Investigations of the determination and transformations of diazinon and malathion under environmental conditions using gas chromatography coupled with a flame ionisation detector

Degradation of two model insecticides, diazinon and malathion, and their degradation products 2-isopropyl-6-methyl-4-pyrimidinol--IMP (diazinon hydrolysis product) and malaoxon (malathion oxidation product) was compared and studied in the environment. The pesticides and their metabolites were extracted from samples (water, soil, chicory) with ethyl acetate and subsequently the extracts were analyzed by GC/FID. It was shown that hydrolysis is the major process in the degradation of these pesticides in water. In fact, 95% of diazinon was degraded, and only 10% of malathion was oxidised. In soil 30% of diazinon exposed to the sunlight was decomposed by photolysis, whereas in soil left in the darkness no degradation products were observed. In soil left under environmental conditions, 90% of diazinon was degraded and 40% from its initial concentration was transformed into IMP. The concentrations of the pesticides after 21 days on chicory were under maximal allowable concentration, which is 0.5 ppm for malathion and for diazinon. The concentration of malaoxon was more than twice as high as the allowable value, which is for the sum of malathion and malaoxon 3 ppm.     

Photodegradation of organophosphorus insecticides - investigations of products and their toxicity using gas chromatography-mass spectrometry and AChE-thermal lens spectrometric bioassay

Four organophosphorus compounds: azinphos-methyl, chlorpyrifos, malathion and malaoxon in aqueous solution were degraded by using a 125 W xenon parabolic lamp. Gas chromatography-mass spectrometry (GC-MS) was used to monitor the disappearance of starting compounds and formation of degradation products as a function of time. AChE-thermal lens spectrometric bioassay was employed to assess the toxicity of photoproducts. The photodegradation kinetics can be described by a first-order degradation curve C=C0e(-kt), resulting in the following half lives: 2.5min for azinphos-methyl, 11.6 min for malathion, 13.3 min for chlorpyrifos and 45.5 min for malaoxon, under given experimental conditions. During the photoprocess several intermediates were identified by GC-MS suggesting the pathway of OP degradation. The oxidation of chlorpyrifos results in the formation of chlorpyrifos-oxon as the main identified photoproduct. In case of malathion and azinphos-methyl the corresponding oxon analogues were not detected. The formation of diethyl (dimethoxy-phosphoryl) succinate in traces was observed during photodegradation of malaoxon and malathion. Several other photoproducts including trimethyl phosphate esters, which are known to be AChE inhibitors and 1,2,3-benzotriazin-4(3H)-one as a member of triazine compounds were identified in photodegraded samples of malathion, malaoxon, and azinphos-methyl. Based on this, two main degradation pathways can be proposed, both result of the (P-S-C) bond cleavage taking place at the side of leaving group. The enhanced inhibition of AChE observed with the TLS bioassay during the initial 30 min of photodegradation in case of all four OPs, confirmed the formation of toxic intermediates. With the continuation of irradiation, the AChE inhibition decreased, indicating that the formed toxic compounds were further degraded to AChE non-inhibiting products. The presented results demonstrate the importance of toxicity monitoring during the degradation of OPs in processes of waste water remediation, before releasing it into the environment.     

Does malaoxon play a role in the geno- and cytotoxic effects of malathion on human choriocarcinoma cells?

This investigation was undertaken to elucidate whether the active metabolite of malathion, malaoxon, has any role in exerting cyto- and genotoxic effects for human choriocarcinoma (JAR) cell line which is an acceptable model for human placental cells. Gas chromatography-mass spectrometry (GC-MS) analysis were separately performed on the cell compartment and supernatant cell culture medium after subjecting the cell line to different malathion concentrations (10–400 μg/mL) and for various incubation periods (0.5 to 24 hours). GC-MS analysis showed that the sonication performed for the disruption of the cells did not cause the chemical change of malathion. The uptake of malathion by the cells was relatively fast. However, the presence of malaoxon, even in trace amounts, could not be confirmed either in samples originating from disrupted cells or in the cell culture medium. Although the hydrolysis of malaoxon occurred in the culture medium, this degradation process could not be counted as a reason for the absence of malaoxon. Since both malathion and malaoxon standard compounds could be accurately detected and distinguished by the applied liquid-liquid extraction and GC-MS methods, one can conclude that, in the case of JAR cells, the parent compound, (i.e. malathion itself) is responsible for the observed in vitro cyto- and genotoxic effects. Our results indicate that the direct toxicity of malathion contributes to the complications of pregnancy observed for environmental malathion exposure.     

