Relation between the content of organochlorine compounds in Finnish human milk and characteristics of the mothers

Neutral organochlorine pesticide and PCB residues were analyzed by GC-MS technique in 183 human milk samples obtained in 1984-1985 from 165 women living in different parts of Finland. The effect of the donors' age, body mass, place of residence, number of children, dietary habits, smoking habits, occupational history, and weight loss on the organochlorine content of human milk were studied. Of all the milk samples analyzed, p,p'-DDE concentrations were above the detection limit in 99.5%, p,p'-DDD + p,p'-DDT in 57.9%, isomers of HCH in 30.0%, cis-chlordane in 4.9%, oxychlordane in 3.3%, trans-nonachlor in 6.0%, heptachlor in 12.0%, and heptachlor epoxide in 6.6%. Mirex was not found in any of the milk samples, whereas the signals of chlorinated terpenes (toxaphenes) were detected but could not be quantitatively determined. The mean fat adjusted residue levels above the detection limit in Finnish human milk samples of primipara mothers were 0.66 ppm for total DDT compounds, 0.08 ppm for HCB, 0.93 ppm for PCBs, 0.41 ppm for chlordane compounds, 0.20 ppm for isomers of HCH, and 0.10 ppm for heptachlor epoxide. The geometric means were 0.46, 0.06, 0.57, 0.02, 0.02, and 0.01 ppm, respectively. The age of the mothers positively correlated with the DDE concentrations in human milk. The residues of OC compounds in human milk did not differ in women living in plywood industry regions, those actually working in the industry, and other mothers. Small differences were detected in the levels of organochlorine compounds in different parts of Finland. No relation was found between the OC content and the fish consumption, smoking habits, weight loss, or social group of the donors.     

Organochlorine compounds in adipose tissue of Greenlanders and southern Danes.

Abdominal fat tissue samples from the general population of Greenland and from southern Denmark were analyzed for o,p'-DDE and p,p'-DDE [1,1-dichloro-2,2-bis(p-chlorophenyl)ethylene], p,p,-DDT [1,1,1-trichloro-2,2-bis(p-chlorophenyl)-ethane], o,p'-DDT and p,p'-DDD [1,1-dichloro-2,2-bis(p-chlorophenyl)ethane], lindane (1,2,3,4,5,6-hexachlorocyclohexane), aldrin (1,2,3,4,10,10-hexachloro-1,4,4a,5,8,8a-hexahydro-endo-exo-1,4:5,8-dimethanonaphthalene), dieldrin (1,2,3,4,10,10-hexachloro-6,7-epoxy-1,4,4a,5,6,7,8,8a-octahydro-endo-exo-1,4:5,8-dimethanonaphthalene), heptachlor (1,4,5,6,7,8,8-heptachloro-3a,4,7,7a-tetrahydro-4,7-methanoindene), heptachlor epoxide, and polychlorinated biphenyls (PCBs). Fat tissue from Greenlanders contained significantly higher amounts of p,p'-DDE (p less than or equal to 0.05), p,p'-DDT (p less than or equal to 0.01), and total DDT (SIGMA DDT, the sum of DDT and its metabolites) (p less than or equal to 0.01) than southern Danes. Lindane, aldrin-like residue, dieldrin heptachlor-like residue, heptachlor epoxide, and PCBs were present in adipose tissue of both groups and there were no significant differences between the groups. The p,p'-DDE level in Greenland was lower than that in the United States, eastern Europe, and India. Among Greenlanders of different ages the highest sigma DDT was found in the age group of 22-45 yr; the content of PCBs increased with age. Among southern Danes the highest sigma DDT was found in a higher age group than in Greenlanders. Among Greenlanders the content of aldrin-like residue decreased with age and that of dieldrin increased with age. A significant correlation was found (on the basis of wet weight) between p,p'-DDE and PCB content in southern Danes (p less than or equal to 0.02). The correlation between sigma DDT and PCB content was also significant in this population (p less than or equal to 0.01). These two relationships were not significantly correlated for the Greenlanders. The DDE/PCB and sigma DDT/PCB ratios were higher in Greenlanders than in southern Danes. These ratios are age-dependent and are highest in the age group 22-45 yr among the Greenlanders. A low DDE/PCB ratio and a low sigma DDT/PCB ratio have been proposed as markers for industrialization. In Greenlanders p,p'-DDE represented about 70% of sigma DDT. For southern Danes this level was about 90%. The data obtained are presented and discussed on the basis of both lipid and wet weight levels.     

