The accumulation and distribution of pp′DDT and related compounds in an apple orchard .I. residues in the soil

Soils from an orchard sprayed annually (1953-1969) with technical DDT (77% pp′DDT and 22 % pp′DDT) were analysed for residues from 1964 until 1969. The amount of DDT found after 17 years was 21 %pp′DDTout of 27.1 kg/ha applied, and 7 % of pp′DDT out of 7.6 kg/ha. The vertical distribution of residues (pp′DDT, pp′DDE, pp′TDE and pp′DDT) showed a linear relationship between log amount and depth, with approximately 80% in the top 10 cm of soil. At depths from 50 to 210 cm, residue values were too small to be determined (i.e. < 1 ng/g dry wt). The surface distribution in the orchard showed a systematic pattern of circular areas of residues, with maximum values centred at each trunk (7.5 μg/g) and decreasing rapidly to each alley (1.9 ug/g). The levels of pp′DDT had reached a steady state (3-4 kg/ha) and the half-life time was calculated as 3.0 j′ears. pp′DDE (1.8 kg/ha) was the main metabolite of pp′DDT. Small amounts of pp′TDE were also found, pp′DDT was less persistent than pp′DDT, with a half-life time of 1.5 years.     

Accumulation and distribution of pp′-DDT and related compounds in an apple orchard. II. Residues in trees and herbage

DDT residues in or on the roots and leaves of the herbage and the roots, bark, leaves and fruit of the trees are given for an apple orchard sprayed annually (1953–1969). The distribution of DDT in both the grass and the grass roots was in circular areas of residues, with maximum values at each trunk and decreasing radially to each alley. Of the spray applied at the green cluster stage 80% was deposited on the grass sward and very little, if any, directly on the soil surface. The pp′-DDT content of the grass fell rapidly with successive mowings (from which the cuttings remained in situ) from 400 μg/g at spraying to 2 μg/g after nine months. 33 g/ha pp′-DDT was found in the herbage roots (0.87% of the total residues in the soil). The residues in the bark (87.5 g/ha) were much lower than expected after 13 years spray application. There were increased amounts of pp′-DDE, pp′-TDE and pp′-TDEE relative to pp′-DDT, indicating some breakdown on the bark, but the chief losses were attributed to volatilisation and to removal by wind and rain. The residue content of root bark varied from 3 μg/g near the emerging trunk to 0.05 μg/g at a depth of 90 cm. The pp′-DDT content of leaves at leaf fall rose from <1 ng/g after a single spring spray to 8.33 μg/g following an additional spray in late June. There was a large loss of DDT from the canopy between the June spray and leaf fall (440–480 g/ha down to 25 g/ha), attributed to volatilisation. The amount of pp′-DDT on the fruit, after a single spray, was 3 ng/g fresh weight (80.9 mg/ha out of a total of 1.0–1.5 kg/ha used).     

In-vivo and in-vitro studies on DDT uptake and metabolism in susceptible and resistant strains of the mosquito Aedes aegypti L.

Twelve strains of Aedes aegypti have been compared for resistance to pp′-DDT, uptake of pp′-DDT, and dehydrochlorination of pp′-DDT to pp′-DDE in vivo and in vitro both at the larval and adult stages. Resistant larvae were shown to contain significantly more pp′-DDE than susceptible larvae after a standard exposure to pp′-DDT but also substantially more pp′-DDT in an unmetabolised state. There was a small increase in the percentage dehydrochlorinated in vivo in the resistant strains compared with the susceptible strains, but this was not correlated with the level of resistance nor with dehydrochlorination in vitro. However, dehydrochlorination in vitro was correlated with resistance. Adult resistance was correlated positively with dehydrochlorination, both in vivo and in vitro, but the resistant adults did not contain increased levels of unmetabolised DDT. By comparing resistance levels at the two stages, it was found that there were two kinds of resistant strain: four strains of Asian origin and one from West Africa were highly resistant as larvae but showed almost no resistance as adults; five strains from Central and South America were highly resistant at both stages. The different mechanisms of resistance in adults and larvae are discussed in relation to genetic studies.     