Assessing the toxicity of organophosphorous pesticides to indigenous algae with implication for their ecotoxicological impact to aquatic ecosystems

This study aimed to determine the toxicity of three organophosphorous pesticides, chlorpyrifos, terbufos and methamidophos, to three indigenous algal species isolated from local rivers and algal mixtures. The diatom Nitzschia sp. (0.30–1.68 mg L^?1 of EC₅₀ -the estimated concentration related to a 50% growth reduction) and the cyanobacteria Oscillatoria sp. (EC₅₀ of 0.33–7.99 mg L^?1) were sensitive to single pesticide treatment and the chlorophyta Chlorella sp. was the most tolerant (EC₅₀ of 1.29–41.16 mg L^?1). In treatment with the mixture of three pesticides, Chlorella sp. became the most sensitive alga. The antagonistic joint toxic effects on three indigenous algae and algal mixtures were found for most of the two pesticide mixtures. The results suggested that mixture of pesticides might induce the detoxification mechanisms more easily than the single pesticide. The synergistic interactions between terbufos and methamidophos to algal mixtures and between methamidophos and chlorpyrifos to Nitzschia sp. indicated methamidophos might act as a potential synergist. Differential sensitivity of three families of algae to these pesticides might result in changes in the algal community structures after river water has been contaminated with different pesticides, posing great ecological risk on the structure and functioning of the aquatic ecosystem.     

Enhanced visual detection of pesticides using gold nanoparticles

The presence of parts per billion (ppb) levels of chlorpyrifos (O,O-Diethyl-O-(3,5,6-trichloro-2-pyridyl) phosphorothioate) and malathion (S-1,2-bis(ethoxycarbonyl) ethyl O,O-dimethyl phosphorodithioate), two common pesticides found in the surface waters of developing countries, have been visually detected using gold nanoparticles. Visual detection of the presence of pesticide is possible when the color change occurring by the adsorption of pesticides on gold nanoparticles is enhanced by sodium sulfate. The method presented here is simple and there is no need of sample preparation or preconcentration. The response occurs within seconds and the color change is very clear. The detection is possible if chlorpyrifos and malathion are present up to a concentration of 20 and 100 ppb, respectively. The method shows great potential for on-site pesticide monitoring. The method is also applicable as a qualitative technique for the performance evaluation of various household water filters, which claim pesticide removal.     

Heterogeneous reactions of suspended parathion, malathion, and fenthion particles with NO(3) radicals

Organophosphorus pesticides (OPPs) emit into the atmosphere in both gas and particulate phases via spray drift from treatments and post-application emission, but most of their degradations in the atmosphere are not well known. In this study, the heterogeneous reactions of nitrate (NO₃) radicals with three typical OPPs (parathion, malathion, and fenthion) absorbed on azelaic acid particles are investigated using an online vacuum ultraviolet photoionization aerosol time-of-flight mass spectrometer (VUV-ATOFMS). The reaction products observed with the VUV-ATOFMS are identified on the basis of GC/MS analysis of the products in the reaction between NO₃ radicals and the coating of the pesticide. Paraoxon is identified as the only product of parathion; malaoxon and bis(1,2-bis-ethoxycarbonylethyl)disulfide as the products of malathion; fenoxon, fenoxon sulfoxide, fenthion sulfoxide, fenoxon sulfone, and fenthion sulfone as the products of fenthion. The degradation rates of parathion, malathion, and fenthion under the experimental conditions are 5.5 × 10⁻³, 5.6 × 10⁻², and 3.3 × 10⁻² s⁻¹, respectively. The pathways of the heterogeneous reactions between the three OPPs and NO₃ radicals are proposed. The experimental results reveal the possible transformations of these OPPs through the oxidation of NO₃ radicals in the atmosphere.     