The presence of environmental pollutants in the semen of farm animals (bull,ram, goat,and boar)

Organochlorine pesticides and polychlorinated biphenyls are widely used in agriculture and industry, respectively, and may affect male reproduction function. Although several pollutants have been detected in human semen, similar studies in farm animals have not appeared. In the present study, the semen of bulls, rams, goats, and boars was assayed for the organochlorine pesticides hexachlorobenzene (HCB), hexachlorocyclohexane (HCH, alpha-, beta-, and gamma-isomers), dieldrin, and heptachlor epoxide, for DDT-related chemicals (o,p'-DDE, p,p'-DDE, p,p'-DDD, p,p'-DDT), and for the PCBs congeners (PCB-52, -101, -138, -150, and -180). In all species of farm animals, the most frequently detected pollutants were p,p'-DDE (80-100% of samples), HCB (73.9-100%), and gamma-HCH (69.6-100%). Species differences in the concentrations of HCB, alpha-, beta-, and gamma-HCH, dieldrin, p,p'-DDE, p,p'-DDD, and PCBs were noted as well as differences in the concentrations of some isomers of HCH, DDT-related chemicals, and PCB congeners in the same species.     

The presence of environmental pollutants in the follicular fluid of farm animals (cattle,sheep, goats,and pigs)

Organochlorine pesticides and polychlorinated biphenyls are widely used in agriculture and industry, respectively. The present study assessed the burden of environmental pollutants in the follicular fluid of farm animals (cattle, sheep, goats, and pigs). An analytical method combining a solid-phase extraction with (C(18)) for clean-up and GC-electron capture detection using a capillary column was implemented for isolation and determination of organochlorine pesticides and polychlorinated biphenyls (PCBs). Of the organochlorine pesticides, hexachlorobenzene (HCB), hexachlorocyclohexane (HCH, alpha-, beta-, and gamma-isomers), dieldrin, heptachlor epoxide, and the DDT-related chemicals (o,p'-DDE, p,p'-DDE, p,p'-DDD, p,p'-DDT) were detected and of the PCBs, the congeners PCB-52, -101, -138, -153, and -180 were detected. In all species of farm animals, the most frequently detected pollutant was gamma-HCH (90-100% of samples) followed by HCB (80-100%), and p,p'-DDE (75-90.91%). Species differences in the concentrations of HCB, beta-HCH, heptachlor epoxide, and DDT-related chemicals in follicular fluid were noted as well as differences in the concentrations of some pollutants within the same species.     

Assessing the estrogenic potential of organochlorine pesticides in primary cultures of male rainbow trout (Oncorhynchus mykiss) hepatocytes using vitellogenin as a biomarker

