RESUMO
Profenofos insecticide poses risks to nontarget organisms including mammals and hydrobionts, and its effects on crops are not known. This study examined the invisible toxicity of profenofos on pakchoi (Brassica rapa L.), using transcriptome and metabolome analyses. Profenofos inhibited the photosynthetic efficiency and light energy absorption by leaves and severely damaged the chloroplasts, causing the accumulation of reactive oxygen species (ROS). Metabolomic analysis confirmed that profenofos promoted the conversion of ß-carotene into abscisic acid (ABA), as evidenced by the upregulation of the carotenoid biosynthesis pathway genes: zeaxanthin epoxidase (ZEP), 9-cis-epoxycarotenoid dioxygenase (NCED3), and xanthoxin dehydrogenase (XanDH). The inhibitory effects on carotenoid accumulation, photosynthesis, and increased ABA and ROS contents of the leaves led to invisible injury and stunted growth of the pakchoi plants. The findings of this study revealed the toxicological risk of profenofos to nontarget crops and provide guidance for the safe use of insecticides.
Assuntos
Brassica rapa , Carotenoides , Metabolômica , Proteínas de Plantas , Brassica rapa/metabolismo , Brassica rapa/genética , Brassica rapa/química , Carotenoides/metabolismo , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Inseticidas/toxicidade , Inseticidas/metabolismo , Folhas de Planta/metabolismo , Folhas de Planta/química , Folhas de Planta/genética , Transcriptoma , Fotossíntese/efeitos dos fármacos , Regulação da Expressão Gênica de Plantas/efeitos dos fármacos , Espécies Reativas de Oxigênio/metabolismo , Organotiofosfatos/metabolismo , Organotiofosfatos/toxicidadeRESUMO
Organophosphates are the most extensively used class of pesticides to deal with increasing pest diversity and produce more on limited terrestrial areas to feed the ever-expanding global population. Profenofos, an organophosphate group of non-systematic insecticides and acaricides, is used to combat aphids, cotton bollworms, tobacco budworms, beet armyworms, spider mites, and lygus bugs. Profenofos was inducted into the system as a replacement for chlorpyrifos due to its lower toxicity and half-life. It has become a significant environmental concern due to its widespread presence. It accumulates in various environmental components, contaminating food, water, and air. As a neurotoxic poison, it inhibits acetylcholinesterase receptor activity, leading to dizziness, paralysis, and pest death. It also affects other eukaryotes, such as pollinators, birds, mammals, and invertebrates, affecting ecosystem functioning. Microbes directly expose themselves to profenofos and adapt to these toxic compounds over time. Microbes use these toxic compounds as carbon and energy sources and it is a sustainable and economical method to eliminate profenofos from the environment. This article explores the studies and developments in the bioremediation of profenofos, its impact on plants, pollinators, and humans, and the policies and laws related to pesticide regulation. The goal is to raise awareness about the global threat of profenofos and the role of policymakers in managing pesticide mismanagement.
Assuntos
Clorpirifos , Inseticidas , Praguicidas , Animais , Humanos , Acetilcolinesterase , Ecossistema , Organotiofosfatos/metabolismo , Inseticidas/toxicidade , Praguicidas/toxicidade , MamíferosRESUMO
Pesticides are widely used for managing pathogens and pests for sustainable agricultural output to feed around seven billion people worldwide. After their targeted role, residues of these compounds may build up and persist in soils and in the food chain. This study evaluated the efficiency of bacterial strains capable of plant growth promotion and biodegradation of profenofos. To execute this, bacteria were isolated from an agricultural area with a history of repeated application of profenofos. The profenofos degrading bacterial strains with growth-promoting characteristics were identified based on biochemical and molecular approaches through partial 16S ribosomal rRNA gene sequencing. The results revealed that one strain, Enterobacter cloacae MUG75, degraded over 90% profenofos after 9 days of incubation. Similarly, plant growth was significantly increased in plants grown in profenofos (100 mg L-1) contaminated soil inoculated with the same strain. The study demonstrated that inoculation of profenofos degrading bacterial strains increased plant growth and profenofos degradation. Novelty statementPesticides are extensively applied in the agriculture sector to overcome pest attacks and to increase food production to fulfill the needs of the growing world population. Residues of these pesticides can persist in the environment for long periods, may enter the groundwater reservoirs and cause harmful effects on living systems highlighting the need for bioremediation of pesticide-contaminated environments. Microbes can use pesticides as a source of carbon and energy and convert them into less toxic and non-toxic products. Application of profenofos degrading rhizobacteria in interaction with the plants in the rhizosphere can remediate the pesticide-contaminated soils and minimize their uptake into the food chain. Hence, this approach can improve soil health and food quality without compromising the environment.
