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Human serum paraoxonase (PON1) activity in acute organophosphorous insecticide poisoning.

Human serum paraoxonase (PON1) and perhaps other mammalian paraoxonases catalyzes the hydrolysis of certain organophosphorus (OP) insecticides and nerve gases and so may alter significantly an individual's susceptibility to the toxicity of these chemicals. Serum PON1 exhibits a substrate dependent polymorphism and this polymorphism shows great interethnic variability. This study focused on the investigation of PON1, arylesterase and cholinesterase activities in 28 acute OP insecticide poisoning cases. Insecticide analysis were performed by GC-NPD and activities of enzymes were measured by using spectrophotometer. The activity levels for salt stimulated PON1, basal PON1 and arylesterase were found as 78.83 (35.39-186.13), 39.97 (2.49-80.43) micromol/min/l and 126.26 (36.34-288.24) mmol/min/l respectively. On the other hand the activity levels for butyrylcholinesterase (BTC) and acetylcholinesterase (AchE) were found as 797.23 (106.3-3823)U/l and 4.65 (0.21-30.29)U/ml. There was a correlation between percent stimulation of PON1 and BTC activities (r=0.446, P<0.05), but this correlation was lower than in cases who exposed to OP insecticides chronically. As a conclusion, in chronic and acute OP exposure, both PON1 level and phenotype must be taken into consideration.

Adult↗

Household insecticides: evaluation and assessment of inhalation toxicity: a workshop summary.

Particularly in tropical countries household insecticides are used on a day-by-day basis to control mosquitoes, other crawling and flying insects to prevent the spread of vector-borne diseases. The products used most often are spray-cans, oil-sprays, mosquito coils as well as slow-release vaporising systems such as mats and liquid vaporiser. The extent and duration of exposure of humans is highly dependent on the type of product used. The objective of this workshop was to analyse the necessity and feasibility of inhalation studies with household insecticides taking into account the specific constrains associated with each type of end-use product. The standardisation of inhalation studies with regard to the generation of test atmospheres, mode and duration of exposure, and selection of adequate toxicological endpoints were addressed. Due to the complex nature of exposure atmospheres generated by some household insecticides, viz. mosquito coils, it is scientifically challenging to characterise the pathomechanism of most concern, since irritant combustion gases, volatile and semi-volatile organic substances, particulates (soot), condensation aerosols and re-condensed substances onto particulates may act independently, synergistically or mixture specific. It has been concluded that for the comparative safety evaluation and risk assessment of indoor insecticide end-use products generally recognised guidance for harmonised inhalation testing is required: 1) For high-dose release products, such as spray-cans, acute inhalation testing appears to be most relevant. 2) For low-dose, slow-release devices, subchronic inhalation studies of 13-weeks, duration of exposure 6 hours/day for 5 consecutive days per week, should be performed on rats preferably with the end-use product. A dose-range finding study of 2-weeks duration, daily exposure, should be available for the justification of dose selection and to demonstrate that the findings of 5 days/week exposure is not different from that utilising a daily exposure. 3) In certain instances, inhalation testing of shorter duration, for example 2 or 4 weeks, may be appropriate if the inhalation toxicity of the basic formula has already been characterised or the major constituents contained in the formulation have been examined in extant studies. The duration of study should also depend on actual use and exposure pattern, the accumulation potential and the mode of action of the ingredients of concern. 4) Particularly for mosquito coil smokes, the possible physico-chemical interactions of particulates, potentially irritant water soluble or nonsoluble vapours and aerosol or vapours of the active ingredient(s) are virtually impossible to predict without appropriate inhalation testing. 5) Physiological measurements may be useful for the detection and quantification of the respiratory tract sensory irritation potency. Biochemical endpoints in lung lavage and conventional histopathology of the lungs and extrathoracic airways are considered to be important. 6) It has been agreed that the nose-only exposure technique provides the most cost-effective and controlled means to expose small laboratory animals to well defined atmospheres as long as the limitations of this technique are taken into account.

Air Pollution, Indoor↗

Insecticides: their effect on microorganisms and persistence in rice soil.

