Chlorinated Pesticides (2,4-D and DDT) Biodegradation at High Concentrations Using Immobilized Pseudomonas Fluorescens

Abstract

Degradation of two chlorinated pesticides (2,4-D and DDT) using a 54-mL glass column packed with tezontle (a low-cost basaltic scoria) was tested. Bacteria were cultured in YPG (yeast, peptone, and glucose) liquid medium at 32°C. The rich medium was pumped during 24 h through the column to inoculate it. Later, the wasted medium was discharged and the pesticide added. Optical densities, TOC, and pesticide concentration were determined. Pesticide removals for 2,4-D (with initial concentration between 100 and 500 mg/L) were about 99%. DDT removal (at initial concentration of up to 150 mg/L) was as high as 55–99%. TOC removals for 2,4-D was in the 36-87% interval, whereas for DDT they were as high as 36–78%.     

Soil column experiments used as a means to assess transport, sorption, and biodegradation of pesticides in groundwater

Soil column experiments are used to investigate the fate of three pesticides of high, intermediate, and low solubility in groundwater: N- phosphonomethyl glycine (glyphosate); O,O-diethyl-S-[(ethylthio)methyl]phosphorodithioate (phorate); (2,4-dichlorophenoxy)acetic acid (2,4-D). Feed solutions are prepared by adding each pesticide (100 mg/L glyphosate, 50 micro g/L phorate, 50 mg/L 2,4-D) along with conservative tracer, KBr, in synthetic groundwater. The concentration of the pesticides in effluents is detected by ion chromatography (glyphosate, 2,4-D) and GC-FID (phorate). The Br(-) breakthrough curves are employed to estimate the dispersion coefficient and mean pore velocity in each column. Solute transport and reactive models accounting for equilibrium/non-equilibrium sorption and biodegradation are coupled with inverse modeling numerical codes to estimate the kinetic parameters for all pesticides.     

Biodegradation behavior of agricultural pesticides in anaerobic batch reactors

This study explored the biodegradation potential of two agricultural pesticides (2,4-D and isoproturon) as well as their effect on the performance of the anaerobic digestion process. Three 3.5 L batch reactors were used, having the same initial isoproturon concentration (25 mg/L) and different 2,4-D concentrations (i.e. 0, 100, or 300 mg/L, respectively). All systems were fed with equal amounts of primary sludge and digested sludge and operated at the low mesophilic range (32 +/- 2 degrees C). Following an acclimation period of approximately 30 days, complete 2,4-D removal was achieved, whereas isoproturon biodegradation was practically negligible. The presence of 2,4-D did not have a direct effect on acidogenesis since soluble organic carbon [expressed either as volatile fatty acids (VFAs) or as total organic carbon (TOC)] peaked within the first 10 days of operation in all bioreactors. Utilization of VFAs however appeared to follow two distinct patterns: one pattern was represented by acetate and butyrate (i.e. no acid accumulation) while the other was followed by propionate, isobuturate, valerate and isovalerate (i.e. acid accumulation, duration of which was related to the initial 2,4-D concentration). On the whole, all reactors exhibited a successful digestion performance demonstrated by complete VFAs utilization, considerable gas production (containing 45 to 65% methane by volume), substantial volatile suspended solids (VSS) reduction (42 to 50%), as well as pH and alkalinity recovery.     

Biodegradation behavior of agricultural pesticides in anaerobic batch reactors

This study explored the biodegradation potential of two agricultural pesticides (2,4-D and isoproturon) as well as their effect on the performance of the anaerobic digestion process. Three 3.5 L batch reactors were used, having the same initial isoproturon concentration (25 mg/L) and different 2,4-D concentrations (i.e. 0, 100, or 300 mg/L, respectively). All systems were fed with equal amounts of primary sludge and digested sludge and operated at the low mesophilic range (32 ± 2°C). Following an acclimation period of approximately 30 days, complete 2,4-D removal was achieved, whereas isoproturon biodegradation was practically negligible. The presence of 2,4-D did not have a direct effect on acidogenesis since soluble organic carbon [expressed either as volatile fatty acids (VFAs) or as total organic carbon (TOC)] peaked within the first 10 days of operation in all bioreactors. Utilization of VFAs however appeared to follow two distinct patterns: one pattern was represented by acetate and butyrate (i.e. no acid accumulation) while the other was followed by propionate, isobuturate, valerate and isovalerate (i.e. acid accumulation, duration of which was related to the initial 2,4-D concentration). On the whole, all reactors exhibited a successful digestion performance demonstrated by complete VFAs utilization, considerable gas production (containing 45 to 65% methane by volume), substantial volatile suspended solids (VSS) reduction (42 to 50%), as well as pH and alkalinity recovery.     