Effects of atrazine on acetylcholinesterase activity in midges (Chironomus tentans) exposed to organophosphorus insecticides

Acetylcholinesterase activity was determined for midge larvae (Chironomus tentans) exposed to either organophosphorus insecticides (OPs) alone or OP insecticides in binary combination with atrazine (200 μg/l). Although atrazine by itself did not reduce the level of acetylcholinesterase activity, atrazine in combination with chlorpyrifos significantly decreased acetylcholinesterase activity as compared to chlorpyrifos only treatments. Although similar trends existed for malathion and methyl parathion, differences were not statistically significant. These results match previously published toxicity data where atrazine, although not acutely toxic even at much higher levels, decreased EC₅₀ values for chlorpyrifos by a magnitude of 4, decreased methyl parathion values by a magnitude of 2, and did not decrease values for malathion.     

Effects of combinations of malathion and cypermethrin on survivability and time of metamorphosis of tadpoles of Indian cricket frog (Fejervarya limnocharis)

This study was undertaken to determine the effect of environmentally realistic concentrations of two commonly used pesticides viz., malathion and cypermethrin, using a fully 3 × 3 factorial experiments on the survivability and time of metamorphosis in a common rice paddy field frog (cricket frog) Fejervarya limnocharis under laboratory conditions. The results suggest that cypermethrin is more toxic than malathion and combinations of higher concentrations of cypermethrin (50 μg/L) with malathion (250 and 500 μg/L) are more deleterious to the survivability of tadpoles. With increasing cypermethrin concentration, the survivability of tadpole decreased (r = ?0.986, P = 0.108). But cypermethrin alone induced early metamorphosis among the surviving tadpoles. However, there was a delay in the time required for metamorphosis induced by malathion and its combination with cypermethrin. The delay in metamorphosis may indicate the altered physiological fitness of the individual. The emergent froglets will be subjected to environmental stressors like high temperature and less humidity of post-monsoon tropical climate that could enhance negative influence triggered by pesticides.     

Does malaoxon play a role in the geno- and cytotoxic effects of malathion on human choriocarcinoma cells?

This investigation was undertaken to elucidate whether the active metabolite of malathion, malaoxon, has any role in exerting cyto- and genotoxic effects for human choriocarcinoma (JAR) cell line which is an acceptable model for human placental cells. Gas chromatography-mass spectrometry (GC-MS) analysis were separately performed on the cell compartment and supernatant cell culture medium after subjecting the cell line to different malathion concentrations (10-400 μg/mL) and for various incubation periods (0.5 to 24 hours). GC-MS analysis showed that the sonication performed for the disruption of the cells did not cause the chemical change of malathion. The uptake of malathion by the cells was relatively fast. However, the presence of malaoxon, even in trace amounts, could not be confirmed either in samples originating from disrupted cells or in the cell culture medium. Although the hydrolysis of malaoxon occurred in the culture medium, this degradation process could not be counted as a reason for the absence of malaoxon. Since both malathion and malaoxon standard compounds could be accurately detected and distinguished by the applied liquid-liquid extraction and GC-MS methods, one can conclude that, in the case of JAR cells, the parent compound, (i.e. malathion itself) is responsible for the observed in vitro cyto- and genotoxic effects. Our results indicate that the direct toxicity of malathion contributes to the complications of pregnancy observed for environmental malathion exposure.     

