Biodegradation of the Organophosphate Insecticide Coumaphos in Highly Contaminated Soils and in Liquid Wastes

Approximately 400000 litres of cattle dip wastes containing approximately 1500 mg litre⁻¹ of the organophosphate insecticide coumaphos are generated yearly along the Mexican border from a USDA program designed to control disease-carrying cattle ticks. Use of unlined evaporation pits for the disposal of these wastes has resulted in highly contaminated soils underlying these sites. Previous work has shown that microbial consortia present in selected dip wastes can be induced to mineralize coumaphos. Our results demonstrate that similar microbial consortia are present in coumaphos-contaminated soils from eight waste sites and that these organisms are capable of mineralizing cou-maphos in these soils using soil slurries to less than 1 mg litre⁻¹ in 7–10 days at 28°C. In addition, our results show that these consortia are able to colonize pea gravel in trickling gravel filters and can be used in these filters to metabolize coumaphos from dip wastes to less than 0·1 mg litre⁻¹ in 7–10 days at 28°C. These simple systems offer potential low cost means to detoxify coumaphos-containing wastes and to bioremediate soils contaminated with this organophosphate compound.     

Breakdown rates of dimethoate in fruit and vegetable dips

In order to determine the effect of pH and temperature on post-harvest dip solutions of dimethoate (500 mg litre^?1), the half-lives and pseudo first-order rate constants were calculated from measurements at pH 4, 6, 8, 10, 11.5, and at two temperatures 25 and 52°C. The half-lives ranged from 206 days to 39.3 min at 25°C, and from 5.6 days to 205s at 52°C; the rate constants ranged from 3.9 × 10^?8 s^?1 to 2.9 × 10^?4 s^?1 at 25°C, and from 1.4 × 10^?6 s^?1 to 3.4 × 10^?3 s^?1 at 52°C. The results show that the water used in dips should have a pH≤7. The addition of benomyl to the dip solutions at two concentrations (0.5 and 1.0 g litre^?1) had no effect on the half-lives and rate constants. The use of hard and salted waters in dips also showed no major effect. A formula was developed that gives the half-life of the dimethoate as a function of the pH and temperature.     

Effect of temperature on the toxicity of low concentrations of methyl bromide to diapausing larvae of the warehouse moth Ephestia elutella (Hübner)

The performance of low concentrations of methyl bromide against diapausing larvae of Ephestia elutella at 15 and 25°C was assessed in extended exposure periods. At concentrations of 1.9 mg litre^?1 and below, test batches required higher concentration-time (ct) products for 100% kill at 25°C than at 15°C. The minimum concentration at which the concentration: time relationship still applied was between 1.3 and 1.9 mg litre^?1 at 15°C, whereas at 25°C it was between 2.7 and 4.0 mg litre^?1. For many individuals within each population sample, however, lower concentrations at moderate dosage levels remained lethal. At 25°C, a ct product of about 90 mg litre^?1 h gave between 53 and 77% kill at 6.1, 4.0, 2.7 and 1.9 mg litre^?1. The trends observed suggest that the most tolerant members of the population have an enhanced ability to detoxify methyl bromide at the higher temperature. The implications of the results for the build-up of resistance and for practical control measures are discussed.     

Depression of the minimum concentration of methyl bromide effective against diapausing warehouse moth larvae at 15°c by prior exposure at a moderate concentration

Warehouse moth (Ephestia elutella) larvae in diapause were exposed at 15°C to methyl bromide at 8 mg litre^?1 for 14.5 h and then immediately exposed at a lower concentration. The exposure at 8 mg litre^?1 killed 44–69% of the larvae treated. Subsequent concentrations down to 1.1 mg litre^?1 obeyed Haber's rule (concentration × time= k, a constant for mortality), but a higher concentration-time product (ct) was required for over 90% kill at 0.8 mg litre^?1. Only concentrations down to 1.9 mg litre^?1 obey Haber's rule if there is no prior exposure at a higher concentration. Although minimum effective concentrations are lower at 15°C than at 25°C, exposure at a higher concentration depresses the subsequent level to a similar extent at each temperature. The contribution to the efficacy of a treatment, of low concentrations persisting at the end of fumigation, is thus likely to be even greater at moderate to low temperatures than at 25°C. The implications for the development of resistance to methyl bromide are discussed.     

