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1,1-Dichloro-2,2-bis (4'-chlorophenyl)ethane (DDD), 1,2-dibromoethane (DBE) and trichloroethylene are three halogenated hydrocarbons that selectively bind to pulmonary epithelial cells and that may be pneumotoxic. The susceptibility of pulmonary cells and the mechanisms of cytotoxicity of these compounds were evaluated using enriched subpopulations of isolated rabbit lung cells incubated with DDD, DBE, and trichloroethylene. These chlorinated and brominated hydrocarbons were studied to evaluate their ability to induce selective pneumotoxicity by their bioactivation in three cell types, i.e. Clara cells, alveolar type II cells, and alveolar macrophages. Evidence of cytochrome P-450 bioactivation was assessed by utilizing the suicide inhibitor, 1-aminobenzotriazole (ABT) to ameliorate cytotoxicity. DDD, DBE and trichloroethylene were cytotoxic to Clara cells, type II cells and alveolar macrophages and the order of cell susceptibility to DDD was Clara > type II > macrophages. DBE and trichloroethylene were nonselectively cytotoxic. ABT reduced the cytotoxic effects of DDD and DBE in Clara cells. These studies indicated that all three compounds were toxic to isolated lung cells and that bioactivation of DDD and DBE in rabbit Clara cells to a cytotoxic intermediate was mediated, at least in part, by cytochrome P-450 oxidation.

The transfer of p,p'-DDT (1,1,1-tricholoro-2,2-bis(4-chlorophenyl)ethane) and its metabolites to infants via breast-feeding was studied in an area of KwaZulu, South Africa, where DDT is used to interrupt malaria transmission. Samples of whole blood were collected from 23 infants, together with samples of breast milk from their respective mothers. The mean sigma DDT (total DDT) in the whole blood was 127.03 micrograms.l-1 and that in the breast milk, 15.06 mg.kg-1 (milk fat). The % DDT (% DDT of sigma DDT) was significantly higher in the infant blood than in the breast milk (P less than 0.05). A multiplicative regression analysis indicated that sigma DDT increased significantly (P less than 0.01) in infant whole blood with infant age. Multiple regression showed that 70.0% of the variation in sigma DDT was due to the variation in parity of the mother, age of the infant, and the sigma DDT in breast milk. These variables accounted also for 76.3% of the variation in p,p'-DDE but only for 38.2% of that in p,p'-DDT. Organochlorines were therefore largely transferred to the infant from the mother, with DDT in the environment playing a secondary role.

Male and female chickens of a broiler-type strain were fed, from 1 day old to 5 weeks of age, diets containing 0, 2.5, or 15.0 p.p.m. (mg/kg) 1,1,1-trichloro-2,2-bis(p-chlorophenyl)ethane (p,p'-DDT). Then the diets with pesticide were withdrawn and the chickens were fed dietary levels of diphenylhydantoin (DPH) at 0, 100, or 250 p.p.m. Adipose-tissue and liver samples were obtained on days 0, 10, 20, and 30 following withdrawal of diets with pesticides to determine DPH effect on DDT, 1,1-dichloro-2,2-bis(p-chlorophenyl)ethylene (DDE), and 1,1-dichloro-2,2-bis(p-chlorophenyl)ethane (DDD) levels. DPH had no effect on the concentration of DDT and DDE in adipose tissue; their levels declined at a rate having a half-life value of 16 days. DDD was not detected in adipose tissue. DDT accounted for 87% of the adipose residues on day 0, but 66% of the residues at day 30. DPH had no effect on the concentrations of DDT and DDE in livers of chickens fed 15.0 p.p.m. DDT, but did significantly reduce the levels of DDD by 28 and 54% for levels of 100 and 250 p.p.m. DPH, respectively. The similarity of these data to studies on dairy cows and humans, and the dissimilarity to data from rat studies were discussed.