Mechanism of DDT resistance in larvae of the mosquito Aedes aegypti L.; The effect of DDT selection

Selection with pp'-DDT was applied to fourth-stage larvae of Aedes aegypti along four lines, starting with larvae of the F₂ generation from crosses between a susceptible strain and each of four resistant strains (two of Trinidad origin and two of Bangkok origin). Larval resistance increased substantially along each line but there was little or no change in the percentage breakdown of DDT to 1,1-dichloro-2,2-bis(4-chloro-phenyl)ethylene (pp'-DDE) in vivo and in no line were these two variables significantly correlated. Percentage breakdown was generally higher at 10 mg than at 50 mg litre^?1. DDT uptake (defined as content of DDT+pp'-DDE) was generally higher after exposure to 50 mg than to 10 mg litre^?1. It increased significantly with selection in the TE×NS line; it remained unchanged in the T8 × NS line; and in the other two lines (BSJ × NS and B51 × NS), it increased initially but dropped as selection progressed, the reduction being highly significant in the second of these lines. The amount of internal residual (unmetabolised) DDT tolerated by larvae of the TE × NS line also increased significantly with resistance. The levels in the other lines followed the pattern of uptake, remaining steady in T8 × NS despite the increase in survival, rising at first and then declining in the two Bangkok lines. Thus selection produced a higher tolerance to internal unmetabolised DDT in the two Trinidad lines but led ultimately to a lower content of DDT+pp'-DDE in the two Bangkok lines. The reasons for this behaviour are discussed.     

Mechanisms of DDT resistance in larvae of the mosquito Aedes aegypti L

Larvae of eight strains of Aedes aegypti were exposed to DDT and compared for resistance, DDT uptake, in-vivo breakdown of DDT and residual unmetabolised DDT. Resistance varied widely between strains, three being fully susceptible, two almost immune and three of intermediate resistance. Breakdown of DDT by dehydrochlorination to 1,1-dichloro-2,2-bis(4-chlorophenyl)ethylene (pp'-DDE) occurred in all strains and was greater in the five resistant types, but there was no significant correlation between the extent of breakdown in the resistant strains and the level of resistance. Moreover the overall difference between susceptible and resistant strains disappeared when they were compared at a low, almost sublethal, concentration of DDT. Larvae of resistant strains carried a greater absolute quantity of unmetabolised DDT in the body and were able to tolerate levels of DDT that were lethal to susceptible larvae. However the two most resistant strains (T8 and B51) contained significantly less DDT plus pp'-DDE than strains of intermediate resistance (T30 and BSJ) from which they had been derived. Addition of the synergist chlorfenethol to DDT increased its knockdown effect on all resistant strains, suggesting that dehydrochlorination was a factor in resistance. Three strains, two DDT-resistant and one DDT-susceptible, were tested with 1,1-bis(4-ethoxyphenyl)-2,2-dimethylpropane (I), an insecticide that cannot be dehydrochlorinated. All the strains were relatively tolerant to it although the DDT-susceptible strains were less tolerant. Addition of the synergist sesamex decreased the level of tolerance to I in all strains which suggested that microsomal oxidation made some contribution to it. It is concluded that three factors contribute to larval DDT resistance in A. aegypti; (a) increased metabolism to pp'-DDE; (b) increased tolerance to unmetabolised internal DDT; and (c) reduced content of DDT+pp'-DDE (only in the most resistant strains and due either to reduced absorption or increased excretion). These factors are discussed in relation to known larval resistance genes R^DDT1 and y.     

Residues of organochlorine insecticides in some fishes and birds in the gezira of Sudan

Twenty-eight specimens representing five types of fish and five types of birds from the Gezira Research Farm, Wad Medani, Sudan, were analysed for residues of organochlorine insecticides. All fish muscle samples were found to contain pp′-DDE, pp′-TDE and pp′-DDT in total concentrations ranging from 0.27 to 16.0 mg/kg. In addition to DDT-type residues, dieldrin (HEOD) was also found in the majority of bird samples. Concentrations in bird breast muscle ranged from 0.07 to 5.5 mg/kg. When bird liver samples were examined they were found to contain higher residues than the breast muscle in nearly all cases.     