Determination of organochlorine and organophosphorus pesticide residues in low moisture,nonfatty products using a solid phase extraction cleanup and gas chromatography

Abstract

A multiresidue solid‐phase extraction (SPE) method for the isolation and subsequent gas Chromatographie determination of organochlorine and organophosphorus pesticide residues in low‐moisture, nonfatty products is described. Residues are extracted from samples with an acetonitrile/water mixture. Cleanup of the extract is performed using graphitized carbon black and anion exchange SPE columns, and analysis is performed by gas chromatography with Hall electrolytic conductivity and flame photometric detection. Recovery data was obtained by fortifying corn, oats and wheat with pesticides. The average recoveries were 79–123% for eight organochlorine and 51–122% for 28 organophosphorus pesticide residues. The limit of quantitation for chlorpyriphos was 0.05 ppm using the Hall electrolytic conductivity detector and <0.005 ppm using the flame photometric detector.     

Toxicological properties of trialkyl phosphorothioate and dialkyl alkyl- and arylphosphonothioate esters

Impurities such as O,S,S-trimethyl phosphorodithioate (TMPD) and the S-methyl isomer of malathion (isomalathion) strongly potentiated the mammalian toxicity of malathion. In contrast, impurities present in the phosphoramidothioate insecticide acephate had an antagonizing effect on its mammalian toxicity. The potentiation of the toxicity of malathion was attributed to inhibition of mammalian liver and serum carboxylesterase. O,O,S-Trimethyl phosphorothioate (TMP), another impurity present in technical malathion and in other organophosphorus insecticides, proved to be highly toxic. Rats given a single oral dose of TMP at a level as low as 20 mg/kg died over a period of three weeks, with death occurring with non-cholinergic signs of poisoning. TMPD also caused similar delayed death in rats. O,O,O-Trimethyl phosphorothioate (TMP=S), also another impurity in technical malathion and a structural isomer of TMP, was a potent antagonist to the delayed toxicity of TMP. Examination of a number of related trialkyl phosphorothioate and dialkyl alkylphosphonothioate esters revealed several of these compounds to be highly toxic to rats.     

Toxic and developmental effects of organophosphorus insecticides in embryos of the South African clawed frog

Four organophosphorus insecticides and the active metabolites of two phosphorothionate insecticides were tested for their toxic and teratogenic effects on embryos of Xenopus laevis. All compounds caused dose-dependent developmental defects, such as abnormal pigmentation, abnormal gut development, notochordal defects and reduced growth. Malathion, malaoxon, parathion, and paraoxon produced severe defects, while monocrotophos and dicrotophos produced considerably milder defects. All compounds reduced NAD+ levels to a similar extent, regardless of the severity of the defects induced. Thus some commonly used organophosphorus insecticides and their metabolites are teratogenic to Xenopus embryos, but reduced NAD+ level does not appear to be important in causing the developmental defects.     

Protective effect of Syzygium cumini against pesticide-induced cardiotoxicity

Pesticide-induced toxicity is a serious issue which has resulted in plethora of diseases all over the world. The organophosphate pesticide malathion has caused many incidents of poisoning such as cardiac manifestations. The present study was designed to evaluate the effect of Syzygium cumini on malathion-induced cardiotoxicity. Dose optimization of malathion and polyphenols such as curcumin, (?)-epicatechin, gallic acid, butylated hydroxyl toluene, etc. was done by MTT cell proliferation assay. Nuclear deformities, ROS production, and integrity of extra cellular matrix components were analyzed by different techniques. S. cumini methanolic pulp extract (MPE), a naturally derived gallic acid-enriched antioxidant was taken to study its effect on malathion-induced toxicity. Nuclear deformities, ROS production, and integrity of extra cellular matrix components were also analyzed. Twenty micrograms per milliliter LD50 dose of malathion was found to cause stress-mediated responses in H9C2 cell line. Among all the polyphenols, gallic acid showed the most significant protection against stress. Gallic acid-enriched methanolic S. cumini pulp extract (MPE) showed 59.76 %?±?0.05, 81.61 %?±?1.37, 73.33 %?±?1.33, 77.19 %?±?2.38 and 64.19 %?±?1.43 maximum inhibition for DPPH, ABTS, NO, H₂O₂ and superoxide ion, respectively, as compared to ethanolic pulp extract and aqueous pulp extract. Our study suggests that S. cumini MPE has the ability to protect against the malathion-mediated oxidative stress in cardiac myocytes.     