Many organochlorine pesticides are suspected of impairing natural hormonal function in organisms by mimicking endogenous estrogen. The aim of this study was to assess the estrogenic activity of the organochlorine pesticides o,p'-DDT, dieldrin, aldrin, heptachlor, mirex and DDT in rainbow trout hepatocyte cultures using vitellogenin (Vtg) as the biomarker. A wide range of pesticide concentrations (0.0001-100 microM) was evaluated. Among the pesticides tested, o,p'-DDT was the most potent inducer of Vtg. The lower potency of technical grade DDT relative to o,p'-DDT could be explained by the fact that this pesticide is a mixture of two different pesticides (18% o,p'-DDT and 77% p,p'-DDT). This suggests that o,p'-DDT is a stronger inducer of Vtg than p,p'-DDT. A simple hypothesis could be that pesticides mixed together competed for the same receptor to favor the formation of a complex with reduced activity towards EREs. If these compounds are classified according to the level of Vtg secreted, we observed the following decreasing order: 17beta-estradiol (E(2))>o,p'-DDT>dieldrin>aldrin>DDT. Non-toxic levels of these compounds competed with E(2) for binding to the estrogen receptor. Heptachlor and mirex did not induce Vtg. Since the latter compounds failed to stimulate Vtg production, the possibility that they could interfere with the estrogenic response by inhibiting E(2) action was tested. In the presence of heptachlor, Vtg production triggered by E(2) significantly decreased. The EC50 value for inhibition of ER binding by heptachlor was cytotoxic for hepatocytes in culture, and this could in part explain the lack of Vtg response observed with this compound at the concentrations tested. Our results indicate that organochlorine pesticides can act as positive or negative modulators of estrogenic function in rainbow trout.     

Age at natural menopause and exposure to organochlorine pesticides in Hispanic women

A cross-sectional study was conducted to evaluate the relationship between exposure to selected organochlorine pesticides (OCP) (p,p'-DDT, p',p'-DDE, dieldrin, hexachlorobenzene, beta-hexachlorocyclohexane [beta-HCH], oxychlordane, trans' nonachlor) and age at natural menopause in a sample of 219 menopausal women participating in the Hispanic Health and Nutrition Examination Survey in 1982-1984. Information on age at menopause, reproductive history, demographic variables, and potential confounding variables was collected via interview. Analysis of variance was employed to compare adjusted mean age at natural menopause among women by category of serum OCP level. Serum levels of p,p'-DDT, p,p'-DDE, beta-HCH, and trans-nonachlor were associated with a younger age at menopause. In particular, women with exposure levels in the highest exposure categories (serum p,p'-DDT > or = 6ppb, beta-HCH > or = 4ppb, or trans-nonachlor > or = 2ppb) had an adjusted mean age at menopause on average 5.7, 3.4, and 5.2 yr earlier, respectively, than women with serum levels of these pesticides below the detection limit. Women with serum p,p'-DDE levels greater than 23.6 ppb (highest quintile) had an adjusted mean age at menopause 1.7 yr earlier than women with serump,p'-DDE levels less than 5.5 ppb (lowest quintile). However, no consistent dose-response effect was apparent across low, medium, and high exposure categories. Interactions were detected for p,p'-DDT in combination with beta-HCH, trans-nonachlor, or oxychlordane, as well as beta-HCH in combination with oxychlordane.     

Determination of chlorinated hydrocarbon pesticides by solid-phase extraction and capillary GC with electron capture detection

Pesticides and their metabolites are extracted and concentrated from serum using C18 solid-phase extraction cartridges. The internal standard aldrin is added to 4 mL of serum or plasma, treated with 2 mL of methanol, and the resultant supernatant applied to the C18 cartridge. After several washes, pesticides are eluted from the column with isoctane and the eluate quantified with capillary gas chromatography using electron capture detection. Extraction efficiency is from 70 to 75% and the method detection limit ranges from 0.1-0.7 ng/mL depending on the analyte. Precision studies demonstrate that CVs range from 3.5 to 25.2%. Standard curves are linear to at least 7 ng/mL for lindane and chlordane isomers, heptachlor, heptachlor epoxide, oxychlordane, trans-nonachlor, dieldrin, dichlorodiphenyldichloroethylene (p,p'-DDE), dichlorodiphenyldichloroethane (p,p'-DDD), and dichlorodiphenyltrichloroethylene (p,p'-DDT).     

Increased ErbB-2 tyrosine kinase activity, MAPK phosphorylation, and cell proliferation in the prostate cancer cell line LNCaP following treatment by select pesticides.