Residues of DDT, DDD, DDE, and dieldrin in 7 species of ducklings from Alberta, Canada, were measured. Levels of DDT and metabolites ranged from 0 to 36.48 ppm in fat of 96% of the birds and from 0 to 0.97 ppm im muscle from 67% of the birds. Residue levels of dieldrin ranged from 0 to 2.62 ppm in fat and from 0 to 0.11 ppm in muscle of 43 and 19% of the birds, respectively. Growth dilution was considered to be the most significant factor in reducing the insecticide residue levels in the ducklings. DDT (total DDT, DDE, and DDD) residue levels were decreased with increased weight of pintail (A. acuta), baldpate (Mareca americana), and gadwall (A. strepera) ducklings. Dieldrin used in grasshopper control did not contribute to residue levels in the ducklings. Contamination by DDT was considered to have originated from outside the habitat. No dieldrin or DDT residues were found in ducklings' food sources analysed.

The metabolism of o,p'-DDD (mitotane), a well-known inhibitor of adrenal steroidogenesis in man and animal, is reviewed. Following oral administration, about 65% of the ingested drug were found to pass in the stool. The drug appeared in the urine in metabolized forms: o,p'-DDA and mono-and dihydroxylated derivatives of o.p'-DDA. These latters were found as well in the stools. An unsaturated metabolite, o,p'-DDE was described in plasma and tissues in man. Serum specimens of treated patients were analyzed for o,p'-DDD during various phases of therapy: the levels and the rate of rise during treatment were very variable (5 to 90 microng/ml). Tissue levels were obtained from animals (rats, dogs) or men (biopsy as well as autopsy): o,p'-DDD was primarily found stored in adipose tissue and fat-containing tissues, essentially adrenals. Practical conclusions can be drawn from these results: there is no correlation between the dose of o,p'-DDD administered and its blood level; there is no correlation between blood levels and the patient's responsiveness to the drug; there is a possibility that the molecule transformed in an active metabolite through its metabolism.

DDT orally given to rats is converted into DDD by reductive dechlorination in the liver within 30 min. 6 h after a single application of 10 mg DDT per rat a quantity of 60 mg DDD/kg was indicated in the liver. DDT is stored in the adrenal glands; in the other organs an equilibrium comes up obviously, even with further application. The quantitative ratios are reported. Information on the experimental details of DDT determination and--anticipatory to the following publications--on the analysis of the most relevant metabolites is given.

Extensive biodegradation of 1,1,1-trichloro-2,2-bis(4-chlorophenyl)ethane (DDT) by the white rot fungus Phanerochaete chrysosporium was demonstrated by disappearance and mineralization of [14C]DDT in nutrient nitrogen-deficient cultures. Mass balance studies demonstrated the formation of polar and water-soluble metabolites during degradation. Hexane-extractable metabolites identified by gas chromatography-mass spectrometry included 1,1,-dichloro-2,2-bis(4-chlorophenyl)ethane (DDD), 2,2,2-trichloro-1,1-bis(4-chlorophenyl)ethanol (dicofol), 2,2-dichloro-1,1-bis(4-chlorophenyl)ethanol (FW-152), and 4,4'-dichlorobenzophenone (DBP). DDD was the first metabolite observed; it appeared after 3 days of incubation and disappeared from culture upon continued incubation. This, as well as the fact that [14C]dicofol was mineralized, demonstrates that intermediates formed during DDT degradation are also metabolized. These results demonstrate that the pathway for DDT degradation in P. chrysosporium is clearly different from the major pathway proposed for microbial or environmental degradation of DDT. Like P. chrysosporium ME-446 and BKM-F-1767, the white rot fungi Pleurotus ostreatus, Phellinus weirii, and Polyporus versicolor also mineralized DDT.