Soil column experiments used as a means to assess transport,sorption, and biodegradation of pesticides in groundwater

Soil column experiments are used to investigate the fate of three pesticides of high, intermediate, and low solubility in groundwater: N- phosphonomethyl glycine (glyphosate); O,O-diethyl-S-[(ethylthio)methyl]phosphorodithioate (phorate); (2,4-dichlorophenoxy)acetic acid (2,4-D). Feed solutions are prepared by adding each pesticide (100 mg/L glyphosate, 50 μ g/L phorate, 50 mg/L 2,4-D) along with conservative tracer, KBr, in synthetic groundwater. The concentration of the pesticides in effluents is detected by ion chromatography (glyphosate, 2,4-D) and GC-FID (phorate). The Br^? breakthrough curves are employed to estimate the dispersion coefficient and mean pore velocity in each column. Solute transport and reactive models accounting for equilibrium/non-equilibrium sorption and biodegradation are coupled with inverse modeling numerical codes to estimate the kinetic parameters for all pesticides.     

Phenol removal from high salinity effluents using Fenton's reagent and photo-Fenton reactions

The removal of pollutants in saline medium by the Fenton's reagent needs a more detailed investigation, since the presence of chloride may inhibit or retard degradation. Phenol was used as a model pollutant and the influence of some important process variables for the removal of total organic carbon and phenol were investigated, such as FeSO4 and H2O2 concentrations, pH and salinity. The reactivity of iron cations and alternative procedures of applying UV radiation (photo-Fenton) were evaluated. Phenol was fast and completely removed by the Fenton's process even in a high saline medium (50,000mg NaCll(-1)). However, TOC was only moderately or poorly removed in saline media, depending on the salt concentration. When the photo-Fenton process was used, mineralization was improved and high TOC removals were observed in moderately saline media (NaCl concentration < or =10,000mgl(-1)). For the highest NaCl concentration tested (50,000mgl(-1)) only a moderate TOC removal was observed (50%).     

Degradation of 2,4-dichlorophenol by combining photo-assisted Fenton reaction and biological treatment.

The photo-Fenton reaction effect on the biodegradability improvement of 100 mg/L solution of 2,4-dichlorophenol (DCP) has been investigated. Biochemical oxygen demand (BOD) at 5 and 21 days, BODn/ chemical oxygen demand (COD) and BODn/total organic carbon (TOC) ratios, average oxidation state, and inhibition on activated sludge were monitored. For 50 mg/L hydrogen peroxide and 10 mg/L iron(II) initial concentrations and 40 minutes of reaction time in the photo-Fenton process, the biodegradability of the pretreated solution, measured as BOD5/COD ratio, was improved from 0 for the original DCP solution up to 0.18 (BOD21/COD = 0.24). At that point, all DCP was eliminated from the solution. To study the effect of the pretreatment step, the biological oxidation of pretreated solutions was tested in two semicontinuous stirred tank reactors, one operated with activated sludge and one with biomass acclimated to phenol. Results showed that more than 80% TOC removal could be obtained by codigestion of the pretreated solution with municipal wastewater. Total organic carbon removals of approximately 60% were also obtained when the sole carbon source for the aerobic reactors was the pretreated solution. The hydraulic retention times used in the bioreactors were of the same order of magnitude as those used at domestic wastewater treatment plants (i.e., between 12 and 24 hours). Kinetic studies based on pseudo-first-order kinetics have also been carried out. Constants were found to be in range 0.67 to 1.7 L x g total volatiles suspended solids(-1) x h(-1).     