The oncogene erbB-2 codes for a receptor tyrosine kinase that functions as a key mitotic signal in a variety of cell types. Amplification or overexpression of erbB-2 occurs in many forms of cancer, such as of the breast, colon, and prostate, and is an indicator of poor prognosis in those diseases. In the human prostate cancer cell lines LNCaP and PC-3, erbB-2 kinase was activated by pesticides of different chemical classes: (1) the organochlorine insecticides beta-hexa-chlorocyclohexane (beta-HCH), o,p'-dichlorodiphenyltrichloroethane (o,p'-DDT), and heptachlor epoxide; (2) the pyrethroid insecticide trans-permethrin, and (3) the fungicide chlorothalonil. o,p'-DDT also causes phosphorylation of mitogen-activated protein kinase (MAPK) and cellular proliferation of the androgen-dependent LNCaP line. However, no proliferative effect was observed in the androgen-independent PC-3 line. The proliferative effect of o,p'-DDT in LNCaP could not be blocked by the androgen receptor antagonist p,p'-dichlorodiphenyldichloroethene (p,p'-DDE), indicating that this effect of o,p'-DDT does not occur through direct interaction with the androgen receptor. Together these data demonstrate a putative mechanism for the action of certain pesticides in hormonal carcinogenesis.     

Effects of two isomers of DDT and their metabolite DDE on CYP1A1 and AhR function in human placental cells

The aim of this study was to investigate the actions of two isomers of DDT (p,p'-DDT, o,p'-DDT) and DDE (p,p'-DDE, o,p'-DDE) on the human placenta. We studied the effects of DDT and its metabolite DDE on CYP1A1 activity and on CYP1A1 and aryl hydrocarbon receptor (AhR) protein expression in placental cells. We used explants from third-trimester human placental tissue and JEG-3 cells, which are first-trimester human placenta cells. The main finding of this study was that the activity of CYP1A1 in the human placenta, measured in terms of ethoxyresorufin-O-deethylase (EROD) activity, was suppressed by treatment of 1, 10, and 100 ng/ml p,p'-DDT, o,p'-DDT, p,p'-DDE and o,p'-DDE. Immunoblot analyses indicated that both isomers of DDT and DDE inhibited the expression of CYP1A1 most effectively at 48 h and/or 72 h after the treatment. Because CYP1A1 activity is mediated by AhR, we evaluated the expression of AhR in placental tissue exposed to DDT and DDE for 1 h to 72 h. Our data showed that DDT and DDE gradually decreased the level of AhR protein, starting at 3 h or 24 h after the start of the experiment. Our results strongly support the involvement of the AhR/CYP1A1 signaling pathway in the mechanism of action of DDT and DDE in the human placenta.     

Organochlorine pesticide residues in chicken eggs: a survey

One hundred and five chicken egg samples were taken from seven geographical locations in Kenya and analyzed for organochlorine pesticide residues using gas-liquid chromatography. Nine organochlorine compounds were detected: alpha- and gamma-HCH/BHC (hexachlorocyclohexane/benzene hexachloride), aldrin, dieldrin, p,p'-DDE [1,1-dichloro-2,2-bis(p-chlorophenyl)ethylene], p,p'-DDD (TDE) [2,2-bis(p-chlorophenyl)-1,1-dichloroethane] (tetrachlorodiphenylethane), o,p'-DDD [2-(o-chlorophenyl)-2-(p-chlorophenyl)-1,1-dichloroethane], o,p'-DDT [2-(o-chlorophenyl)-2-(p-chlorophenyl)-1,1,1-trichloroethane], and p,p'-DDT. Eighteen of the samples contained DDT levels exceeding the practical residue limit (0.5 ppm). The mean DDT residue content was 0.68 ppm, a result influenced greatly by the high contamination in the 18 samples. Both individual and mean residue levels for other organochlorine pesticides detected were well within the respective practical residue limits.     