A field experiment was conducted to investigate the effect of four insecticides, HCH, phorate, carbofuran and fenvalerate, at recommended doses on the preponderance of bacteria, actinomycetes and fungi. We also measured the persistence of the insecticides in the rhizosphere soil of rice. HCH and fenvalerate stimulated the proliferation of all of the microorganisms significantly. Phorate increased the population of bacteria and actinomycetes. Carbofuran accentuated the preponderance of actinomycetes in soil. Insecticides, in general, did not have marked influence on the proliferation of Bacillus, Streptomyces, Aspergillus and Fusarium in soil. However, we observed a stimulation of growth of Staphylococcus, Proteus and Sarcina with HCH, Pseudomonas, Corynebacterium, Erysipelothrix and Rhizopus with phorate, Serratia, Corynebacterium, Klebsiella, Escherichia, Rhizopus and Humicola with carbofuran, and Staphylococcus, Sarcina, Klebsiella and Nocardia with fenvalerate. On the other hand, there was an inhibition in growth of Pseudomonas, Micrococcus, Nocardia and Penicillium with HCH, of Pseudomonas, Micrococcus and Penicillium with carbofuran, and of Pseudomonas, Micrococcus and Micromonospora with fenvalerate. Different types of insecticides exhibited differential patterns of dissipation in soil. HCH had the highest persistence followed by phorate, carbofuran and fenvalerate, respectively.

Actinomycetales↗

The molecular basis of two contrasting metabolic mechanisms of insecticide resistance.

The esterase-based insecticide resistance mechanisms characterised to date predominantly involve elevation of activity through gene amplification allowing increased levels of insecticide sequestration, or point mutations within the esterase structural genes which change their substrate specificity. The amplified esterases are subject to various types of gene regulation in different insect species. In contrast, elevation of glutathione S-transferase activity involves upregulation of multiple enzymes belonging to one or more glutathione S-transferase classes or more rarely upregulation of a single enzyme. There is no evidence of insecticide resistance associated with gene amplification in this enzyme class. The biochemical and molecular basis of these two metabolically-based insecticide resistance mechanisms is reviewed.

Animals↗

Modulation of the mutagenicity of heterocyclic amines by organophosphate insecticides and their metabolites.

People are commonly exposed to organophosphorus ester (OP) insecticides through the treatment of pets, homes, lawns, gardens, workplaces and in commercial agriculture. Aromatic amines are another chemical class with wide human exposure particularly dietary heterocyclic aromatic amines (HAAs). Previously, we reported that specific aromatic amines and ethyl paraoxon (the metabolite of the insecticide ethyl parathion) induced enhanced mutagenic responses in Salmonella typhimurium. In the present study, we demonstrated that the mutagenicity of 2-acetoxyacetylaminofluorene (2AAAF) and the heterocyclic dietary carcinogen 2-amino-1-methyl-6-phenylimidazo(4,5-b)pyridine (PhIP) was enhanced in the presence of the OP insecticides, ethyl parathion or methyl parathion or a metabolite (methyl paraoxon). The mutagenicity of 2-amino-3-methylimidazo-(4,5-f)quinoline (IQ) was increased by methyl parathion and methyl paraoxon but not by ethyl parathion. This mutagenic synergy was expressed in S. typhimurium strain YG1024. Mammalian microsomal activation was required for PhIP and IQ to express mutagenic synergy. Synergistic responses are rarely incorporated in risk assessment models, yet such responses are important in establishing accurate toxicological characteristics of agents. Under real world conditions where people are exposed to a multitude of agents, the results of this study raise a concern about the environmental and public health impacts of OP insecticides.

Acetoxyacetylaminofluorene↗

Insecticidal activity of essential oils: octopaminergic sites of action.

UNLABELLED: A study was conducted to determine the insecticidal activity and mechanism of action of three essential oils (eugenol, alpha-terpineol and cinnamic alcohol) and an equal part mixture (3-blend) against American cockroaches (Periplaneta americana). To address species differences in response to treatment with the test oils, Carpenter ants (Camponotus pennsylvanicus De Geer), and German cockroaches (Blattella germanica) were included in this study. Exposed American cockroaches demonstrated hyperactivity followed by hyperextension of the legs and abdomen, then fast knockdown or quick immobilization followed by death. Ants and German cockroaches showed fast immobilization/knockdown followed by mortality. The 1:1:1 mixture (3-blend) was substantially effective against all test insects. One of the most remarkable observations was the increased frequency of heartbeats of American cockroaches in response to topical application of test oils. The changes in the pattern of cAMP level was biphasic. A significant increase in the cAMP level was found in response to 1 nmol/ml of eugenol, or 3-blend or 10 nmol/ml of alpha-terpineol. At higher concentrations a significant decrease in cAMP level was found. Blockage of octopamine receptors binding sites was also illustrated at lower concentrations of the test chemicals as judged by the decreased binding activity of [3H]octopamine to its receptors. IN CONCLUSION: (1) test oils are neuro-insecticides and their insecticidal activity is species-dependent; (2) a synergistic effect of the three oils was found when they were equally mixed (3-blend); and (3) the octpaminergic system mediates the insecticidal activity of eugenol, alpha-terpienol and the 3-blend.