Comparative excretion of DDT analogues into milk of indian buffalo,Bubalus bubalis L. following oral administration

Four groups of Indian buffaloes were fed daily with 25 mg of p,p′-DDT p,p′-TDE p,p′-DDE or o,p′-DDT for 100 days. Milk was analysed for organochlorine residues during this period and also for 100 days after pesticide administration had been discontinued. For the period showing ‘plateau level’ residues, 17.2% of p,p′-DDE, 17% of p,p′-TDE, 14% of p,p′-DDT as p,p′-DDT (3.5%); p,p′-TDE (10.5%); 3.2% of p,p′-DDT as o,p′-DDT (1.3%) and o,p'-TDE (1.9%) of their administered amounts were excreted in the milk. Since these compounds were excreted at different rates, the residue levels in the milk expected from a given feed would depend on their concentration and proportional distribution in the feed. The maximum tolerable content of DDT analogues in feed was derived to be 0.1 mg kg^?1 (dry weight basis) by using the maximum accumulation coefficient and incorporation of the necessary safety margin. It is concluded that Indian buffaloes fed with rations contaminated with a total of DDT analogues below this limit will yield milk of acceptable quality. Following the termination of feeding with contaminated rations, the elimination of p,p′-DDE in the milk took the longest time and that of o,p′-DDT the shortest. These results suggest that the time required for the initial high residue concentration to decline to less than the legal limit would be determined by the relative amounts of DDE, TDE and DDT in the milk, after elimination of the potent source of contamination.     

Pesticide residues in soil and foodstuff I. Chlorinated pesticides in cattle feed and milk produced in orchard and non-orchard areas

Milk, hay and silage produced in orchard and non-orchard areas have been analysed for their content of chlorinated pesticides. The residue levels in milk produced in orchard areas were about double those in milk produced in non-orchard areas. The levels in hay were much higher (30% up to sevenfold) in the hay produced in orchard areas and the levels in silage from the two areas showed small differences. Milk collected after cattle had been grazing in an orchard for 3 to 4 days contained 10% more pp′-DDT and 2-, 3-, 7- and 15-fold morepp′-DDE, pp′-TDE and heptachlor epoxide, gamma-BHC and pp′-TDE olefin [1-chloro-2,2-bis(p-chlorophenyl)ethylene], respectively, than did the samples collected one day before the cattle grazed in the orchard.     

The metabolism of DDT in vivo by the Japanese quail (Coturnix coturnix japonica)

Male and female Japanese quail (Coturnix coturnix japonica) were given intraperitoneal injections of [¹⁴C]DDT in ethanol at a rate of 13.4 mg/kg body wt. Fifty-six days later the tissues and droppings were analysed for total ¹⁴C and metabolites. The rate of loss of ¹⁴C in droppings was very similar in males and females. The maximal rate was reached on the third day, and 65–66% of the injected dose was voided by the fifty-sixth day. Ninety-three to ninety-four percent of the ¹⁴C in droppings and 83–90% of the ¹⁴C in tissues were extracted by solvents. Combined extracts from males and females were used for determination of DDT and its metabolites. Expressing all results as percentages of injected dose, the following were isolated from droppings: DDA (24%), DDT (3%), DDD (5.1%), DDE (11%), and uncharacterised polar metabolites (17%). Twenty-five percent of the dose was retained in the tissues and this was largely accounted for as DDT (10.4%) and DDE (10.5%). Of the total metabolites found 31% was DDE (almost equally divided between tissues and droppings) and 35% was DDA (almost entirely in droppings). Since DDD was not found in significant quantities in tissues, the substantial quantities in droppings were probably produced from DDT by the action of microorganisms.     