Demographic and seasonal influences on human serum pesticide residue levels

This study was intended to characterize more fully the distribution of serum concentrations of 16 pesticide residues with regard to key demographic and seasonal variables in a subsample of persons from the Second National Health and Nutrition Examination Survey between the ages of 12 and 74 yr old. Blood sera in 2-ml aliquots were analyzed, and the results were confirmed for 5994 persons. Almost all participants (99.5%) had p,p'-dichlorodiphenyldichloroethylene (p,p'-DDE) concentrations greater than or equal to 1 ppb, ranging as high as 378.6 ppb. For the other pesticide residues, only beta-benzene hexachloride (beta-BHC) (quantified in 17.2% of the sera), dieldrin (10.6%), and another DDT-related residue, p,p'-dichlorodiphenyltrichloroethane (p,p'-DDT) (35.7%) were found at quantifiable levels in more than 10% of the serum specimens. Of the remaining analytes, hexachlorobenzene (HCB), trans-nonachlor (TNC), and heptachlor epoxide (HE) were found at quantifiable concentrations in 1-10% of the specimens, whereas o,p'-DDT, o,p'-DDE, p,p'-DDD, mirex, alpha-BHC, gamma-BHC, heptachlor, delta-BHC, and aldrin were found in less than 1% of the serum specimens. Results showed that increasing age, residing on a farm, or being a male conferred increased risks of exposure to most of the pesticide residues, independent of all other demographic and seasonal factors. In a pattern less consistent across the different pesticide residues and for fewer of the pesticides, persons who lived below the national poverty level, were nonwhite, resided in the South or West, or were examined in the spring or winter also seemed to have an increased likelihood of having quantifiable serum levels.     

Dichlorodiphenyltrichloroethane suppresses neurite outgrowth and induces apoptosis in PC12 pheochromocytoma cells

It is well known that dichlorodiphenyltrichloroethane (DDT) is used as an insecticide and prevents many people in the tropical zone from devastating malaria. On the other hand, a number of reports have indicated that it may act as an endocrine disruptor and also has possible carcinogenic effects. However, the effects of DDT on the neural cells remain to be investigated. In this study, therefore, we observed the effects of p,p'-DDT, o,p'-DDT and its major metabolite p,p'-DDE on the differentiation and survival of PC12 pheochromocytoma cells. After stimulation with nerve growth factor, PC12 cells exhibited remarkable neurite outgrowth, suggesting that neuronal differentiation was induced by this growth factor. p,p'-DDT and o,p'-DDT suppressed this neurite outgrowth dose dependently, and p,p'-DDE also revealed a similar effect but to a lesser extent. Apoptotic cell death was induced within 3-6 h after treatment with p,p'-DDT and o,p'-DDT. Again p,p'-DDE showed a weaker apoptosis-inducing effect. In the organochlorine-treated PC12 cells phosphorylation of p44/42 mitogen-activated protein kinase (MAPK) was upregulated, whereas phosphorylation bands were not detected in any kinases of other MAPK groups such as p38 MAPK and SAPK/JNK. A kinase assay on p44/42 MAPK revealed that the extent of phosphorylation of Elk-1 substrates well correlated with the suppressive effect on neuronal differentiation and apoptosis-inducing activity. These results suggest that p,p'-DDT and o,p'-DDT exerted their effects on neuronal cells by the stimulation of p44/42 MAPK, and p,p'-DDE had less effects than the other two organochlorines.     