Effect of ABT on the activity and rate of degradation of isoproturon in susceptible and resistant biotypes of Phalaris minor and in wheat

The effect of the monooxygenase inhibitor, 1-aminobenzotriazole (ABT) on isoproturon phytotoxicity and metabolism was studied in resistant (R) and susceptible (S) biotypes of Phalaris minor and in wheat (Triticum aestivum). Addition of ABT (2·5, 5 and 10 mg litre^-1) to isoproturon (0·25, 0·5, 1, 2 and 4 mg litre^-1) in the nutrient solution significantly enhanced the phytotoxicity of isoproturon against the R biotype. Isoproturon at 0·25 mg litre^-1 reduced the dry weight (DW) of the S biotype by 77%, whereas the R biotype required 4·0 mg litre^-1 for similar reduction. Addition of 10 mg litre^-1 of ABT to the 0·25 mg litre^-1 isoproturon caused 71 and 82% reduction in DW of R and S biotypes, respectively. Wheat was more sensitive to the mixture of isoproturon and ABT than the R biotype of P. minor. Reduced concentrations of ABT in the mixture from 10 to 2·5 mg litre^-1 increased the DW of the R biotype more than that of the S biotype. The R biotype metabolised [¹⁴C]isoproturon at a faster rate than the S biotype. ABT (5 mg litre^-1) inhibited the degradation of [¹⁴C]isoproturon in both biotypes of P. minor and in wheat. In the presence of ABT, about half of the applied [¹⁴C]isoproturon remained as parent herbicide in all the three species after two days. The metabolites were similar in the R and S biotypes and wheat as determined by co-chromatography with reference standards and mass spectroscopy (MS). ABT inhibited the appearance of the hydroxy and monomethyl metabolites and their conjugates in all the test plants. These results suggest that the activity of the enzymes responsible for the degradation of isoproturon is greater in the R than in the S biotype of P. minor, resulting in its rapid detoxification. Incorporation of the monooxygenase inhibitor ABT into the nutrient solution greatly inhibited the degradation of [¹⁴C]isoproturon in the R biotype and increased its phytotoxicity. Both hydroxylation and N-dealkylation reactions were found to be sensitive to ABT; inhibition of hydroxylation was greater than that of demethylation. Since ABT could not completely suppress isoproturon degradation, it is possible that more than one monooxygenase is involved. © 1998 SCI     

Minimum concentration levels of methyl bromide required for full efficacy against seven species of stored-product beetle at two temperatures

The dosages of methyl bromide required to control insect pests can be expressed as concentration, time (Ct) products for concentrations down to a certain minimal level, here defined as the efficacy threshold, below which Habers Ct rule (C×t=k,a constant for mortality) no longer applies. Twelve strains of seven species of stored-product beetle were tested to identify efficacy threshold concentrations at 15°C and 25°C. For each strain the efficacy threshold was higher at 25°C than at 15°C. Different species differed widely in the threshold levels identified. Strains with a higher than usual tolerance to methyl bromide than their standard stock counterparts had slightly higher efficacy thresholds at 25°C but not at 15°C. The order of tolerance of the standard stocks to methyl bromide at 15°C was Tribolium confusum> T. castaneum> Oryzaephilus surinamensis> Rhyzopertha dominica> Sitophilus granarius> S. zeamais> S. oryzae, and at 25°C Tribolium castaneum > T. confusum> R. dominica> O. surinamensis> S. granarius> S. zeamais> S. oryzae. The order of efficacy thresholds largely reflected the order of susceptibility, S. oryzae and S. zeamais having the lowest efficacy thresholds of about 0.6 mg litre^?1 at 15°C and 1.3-2.0 mg litre^?1 at 25°C, while Tribolium spp. had the highest, 1.3-2.0 mg litre^?1 at 15°C and 2.5-4.0 mg litre^?1 at 25°C.     