Assuntos
Poluentes do Solo , Solanum lycopersicum , Biodegradação Ambiental , Humanos , Organotiofosfatos/metabolismo , Rizosfera , Microbiologia do Solo , Poluentes do Solo/metabolismoRESUMO
In the current scenario of overuse of pesticides (resulting in soil and water pollution and ultimately leading to biomagnification), a research project was carried out to study biodegradation of Triazophos. For this purpose, three bacterial strains (Pseudomonas kilonensis MB490, Pseudomonas kilonensis MB498 and Pseudomonas sp. MB504), isolated from cotton fields of Mianwali, Pakistan were investigated for Triazophos degradation and metabolite formation in M-9 broth, soil slurry and soil microcosm after incubation for 9 days. There was 88.4-95.8% Triazophos degradation in M-9 broth, 99.90% degradation in soil slurry and 92.74 to 96% Triazophos degradation in soil microcosm by these bacteria after 9 days. While there was negligible Triazophos degradation (upto 7%) in the controls without bacteria. According to GCMS analysis, 7 unique and novel metabolites (1, 2, 4-Triazole-4-amine, N-(2-Thienylmethyl), Benzene sulfonic acid hydrazide, Benzene sulfonic acid methyl ester, 4H-1,2,4-Triazole-4-benzenesulfonamide, 4, 5 dihydro-N-(O-toyl)-3-furamide, Ethyl 4-phenyldiazenylbenzoate and Dibutyl methanephosphonate) of Triazophos were revealed. Current results strongly suggest the potential of these bacterial strains for the remediation of Triazophos contaminated agricultural soils.
Assuntos
Organotiofosfatos/metabolismo , Praguicidas/metabolismo , Pseudomonas/metabolismo , Poluentes do Solo/metabolismo , Triazóis/metabolismo , Agricultura , Biodegradação Ambiental , Recuperação e Remediação Ambiental/métodos , Gossypium , Paquistão , Pseudomonas/efeitos dos fármacos , Pseudomonas/isolamento & purificação , Microbiologia do SoloRESUMO
Organophosphorus insecticides account for approximately 28% of the global commercial insecticide market, while 40% of them are chiral enantiomers. Chiral enantiomers differ largely in their toxicities. Enantiomers that are less active or inactive do not offer the needed efficacy but pollute the environment and cause toxicities to non-target species. Cupriavidus nantongensis X1T, a recently isolated bacterial strain, could degrade S-profenofos 2.3-fold faster than R-profenofos, while the latter is the active enantiomer potently against pest insects and has greater mammalian safety. The degradation enzyme encoded by opdB was expressed via Escherichia coli and purified. The degradation kinetics of R- and S-profenofos showed that both the purified OpdB and crude enzyme extracts had no enantiomer degradation selectivity, which strongly indicated that the degradation selectivity occurred in the uptake process. Metabolite analyses suggested a novel dealkylation pathway. This is the first report of bacterial selective uptake of organophosphates. Selective degradation of S-profenofos over R-profenofos by the strain X1T suggests a concept of co-application of racemic pesticides and degradation-selective bacteria to minimize contamination and non-target toxicity problems.