Distribution of organochlorines and PCB congeners in Korean human tissues

In order to investigate the residual amounts of organochlorines and polychlorinated biphenyls (PCBs) in Korean human tissues (blood, adipose tissue, liver, kidney cortex, and lung), the samples were collected from the autopsied cadavers of 40 men and 40 women (from teens to seventies of age). Alpha-BHC, beta-BHC, gamma-BHC, delta-BHC, p,p'-DDT, p,p'-DDD, p,p'-DDE, endrin, dieldein, aldrin, and 7 marker PCBs (28, 52, 101, 118, 138, 153, and 180) were determined in human tissues. The levels of organochlorines and PCB congeners indicated that they have been widely distributed in Korean human body. Positive correlations in terms of age were observed for the following cases: p,p'-DDE, p,p'-DDT, Sigma-DDT, PCB 118, PCB 138, PCB 153, and Sigma-PCB in the adipose tissue, and p,p'-DDE in the lung. Concentration of these compounds showed a significant age-related increase. Accumulation of these compounds in aged people revealed that these compounds were more slowly eliminated in our environment and risk assessment was necessary for further proper action. Significant differences in the levels of PCBs between genders were found for PCB 118 in the adipose tissue and PCB 138 in the liver. Positive correlation coefficients between tissues were detected with p,p'-DDE and p-BHC.     

DDT and its metabolite DDE alter steroid hormone secretion in human term placental explants by regulation of aromatase activity

Placental explants were used to compare the effects of two isomers of DDT (1,1,1,-trichloro-2,2-bis(p-chlorophenyl)ethane), p,p'-DDT and o,p'-DDT and their metabolites p,p'-DDE and o,p'-DDE (1,1,-dichloro-2,2-bis(p-chlorophenyl)ethylene) on steroid hormone secretion (estradiol (E2) and progesterone (P4)). Explants were treated with 1, 10, 100ng/ml or 1microg/ml of each compound for 24h. We found that all investigated compounds at all doses caused reductions of estradiol secretion. Moreover, it was shown that the inhibition of estradiol secretion was due to direct action on aromatase activity. Twenty-four-hour exposure to p,p'-DDE, o,p'-DDT or o,p'-DDE at doses of 100ng/ml or 1microg/ml increased P4 secretion, suggesting that these compounds act on P450scc. The fluorometric assay confirmed that all investigated compounds inhibited aromatase activity at a concentration of 100ng/ml. Our findings suggest that by decreasing estradiol secretion with concomitant stimulation of progesterone secretion, DDT could be a factor that influences the outcome of pregnancy.     

Organochlorine pesticides in human milk from different areas of Kenya 1983-1985

Residue levels of the chlorinated hydrocarbons p,p'-DDT (2,2-bis(p-chlorophenyl)-1,1,1-trichloroethane), p,p'-DDE (2,2-bis(p-chlorophenyl)-1,1-dichloroethane), hexachlorobenzene (HCB), alpha-, beta-, and gamma-hexachlorocyclohexane (HCH), aldrin, dieldrin, and polychlorinated biphenyls (PCBs) were determined in human milk of Kenyan mothers living in different areas of Kenya. The main organochlorine contaminants found in all the milk samples analyzed were p,p'-DDT and p,p'-DDE. Great regional differences were found, and mean levels of sum DDT and DDT/DDE ratio ranged from 1.1 to 18.7 mg/kg milk fat and from 0.7 to 5.7, respectively. In general, relatively low residue levels of HCB, alpha-HCH, beta-HCH, aldrin, and dieldrin were detected in 59, 37, 27, 37, and 19%, respectively, of all the milk samples analyzed. Quantifiable residue levels of PCBs and alpha-HCH were not found. The results were examined in relation to differences in living conditions with regard to agricultural activities, dietary habits, and reported use of pesticides in the various sampling areas.     

Factors influencing activities of biotransformation enzymes,concentrations and compositional patterns of organochlorine contaminants in members of a marine food web