A monoclonal antibody‐based ELISA for the analysis of the insecticide flucythrinate in environmental and crop samples

A competitive enzyme‐linked immunosorbent assay (ELISA) has been developed for the detection of the insecticide flucythrinate in environmental and food samples. Two types of haptens, the acid moiety that is the hydrolyzed product of flucythrinate, and the carboxylated propyl derivative of the alcohol moiety, were used to prepare monoclonal antibodies (MAbs). Five MAbs, which raised against the former hapten, were reactive with flucythrinate. Among them, MAb F1A27‐4 showed the highest activity toward flucythrinate, and did not cross‐react with other pyrethroids such as cycloprothrin, fenvalerate, fluvalinate, etofenprox and silafluofen. The assay conditions of indirect competitive ELISA with MAb F1A27‐4 were studied to optimize the detection of flucythrinate in environmental and food samples. Incubation at 4 °C in the assay buffer, pH 8, with 300 mM sodium chloride improved the sensitivity. The addition of rabbit serum albumin or rabbit antiserum and the presence of 50 ml litre^?1 of methanol reduced matrix effects of the samples. Under optimized conditions, the ELISA detected flucythrinate spiked in water, soil, and extracts of apple and tea samples down to 10 mg litre^?1, 0.2 mg litre^?1, 0.3 mg litre^?1 and 0.3 mg litre^?1, respectively. The mean recovery and CV ranged from 91% to 120% and from 5% to 12%, respectively. The ELISA results in apple samples correlated well with those from LC–MS analysis (r² = 0.99, n = 12). © 2001 Society of Chemical Industry     

Effect of short exposures to a high concentration on the subsequent toxicity of low concentrations of methyl bromide to diapausing larvae of the warehouse moth,Ephestia elutella (Hübner)

Ephestia elutella larvae in diapause were exposed at 25°C to methyl bromide at 12 mg litre^?1 for 3.5 or 7.5 h and then immediately exposed to a lower concentration. The minimum effective concentration (that at which Haber's rule, concentration × time = k, a constant for mortality, still applied) was about 3 mg litre^?1 in tests with no previous exposure toa high concentration, but it was about 2.5 mg litre^?1 for individuals surviving a 3.5 h exposure to 12 mg litre^?1, and was about 1.6 mg litre^?1 for those surviving a 7.5 h exposure to 12 mg litre^?1. These exposures to 12 mg litre^?1, respectively, killed 2–20% and 50–75% of larvae exposed, and hence the smaller the proportion of survivors of exposure to a high concentration, the lower the minimum effective concentration needed against them. Thus the low concentration persisting at the end of a practical fumigation should contribute significantly to the success of the treatment and be much more effective than any similar low concentration present soon after the introduction of gas.     

Degradation and adsorption of 14C-MCPA in soil—influence of concentration, temperature and moisture content on degradation

MCPA was weakly absorbed in soils with 2.4, 3.0 and 2.9% humus. K_d-values were 0.7, 0.9 and 1.0, respectively. In soil, not previously treated with MCPA, the degradation of 0.05 mg kg^?114C-MCPA followed first-order reaction kinetics whereas degradation of 5 mg kg^?1 was only first-order for 2 weeks; exponentially increasing degradation rates followed indicating enrichment of the soil with MCPA decomposers. Degradation was monitored by evolution of ¹⁴CO₂. The influence of temperature on degradation of MCPA (4 mg kg^?1) could initially be described by Q₁₀ values or by the Arrhenius equation. After 1 day of incubation in two field soils Q₁₀ values were 3.3 and 2.9, respectively, between 0°C and 29°C; the activation energies were 87 and 76 kj mol^?1. Exponentially increasing degradation rates followed with doubling times of about 4.0, 1.8, 1.2 and 0.6 days at 6,10, 15 and 21°C, respectively. After 51 days of incubation, at temperatures between 6°C and 29°C, about 60%¹⁴C was evolved in CO₂ and only traces of MCPA were left in the soil. At 0°C and at 40°C only 1% and 10%¹⁴C, respectively, were evolved as CO₂ after 51 days. ¹⁴C-MCPA (4 mg kg^?1) was incubated at moisture contents from that in air-dried soil to 2.3 times field capacity. Optimum for degradation was from 0.6 to 1.2. field capacity. Degradation was very slow where water contents were below the level of wilting point and was nil in air-dried soil. In wet soil degradation was delayed, but even in water-logged soil (2.3 times field capacity) MCPA was decomposed after 4 to 5 weeks at 10°C.     