Assuntos
Cupriavidus/metabolismo , Inseticidas/química , Inseticidas/metabolismo , Organotiofosfatos/química , Organotiofosfatos/metabolismo , Biodegradação Ambiental , Cupriavidus/química , Cinética , EstereoisomerismoRESUMO
The widespread use of pesticides has been one of the major anthropogenic sources of environmental pollution. Organophosphorus (OP) pesticides are predominantly used in agriculture due to their broad-spectrum insecticidal activity and chemical stability. The study was focused on the biodegradation of OP pesticides, Profenofos (PF) and Quinalphos (QP) in culture media using bacterium isolated from wetland paddy rhizosphere. The strain VITPSCQ3 showed higher pesticide tolerance, efficient biofilm formation and was capable of synthesizing organophosphate degrading enzymes. Based on the 16S rRNA gene sequencing the isolate exhibited maximum sequence similarity with Kosakinia oryzae (GenBank accession number: KR149275). Biodegradation assay with various concentrations of PF and QP (200, 400, 600 and 800 mg L-1) showed maximum degradation up to 82% and 92% within 48 h. The kinetic studies revealed the biodegradation rates (k) to be 0.0844 min-1 and 0.107 min-1 with half-lives (h) of 18 h and 14.8 h for PF and QP. The degradation products were identified by GCMS and possible degradation pathways were proposed using Insilico techniques. To the best of our knowledge, this is the first report on the biodegradation of PF and QP using Kosakonia oryzae. Bioremoval of PF and QP from aqueous solution was performed using the biofilm of VITPSCQ3 developed on selected substrates in a circulating Vertical-flow packed-bed biofilm (VFPBB) bioreactor. Charcoal, gravel and mushroom (Agaricus bisporus) were used as biofilm carriers. Mushroom showed strong biofilm formation with optimum biodegradation capacity of up to 96% for PF and 92% for QP within 120 min reaction time.
Assuntos
Biofilmes/crescimento & desenvolvimento , Reatores Biológicos/microbiologia , Enterobacteriaceae/enzimologia , Poluentes Ambientais/metabolismo , Inseticidas/metabolismo , Organotiofosfatos/metabolismo , Compostos Organotiofosforados/metabolismo , Biodegradação Ambiental , Enterobacteriaceae/genética , Enterobacteriaceae/fisiologia , Cinética , RizosferaRESUMO
Profenofos insecticide is one of the most broadly used organophosphorus pesticides causing the contamination of soil and groundwater. Since dissolved oxygen concentration in groundwater is limited, this study aimed to investigate profenofos biodegradation and detoxification under aerobic and anoxic conditions using the profenofos-degrading Pseudomonas plecoglossicida strain PF1 (PF1). Anoxic biodegradation under the presence of nitrate was the focus. The results showed that profenofos at 10-150 mg/L was degraded under aerobic and anoxic conditions with removal efficiencies of 38-55% and 27-45%, respectively. Kinetic analysis following the Michaelis-Menten model revealed that the maximum substrate degradation rates and the Michaelis constants were 13.07 and 8.92 mg/L/d and 92.07 and 84.76 mg/L under aerobic and anoxic conditions, respectively. The culture preferred an aerobic environment resulting in better biodegradation performance. During the degradation experiment, 4-bromo-2-chlorophenol and 1,1-dimethylethylphenol were detected as profenofos biodegradation intermediate products. Microbial toxicity, phytotoxicity, and cytogenotoxicity assays showed that the toxicity of the contaminated water significantly decreased after both aerobic and anoxic biodegradation by PF1. The results from this study indicated that PF1 has the potential for bioremediation in a profenofos-contaminated environment under the presence or absence of oxygen.
Assuntos
Biodegradação Ambiental , Inseticidas/metabolismo , Organotiofosfatos/metabolismo , Pseudomonas/metabolismo , Animais , Clorofenóis , Besouros/metabolismo , Água Subterrânea , Inativação Metabólica , Cinética , Nitratos , Oxigênio/metabolismo , PraguicidasRESUMO
Organophosphorus insecticides (OPs) have been widely used to control agricultural pests, which has raised concerns about OP residues in crops and the environment. In this study, we investigated the degradation kinetics and pathways of 8 OPs by Cupriavidus nantongensis X1T and identified the enzyme via gene cloning and in vitro assays. The degradation half-life of methyl parathion, triazophos, and phoxim was only 5, 9, and 43 min, respectively. It was 46 fold faster than that of triazophos by Bacillus sp. TAP-1, a well-studied triazophos-degrader. Strain X1T completely degraded not only chlorpyrifos, methyl parathion, parathion, fenitrothion, triazophos, and phoxim at 50 mg/L within 48 h but also the phenolic metabolites. This was the fastest degradation of OPs by bacterial whole cells reported thus far. The OPs were first hydrolyzed by an OP hydrolase encoded by the opdB gene in strain X1T, followed by further degradation of the metabolites. The crude enzyme maintained a full activity.