Administration, Topical↗

Insecticide susceptibility and detoxication enzyme activities among Coptotermes formosanus Shiraki workers sampled from different locations in New Orleans.

Coptotermes formosanus Shiraki worker termites were sampled from 20 locations in the City Park area of New Orleans, LA. The termites were subsequently assayed to determine their susceptibility to cypermethrin, chlordane and chlorpyrifos, and detoxication enzyme activity. Cypermethrin was most toxic against Formosan subterranean termite workers, chlorpyrifos exhibited intermediate toxicity and chlordane was least toxic. A comparison of insecticide susceptibility between the most and least tolerant colonies revealed 1.9-, 1.7- and 1.8-fold differences in susceptibility for cypermethrin, chlorpyrifos and chlordane, respectively. As with the bioassay data, although significant differences were noted, a great deal of overlap was observed among the colonies for total cytochrome P450 content (difference of 2.2-fold between high and low value) aldrin epoxidation (3.6-fold) and cytosolic esterase (3.9-fold) activity. No significant differences were observed among the colonies for methoxyresorufin O-demethylase or glutathione S-transferase activity. Conversely, microsomal esterase activity varied greatly; a 38-fold difference was observed between the most (Cf1776) and least (Cf1387) active colonies. However, no significant correlation was observed between insecticide susceptibility and microsomal esterase activity. In fact, no significant correlations were observed between any of the enzyme activities measured and insecticide susceptibility. These results are discussed in the context of insecticide selection and future control effectiveness.

Animals↗

Persistence of the insecticide Dimilin 45 ODC on conifer forest foliage in an Atlantic-climate ecosystem.

The oil formulation of diflubenzuron (Dimilin 45 ODC) persisted for 10-12 weeks on the foliage of a conifer forest in an Atlantic-climate ecosystem. Within 22-30 days following treatment, 55-80% of the insecticide had been removed from the foliage. During this period, the concentration of diflubenzuron was higher than 370 ng g(-1). Aerial application at 56.3 g of Al ha(-1) resulted in deposition levels of the insecticide ranging from 867.5 to 1824.4 ng g(-1), depending upon forest characteristics. The results showed that aerial application is only a suitable technique for the treatment of forest areas with dense foliage and/or high tree density and no more than 15% of tree-free area. The only metabolite detected was 2,6-difluorobenzamide, and this persisted on foliage until the first rainfalls occurred. An empirical mathematical correlation was found to express the influence of meteorological variables--rainfall, solar radiation and temperature--on the persistence of the insecticide. These results suggested that degradation of diflubenzuron on foliage could be due to photodegradation. Some recommendations were made to optimize the deposition of the insecticide on foliage and to minimize its persistence and the off-site spray drift.

Diflubenzuron↗

Insecticides in Chinese medicinal plants: survey leading to jacaranone, a neurotoxicant and glutathione-reactive quinol.

Sixty-two plant species from central China were purported to have insecticidal activity. Adult house fly (Musca domestica) toxicity assays were used to identify the two most active plants and guide the chromatographic isolation of the insecticidal components. The active ingredient of Senecio palmatusPall. (Asteraceae) was characterized as methyl (1-hydroxy-4-oxocyclohexa-2,5-dien-1-yl)acetate (jacaranone) (1), previously known to have insect antifeedant activity. Mono- and bisglutathione (GSH) adducts are formed on incubation of 1 with GSH and rat liver GSH S-transferase. The toxic action of 1 in mice (intraperitoneal LD(50) = 150-200 mg/kg) is associated with both neurological signs and GSH depletion in liver 90 min after treatment. Paeonia suffruticosa var. papaveracea (Andr.) Kerner (Paeoniaceae) was the other active plant found here to have 2'-hydroxy-4'-methoxyacetophenone (paeonol) (2) as an insecticidal ingredient in the root and in one case, for the whole plant, contaminated with S-tert-butylthiomethyl O,O-diethyl phosphorodithioate (terbufos) (3), a synthetic anticholinesterase insecticide.