The accumulation of polychlorinated biphenyls (PCBs; 34 congeners), sigmaDDT (p,p'-DDT, o,p'-DDT, p,p'-DDD, o,p'-DDD and p,p'-DDE), chlordanes (sigmaCHL; trans-chlordane, cis-chlordane, trans-nonachlor, cis-nonachlor and oxychlordane), hexachlorocyclohexanes (sigmaHCH; alpha-, beta- and gamma-isomers), hexachlorobenzene (HCB) and mirex was investigated in members of a marine food web from the Hvaler and Torbj?rnskaer archipelago, south-eastern Norway. The species studied were bullrout (Myoxocephalus scorpius), cod (Gadus morhua), herring gull (Larus argentatus) and harbour seal (Phoca vitulina). Furthermore, hepatic biotransformation enzyme activities (ethoxyresorufin-O-deethylase (EROD), pentoxyresorufin-O-depentylase (PROD) and glutathione S-transferase (GST)) were measured in all species. The objectives of the study were to investigate factors causing intraspecies variation in activities of biotransformation enzymes, as well as in concentrations and compositional patterns of the organochlorines (OCs). High correlations between EROD and PROD activities were found in all species, suggesting a single, common catalyst, CYP1A, and render the PROD assay questionable as biomarker for CYP2B inducers in marine wildlife. Furthermore, GST activities are shown to be dependent on biological factors, such as age (in harbour seal) and sex (in bullrout). In fish, the OC concentrations vary between the sexes, likely due to differences in fat deposition strategies and possibly sex dimorphism. In seals, concentrations and compositional patterns of the OCs vary with age, owing to selective transfer from mother to pup in utero and mainly through lactation, but likely also due to age specific xenobiotic metabolising capacity.     

Association of maternal blood pressure and hemoglobin level with organochlorines in human milk

Milk samples of women from the general population in Lucknow were analysed by gas-liquid chromatography to determine the concentration of organochlorine pesticides and their possible association with maternal characteristics. p,p'-DDT (dichloro diphenyltrichloroethane), p,p-DDE (dichloro diphenyltrichloroethylene), p,p'-DDD (dichloro diphenyl dichloroethane), alpha, beta gamma and delta isomers of hexachlorocyclohexane (HCH) and aldrin were detected. The calculated daily intake of DDT and HCH by the neonates exceeded their acceptable daily intake (ADI) set by the WHO. A statistically significant association was noted between alpha-HCH residues in milk and hemoglobin (Hb) level in maternal blood (P < 0.001). Also, there was a significant association between gamma-HCH and p,p'-DDT levels in milk and diastolic blood pressure of mother (P < 0.05). An apparent negative relationship between p,p'-DDE level in mother's milk and birthweight of the baby was also noticed (P < 0.05). These results highlight the need for periodic monitoring to determine organochlorine contamination in human milk and assess their impact on mother and on neonatal health.     

Effect of single and repeated in vitro exposure of ovarian follicles to o,p'-DDT and p,p'-DDT and their metabolites

The aim of the presented study was to compare the effect of o,p'-DDT [1,1-dichloro-2,2-bis-(p,p'-chlorophenyl)-ethylene] and p,p'-DDT [1,1,1-trichloro-2,2-bis-(p-chlorophenyl)-ethane] and their metabolites DDE and DDD on estradiol secretion by ovarian follicles, the target organs of environmental estrogens. Theca interna (Tc) and granulosa cells (Gc) were collected from medium size porcine follicles and cultured as a monolayer. The cells were initially cultured for 24 h to allow attachment to the plates and then media were changed for the new ones and o,p'-DDT and p,p'-DDT and their metabolites: o,p'-DDE, p,p'-DDE and o,p'-DDD were added at doses of 4, 40, 400 ng and 4 microg/ml medium to investigate dose-dependent effects. Media were collected after 24 h and frozen for estradiol content determination. When the effect of single and repeated exposure was investigated, the lowest dose of 4 ng/ml and the highest one of 4 microg/ml were chosen on the basis of the results of Experiment 1. o,p'-DDT exerted antiestrogenic action at all doses used while its metabolites and p,p'-DDT and its metabolites decreased estradiol secretion only when present in the medium at a dose of 4 ng/ml. The highest doses caused the increase in estradiol secretion. Parent o,p'-DDT and its metabolites showed antiestrogenic action after single exposure to 4 ng/ml while parent p,p'-DDT and its metabolites caused estrogenic action. All investigated compounds, except o,p'-DDT, increased estradiol secretion after single exposure to the dose of 4 microg/ml. Repeated exposure resulted in a massive antiestrogenic action of all investigated chemicals. In conclusion, our study points to time-dependent effect of DDT and its metabolites on ovarian follicles with the strongest estrogenic properties observed after single exposure and antiestrogenic action caused by repeated exposure. Given the duration of folliculogenesis, one can imagine many different potential mechanisms by which DDT could influence steroidogenesis.     