Uptake,metabolism and effects of DDT,fenitrothion and chlorpyrifos on Tetrahymena pyriformis

The uptake and metabolism of DDT, fenitrothion and chlorpyrifos were studied in cultures of the ciliate protozoan Tetrahymena pyriformis. When cultures were treated with DDT in concentrations varying from 0.01 to 0.5 μg ml⁻¹, concentrations found in T. pyriformis were 3.8 to 335 μg g⁻¹ dry weight. The accumulation of fenitrothion ranged from 28.7 μg g⁻¹ in cultures treated with 1 μg ml⁻¹ to 2260 μg g⁻¹ in cultures treated with 10 μg ml⁻¹. Under similar experimental conditions chlorpyrifos was accumulated from 24.7 to 15400 μg g⁻¹. The patterns of uptake were dependent on the growth cycle, the ability of the organism to metabolise insecticide and the type of the insecticide used. Maximum accumulation of DDT, fenitrothion and chlorpyrifos occurred in 2, 4 and 6 h respectively. Tetrahymena metabolised DDT to DDD and DDE but failed to metabolise fenitrothion and chlorpyrifos. The effects on growth and morphology of T. pyriformis were studied over a period of 5 days. Higher concentrations (10, 50 and 100 μg ml⁻¹) of DDT inhibited only the growth of the organisms and did not change cell morphology. Fenitrothion was extremely toxic to the organisms and at 5 and 10 μg ml⁻¹ cells became more or less spherical and died after 48 h. However, concentrations of 0.5, 1 and 2.5 μg ml⁻¹ fenitrothion caused growth inhibition, but only at 2.5 μg ml⁻¹ was this permanent. Chlorpyrifos inhibited the growth of the organisms at 1, 5 and 10 μg ml⁻¹ but the morphology was affected only at 5 and 10 μg ml⁻¹.     

Distribution and breakdown of DDT in orchard soil

Amounts of DDT and its breakdown products were determined in soil in an apple orchard in Herefordshire. Samples were taken for a number of years (1972–79) after use of the insecticide in the orchard had ceased in 1969. The results were compared with those obtained in an investigation of the same orchard in 1968. From 1968 to 1979, soil residues of pp′-DDT, p′--DDT and pp′--TDE decreased gradually whereas those of pp′--DDE increased, and there were linear relationships between log (concentration) and time. The calculated time for 50% decrease in concentration (Dt₅₀) was 11.7 years for pp′--DDT, 3.3 years for pp′--TDE and 7.1 years for op′--DDT; the time for doubling the concentration for pp′--DDE was 9.1 years. Regression analysis on the two major components (pp′--DDT+pp′--DDE) indicated that the total amount (2.7 mg kg^?1) was not decreasing with time. It was concluded that during a post-spray era, the breakdown of pp′--DDT to pp′--DDE was a significant feature of the persistence of DDT, and that, in contrast to the findings of other workers who sampled when DDT was being used, there were no losses by volatilisation. There was an exponential decrease in the amount of DDT residues with increasing soil depth and approximately 90% was found in the top 10 cm of the undisturbed soil profile.     

DDT and sodium transport in the eel electroplaque

DDT inhibits the ATPase activity of the intact eel electroplaque. At a concentration of 10^?5M, DDT inhibited 46% of the total ATPase activity, and 10^?4M DDT inhibited 62% of the total ATPase activity and 62% of the ouabain-sensitive ATPase activity. The latter concentration of DDT reduced the rate of Na efflux from intact electroplaques and slowed the rate of recovery of the membrane potential following a large depolarization produced by carbamylcholine application. Repetitive direct stimulation of the innervated membrane at 10 Hz during the application of 10^?4M DDT produced a significant irreversible depolarization. Ouabain, 10^?4M, produced similar effects. The possible role of the inhibition of active NaK transport in producing the symptoms of DDT poisoning is discussed.     