Animals↗

Insecticide formulations based on nicotine oleate stabilized by sodium caseinate.

Organic farming and new trends toward the use of safer insecticides for crop protection have created new opportunities for botanical insecticides in the pesticide market. In this study, the botanical insecticide nicotine was formulated as a dispersion (20 vol %) stabilized by sodium caseinate, with nicotine oleate solutions used as the dispersed phase. The formulation showed a phase transition on increasing the nicotine oleate concentration, being an emulsion at 7.5-8.2 wt %, a suspo-emulsion at 8.2-9.7 wt %, and a suspension at 9.7-10.8 wt %. Biological activity, apparent viscosity, dispersion time, and protein surface coverage were dependent on nicotine oleate concentration. The emulsion with 8.2 wt % nicotine oleate and the suspo-emulsion with 8.7 wt % nicotine oleate were found to be the most appropriate formulations for insecticide purposes due to their high bioactivity, low viscosity, and low dispersion time. Nicotine oleate formulations showed good creaming and microbiological stability for at least 4 months without losing their biological activity.

Caseins↗

Receptor assay-guided isolation of anti-GABAergic insecticidal alkaloids from a fungal culture.

The gamma-aminobutyric acid (GABA) receptor bears sites of action for insecticides. To discover GABA receptor-directed insecticides in natural products, fungal culture extracts were screened for their ability to inhibit specific binding of the radiolabeled noncompetitive antagonist [3H]1-(4-ethynylphenyl)-4-n-propyl-2,6,7-trioxabicyclo[2.2.2]octane to housefly head membranes. The screening efforts led to the isolation of two alkaloids from Aspergillus terreus: PF1198A (alantrypinone) and PF1198B (serantrypinone), which had IC50 values of 0.34 and 2.1 microM, respectively, in this assay. These compounds were ca. 47-61-fold selective for housefly vs rat GABA receptors. Both compounds showed insecticidal activity against Myzus persicae in the range of 100-500 ppm. Binding assay-guided screening should provide significant opportunities for the identification of novel and selective insecticides.

Alkaloids↗

Insecticidal and fumigant activities of Cinnamomum cassia bark-derived materials against Mechoris ursulus (Coleoptera: attelabidae).

The insecticidal and fumigant activities of Cinnamomum cassia (Blume) bark-derived materials against the oak nut weevil (Mechoris ursulus Roelofs) were examined using filter paper diffusion and fumigation methods and compared to those of the commercially available Cinnamomum bark-derived compounds (eugenol, salicylaldehyde, trans-cinnamic acid, and cinnamyl alcohol). The biologically active constituent of the Cinnamomum bark was characterized as trans-cinnamaldehyde by spectroscopic analysis. In a test with the filter paper diffusion method, trans-cinnamaldehyde showed 100 and 83.3% mortality at rates of 2.5 and 1.0 mg/filter paper, respectively. At 2.5 mg/paper, strong insecticidal activity was produced from eugenol (90.0% mortality) and salicylaldehyde (88. 9%), whereas trans-cinnamic acid revealed moderate activity (73.3%). At 5 mg/paper, weak insecticidal activity (50.0%) was produced from cinnamyl alcohol. In a fumigation test, the Cinnamomum bark-derived compounds were much more effective against M. ursulus larvae in closed cups than in open ones. These results indicate that the insecticidal activity of test compounds was attributable to fumigant action, although there is also significant contact toxicity. As a naturally occurring insect-control agent, the Cinnamomum bark-derived materials described could be useful as a new preventive agent against damage caused by M. ursulus.

Aldehydes↗

Influence of insecticides on microbial transformation of nitrogen and phosphorus in Typic Orchragualf soil.