Sublethal contaminant exposure alters behavior in a common insect: Important implications for trophic transfer

This study examined the effects of sub-lethal exposure of the ubiquitous pesticide malathion on the behavior of the model orthopteran species, the house cricket (Acheta domesticus). Increasing concentrations of malathion caused male crickets to increase periods of non-directional movement, such as twitching and grooming, directional movement, and to seek out shelter less. These are all behavioral alterations that may increase the cricket's chances of being preyed upon, and thus have the potential for serious ecological consequences through trophic transfer. This study also revealed that female crickets appeared to be less affected by malathion than their male conspecifics, indicating a potential sex-bias in both susceptibility and possible predator attack.     

Isolation and characterization of a Rhodococcus strain with phenol-degrading ability and its potential use for tannery effluent biotreatment

Introduction

Wastewater derived from leather production may contain phenols, which are highly toxic, and their degradation could be possible through bioremediation technologies.

Materials, methods and results

In the present work, microbial degradation of phenol was studied using a tolerant bacterial strain, named CS1, isolated from tannery sediments. This strain was able to survive in the presence of phenol at concentrations of up to 1,000?mg/L. On the basis of morphological and biochemical properties, 16S rRNA gene sequencing, and phylogenetic analysis, the isolated strain was identified as Rhodococcus sp. Phenol removal was evaluated at a lab-scale in Erlenmeyer flasks and at a bioreactor scale in a stirred tank reactor. Rhodococcus sp. CS1 was able to completely remove phenol in a range of 200 to 1,000?mg/L in mineral medium at 30 ± 2?°C and pH 7 as optimal conditions. In the stirred tank bioreactor, we studied the effect of some parameters, such as agitation (200?C600 rpm) and aeration (1?C3?vvm), on growth and phenol removal efficiency. Faster phenol biodegradation was obtained in the bioreactor than in Erlenmeyer flasks, and maximum phenol removal was achieved at 400?rpm and 1 vvm in only 12?h. Furthermore, Rhodococcus sp. CS1 strain was able to grow and completely degrade phenols from tannery effluents after 9?h of incubation.

Conclusion

Based on these results, Rhodococcus sp. CS1 could be an appropriate microorganism for bioremediation of tannery effluents or other phenol-containing wastewaters.     

Genotoxicity associated with oxidative damage in the liver and kidney of mice exposed to dimethoate subchronic intoxication

Background and aims  

Because of the widespread use of pesticides for domestic and industrial applications, the evaluation of their toxic effects is of major concern to public health. The aim of the present study was to investigate the propensity of dimethoate (DM), an organophosphorus pesticide, to cause oxidative damage in the liver and kidney of mice and its associated genotoxic effect.     

Binding and detoxification of chlorpyrifos by lactic acid bacteria on rice straw silage fermentation

This investigation examined the reduction of pesticide residues on straw inoculated with lactic acid bacteria (LAB) during ensiling. Lactobacillus casei WYS3 was isolated from rice straw that contained pesticide residues. Non-sterilized rice straw, which was inoculated with L. casei WYS3, showed increased removal of chlorpyrifos after ensiling, compared with rice straw that was not inoculated with L. casei WYS3 or sterilized rice straw. In pure culture, these strains can bind chlorpyrifos as indicated by high-performance liquid chromatography analysis. Viable L. casei WYS3 was shown to bind 33.3–42% of exogenously added chlorpyrifos. These results are similar to those of acid-treated cells but less than those of heat-treated cells, which were found to bind 32.0% and 77.2% of the added chlorpyrifos respectively. Furthermore, gas chromatography–mass spectrometry analysis determined that L. casei WYS3 detoxified chlorpyrifos via P-O-C cleavage. Real-time polymerized chain reaction analysis determined that organophosphorus hydrolase gene expression tripled after the addition of chlorpyrifos to LAB cultures, compared with the control group (without chlorpyrifos). This paper highlights the potential use of LAB starter cultures for the detoxification and removal of chlorpyrifos residues in the environment.     