Assuntos
Clorpirifos/metabolismo , Cupriavidus/metabolismo , Inseticidas/metabolismo , Bacillus/metabolismo , Biodegradação Ambiental , Clorpirifos/química , Cupriavidus/química , Inseticidas/química , Cinética , Organotiofosfatos/metabolismo , Triazóis/metabolismoRESUMO
Triazophos (TAP) was a widely used organophosphorus insecticide in developing countries. TAP could produce specific metabolites triazophos-oxon (TAPO) and 1-phenyl-3-hydroxy-1,2,4-triazole (PHT) and non-specific metabolites diethylthiophosphate (DETP) and diethylphosphate (DEP). The objective of this study involved computational approaches to discover potential mechanisms of molecular interaction of TAP and its major metabolites with endocrine hormone-related proteins using molecular docking in silico. We found that TAP, TAPO and DEP showed high binding affinity with more proteins and enzymes than PHT and DETP. TAP might interfere with the endocrine function of the adrenal gland, and TAP might also bind strongly with glucocorticoid receptors and thyroid hormone receptors. TAPO might disrupt the normal binding of androgen receptor, estrogen receptor, progesterone receptor and adrenergic receptor to their natural hormone ligands. DEP might affect biosynthesis of steroid hormones and thyroid hormones. Meanwhile, DEP might disrupt the binding and transport of thyroid hormones in the blood and the normal binding of thyroid hormones to their receptors. These results suggested that TAP and DEP might have endocrine disrupting activities and were potential endocrine disrupting chemicals. Our results provided further reference for the comprehensive evaluation of toxicity of organophosphorus chemicals and their metabolites.
Assuntos
Disruptores Endócrinos/metabolismo , Enzimas/metabolismo , Organofosfatos/metabolismo , Organotiofosfatos/metabolismo , Praguicidas/metabolismo , Proteínas/metabolismo , Triazóis/metabolismo , Animais , Sítios de Ligação , Disruptores Endócrinos/química , Enzimas/química , Humanos , Ligantes , Simulação de Acoplamento Molecular , Organofosfatos/química , Organotiofosfatos/química , Praguicidas/química , Ligação Proteica , Proteínas/química , Triazóis/química , TurquiaRESUMO
In recent decades, biodegradation has been considered a promising and eco-friendly way to eliminate organophosphorus pesticides (OPs) from the environment. To enrich current biodegrading-enzyme resources, an alkaline phosphatase (AP3) from Bacillus amyloliquefaciens YP6 was characterized and utilized to test the potential for new applications in the biodegradation of five broad-spectrum OPs. Characterization of AP3 demonstrated that activity was optimal at 40 °C and pH 10.3. The activity of AP3 was enhanced by Mg2+, Ca2+, and Cu2+, and strongly inhibited by Mn2+, EDTA, and L-Cys. Compared to disodium phenyl phosphate, p-nitrophenyl phosphate (pNPP) was more suitable to AP3, and the Vm, Km, kcat, kcat/Km values of AP3 for pNPP were 4,033 U mg-1, 12.2 mmol L-1, 3.3 × 106 s-1, and 2.7 × 108 s-1mol-1L, respectively. Degradation of the five OPs, which included chlorpyrifos, dichlorvos, dipterex, phoxim, and triazophos, was 18.7%, 53.0%, 5.5%, 68.3%, and 96.3%, respectively, after treatment with AP3 for 1 h. After treatment of the OP for 8 h, AP3 activities remained more than 80%, with the exception of phoxim. It can be postulated that AP3 may have a broad OP-degradation ability and could possibly provide excellent potential for biodegradation and bioremediation in polluted ecosystems.