Inhibition of acetyl-coenzyme a carboxylase by coenzyme a conjugates of grass-selective herbicides

A total of 146 samples of different kinds of cheeses produced in Spain were analysed in order to ascertain the specific contamination pattern. The organochlorine compounds studied were those most commonly investigated in previous surveys: α-HCH, β-HCH, γ-HCH (lindane), γ-HCH, chlordane, aldrin, dieldrin, endrin, heptachlor, heptachlor epoxide, and the isomers and metabolites of DDT. α-HCH, β-HCH, γ-HCH, chlordane, p,p′, DDT, and p,p′-DDE were found in more than 76% of samples; p,p′-DDE and γ-HCH were the most frequently detected, with frequencies of 100 and 97.9% respectively. γ-HCH, aldrin, dieldrin, heptachlor, heptachlor epoxide, o,p′-DDT, p,p′-DDD and o,p′-DDD were observed at lower frequencies. No residues of endrin were detected in any sample. Insecticides exceeding the maximum residue limits (MRLs) were chlordane, β-HCH, α-HCH and γ-HCH, with 42, 20, eight and six samples respectively. Mean residues of organochlorines found were as follows (μ kg^?1 butterfat): α-HCH = 46.3; β-HCH = 46.5; γ-HCH = 54.2; δ-HCH = 16.9; aldrin = 16.7; dieldrin = 9.7; heptachlor = 15.9; heptachlor epoxide = 14.8; chlordane = 50.2; o,p′-DDT = 5.1; p,p′-DDT = 12.4; o,p′-DDT = 19.6; p,p′-DDD = 46.7; o,p′-DDE = 6.9; p,p′-DDE = 40.7 (.DDT = 55.0). Estimated dietary intakes (EDIs) from cheese consumption were compared to acceptable daily intakes (ADIs) for the pesticides where residues exceeded the MRL. EDIs calculated were in all cases below ADIs, and, therefore, based on the ADIs, there is no health risk involved in the consumption of cheese from Spain arising from organochlorine residues.     

Binding of [14C]DDT and [14C]dicyclohexylcarbodi-imide to a lipoprotein of the membrane sector of mitochondrial ATpase

Intact mitochondria, isolated from red coxal muscle of the American cockroach (Periplaneta americana L.), were incubated in the presence of 1,1,1-trichloro-2,2-bis(4-chloro[¹⁴C]phenyl)ethane ([¹⁴C]DDT) to isolate a suspected binding site for DDT in the membrane sector of the mitochondrial ATPase. The requirements for the binding of DDT were compared with those for the binding of dicyclohexyl[¹⁴C]carbodi-imide([¹⁴C]DCCD), a potent inhibitory probe of mitochondrial ATPase activity. [¹⁴C]DDT appeared to bind to a proteolipid of the membrane sector, which also binds [¹⁴C]DCCD. Exchange experiments, with [¹⁴C]DCCD, [¹⁴C]DDT and unlabelled DDT at different concentrations, indicated that DDT and DCCD may be acting on a similar protein. This protein may act as the energy transducing protonophore required for the synthesis and hydrolysis of ATP in coupled mitochondria. Inhibition of mitochondrial ATPase activity may be a consequence of DDT and DCCD binding to this proteolipid protonophore, resulting in the disruption of energy transduction in muscle and nerve.     

Excretion of DDT residues into milk of the Indian buffalo,Bubalus bubalis (L.) after oral and dermal exposure

Three groups of buffaloes were fed with 20, 100 and 400 mg of p,p′-DDT in their daily rations. The DDT residues in the milk fat of the treated animals showed an initial rapid rise but soon attained a dose-dependent equilibrium. The transfer coefficient of DDT residues in milk at ‘plateau’ levels showed an average value of about 12%. Half-life values for the rate of decline of DDT residues during the post-dosing period were computed according to a two-open-compartment model. Dermal application of p,p′-DDT to buffaloes also resulted in excretion of a significant amount of its residues in milk. TDE was the predominant compound present in milk when buffaloes had ingested p,p′-DDT, whilst p,p′-DDT itself was present in greater quantity than its metabolites when animals were treated dermally.     