The microbial transformation of DDT, DDD and DDE was studied in Gram-negative strain B-206 and a number of phenolic metabolites were identified as the trimethylsilyl derivatives in the bacterial extracts by gas chromatography/mass spectrometry. The major metabolites of DDT were DDD, DDE, DDMU, 1,1,1-trichloro-2-(2-hydroxy-4-chlorophenyl)-2-(4'-chlorophenyl) ethane, 1,1,1-trichloro-2-(2-hydroxy-4-chlorophenyl)-2-(4'-hydroxyphenyl) ethane, and 1,1,1-trichloro-2,2-bis-(2-hydroxy-4-chlorophenyl) ethane. Conversely, DDD was mainly degraded into DDE, 1,1-dichloro-2-(2-hydroxy-4-chlorophenyl)-2-(4'-chlorophenyl) ethane and 1,1-dichloro-2-(2-hydroxy-4-chlorophenyl)-2-(4'-hydroxyphenyl) ethane. Finally, DDE was transformed into DDMU, 1,1-dichloro-2-(2-hydroxy-4-chlorophenyl)-2-(4'-chlorophenyl) ethylene, 1,1-dichloro-2-(2-hydroxy-4-chlorophenyl)-2-(4'hydroxyphenyl) ethylene and 1-chloro-2-(2-hydroxy-4-chlorophenyl)-2-(4'-chlorophenyl) ethylene. The phenolic metabolites exhibited [M - TMSCl]+., [M - HCl - TMSCl]+. and/or [M - HCl - TMSCl - Me]+ fragment ions which reflect the presence of an ortho hydroxyl group in these molecules. Other mass spectral features used to determine their structure are presented and a metabolic scheme accounting for their formation is proposed.

DDD is formed from DDT by reductive dechlorination. After application of DDD to rats the dechlorination product DDMU could be detected only after short exposures in the organs, in high concentrations, however, permanently in the faeces. The distribution of DDD in the organism of the rats is studied.

pp'DDT was converted to DDD 1-1 dichloro-2,2 bis (p-chlorophenyl) ethane, by treated 12 000 x g supernatant preparations from grey mullet livers. The anaerobic reductive dechlorination occurs whether or not exogenous NADPH and riboflavin is added after a heat pretreatment, otherwise the dechlorination is lowered when the cofactors are not added.

The metabolism and excretion of 14C-DDT, 14C-DDD, 14C-DDE and 14C-DDMU were compared in male rats and male Japanese quail after i.p. injection. The rate of excretion of radioactivity was greater in the rat than in the quail for all compounds except DDMU. Skin and fat were major sites of residual radioactivity in both species, the compound administered and DDE accounting for most of the radioactivity except in the case of DDMU. The four radiolabelled compounds were all present in significant quantities in excreta of both species in unchanged forms. The metabolic patterns for DDT and DDD were similar in rat and quail, except that the rat formed 2,2-di(4-chlorophenyl)ethanol (DDOH) from DDT while the quail did not. Rat and quail metabolized DDE and DDMU differently. Ring-hydroxylated derivatives of DDE were formed only in the rat and analogous metabolites of DDMU were produced only by quail. Both species produced di(4-chlorophenyl)acetic acid as a metabolite of all four compounds administered, although the formation was generally less and slower in quail than rat. Comparative metabolism of DDMU with the other compounds indicated that this compound is not a metabolic intermediate in the metabolism of DDT in either rat or quail.

Previous studies have shown that o,p'-DDD is activated and covalently bound in the mouse lung. In order to examine the structure dependency of the selective lung binding, the 14C-labelled DDD isomers p,p'-DDD, m,p'-DDD and o,m'-DDD were injected intravenously into female C57B1 mice and covalent binding was measured. Autoradiography of solvent-extracted tape-sections showed that all isomers were selectively and covalently bound in the lung alveolar region. As determined by exhaustive extraction of homogenized tissue, maximal binding was observed 4 hr after injection, although the lung/liver concentration ratio increased for 12 days. Covalent protein binding was also observed in vitro, implying that the activation of DDD to a reactive metabolite takes place in the target organ. Since the aryl-chlorine substitution pattern did not change the selective lung binding, bioactivation of DDD may take place at the ethane side-chain.