Treatment and reuse of wastewater in pesticide 2,4-D production

Waste-water from the production of pesticide 2,4-D often contains high concentrations of 2,4-DCP and 2,6-D as the primary pollutants. Treatment of waste-water collected from a 2,4-D manufacturer was carried out using a technology combining acidification with hyper-crosslinked resin NDA-150 adsorption process. The overall process recovered 5.4 kg of 2,4-DCP and 0.6 kg of 2,6-D per cubic metre of the wastewater. The treatment reduced the concentration of 2,4-DCP in the wastewater from >6000 mg/L to <0.5 mg/L. The optimal operation parameters of adsorption and desorption were determined. The hyper-crosslinked resin adsorbent can be re-used after regeneration by NaOH aqueous solution. The recovered 2,4-DCP with a sufficiently high purity may be re-used in the production of 2,4-D. The technology may thus be applied to the treatment of waste-water for reclamation of chemicals for 2,4-D production while minimising the environmental nuisances and hazards that may be caused by these chemicals.     

Coagulation and precipitation as post-treatment of anaerobically treated primary municipal wastewater.

The main objective of this study was to investigate the feasibility of coagulation as a post-treatment method of anaerobically treated primary municipal wastewater. Both mesophilic and ambient (20 degrees C) temperature conditions were investigated in a laboratory-scale upflow anaerobic sludge bed (UASB) reactor. In addition, optimization of the coagulant, both in terms of type and dose, was performed. Finally, phosphorus removal by means of aluminum and iron coagulation and phosphorus and ammonia nitrogen removal by means of struvite precipitation were studied. Anaerobic treatment of primary effluent at low hydraulic retention times (less than 15 hours) resulted in mean chemical oxygen demand (COD) removals ranging from 50 to 70%, while, based on the filtered treated effluent, the mean removals increased to 65 to 80%. Alum coagulation of the UASB effluent gave suspended solids removals ranging from approximately 35 to 65%. Turbidity removal reached up to 80%. Remaining COD values after coagulation and settling were below 100 mg/L, while remaining total organic carbon (TOC) levels were below 50 mg/L. Filterable COD levels were generally below 60 mg/L, while filterable TOC levels were below 40 mg/L. All coagulants tested, including prepolymerized aluminum and iron coagulants, demonstrated similar efficiency compared with alum for the removal of suspended solids, COD, and TOC. Regarding struvite precipitation, optimal conditions for phosphorus and nitrogen removal were pH 10 and molar ratio of magnesium: ammonia-nitrogen: phosphate-phosphorus close to the stoichiometric ratio (1:1:1). During struvite precipitation, removal of suspended solids reached 40%, while turbidity removal reached values up to 80%. The removal of COD was approximately 30 to 35%; yet, when removal of organic matter was based on the treated filterable COD, the removal increased to approximately 65%. In addition, nitrogen was removed by approximately 70%, while phosphorus removal ranged between approximately 30 and 45% on the basis of the initial phosphorus concentration. Finally, size fractionation of the organic matter (COD) showed that the various treatment methods were capable of removing different fractions of the organic matter.     

Multivariate analysis of photo-Fenton degradation of the herbicides tebuthiuron, diuron and 2,4-D

The degradation of herbicides in aqueous solution by photo-Fenton process using ferrioxalate complex (FeOx) as source of Fe2+ was evaluated under blacklight irradiation. The commercial products of the herbicides tebuthiuron, diuron and 2,4-D were used. The multivariate analysis, more precisely, the response surface methodology was applied to evaluate the role of FeOx and hydrogen peroxide concentrations as variables in the degradation process, and in particular, to define the concentration ranges that result in the most efficient degradation of the herbicides. The degradation process was evaluated by the determination of the remaining total organic carbon content (TOC), by monitoring the decrease of the concentrations of the original compounds using HPLC and by the chloride ion release in the case of diuron and 2,4-D. Under optimized conditions, 20 min were sufficient to mineralize 93% of TOC from 2,4-D and 90% of diuron, including oxalate. Complete dechlorination of these compounds was achieved after 10 min reaction. It was found that the most recalcitrant herbicide is tebuthiuron, while diuron shows the highest degradability. However, under optimized conditions the initial concentration of tebuthiuron was reduced to less than 15%, while diuron and 2,4-D were reduced to around 2% after only 1 min reaction. Furthermore, it was observed that the ferrioxalate complex plays a more important role than H2O2 in the photodegradation of these herbicides in the ranges of concentrations investigated.     