A field unit for determining dimethoate in fruit and vegetable dips

The concentration of dimethoate in post-harvest dips needs to be maintained at 400 or 500 mg litre^?1 to ensure that fruit fly are killed and also to keep residue levels below the MRL. A field unit which contains a colorimeter, a heating block and cooling positions has been evaluated. The diluted dip sample produces a colour with 4-(p-nitrobenzyl)pyridine after heating and the addition of tetraethylenepentamine. Precision of the unit was acceptable for a field test (repeatability of ± 10.1%) and there was reasonable agreement between determinations made on the unit and by an HPLC method for a range of dimethoate concentrations from 300 to 700 mg litre^?1. The unit can be used at the dip site and the analysis is completed in 30 minutes.     

Determination of residual 2,4,5-T in vegetables by high performance liquid chromatography with ultraviolet and fluorometric detection after derivatization with 2-(2,3-naphthalimino)ethyl trifluoromethanesulfonate (NE-OTf)

2,4,5-T was extracted with acetone at below pH 1·0 and the extract was concentrated. After adding 100 g litre^-1 sodium chloride solution to the residual solution, 2,4,5-T was extracted with ethyl acetate+hexane (20+80 by volume). The extract was evaporated to dryness and the residue was dissolved in acetonitrile. 18-crown-6, potassium fluoride and NE-OTf were added to the acetonitrile solution and then allowed to react at 50°C for 20 min. The product was injected to a HPLC with ultraviolet detection operated at 259 nm and fluorometric detection at 394 nm emission and 259 nm excitation. The determination limits of the 2,4,5-T derivative in the sample were 20 μg litre^-1 with UV detection and 10 μg litre^-1 with fluorometric detection. © 1997 SCI.     

Susceptibility pattern and development of resistance in the diamondback moth,Plutella xylostella L,to Bacillus thuringiensis Berl var kurstaki in India

Base‐line susceptibility for six‐day‐old larvae of the diamondback moth, Plutella xylostella, against Bacillus thuringiensis var kurstaki (Biobit^®) was studied by a cabbage leaf disc dip bioassay technique. Diamondback moth from 13 locations in seven different states spread over a distance of about 3000 km longitudinally was used for these studies. Forty‐eight‐hour LC₅₀ values varied from 1.0 to 10.97 mg AI litre⁻¹. Further investigations on the development of resistance under laboratory conditions showed an increase in LC₅₀ from 2.76 (for unselected F₁ generation) to 5.28 mg AI litre⁻¹ (for selected F₉ generation), using a selection concentration of 6.4 mg AI litre⁻¹. This suggested a possibility of the development of resistance under field conditions if there were to be extensive and indiscriminate use of B thuringiensis. These findings are discussed in relation to integrated pest management and the mechanisms of resistance in resistance management tactics. © 2000 Society of Chemical Industry     

Laboratory Evaluation of the Novel Naturally Derived Compound Spinosad against Ceratitiscapitata

Laboratory studies were conducted to determine the effect of the naturally derived compound spinosad on Ceratitis capitata Wied. (Diptera, Tephritidae). The organophosphate fenthion was used as a standard. Direct dose-dependent mortality and reduced fecundity were observed in oral treatment of adults with spinosad. The LC₉₀ values 14 h and seven days after treatment were 19·50 and 0·49 mg litre⁻¹ respectively. Fenthion was less active (the LC₅₀ eight days after treatment was 1·17 mg litre⁻¹) and did not affect the fecundity of the fly. Adults were also very susceptible to spinosad and fenthion via residual contact. For spinosad, 100% mortality was recorded 48 h after treatment for a dose of 10 mg litre⁻¹. Spinosad was more effective than fenthion in suppressing larval development when neonate larvae were reared on treated diet supplemented with a range of concentrations from 0·02 to 0·83 mg kg⁻¹ diet. Last-instar larvae were much less susceptible to spinosad or fenthion when exposed via dipping or when they pupated in treated medium and both products had similar performance. A lack of ovicidal activity was observed in direct egg-treatments with spinosad but significant reductions from 1 mg litre⁻¹ onwards were recorded for fenthion.     