The postmortem reductive dechlorination of pp′DDT in avian tissues

Pigeons, Bengalese finches, and Japanese quail were dosed with pure pp′DDT, and their tissues were examined for residues immediately after death, or following storage. DDD residues in livers extracted immediately after death were always low (<5% DDT concentration) and sometimes undetectable. Reductive dechlorination of DDT to DDD occurred postmortem, relatively repidly at 20°C (90% after 8 hr), but more slowly at ?10 and ?20°C. The conversion could not be attributed to bacterial action and was evidently due to processes operating within the cell under anaerobic conditions. During storage at ?12 to ?15°C, reductive dechlorination occurred in brain and in embryos, but did not occur to any significant extent in depot fat or in eggs without embryos. These findings raise questions about the importance of DDD as an in vivo metabolite in birds and other vertebrates. The interpretation of DDD residues in tissues from laboratory and field studies is discussed.     

The effect of DDT and its analogs on the fragility of human erythrocytes

The antihemolytic actions of DDT and eight analogs were examined with human erythrocytes. Apparent aqueous concentrations to produce 60% of control hemolysis ranged from 3.7 × 10^?4 to 2.4 × 10^?6M, with DDT being one of the least active. No correlation was found between antihemolytic potency and neurotoxicity, and it was concluded that the findings did not illuminate the toxic or neural actions of these compounds.     

Influence of methyl bromide soil fumigation method on fungicidal efficacy and bromide residues

Highest inorganic bromine residues (30 ppm) were found when soil was fumigated with liquid methyl bromide (MB) introduced by conventional means into evaporating dishes. With preheated (vaporized) MB or injection of MB/chloropicrin (CP) mixtures, bromide concentrations were reduced by 50%. They were uniform throughout the soil (0 to 60 cm) except after MB/CP injection, when larger residues were found in the 30-60 cm layer. Leaching with 2000 m^p3/ha (20 cm) of water always reduced bromide content to 7.5 and 10 ppm at the 0-30 and 30-60 cm depth, respectively. Organic amendments to soils substantially increased bromide levels up to 118 ppm, most of which was found in the upper soil layers; two teachings with 2000 m^p3 /ha water were required to return the soils to their normal state.Sclerotium rolfsii andFusarium oxysporum f. sp.dianthi cultures buried in soil were eliminated from the upper 30 cm with MB applied either conventionally or preheated. At 50 cm, 500 kg/ha of the preheated gas was superior to 1000 kg/ha of the cold gas. All MB fumigations suppressed carnation flower yields compared with CP alone but were superior to no treatment. After leaching, MB-fumigated soils yielded the highest number of flowers.     

The in vivo sensitivity of ATPases to DDT,DDE, and physical immobilization in American cockroaches

American cockroaches injected with sublethal doses of DDT (0.75 μg/roach) at 5-day intervals showed a 40% reduction in oligomycin-sensitive Mg²⁺ATPase from muscle homogenates, and a 23% reduction of Na⁺-K⁺ATPase from nerve cords. Thus, the maximum effect measured occurred with the same enzyme and tissue as determined from in vitro studies. The metabolite, DDE, used at 15 μg per roach, gave no significant change in activity of the ATPase system following injection. In contrast, high single doses of DDT (7.5 μg/roach) and 100 μg DDE and dicofol per roach caused over 30% increase in oligomycin-sensitive Mg²⁺ATPase of muscle and a 10–15% increase in Na⁺-K⁺ATPase of nerve cords measured 24 and 48 hr later. While a similar response was observed for Mg²⁺ATPase activities in cockroaches that were immobilized, the increase in enzyme activities were much greater than that caused by the pesticides.     

Difenzoquat, 1,2-dimethyl-3,5-diphenylpyrazolium ion,a selective herbicide for the control of wild oats (Avena spp.) in wheat and barley

The results of a series of replicated trials and of a series of commercial grower trials conducted in the United Kingdom during 1972 and 1973, established the effectiveness of difenzoquat (1,2-dimethyl-3,5-diphenylpyrazolium ion) as the methyl sulphate as a selective post-emergence herbicide for the control of wild oats in wheat and barley. Doses of 0.75 to 1.0 kg cation/ha applied at 200 or 400 litre/ha and at 2.1 or 3.5 kg/cm² gave good to excellent results when applied at crop growth stages 3 to 5 (Feekes—Large scale) for spring crops and at crop growth stages 4 to 6 for winter crops. In these experiments wild oats over a range of stages from 1st leaf to tillering were well controlled.