Effects of culture parameters on the degradation of 2,4-dichlorophenoxyacetic acid (2,4-D) and 2,4-dichlorophenol (2,4-DCP) by selected fungi. Groupe pour l'Etude du Devenir des Xénobiotiques dans l'Environnement (GEDEXE).

In order to enhance 2,4-D and 2,4-DCP degradation by four selected fungi (Cunninghamella elegans, C. echinulata, Rhizoctonia solani and Verticillium lecanii), three culture parameters (initial chemical concentration, amounts of glucose and nitrogen) were varied. The levels of both xenobiotics in the culture media were monitored by HPLC analysis after five days of cultivation. The best results were obtained at low initial concentration (20 mg.L-1 vs 100) and with low amounts of glucose (5 g.L-1 vs 10) and nitrogen (2.4 mM vs 24). When these two elements were lacking from the culture media, biodegradation was not suppressed, but took place to a lesser extent. Thus, initial chemical concentration and amounts of carbon and nitrogen, in the culture medium, were shown to strongly influence the extent of 2,4-D and 2,4-DCP removal by fungi.     

Synergy of ozonation and photocatalysis to mineralize low concentration 2,4-dichlorophenoxyacetic acid in aqueous solution

Concentration of 2,4-dichlorophenoxyacetic acid (2,4-D) may affect its degradation kinetics in advanced oxidation systems, and combinations of two or more systems can be more effective for its mineralization at low concentration levels. Degradations and mineralizations of 0.045mM 2,4-D using O(3), O(3)/UV, UV/TiO(2) and O(3)/UV/TiO(2) systems were compared, and influence of reaction temperature on the mineralization in O(3)/UV/TiO(2) system was investigated. 2,4-D degradations by O(3), O(3)/UV and UV/TiO(2) systems were similar to the results of earlier investigations with higher 2,4-D concentrations. The degradations and total organic carbon (TOC) removals in the four systems were well described by the first-order reaction kinetics. The degradation and removal were greatly enhanced in O(3)/UV/TiO(2) system, and further enhancements were observed with larger O(3) supplies. The enhancements were attributed to hydroxyl radical (()OH) generation from more than one reaction pathway. The degradation and removal in O(3)/UV/TiO(2) system were very efficient with reaction temperature fixed at 20 degrees C. It was suspected that reaction temperature might have influenced ()OH generation in the system, which needs further attention.     

Adsorption characteristics of 2,4-dichlorophenoxyacetic acid (2,4-D) from aqueous solution on powdered activated carbon

The removal of 2,4-dichlorophenoxyacetic acid (2,4-D), one of the most commonly used phenoxy acid herbicides, from aqueous solution was studied by using acid-washed powdered activated carbon (PAC) as an adsorbent in a batch system. Adsorption equilibrium, kinetics, and thermodynamics were investigated as a function of initial pH, temperature, and initial 2,4-D concentration. Powdered activated carbon exhibited the highest 2,4-D uptake capacity of 333.3 mg g(-1) at 25 degrees C and an initial pH value of 2.0. Freundlich, Langmuir, and Redlich-Peterson isotherm models were used to express the equilibrium data of 2,4-D depending on temperature. Equilibrium data fitted very well to the Freundlich equilibrium model in the studied concentration range of 2,4-D at all the temperatures studied. Three simplified models including pseudo-first-order, pseudo-second-order, and saturation-type kinetic models were used to test the adsorption kinetics. It was shown that the adsorption of 2,4-D on PAC at 25, 35, and 45 degrees C could be best fitted by the saturation-type kinetic model with film and intraparticle diffusions being the essential rate-controlling steps. The activation energy of adsorption (EA) was determined as--1.69 kJ mole(-1). Using the thermodynamic equilibrium coefficients obtained at different temperatures, the thermodynamic constants of adsorption (deltaG degrees, deltaH degrees, and deltaS degrees) were also evaluated.     