Assuntos
Fosfatase Alcalina/metabolismo , Bacillus amyloliquefaciens/enzimologia , Organofosfatos/metabolismo , Praguicidas/metabolismo , Compostos de Anilina/metabolismo , Biodegradação Ambiental , Clorpirifos/metabolismo , Nitrofenóis/metabolismo , Compostos Organofosforados/metabolismo , Organotiofosfatos/metabolismo , Triazóis/metabolismoRESUMO
Regiospecific replacement of a single phosphate (PO2) by a dithiophosphate (PS2) group in an RNA can dramatically increase its binding affinity for a target protein. Thus, complexes between antithrombin and anti-VEGF RNA aptamers with single dithiophosphate moieties and thrombin and VEGF, respectively, display equilibrium dissociation constants KD of ca. 1 pM, 1000-fold tighter than the native RNA complexes (ca. 1 nM). Inspection of crystal structures of the native and PS2-RNA aptamer:thrombin complexes reveals an RNA-induced fit in the latter. This leads to a close approach between PS2 and the phenyl ring edge of Phe-232 that is surrounded by pairs of lysines and arginines. To better understand the origins of the tighter binding and individual contributions to the interaction energy, we carried out QM calculations with phosphate- and dithiophosphate-benzene and dimethyl phosphate- and dimethyl dithiophosphate-benzene model systems. These calculations demonstrate that the dithiophosphate-benzene interaction is much stronger than the corresponding interaction with phosphate. QM/MM calculations with the full complexes confirmed this finding and support the hypothesis that the electric field generated by basic residues surrounding Phe-232 is key to the polarization of the PS2 moiety. Thus, disparate polarization and dispersion energies between the PO2 and PS2 complexes contribute critically to the difference in binding affinity. By comparison, easier desolvation of the dithiophosphate group compared to phosphate does not contribute decisively to the observed difference in binding affinity. Favorable polarization and dispersion energies may be a general feature of the dramatic affinity gains seen for complexes between RNAs carrying dithiophosphate groups and their binding proteins.
Assuntos
Aptâmeros de Nucleotídeos/metabolismo , Organotiofosfatos/metabolismo , Trombina/metabolismo , Aptâmeros de Nucleotídeos/química , Humanos , Modelos Moleculares , Conformação de Ácido Nucleico , Organotiofosfatos/química , Ligação Proteica , Teoria Quântica , Fator A de Crescimento do Endotélio Vascular/metabolismoRESUMO
Acetylcholinesterase-1 (AChE1) is a vital enzyme involved in neurotransmission and represents an attractive insecticide-target for organophosphates and carbamates in Plutella xylostella (Linneaus), an important pest of cruciferous crops worldwide. However, insecticide-resistance often occurs due to mutations, making many organophosphates and carbamates ineffective. In particular, A298S and G324A mutations in AChE1 significantly lower the binding affinity of insecticides. In the present study, the wild-type and mutant AChE1 structures were constructed and their structural stabilities, residual flexibilities were investigated through molecular dynamics simulations. Subsequently, the structural and energetic changes responsible for the insecticide-resistance in AChE1 were analyzed using molecular docking. The results of molecular dynamics simulation showed that the mutant AChE1 shows little structural deviation than the wild-type, indicate the structural instability. Furthermore, the docking results demonstrated that these mutations break the intermolecular hydrogen bonding interactions and thereby affect the prothiofos as well as all insecticide binding. Hence, the results could provide some insights into the resistance mechanism of AChE1 in insecticides binding and helpful in the development of novel insecticides that are less susceptible to insecticide-resistance.
Assuntos
Acetilcolinesterase/genética , Acetilcolinesterase/metabolismo , Inseticidas/metabolismo , Mariposas/enzimologia , Mariposas/genética , Acetilcolinesterase/química , Substituição de Aminoácidos , Animais , Brassicaceae/parasitologia , Estabilidade Enzimática , Ligação de Hidrogênio , Resistência a Inseticidas , Simulação de Acoplamento Molecular , Simulação de Dinâmica Molecular , Mariposas/química , Mariposas/efeitos dos fármacos , Organotiofosfatos/metabolismo , Mutação Puntual , Ligação ProteicaRESUMO
The use of insecticides has been a primary tool to manage Bactrocera dorsalis in Pakistan; however, recent reports of field control failures necessitate mapping out the insecticide resistance problem. Therefore, eight field strains from Pakistan, were evaluated for their resistance against trichlorfon and spinosad. Compared with a reference strain, six field strains showed high levels of resistance to trichlorfon, while two field strains expressed intermediate resistance. In case of spinosad, five field strains fell in the susceptible range, whereas, the rest of the strains represented minor resistance. Correlation analysis between LD50 values of trichlorfon and spinosad of all the field strains revealed non-significant association, suggesting the possibility of lack of cross-resistance between both insecticides. Synergism bioassays implementing S,S,S-tributylphosphorotrithioate (DEF) and piperonyl butoxide (PBO) revealed that the LD50 values of trichlorfon in the presence of either DEF or PBO in seven field strains were significantly reduced. However, DEF and PBO had a non-significant effect on synergizing spinosad toxicity. The results revealed resistance to trichlorfon in field strains of B. dorsalis, which might be metabolic-based. Absence or minor resistance to spinosad and lack of cross-resistance to trichlorfon, suggest that spinosad could be a potential candidate for managing B. dorsalis.