Heme compounds, in combination with a reducing agent and oxygen, can express various activities of cytochrome P-450 enzymes. In the present study it was found that a mixture of hemin and excess cysteine was able to degrade the insecticide DDT partially. The major products were three water-soluble, non-toxic conjugates of DDT metabolites with cysteine which had lost two or three of the five chlorine atoms of DDT per molecule and whose structures were elucidated by gas chromatography/mass spectrometry. In 0.05 M NH4HCO3, pH 7.7/ethanol (5:6, by vol.), the degradation reaction catalyzed by the hemin-cysteine model system was at least 8 x 10(4) times faster than the uncatalyzed reaction. In the presence of a designed 24-residues polypeptide or beta-casein, two DDT-binding proteins, an additional fourfold increase in the rate of DDT degradation was observed. Although the concentrations of DDT and cysteine occurring in an organism would be expected to be lower than those in the experiments described, the formation of water-soluble conjugates of DDT metabolites with cysteine (and other amino acids) could also play a role in metabolism and excretion of DDT in vivo.

Four-week old broiler chickens were fed rations with varying levels of technical grade DDT for a 4-week period. DDT and metabolite concentrations were measured in adrenal, liver, and brain. Accumulations per gram of tissue were greatest in the adrenals, followed by those in the liver, then in the brain. All birds fed 2700 p.p.m. of DDT died with 12 days after beginning feeding, and moderate signs of toxicity, such as ataxia and trembling, appeared in those fed 900 p.p.m. Although both DDT metabolites, p,p'-DDE and p,p'-DDD, were found in the tissues, the higher levels of the former indicated that the aerobic conversion of DDE occurs more readily than the anaerobic conversion. Significant quantities of o,p'-DDT appear only in tissues of birds fed high levels of the technical grade DDT.

The distribution of residues of DDT and its metabolites (DDD and DDE) in tissues of Triaenophorus nodulosus and its host was studied. It has been noted that cestodes can accumulate chlororganic substances in great quantity. The ability of Bothriocephalus gowkongensis has been established to dechlorate pp-DDT and to excrete pp-, op- and mp-DDD from the organism. Pp-DDT decreases the glycogene content and causes the rise of amylolytic activity in the above cestodes in acute tests. The effect of pp-DDT on the locomotor activity of cestodes is of two-phase character. The first one is associated with the muscular influence, the second one-- with neurotrophic effect. A high toxicity of mp-DDD )0.1 mg/l) for the locomotor activity of cestodes as compared with the effect of pp- and op-DDD was revealed.

The urinary metabolites of 1,1,1-trichloro-2,2-bis(p-chlorophenyl)ethane (DDT), 1,1-dichloro-2,2-bis(p-chlorophenyl)ethane (DDD), and 1-chloro-2,2-bis(p-chlorophenyl)ethene in female hamsters are reported. The principal metabolite of both DDT and DDD is 2,2-bis(p-chlorophenyl) acetic acid. DDT- and DDD-treated animals also excreted small amounts of DDD, 1-chloro-2,2-bis(p-chlorophenyl)ethene, 1,1-dichloro-2,2-bis-(p-chlorophenyl)ethene, 2-hydroxy-2,2-bis(p-chlorophenyl)acetic acid, and 2,2-bis(p-chlorophenyl)ethanol. 1-Chloro-2,2-bis(p-chlorophenyl)ethene is metabolized to afford significant amounts of 2,2-bis(p-chlorophenyl)acetic acid, 2,2-bis(p-chlorophenyl)-ethanol, 2-hydroxy-2,2-bis(p-chlorophenyl)acetic acid, 2,2-bis-(p-chlorophenyl)acetaldehyde, and 1,1-bis(p-chlorophenyl) ethan-1,2-diol. These results indicate that the metabolic disposition of DDT in the hamster, a species refractory to DDT tumorigenicity, is very similar to that observed previously in the mouse, a species sensitive to DDT tumorigenicity. The one exception is that the hamster is not nearly as efficient as the mouse in converting DDT to 1,1-dichloro-2,2-bis(p-chlorophenyl)ethene, a metabolite that is tumorigenic in both species.

