in dust for three worker groups (16 pesticide handlers, 15 green fruit thinners, 15 organic orchard workers) ranged from 0.027-1.5 µg/g, with levels in vehicle dust higher than in house dust, and levels in house dust from handlers' homes higher than levels from tree fruit thinners' homes. Vehicle and house dust concentrations of azinphosmethyl were highly associated (R(2) = 0.44, P < 0.001). Significant differences were found across worker groups for availability of laundry facilities, work boot storage, frequency of hand washing, commuter vehicle use, parking location, and safety training. CONCLUSIONS: These findings support a focus on intervention activities to reduce take home pesticide exposure closer to the source of contamination; specifically, the workplace and vehicles used to travel to the workplace.

Evaluation of in vitro and in vivo toxic effects of newly synthesized benzimidazole-based organophosphorus compounds.

This paper reports the toxic properties of eight newly synthesized benzimidazole-based organophosphorus (OP) compounds in Xenopus laevis in both in vivo and in vitro conditions. For both experiments, a commercial solution of azinphos methyl (AzM, Gusathion M WP25) was used as a reference compound. The 24-h median lethal concentrations (LC50) of all tested compounds were determined for 46th stage tadpoles in the range of 9.54-140.0 µM. For evaluation of the lethality of the compounds, the activity of the enzyme biomarkers acetylcholinesterase (AChE), carboxylesterase, glutathione S-transferase (GST), glutathione peroxidase, glutathione reductase, lactate dehydrogenase, aspartate aminotransferase, and alanine aminotransferase were determined in vivo in X. laevis tadpoles exposed to three concentrations (LC50, LC50/2, and LC50/4) of tested compounds. All exposure concentrations of AzM and seven of eight tested compounds caused CaE inhibition in in vivo conditions. Furthermore, the AChE inhibition capacity of tested compounds in commercial electric eel AChE and in X. laevis homogenates and also CaE inhibition capacity in only X. laevis homogenates were assayed for a 30-min in vitro exposure period. Eight OP compounds did not inhibit AChE activity more than 23 percent, but AzM exposure inhibited AChE activity by 26 percent for X. laevis homogenates and 97 percent for electric fish AChE in in vitro conditions. Also, CaE inhibition levels in X. laevis tadpole homogenates were 46 percent for AzM and between 8 percent and 33 percent for other compounds in in vitro conditions.

Evaluation of cytochrome P450 activity in field populations of Cydia pomonella (Lepidoptera: Tortricidae) resistant to azinphosmethyl, acetamiprid, and thiacloprid.

The Río Negro and Neuquén Valley is the most important apple and pear growing region in Argentina. Cydia pomonella L. (Lepidoptera: Tortricidae), the main fruit-tree pest is being controlled by azinphosmethyl (AzMe), acetamiprid (Acet), and thiacloprid (Thia) among other insecticides. The objective of this study was to evaluate the response of neonate larvae of codling moth to these three insecticides and on the role of cytochrome P450 monooxygenase in their toxicity. All field populations presented significantly lower mortality to a discriminating concentration (DC) of Acet and AzMe. In addition, 13 of the 14 populations showed significantly lower mortality to DC of Thia. Most of the field populations (71%) showed significantly higher 7-ethoxycoumarine O-deethylase activity compared with the laboratory-susceptible strain. While positive significant correlation (gamma = 0.59) was found between Thia and AzMe mortalities at the DC level, no significant correlations were detected between Acet and Thia (gamma = 0.35) or Acet and AzMe (gamma = 0.12). However, Acet and Thia mortalities were significantly correlated to the percentage of individuals exhibiting 7-ethoxy-coumarine O-deethylase activity activities higher than the mean upper 95% confidence limit of the susceptible strain (gamma = -0.52 and gamma = -0.63, respectively).

A high-performance Calix@ZnO based bifunctional nanomaterial for selective detection and degradation of toxic azinphos methyl in environmental samples.

One of the key tenets of sustainable agriculture and food safety is the removal of toxic pesticides from the environment. However, developing reliable, affordable, and efficient methods for detecting and degrading pesticides into non-toxic degradable products remains an immediate matter of concern. Herein, we attempt to develop a strategy for the detection as well as degradation of highly toxic phosphorodithioate pesticide, Azinphos methyl (AZM), using hybrid zinc oxide nanoparticles (ZnO NPs). Considering the non-selectivity of bare ZnO and receptor R1, we have fabricated the heterocalixarene-based Calix (R1) over zinc oxide (ZnO) surface in situ via the sol-gel process. The synthesized heterocaliaxrene-modified ZnO (R1@ZnO) NPs show an excellent affinity for the selective and sensitive detection of AZM with a tremendously low limit of detection (68 mg L-1) and no interference from other pesticides. Degradation of AZM was fully supported by fluorescence spectroscopy, scanning electron microscopy (SEM), 1H NMR titrations, FTIR spectroscopy, cyclic voltammetry, and mass spectroscopy, which unequivocally confirmed the formation of non-toxic products. According to our findings, R1@ZnO NPs are sustainable nanomaterials that can be employed for environmental remediation since they operate in an aqueous medium.