Gas-liquid chromatographic determination of hydrazine in maleic hydrazide formulations and in samples stored at an elevated temperature

A method is described for the analysis of small amounts of hydrazine in maleic hydrazide formulations. Following derivative formation with pentafluorobenz-aldehyde, the pentafluorobenzaldehyde azine was extracted with hexane and determined by gas-liquid chromatography with electron-capture detection. Recoveries of 72-80% were obtained from samples fortified with 1 and 10 μg of hydrazine. The limit of detection was 0.05 mg kg^?1. Fourteen commercial formulations with maleic hydrazide concentrations ranging from 180-360 g litre^?1 were investigated. The hydrazine content of the maleic hydrazide used in these formulations ranged from less than 0.05 to 53 mg kg^?1. During the storage of two samples at 50°C for 10 weeks, the hydrazine contents increased from 2.2 to 124 and 0.4 to 54 mg litre^?1, respectively.     

Availability and biodegradation of metribuzin in alluvial soils as affected by temperature and soil properties

Herbicide degradation in soils is highly temperature‐dependent. Laboratory incubations and field experiments are usually conducted with soils from the temperate climatic zone. Few data are available for cold conditions and the validation of approaches to correct the degradation rate at low temperatures representative of Nordic environments is scarce. Laboratory incubation studies were conducted at 5, 15 and 28°C to compare the influence of temperature on the dissipation of metribuzin in silt/sandy loam soils in southern and northern Norway and in a sandy loam soil under temperate climate in France. Using ¹⁴C‐labelled metribuzin, sorption and biodegradation were studied over an incubation period of 49 days. Metribuzin mineralisation and total soil organic carbon mineralisation rates showed a positive temperature response in all soils. Metribuzin mineralisation was low, but metabolites were formed and their abundance depended on temperature conditions. The rate of dissipation of ¹⁴C‐metribuzin from soil pore water was strongly dependent on temperature. In Nordic soils with low organic content, metribuzin sorption is rather weak and biodegradation is the most important process controlling its mobility and persistence.     

Residues of the algicide endothal in water,soil and rice,after paddy field applications

In order to obtain residue data from the application of the algicide endothal in Italian rice paddy fields, two experiments were carried out using a 50 g kg^?1 granular formulation in a small pond and the same granular and two liquid formulations in actual paddy fields of the Italian rice growing area. Endothal decay in the pond water was very rapid, reaching residue levels of 0·01-1·02 mg litre^?1 in two days and 0·004-0·01 mg litre^?1 at the third day. The muddy soil of the pond was free from measurable endothal residues( <0·02 mg kg^?1). In the paddy-field waters, the endothal decay was slower, with an average half-life time of 3·3 days, independently of the type of formulation. The actual residues in water after 6 days ranged from 0·3 to 1·3 mg litre^?1 according to the initial amount of product applied, and, consequently, to the initial concentration in water. Rice samples collected at the normal harvest time from the two paddy fields, treated with three different formulations, showed no endothal residue at the minimum detectable level of 0·01 mg kg^?1.     

Transformation of aldicarb sulfoxide and aldicarb sulfone in four water-saturated sandy subsoils

The transformation of aldicarb sulfoxide and aldicarb sulfone was studied in incubations with water-saturated subsoils under simulated field conditions at 10°C. The subsoils were collected at four locations from beneath the water table at a depth of 2.5 to 3.5 m. In three of the subsoils, the half-life of sulfoxide, incubated at concentrations of 0.14-0.17 mg litre^?1, ranged from 0.7 to 2.8 years. At higher concentrations (8-13 mg litre^?1), its half-life ranged from 3.4 to 6.4 years. At the lower concentration, a large fraction of sulfoxide was transformed into sulfone. The rates of transformation of the sulfone at the lower concentration in the three subsoils corresponded to half-lives of 3.3 to 8.1 years, but in only one subsoil was a significant transformation rate (half-life 6.7 years) measured at the higher concentration during the 2.3-year incubation period. The half-lives at the lower concentrations were more like those in field studies, and perhaps would still underestimate transformation rates under field conditions. After a year, 2.5-15% of the higher sulfoxide and sulfone doses had been trapped as [¹⁴C] carbon dioxide. In the fourth subsoil, with more anaerobic conditions, the half-life of sulfoxide at both concentrations was less than 0.02 year and that of sulfone was about 0.04 year. Four or five radio-labelled transformation products could be traced in this subsoil and about half of the dose of both compounds was trapped as [¹⁴C] carbon dioxide.     