Parameter optimization of the fungicide (Vapam) sorption onto soil modified with clinoptilolite by Taguchi method

This study employs the Taguchi optimization methodology to optimize the effective parameters for the pesticide (Vapam) sorption onto soil modified with natural zeolite (clinoptilolite). The experimental factors and their ranges chosen for determination of the effective parameters were: initial Vapam concentration (0.4-1.6 mg/L), initial pH of the pesticide solution (2-12), the percentage of clinoptilolite in the modified soil (0-6 %), temperature (15-35°C) and shaking time (2-24 h). The orthogonal array (OA) L(16) and the bigger the better response category of the Taguchi method were selected to determine the optimum conditions: initial Vapam concentration (1.2 mg/L), initial pH of the pesticide solution (2), the percentage of clinoptilolite in the modified soil (4 %), temperature (15°C) and shaking time (2 h). The results showed that in comparison with other parameters, the initial Vapam concentration was the most effective one for the sorption of this pesticide onto soil, modified with clinoptilolite. Moreover, after determining the optimum levels of the sorption process parameters, confirmation experiments were performed to prove the effectiveness of the Taguchi's experimental design methodology.     

Dicamba and 2,4-D residues following applicator cleanout: A potential point source to the environment and worker exposure

This paper presents a survey of pesticide residues in tanks following application and throughout the cleanout procedure as conducted by 46 volunteer operators across Colorado. While many pesticides were detected, this paper focuses on dicamba and 2,4-D, which were detected by liquid chromatography/tandem mass spectroscopy (LC-MS/MS). An exponential decrease in concentration was observed with sequential rinses, although this decrease may be more rapid for more water-soluble pesticides. More than 95% of the pesticide in the prerinse solution was removed by the end of the third rinse in all but three operator samples. Concentrations after three rinses were 0.41 ± 0.25 and 3.3 ± 1.1 mg/L for dicamba and 2,4-D, respectively. These concentrations suggest that the recommended practice of three rinses may not be adequate to eliminate off-target effects or point sources of pesticide waste, and that the recommended standard of personal protective equipment is essential to prevent worker exposure to the chemicals.

Implications:?This paper demonstrates that the waste generated during cleanout of pesticide application devices constitutes a potential source of pollution and worker exposure. In particular, while the first rinse of pesticide containers is often treated as hazardous waste and reapplied to crops, the remaining rinses are not. This work demonstrates that the wastewater generated in subsequent rinses can have high enough concentrations to impact worker health, cause off-target effects on crops, and potentially constitute a point source of pesticides. The practical implication is for improved recommendations and regulations regarding pesticide applicators and their cleanout process.     

Reductive transformation of 2,4-dichlorophenoxyacetic acid by nanoscale and microscale Fe3O4 particles

Reductive transformation of 2,4-dichlorophenoxyacetic acid (2,4-D) by nanoscale and microscale Fe₃O₄ was investigated and compared. Disappearance of the parent species and formation of reaction intermediates and products were kinetically analyzed. Results suggest that the transformation of 2,4-D followed a primary pathway of its complete reduction to phenol and a secondary pathway of sequential reductive hydrogenolysis to 2,4-dichlorophenol (2,4-DCP), chlorophenol (2-CP, 4-CP) and phenol. About 65% of 2,4-D with initial concentration of 50 μ M was transformed within 48 h in the presence of 300 mg L^?1 nanoscale Fe₃O₄, and the reaction rates increased with increasing dosage of nanoscale Fe₃O₄. The decomposition of 2,4-D proceeded rapidly at optimum pH 3.0. Chloride was identified as a reduction product for 2,4-D in the magnetite–water system. Reductive transformation of 2,4-D by microscale Fe₃O₄ was slower than that by nanoscale Fe₃O₄. The reactions apparently followed pseudo-first-order kinetics with respect to the 2,4-D transformation. The degradation rate of 2,4-D decreased with the increase of initial 2,4-D concentration. In addition, anions had a significant adverse impact on the degradation efficiency of 2,4-D.     