Assuntos
Resistência a Inseticidas/efeitos dos fármacos , Inseticidas/efeitos adversos , Tephritidae/efeitos dos fármacos , Animais , Combinação de Medicamentos , Humanos , Macrolídeos/farmacologia , Organotiofosfatos/metabolismo , Paquistão , Butóxido de Piperonila/metabolismo , Tephritidae/patogenicidade , Triclorfon/farmacologiaRESUMO
This study was performed to identify the degradation products of profenofos "a phenyl organothiophosphate insecticide" in raw water (RW) collected from the entry point of Metropolitan Water Works Authority "Bangkaen, Thailand" and ultrapure water (UPW) with and without TiO2 under simulated sunlight irradiation. Degradation of profenofos was followed with ultrahigh performance liquid chromatography (UHPLC) and follows pseudo first-order kinetic. Accordingly, high-field FTICR mass spectrometry coupled to an electrospray ionization source was used to reveal the degradation routes of profenofos and the isotopic fine structures (IFS) elucidations to approve the chemical structures of its degradation products. More degradation products were detected in UPW as compared to RW. Consequently, two main degradation pathways namely (i) interactive replacements of bromine and hydrogen by hydroxyl functional groups and (ii) rupture of PO, PS, CBr and CCl bonds were observed. None interactive replacement of chlorine by hydroxyl functional group was detected. Accordingly, mechanistical pathways of the main degradation products were established.
Assuntos
Cromatografia Líquida/métodos , Inseticidas/análise , Espectrometria de Massas/métodos , Organotiofosfatos/análise , Poluentes Químicos da Água/análise , Concentração de Íons de Hidrogênio , Inseticidas/metabolismo , Cinética , Organotiofosfatos/metabolismo , Fotólise , Luz Solar , Tailândia , Titânio/químicaRESUMO
Pesticides play an important role in the protection of different crops. Among the diverse sets of pesticides used all over the world, the organophosphates are the most widely used group. Profenofos [O-(4-bromo-2-chlorophenyl) O-ethyl S-propyl phosphorothioate] is one of the most largely used organophosphate insecticides on field crops, vegetables, and fruit crops. The World Health Organization classifies this compound as moderately hazardous (Toxicity Class II), and its residues have been found in vegetables like okra [ (L.) Moench], gooseberries ( sp.), green chilies [ (L.)], curry leaves [ (L.) Spreng], mint leaves [ (L.)], and coriander leaves [ (L.)]. Dietary intake of profenofos (PFF) is the major exposure pathway for humans. When applied to agricultural fields, PFF residues spread into every part of the environment: ambient air, surface water, and soil. In this review, we discuss the worldwide usage of PFF pesticide, its toxic effects on humans and other living organisms in the environment, and biodegradation of this chemical by various microbial strains. To date, no complete biodegradation pathway has been established for PFF pesticide, calling for a study of this nature.