The lymphatic absorption of a series of 14C-labelled compounds structurally related to p,p'-DDT has been examined in thoracic duct-cannulated rats. The compounds (p,p'-DDT, p,p-DDT, P,P'-DDD, p,p-DDD, DDE, DDA and 2,4-D) varied markedly in lipid solubility, as well as in extent of lymphatic absorption, urinary and biliary excretion, and localization in body fat. More than 12% of the dose was recovered in 24-hour thoracic duct lymph of rats treated with the p,p'- and o,p-isomers of DDT and DDD and with DDE, and this percentage was increased by concomitant fat absorption. A strict parallel between lipid solubility and lymphatic absorption was not observed in this series of compounds, possibly because of differences in their rates and routes of excretion.

Dichlorodi[U-14C]phenyltrichloroethane ( [14C]DDT), incubated with rat hepatic microsomes and NADPH, produced reactive intermediates which covalently bound to microsomal protein and lipids. In atmospheric oxygen, DDT bound to microsomal protein; however, binding was increased up to approximately 70% by oxygen depletion. Low levels of [14C]DDT binding to microsomal lipids occurred under atmospheric oxygen but, in contrast to protein binding, DDT-phospholipid binding was increased up to 20-fold by oxygen depletion. Dichlorodiphenyldichloroethane (DDD) was rapidly formed from DDT under anaerobic conditions, although when DDD was utilized as substrate, binding to microsomal protein occurred only in the presence of oxygen. Sodium dithionite, added to microsomes, produced [14C]DDT phospholipid and protein binding, and DDD formation, but failed to support DDD metabolism or binding. The data are consistent with the reductive formation of a DDT free-radical intermediate that led to the formation of DDD and that was bound preferentially to microsomal lipids.

The metabolites of 1,1,1-trichloro-2,2-bis(p-chlorophenyl)ethane (DDT) and 1,1-dichloro-2,2-bis(p-chlorophenyl)ethane (DDD) found in the urine of female Swiss mice are reported. The metabolites of DDT are DDD, 1-chloro-2,2-bis(p-chlorophenyl)ethene (DDMU), 1,1-dichloro-2-bis(p-chlorophenyl)ethene (DDE), 2,2-bis(p-chlorophenyl)acetic acid (DDA), 2-hydroxy-2,2-bis(p-chlorophenyl)acetic acid (alpha OH-DDA) and 2,2-bis(p-chlorophenyl)ethanol (DDOH), while DDD afforded DDMU, DDE, DDA, alpha OH-DDA and DDOH. The relative excreted levels of DDA and DDOH and the absence of 2,2-bis(p-chlorophenyl)acetaldehyde (DDCHO) are not consistent with the generally accepted pathway for DDA formation, which involves sequential metabolism of DDT and DDD via DDOH to afford DDA. The quantitative results are interpreted to mean that DDA is formed by hydroxylation at the chlorinated sp3-side chain carbon of DDD to give 2,2-bis(p-chlorophenyl)acetyl chloride (DDA-Cl), which in turn is hydrolyzed to DDA. The excretion of alpha OH-DDA from both DDT- and DDD-treated mice has never been previously observed. It is suggested that this metabolite arises from the initial epoxidation of DDMU, a metabolite of DDT and DDD, to yield 1,2-epoxy-1-chloro-2,2-bis(p-chlorophenyl)ethane (DDMU-epoxide). This chloroepoxide is then hydrolyzed and oxidized to produce the alpha OH-DDA.