Enzyme immunoassay for atrazine analysis in water and soil

Enzyme immunoassay (EIA) has been tested for the detection of atrazine in soil and water. EIA kits and atrazine-fortified samples were received from the International Atomic Energy Agency. Atrazine concentrations of about 0·01 μg litre^-1 could be detected and the central detection point was found at about 0·15 μg litre^-1 which is a reasonably sensitive region for atrazine. A validation study with spiked local water samples yielded acceptable results. No treatment was required for water samples. Extraction of atrazine from soil was done by simple shaking with methanol without any clean-up steps. Detection limits of 1×10^-2 μg litre^-1 for water and 5×10^-3 μg kg^-1 for soil were achieved. © 1998 SCI.     

Synthesis and Evaluation of New Ethyl 2,3- Dihydro-3-oxoisothiazolo[5,4-b]pyridine- 2-alkanoate Derivatives as Potential Herbicides

Several ethyl 2,3-dihydro-3-oxoisothiazolo[5,4-b]pyridine-2-alkanoate derivatives were synthesized as herbicides. Only 5-methyl derivatives inhibited both hypocotyl and root growth in the lettuce (Lactuca sativa L.) seedling test at 100 mg litre^-1. Only ethyl propionate and valerate derivatives showed significant inhibition at 0·1 mg litre^-1, whereas ethyl acetate or butyrate derivatives were inactive. Contrary to unoxidized derivatives, the inhibitory effect of 1-oxide and 1,1-dioxide derivatives was strongly dependent on concentration; ethyl 2,3-dihydro-5-methyl-3-oxoisothiazolo[5,4-b]pyridine-2-propionate 1,1-dioxide inhibited 100% of germination at 100 mg litre^-1 and 45% of lettuce seedling growth at 0·1 mg litre^-1. Quantitative structure–inhibition of growth relationship analysis carried out by adaptive least-squares (ALS) method gave a good correlation with small and hydrophobic 5-substituents as well as with odd carbon-chain ethyl alkanoates in position 2. Active compounds did not show auxin-like activity from 0·1 to 100 mg litre^-1. © 1997 SCI.     

Relation between volatility rating and composition of phenoxy herbicide ester formulations

A CIPAC/AOAC test with tomato plants is used to specify the volatility ratings of herbicide ester formulations. This work compares the tomato plant test with an alternative chemical one. The concentrations of esters and the effective molecular weight and density of each formulation were used with the ester vapour pressures to calculate its herbicide vapour pressure as complete, and evaporated formulations. The range was from 28.8 mPa (at 257deg;C) for a mixture of 2,4–D esters to 0–07 mPa (at 25°C) for a 2,4,5–T-(iso-octyl) formulation, as complete formulations, and 35-5 and 0–16 mPa (at 25°C) as evaporated ones. A value of 0–6 mPa (at 25°C) was selected on the basis of the tomato plant test as the cut-off area for low-volatile esters and is recommended to be included in specifications for herbicide esters. Formulations with a herbicide vapour pressure above 3.3 mPa (at 25°C) are high-volatile ones according to the tomato plant test, while between 0–6–3.3 mPa (at 25°C) is a borderline region where the test gives mixed results. Levels of 2,4–D-ethyl and methyl were added to pure 2–ethylhexyl esters of 2,4–D and a 2,4,5–T-(iso-octyl) formulation to find what level of contamination would change the rating of these esters from low to high volatile. Formulations of 2,4–D-(iso-octyl) should not contain more than 11 g litre^?1 2,4–D as methyl ester or 2.0 g litre^?1 2,4–D as ethyl ester. Formulations of 2,4,5–T-(iso-octyl) should not contain more than 26 g litre^?1 2,4–D as methyl ester or 4.7g litre^?1 2,4–D as ethyl ester.