Hydraulic residence time effects on performance of an activated sludge unit treating wastewater containing dichlorophenol.

Wastewaters containing chlorophenol compounds are difficult to treat by biological means because of the toxic effects of those compounds on microorganisms. To investigate the adverse effects of chlorophenols on microorganisms, synthetic wastewater containing 2,4 dichlorophenol (DCP) was biologically treated in an activated sludge unit at different hydraulic residence times (HRTs) between 5 and 40 hours, whereas the feed chemical oxygen demand (COD), DCP concentrations, and sludge age were kept constant at 2500 +/- 50 mg/L, 150 mg/L, and 20 days, respectively. The resazurin method based on dehydrogenase activity was used for assessment of the feed and effluent wastewater toxicity. Percent COD, DCP, and toxicity removals increased, and the effluent COD, DCP, and toxicity levels decreased with increasing HRT. Biomass concentration in the aeration tank increased with increasing HRT because of low levels of DCP at high HRT levels, resulting in high COD, DCP, and toxicity removals. The sludge volume index decreased with increasing HRT, yielding well-settling organisms as a result of low levels of toxicity and high concentrations of active cells. Percent DCP and COD removals decreased with increasing specific DCP loading rate. The rates of DCP and COD removals showed a maximum at a low DCP concentration of 6 mg/L in the aeration tank, corresponding to a 25-hour HRT.     

Solar photocatalytic mineralization of commercial pesticides: methamidophos

Aqueous solutions of methamidophos (O,S-dimethyl phosphoramidothioato) are mineralised in sunlight in the presence of dispersed particles of TiO2 in a pre-industrial pilot plant. A commercial pesticide (Tamaron 50, 50% Methamidophos) is used as a model to demonstrate the application of the treatment. Total Organic Carbon (TOC) and PO4(3-) are analysed to confirm the mineralization of the contaminants. The concentration of organics in water was set at TOC approximately 100 mg/L, but with only illuminated TiO2, mineralization was slow. Oxidation of the organics present in the formulation is enhanced 18 times by use of an appropriate electron scavenger (peroxydisulphate) compared to TiO2 alone.     

Biodegradation of nicosulfuron by the bacterium Serratia marcescens N80

By enrichment culturing of the sludge collected from the industrial wastewater treatment pond, we isolated a highly efficient nicosulfuron degrading bacterium Serratia marcescens N80. In liquid medium, Serratia marcescens N80 grows using nicosulfuron as the sole nitrogen source, and the optimal temperature, pH values, and inoculation for degradation are 30-35°C, 6.0-7.0, and 3.0% (v/v), respectively. With the initial concentration of 10 mg L?1, the degradation rate is 93.6% in 96 hours; as the initial concentrations are higher than 10 mg L?1, the biodegradation rates decrease as the nicosulfuron concentrations increase; when the concentration is 400 mg L?1, the degradation rate is only 53.1%. Degradation follows the pesticide degradation kinetic equation at concentrations between 5 mg L?1 and 50 mg L?1. Identification of the metabolites by the liquid chromatography/mass spectrometry (LC/MS) indicates that the degradation of nicosulfuron is achieved by breaking the sulfonylurea bridge. The strain N80 also degraded some other sulfonylurea herbicides, including ethametsulfuron, tribenuron-methyl, metsulfuron-methyl, chlorimuron-ethyl,and rimsulfuron.     

The effects of various soil factors and amendments on the degradation of pesticide mixtures

Soil disposal is the most common method of handling dilute pesticide wastes. The effects of several soil factors on the degradation of a pesticide mixture were investigated. Atrazine, captan, carbaryl, 2,4-D, diazinon, fenitrothion, and trifluralin were studied under varying laboratory conditions of soil type, pH, moisture content, organic matter content, microbial activity, and pesticide concentration. Of the variables investigated, pesticide concentration was the single most important factor with degradation rates much slower at high (1000 ppm) levels of fortification than moderate (100 ppm) levels. Under most conditions, chemical degradation was at least as significant as microbial degradation. Effects of other factors varied considerably and were pesticide dependent.