Four insecticides, viz., BHC, phorate, carbofuran, and fenvalerate, were applied at the rate of 7.5, 1.5, 1.0, and 0.35 kg a.i. ha(-)(1), respectively, to investigate their effects on the growth and activities of N(2)-fixing and phosphate-solubilizing microorganisms in relation to the availability of N and P in laterite (Typic Orchragualf) soil. Insecticides in general, and BHC and phorate in particular, stimulated the proliferation of aerobic nonsymbiotic N(2)-fixing bacteria and phosphate-solubilizing microorganisms and also their biochemical activities, such as nonsymbiotic N(2)-fixing and phosphate-solubilizing capacities, which resulted in greater release of available N (NH(4)(+) and NO(3)(-)) and P in soil. All the insecticides were persistent in soil for a short period of time, and the rate of dissipation was highest for fenvalerate followed by phorate, carbofuran, and BHC, depicting the half-lives (T(1/2)) 8.8, 9.7, 16.9, and 20.6 days, respectively. The insecticides followed first-order reaction kinetics during their dissipation in soil.

Insecticides↗

Exposure of California quail to organophosphorus insecticides in apple orchards in the Okanagan Valley, British Columbia.

We studied the exposure and effect of the organophosphate insecticides azinphos-methyl and diazinon on adult California quail (Callipepla californica) in an apple orchard in the Okanagan Valley, British Columbia. Cholinesterase activity was measured in plasma samples (n = 65) collected from 54 individuals either prior to spraying, immediately (< 24 hours) or 10 days after three spray events. Mean plasma cholinesterase levels declined significantly (P < 0.05, n = 12) to 61% of pre-spray mean activity (controls) immediately following the first spray event, but by ten days had recovered to 86% of mean control activity. Subsequent spray events caused no significant declines in mean plasma cholinesterase activity. Four of the 26 quail sampled within 24 h of a spray event exhibited plasma-ChE inhibition exceeding 50% inhibition. Radio-tagged quail (n = 25) were monitored throughout the breeding season to determine use of orchards and detect changes in use patterns resulting from the spraying of insecticides. Use of orchards by quail varied over the summer, with the highest use occurring in May, declining to very low use by July. Quail exhibited a diurnal pattern, roosting in sparsely forested uplands at night, travelling to orchard areas to feed early each morning and returning to roosts at dusk. Orchard use by quail differed during spray events compared to non-spray times. During the three hour period immediately after spraying (0530-0800), 14-20% of observed quail were in the orchard, after which use declined to < 4%, and returned to 12% by the next day. During non-spray times, 3-13% of radio-tagged the quail were observed in orchard habitat, with the heaviest use (13%) occurring later in the day (0830-1700 h). Seven radio-tagged quail were predated during the study period. However, no deaths could be attributed to insecticide poisoning as carcasses were not in suitable condition for testing. It was concluded that adult quail using orchard habitat early in the summer may be acutely poisoned by anti-cholinesterase insecticides, but the risk of exposure declined over the summer.

Agriculture↗

Minor structural changes in nicotinoid insecticides confer differential subtype selectivity for mammalian nicotinic acetylcholine receptors.

The major nitroimine insecticide imidacloprid (IMI) and the nicotinic analgesics epibatidine and ABT-594 contain the 6-chloro-3-pyridinyl moiety important for high activity and/or selectivity. ABT-594 has considerable nicotinic acetylcholine receptor (AChR) subtype specificity which might carry over to the chloropyridinyl insecticides. This study considers nine IMI analogues for selectivity in binding to immuno-isolated alpha1, alpha3 and alpha7 containing nicotinic AChRs and to purported alpha4beta2 nicotinic AChRs. Alpha1- and alpha3-containing nicotinic AChRs (both immuno-isolated by mAb 35, from Torpedo and human neuroblastoma SH-SY5Y cells, respectively) are between two and four times more sensitive to DN-IMI than to (-)-nicotine. With immuno-isolated alpha3 nicotinic AChRs, the tetrahydropyrimidine analogues of IMI with imine or nitromethylene substituents are 3-4 fold less active than (-)-nicotine. The structure-activity profile with alpha3 nicotinic AChRs from binding assays is faithfully reproduced in agonist potency as induction of 86rubidium ion efflux in intact cells. Alpha7-containing nicotinic AChRs of SH-SY5Y cells (immuno-isolated by mAb 306) and rat brain membranes show maximum sensitivity to the tetrahydropyrimidine analogue of IMI with the nitromethylene substituent. The purported alpha4beta2 nicotinic AChRs [mouse (Chao & Casida, 1997) and rat brain] are similar in sensitivity to DN-IMI, the tetrahydropyrimidine nitromethylene and nicotine. The commercial insecticides (IMI, acetamiprid and nitenpyram) have low to moderate potency at the alpha3 and purported alpha4beta2 nicotinic AChRs and are essentially inactive at alpha1 and alpha7 nicotinic AChRs. In conclusion, the toxicity of the analogues and metabolites of nicotinoid insecticides in mammals may involve action at multiple receptor subtypes with selectivity conferred by minor structural changes.