Biodegradation of malathion, α- and β-endosulfan by bacterial strains isolated from agricultural soil in Veracruz,Mexico

The objective of this study was to evaluate the capacity of two bacterial strains isolated, cultivated, and purified from agricultural soils of Veracruz, Mexico, for biodegradation and mineralisation of malathion (diethyl 2-(dimethoxyphosphorothioyl) succinate) and α- and β-endosulfan (6,7,8,9,10,10-hexachloro-1,5,5a,6,9,9a-hexahydro-6-9-methano-2,4,3-benzodioxathiepine-3-oxide). The isolated bacterial strains were identified using biochemical and morphological characterization and the analysis of their 16S rDNA gene, as Enterobacter cloacae strain PMM16 (E1) and E. amnigenus strain XGL214 (M1). The E1 strain was able to degrade endosulfan, whereas the M1 strain was capable of degrading both pesticides. The E1 strain degraded 71.32% of α-endosulfan and 100% of β-endosulfan within 24 days. The absence of metabolites, such as endosulfan sulfate, endosulfan lactone, or endosulfan diol, would suggest degradation of endosulfan isomers through non-oxidative pathways. Malathion was completely eliminated by the M1 strain. The major metabolite was butanedioic acid. There was a time-dependent increase in bacterial biomass, typical of bacterial growth, correlated with the decrease in pesticide concentration. The CO₂ production also increased significantly with the addition of pesticides to the bacterial growth media, demonstrating that, under aerobic conditions, the bacteria utilized endosulfan and malathion as a carbon source. Here, two bacterial strains are shown to metabolize two toxic pesticides into non-toxic intermediates.     

The effects of bayluscide and malathion on the survival of Schistosoma mansoni miracidia

Laboratory experiments were conducted to assess the toxic effects of bayluscide and malathion against Schistosoma mansoni miracidia. The results indicate that survival of miracidia varied with times of exposure and concentrations of tested chemicals. Statistical analyses reveal that LC5, LC50 and LC95 for bayluscide were 0.04 ppm, 0.06 ppm and 0.12 ppm after 2 hours of exposure; 0.02 ppm, 0.03 ppm and 0.06 ppm after 4 hours of exposure; and 0.01 ppm, 0.02 ppm and 0.04 ppm after 6 hours of exposure respectively. These data indicate that bayluscide is much more toxic to the first stage larvae of schistosomes than to snail intermediate hosts cited in the literature. Application of lower concentrations of molluscicide in the transmission sites is thus expected to curtail the survival of miracidia; therefore controlling schistosomiasis at relatively low costs. Such applications also reduce the risk of toxicity to non target organisms present in the aquatic environment. Statistical analysis of the results of tests using malathion gave LC5, LC50 and LC95 values of 83.38 ppm, 153.11 ppm and 245.85 ppm after 2 hours of exposure; and 76.86 ppm, 116.48 ppm and 172.04 ppm after 4 hours of exposure respectively. These data indicate that the use of malathion as an insecticide in tropical ecosystems may also affect the survival and viability of schistosome miracidia. Such uses could help reducing the risk of schistosomiasis transmission in these particular locations.     

Effect of some organophosphate insecticides on acetylcholinesterase of adult coccinella septempunctata (coccinellidae)

Abstract

The in vitro enzyme activity of head homogenates of Coccinella septempunctata from different habitats (wheat, barley, rye and set‐aside fields in Belgium and Hungary) and the effect of in vivo surface contact treatments with organophosphorous active ingredients on the same species were investigated. The in vitro studies indicated that the acetylcholinesterase (AChE) of C. septempunctata was less sensitive to inhibition by paraoxon than by malaoxon in the case of each population. The differences found between parathion and malathion treatments in in vivo testing were not significant. The inhibition process of paraoxon suggests that the seven‐spot ladybird may have at least two AChEs responding differently for the paraoxon inhibition.