The effects of PCBs (polychlorinated biphenyls) or a combination of DDT (1,1,1-trichloro-2,2-bis (p-chlorophenyl) ethane), DDD (1,1-dichloro-2,2-bis (p-chlorophenyl) ethane) and DDE (1,1-dichloro-2,2,-bis (p-chlorophenyl ethylene) on organ weights, liver storage of vitamin A and carotene, selected blood chemistry parameters, and serum protein fractions were determined in penned white pelicans (Pelecanus erythrorhynchos) receiving a daily dosage of these compounds. Birds received 100 mg of PCBs or a combination of DDT (20 mg), DDD (15 mg), and DDE (15 mg) injected into the first fish fed each day for ten weeks. A greater percentage of PCB treatment was retained in brain, liver, carcass and feathers than the percentage of DDT + DDD + DDE treatment. Liver weight as percent of body weight decreased (p less than 0.01) in DDT + DDD + DDE-treated birds and increased (p less than 0.01) as a total weight in PCB-treated birds. Spleen weight as percent of body weight was greater (p less than 0.05) in PCB-treated birds. Neither treatment had a significant effect on the weight of the brain, heart, or kidney. Liver vitamin A levels were greater (p less than 0.01) on a mug/g of liver basis in the DDT-treated birds than in controls. Significant lowering of serum potassium and protein values was noted in both the PCB- and the combination of DDT, DDD, DDE-treated birds, while serum calcium values were lowered (p less than 0.01) only in PCB-treated birds. Vaues of serum inorganic phosphorus, uric acid and magnesium were not significantly changed by either treatment. Globulin fractions were unaltered by either treatment, but albumin fractions were lowered (p less than 0.01) in the PCB-treated pelicans.

The possibility of complete degradation of DDT by Pseudomonas aeruginosa 640x was demonstrated in principle. A study of the conditions of degradation of DDT by this culture was made. It was demonstrated that only dechlorination of DDT to DDD is accomplished without addition of a supplementary substrate. The rest of the processes right up to the formation of benzhydrol and phenylacetic acid take place only under conditions of cometabolism. For dechlorination of the aliphatic fragment of DDT and the aromatic rings, anaerobic conditions, nitrates in the form of electron acceptors, and calcium lactate as a cosubstrate are preferred. Degradation of nonchlorinated benzophenone takes place only under aerobic conditions with glycerol as a cosubstrate. Phenylacetic acid and benzhydrol are used by the culture as sole sources of carbon; aerobic conditions are necessary for their degradation. On the basis of analysis of decomposition products of DDT and a study of the pathways of degradation of its metabolites and analogs, a means of converting DDT by P. aeruginosa 640x is proposed.

A study was under-taken to investigate the influence of pretreatments with various hepatic microsomal enzyme inducers on the elimination of DDT residues in rats previously contaminated with p,p' -DDT at 5 mg/kg/day, during 20 days. The following inducers were used : phenobarbital (50 a mg/kg/day, i.p.), 3,4-benzopyrene (20 mg/kg/day, s.c.) and norethandrolone (20 mg/kg/day, s.c.), all given during 14 consecutive days. Each inducer was administered singly or in combination with the other two according to a 2 X 2 X 2 factorial experiment. The animals were then sacrificed for the measurement of p,p' -DDT, p,p' -DDD and p,p' -DDE residues in the blood, brain, abdominal fat, liver and kidneys. The results show that phenobarbital lowers markedly the concentration of total residues in the fat tissue and brain and that norethandrolone brings about a reduction of the residues in the blood, brain and kidneys, but not in the fat tissue. On the opposite, 3,4-benzopyrene produces an increase of the residues in the brain, liver and kidneys. Phenobarbital thus appears to be more efficacious than the other two inducers in facilitating the elimination of DDT residues from the fat tissue. In addition, it appears that under the experimental conditions used during this investigation, the elimination of DDT residues is not further accelerated by combining the inducers one with each other.

The possibility that lead could affect the metabolism of the insecticide 1,1-dichloro-2, 2-bis(p-chlorophenyl)ethane, DDD, was examined by studies of the effects of chronic oral Pb treatment on DDD conversion to 2,2-bis(p-chlorophenyl)acetic acid, DDA. Rats were given either distilled deionized water or 0.05, 0.58, 17 or 352 ppm Pb solutions as drinking water. Systolic blood pressure and body weight were measured weekly. Rats drinking 352 ppm lead chloride manifested a statistically significant increase in blood pressure. Rats drinking 352 ppm and 17 ppm lead chloride showed a significant decrease in the rate of weight gain compared to controls. All groups showed an increase in the excretion of total DDA compared to controls.