Animals↗

The methodology and operation of a pilot randomized control trial of the effectiveness of the Bug Busting method against a single application insecticide product for head louse treatment.

A Department of Health leaflet suggests two treatment methods for head lice: mechanical removal by wet combing; and insecticide lotion/rinses. However, there are no reports in the literature comparing the effectiveness of these two treatment methods and well controlled clinical trials of insecticide treatments are sparse. A pilot randomized control trial of the effectiveness of a specific method of wet combing, 'Bug Busting', against a single application of a proprietary insecticide product is reported. The difficulties of designing a trial are discussed and modifications that would allow a definitive trial to take place are suggested. The pilot study included enzyme analysis of lice for insecticide resistance status assessment.

Animals↗

Molecular characterization of the amplified aldehyde oxidase from insecticide resistant Culex quinquefasciatus.

Primary structural information including the complete nucleotide sequence of the first insect aldehyde oxidase (AO) was obtained from the common house mosquito Culex quinquefasciatus (Say) through cloning and sequencing of both genomic DNA and cDNA. The deduced amino-acid sequence encodes a 150-kDa protein of 1266 amino-acid residues, which is consistent with the expected monomeric subunit size of AO. The Culex AO sequence contains a molybdopterin cofactor binding domain and two iron-sulfur centres. A comparison of the partial sequences of AO from insecticide resistant and susceptible strains of C. quinquefasciatus shows two distinct alleles of this enzyme, one of which is amplified in the insecticide resistant strain on a 30-kb DNA amplicon alongside two resistance-associated esterases. The amplified AO gene results in elevated AO activity in all life stages, but activity is highest in 3rd instar larvae. The elevated enzyme can be seen as a separate band on polyacrylamide gel electrophoresis. The role of AO in xenobiotic oxidation in mammals and the partial inhibition of elevated AO activity by a range of insecticides in Culex, suggest that this AO may play a role in insecticide resistance.

3' Untranslated Regions↗

Aldehyde oxidase is coamplified with the World's most common Culex mosquito insecticide resistance-associated esterases.

The evolution and spread of insecticide resistance is an important factor in human disease prevention and crop protection. The mosquito Culex quinquefasciatus is the main vector of the disease filariasis and a member of a species complex which is a common biting nuisance worldwide. The common insecticide resistance mechanism in this species involves germline amplification of the esterases estalpha21 and estbeta21. This amplification has arisen once and rapidly spread worldwide. Less common and more variable resistance phenotypes involve coamplification of estalpha3 and estbeta1, or individual amplification of a single estbeta1, different alleles of the same estalpha and estbeta gene loci. Estalpha21 and estbeta21 are on the same large fragment of amplified DNA (amplicon) 2.7 kb apart. We have now shown that this amplicon contains another full-length gene immediately 5' of estalpha21 which codes for a molybdenum-containing hydroxylase, with highest homology to aldehyde oxidase (AO) from other organisms. The full-length putative AO gene is not present on the estalpha3/estbeta1 or estbeta1 amplicons, but multiple truncated 5' ends of this gene are present around the presumed estalpha3/estbeta1 amplicon breakpoint. Polymerase chain reaction (PCR) analysis of insecticide-susceptible genomic DNA demonstrated that a different allele of the putative AO gene in its non-amplified form is immediately 5' of estalpha. The 'AO' gene on the estalpha21/estbeta21 amplicon is expressed and resistant insects have greater AO activity. This AO activity is sensitive to inhibition by an aldehyde-containing herbicide and pesticide. This enzyme may confer a selective advantage to these insects in the presence of insecticide, as AO in mammals is believed to be important in the detoxification process of several environmental pollutants.

Aldehyde Oxidase↗