Assuntos
Biodegradação Ambiental , Organotiofosfatos/toxicidade , Praguicidas/toxicidade , Acetilcolinesterase , Humanos , Organotiofosfatos/metabolismo , Praguicidas/metabolismoRESUMO
To understand the mechanism underlying organophosphate pesticide toxicity, cyanobacterium Anabaena PCC 7120 was subjected to varied concentrations (0, 5, 10, 20 and 30 mg L(-1)) of profenofos and the effects were investigated in terms of changes in cellular physiology, genomic template stability and protein expression pattern. The supplementation of profenofos reduced the growth, total pigment content and photosynthetic efficiency of the test organism in a dose dependent manner with maximum toxic effect at 30 mg L(-1). The high fluorescence intensity of 2', 7' -dichlorofluorescin diacetate and increased production of malondialdehyde confirmed the prevalence of acute oxidative stress condition inside the cells of the cyanobacterium. Rapid amplified polymorphic DNA (RAPD) fingerprinting and SDS-PAGE analyses showed a significant alteration in the banding patterns of DNA and proteins respectively. A marked increase in superoxide dismutase, catalase, peroxidase activity and a concomitant reduction in glutathione content indicated their possible role in supporting the growth of Anabaena 7120 up to 20 mg L(-1). These findings suggest that the uncontrolled use of profenofos in the agricultural fields may not only lead to the destruction of the cyanobacterial population, but it would also disturb the nutrient dynamics and energy flow.
Assuntos
Anabaena/enzimologia , Catalase/metabolismo , DNA de Algas/efeitos dos fármacos , Inseticidas/toxicidade , Malondialdeído/toxicidade , Organotiofosfatos/toxicidade , Fenômenos Fisiológicos/efeitos dos fármacos , Anabaena/efeitos dos fármacos , Catalase/efeitos dos fármacos , Relação Dose-Resposta a Droga , Eletroforese em Gel de Poliacrilamida , Inseticidas/metabolismo , Malondialdeído/metabolismo , Organotiofosfatos/metabolismo , Fotossíntese/efeitos dos fármacos , Técnica de Amplificação ao Acaso de DNA Polimórfico , Superóxido Dismutase/efeitos dos fármacosRESUMO
Triazophos is a broad-spectrum and highly effective insecticide, and the residues of triazophos have been frequently detected in the environment. A triazophos-degrading bacterium, Burkholderia sp. SZL-1, was isolated from a long-term triazophos-polluted soil. Strain SZL-1 could hydrolyze triazophos to 1-phenyl-3-hydroxy-1,2,4-triazole, which was further utilized as the carbon sources for growth. The triazophos hydrolase gene trhA, cloned from strain SZL-1, was expressed and homogenously purified using Ni-nitrilotriacetic acid affinity chromatography. TrhA is 55 kDa and displays maximum activity at 25°C, pH 8.0. This enzyme still has nearly 60% activity at the range of 15°C-50°C for 30 min. TrhA was mutated by sequential error prone PCR and screened for improved activity for triazophos degradation. One purified variant protein (Val89-Gly89) named TrhA-M1 showed up to 3-fold improvement in specific activity against triazophos, and the specificity constants of Kcat and Kcat/Km for TrhA-M1 were improved up to 2.3- and 8.28-fold, respectively, compared to the wild-type enzyme. The results in this paper provided potential material for the contaminated soil remediation and hydrolase genetic structure research.
Assuntos
Burkholderia/genética , Hidrolases/genética , Hidrolases/metabolismo , Organotiofosfatos/metabolismo , Microbiologia do Solo , Triazóis/metabolismo , Biodegradação Ambiental , Burkholderia/enzimologia , Burkholderia/crescimento & desenvolvimento , Burkholderia/isolamento & purificação , Cromatografia de Afinidade , Clonagem Molecular , Recuperação e Remediação Ambiental , Expressão Gênica , Hidrolases/química , Hidrolases/isolamento & purificação , Hidrólise , Cinética , MutaçãoRESUMO
Triazophos (TAP) is a widely used pesticide that is easily accumulated in the environment due to its relatively high stability: this accumulation from agricultural runoff results in potential hazards to aquatic ecosystems. Constructed wetlands are generally considered to be an effective technology for treating TAP polluted surface water. However, knowledge about the biological mechanisms of TAP removal is still lacking. This study investigates the responses of a wetland plant (Canna indica), substrate enzymes and microbial communities in bench-scale horizontal subsurface-flow constructed wetlands (HSCWs) loaded with different TAP concentrations (0, 0.1, 0.5 and 5 mg · L(-1)). The results indicate that TAP stimulated the activities of superoxide dismutase (SOD) and peroxidase (POD) in the roots of C. indica. The highest TAP concentrations significantly inhibited photosynthetic activities, as shown by a reduced effective quantum yield of PS II (ΦPS II) and lower electron transport rates (ETR). However, interestingly, the lower TAP loadings exhibited some favorable effects on these two variables, suggesting that C. indica is a suitable species for use in wetlands designed for treatment of low TAP concentrations. Urease and alkaline phosphatase (ALP) in the wetland substrate were activated by TAP. Two-way ANOVA demonstrated that urease activity was influenced by both the TAP concentrations and season, while acidphosphatase (ACP) only responded to seasonal variations. Analysis of high throughput sequencing of 16S rRNA revealed seasonal variations in the microbial community structure of the wetland substrate at the phylum and family levels. In addition, urease activity had a greater correlation with the relative abundance of some functional microbial groups, such as the Bacillaceae family, and the ALP and ACP may be influenced by the plant more than substrate microbial communities.