Bioaccumulation of DDT pesticide in cultured Asian seabass following dietary exposure

Bioaccumulation and metabolism of p,p'-DDT was studied in the marine carnivorous fish Lates calcarifer, Asian seabass, in a controlled aquaculture experiment. Over a 42-d period, seabass were fed pellets dosed with p,p'-DDT at environmentally realistic levels. Virtually all p,p'-DDT in pellets bioaccumulated in the fish with an uptake efficiency of 98%. The levels of p,p'-DDT and the metabolites p,p'-DDD and p,p'-DDE were analyzed in muscle, liver, visceral fat, brain, and remaining tissues. Partitioning of p,p'-DDT and its metabolites among the control, low-dose, and high-dose exposed seabass were 14.8% in muscle, 3.5% in liver, 37.1% in visceral fat, 0.11% in brain, and 45.5% in remaining tissues, where partitioning between tissues was a function of tissue lipid content. p,p'-DDT bioaccumulation increased linearly with exposure in visceral fat and muscle tissue. The metabolism of p,p'-DDT, which occurs mainly in the liver, resulted in the degradation of 2.5% of p,p'-DDT into p,p'-DDD. These new findings show that bioaccumulation processes at environmentally realistic ingestion exposure levels (ng/g) differ from previous DDT ingestion studies conducted at unrealistically high DDT levels (microg/g), highlighting the need to revise models on the transfer of persistent organic pollutants in the marine environment and aquaculture systems.     

DDT inhibits the functional activation of murine macrophages and decreases resistance to infection by Mycobacterium microti

DDT is still widely used in several parts of the world to control malaria, typhoid and dengue vectors, even though its use was banned in many countries based on toxicity data in wild life species. DDT has been shown to have immunotoxic effects in mice and to increase susceptibility to intracellular pathogens such as Mycobacterium leprae. However, little is known about the mechanisms underlying this effect. Activated macrophages play an important defensive role against intracellular pathogens, therefore our objective was to evaluate the effect of in vitro exposure to technical grade DDT (a mixture of three forms: 1,1,1-thricloro-2,2-bis(p-chlorophenyl)ethane (p,p'-DDT) (85%), o,p'-DDT (15%) and o,o'-DDT (trace amounts)), p,p'-DDT, 1,1-dicloro-2,2-bis(p-chlorophenyl)ethylene (p,p'-DDE) and 1,1-dichloro-2,2-bis(p-chlorophenyl)ethane on the functional activation of J774A.1 macrophages and their capability to limit growth of intracellular pathogens, using Mycobacterium microti as a model. We evaluated cytotoxicity and the effect on cell proliferation of 2.5, 5.0 and 10 microg/ml of DDT compounds. Functional macrophage activity (NO(*) and O(2)(-) production, and mRNA expression of TNF-alpha, IL-1beta and iNO synthase) and the ability of treated cells to limit infection by M. microti in IFN-gamma-activated macrophages were evaluated in cells exposed to 2.5 microg/ml of DDT compounds. Doses of 5 and 10 microg/ml induced direct cytotoxic effects precluding meaningful analysis of the above parameters, whereas 2.5 microg/ml of all DDT compounds inhibited macrophage activity and reduced their ability to limit the intracellular growth of M. microti without inducing cytotoxicity. Technical grade DDT and p,p'-DDE were the more potent compounds. Therefore, exposure to DDT compounds could represent an important risk for infection development by those intracellular pathogens against which NO(*) and/or O(2)(-) production represent the main immune protective mechanism.