Assuntos
Organotiofosfatos/análise , Praguicidas/análise , Triazóis/análise , Eliminação de Resíduos Líquidos/métodos , Áreas Alagadas , Recuperação e Remediação Ambiental , Água Doce/química , Organotiofosfatos/metabolismo , Praguicidas/metabolismo , Triazóis/metabolismo , Movimentos da ÁguaRESUMO
Chlorpyrifos is an organophosphorus pesticide commonly used in agriculture. It is noxious to a variety of organisms that include living soil biota along with beneficial arthropods, fish, birds, humans, animals, and plants. Exposure to chlorpyrifos may cause detrimental effects as delayed seedling emergence, fruit deformities, and abnormal cell division. Contamination of chlorpyrifos has been found about 24 km from the site of its application. There are many physico-chemical and biological approaches to remove organophosphorus pesticides from the ecosystem, among them most promising is biodegradation. The 3,5,6-trichloro-2-pyridinol (TCP) and diethylthiophosphate (DETP) as primary products are made when chlorpyrifos is degraded by soil microorganisms which further break into nontoxic metabolites as CO(2), H(2)O, and NH(3). Pseudomonas is a diversified genus possessing a series of catabolic pathways and enzymes involved in pesticide degradation. Pseudomonas putida MAS-1 is reported to be more efficient in chlorpyrifos degradation by a rate of 90% in 24 h among Pseudomonas genus. The current review analyzed the comparative potential of bacterial species in Pseudomonas genus for degradation of chlorpyrifos thus, expressing an ecofriendly approach for the treatment of environmental contaminants like pesticides.
Assuntos
Clorpirifos/metabolismo , Praguicidas/metabolismo , Pseudomonas/metabolismo , Amônia/metabolismo , Biotransformação , Dióxido de Carbono/metabolismo , Redes e Vias Metabólicas , Organotiofosfatos/metabolismo , Piridonas/metabolismo , Água/metabolismoRESUMO
Profenofos is an organophosphate pesticide used extensively in agriculture to control pests. A bacterium capable of degrading profenofos was isolated from pesticide-contaminated soil samples and identified as Pseudoxanthomonas suwonensis strain HNM based on its morphological and biochemical characteristics and phylogenetic analysis of 16S rRNA gene sequences. 4-Bromo-2-chlorophenol was identified as a metabolite of profenofos degradation by HPLC and GC-MS analysis. The organism degraded profenofos by hydrolysis to yield 4-bromo-2-chlorophenol which was further utilized as carbon source for growth. The organism utilized various organophosphate pesticides such as temephos, quinalphos, and chloropyrifos as carbon sources. The optimum conditions for degradation of profenofos by P. suwonensis strain HMN were found to be at pH 7 and 30 °C. We have investigated the rate of degradation of profenofos by the free and immobilized cells of P. suwonensis strain HNM in various matrices such as sodium alginate (SA), sodium alginate-polyvinyl alcohol (SA-PVA), and SA-bentonite clay. The rate of degradation of 3 and 6 mM profenofos by the freely suspended cells were compared with that by immobilized cells in batches and semi-continuous with shaken cultures. The SA-bentonite clay-immobilized cells showed higher rate of degradation of 3 and 6 mM profenofos then freely suspended cells and cells immobilized in SA and SA-PVA. The SA-bentonite clay-immobilized cells of P. suwonensis strain HNM could be reused for more than 32 cycles without losing their degradation capacity. Thus, the immobilized cells are more efficient than freely suspended cells for the degradation of organophosphate pesticide contaminated water.