Effects of dioxins and polychlorinated biphenyls (PCBs) on thyroid function in infants born in Japan – the second report from research on environmental health

Dioxins (PCDD + PCDF) and polychlorinated biphenyl (PCB) are potentially hazardous compounds and have structural similarity to thyroid hormones. Our research group on “Dioxins and PCB in Human Milk” was organized in 1997 and has been active for the past three years. We collected breast milk from 80 mothers living in Tokyo, Saitama, Ishikawa and Osaka Prefecture in Japan at 5, 30, 150, 300 postpartum days and measured PCDDs, PCDFs and PCBs during 1998–1999. We added another 20 prefectures and cities during 1999–2000. Breast milk was obtained from 415 mothers at 30 postpartum days (breast-feeding group). Blood was taken from infants who were breast-fed at the age of 1 year for evaluation of thyroid and immune functions. Blood was also taken from 53 infants who were bottle-fed (bottle-feeding group) as a control. Serum T4, T3, FT4 and TSH levels in the breast-feeding (n=337) and bottle-feeding (n=53) groups were within normal ranges and were not significantly different between the two groups. Although there were a large geographic differences of dioxins and co-PCB content in breast milk, there were no differences in serum T4, T3, FT4, TSH levels and no significant correlation between the mean serum levels of TSH and TEQ in breast milk. There was a significant correlation between serum TSH at 1 year of age and that from blood on dried filter paper TSH at 5 days of age. We concluded that dioxin intake from breast milk in Japanese infants did not impair thyroid function. However, long-term effects remain to be evaluated.     

Infant exposure to polychlorinated dibenzo-p-dioxins, dibenzofurans and biphenyls (PCDD/Fs, PCBs)--correlation between prenatal and postnatal exposure

Polychlorinated dibenzo-p-dioxins, dibenzofurans (PCDD/Fs) and polychlorinated biphenyls (PCBs) are recognized environmental endocrine disruptors, which are environmentally persistent and may bio-accumulate in human bodies. Pregnant and nursing women may pass these pollutants to their babies both trans-placentally and lactationally. We measured and examined correlations of the levels of PCDD/Fs and PCBs in perinatal venous serum, placenta, umbilical-cord serum (representing prenatal exposure), and breast milk (postnatal exposure). Subjects included pregnant women without clinical complication between the ages of 25 and 35, who delivered their babies during 2000.12.01 and 2001.11.30 in central Taiwan. A total of 20 participants were randomly selected from those provided the four biological specimens for analysis of 17 PCDD/Fs, and 12 dioxin-like PCBs and six indicator PCBs using high-resolution gas chromatography/high-resolution mass spectrometry. Higher PCDD/F levels were found in placenta (10.3 TEq-pg/g lipid) and venous serum (9.1 TEq-pg/g lipid) compared to those in breast milk (7.6 TEq-pg/g lipid). Pearson and Spearman correlation analyses showed well association of PCDD/F and PCB levels between different specimens. The total dioxin/PCB level in milk, venous and cord serum can be well predicted by that in placenta through regression functions. This first study of multi-specimen correlation further established rather consistent ratios of these chemical concentrations in various specimens relative to those in venous serum.     

The effect of copper sulphate on some biochemical parameters of rainbow trout

Copper sulphate has an adverse effect on some biochemical parameters of trout Salmo gairdneri. The main targets of damage are: (a)Tissue necrosis as demonstrated by increased ASAT and ALAT levels in blood plasma. (b)Inhibition of acetylcholinesterase activity. There is also a general stress effect as reflected by an elevated blood glucose level.     

Perinatal dioxin exposure and later effects--a review

Negative effects of perinatal exposure to background levels of dioxins and PCBs in Europe and the USA have been documented. Four facets of development are reviewed in this paper: 1. Brain development and thyroid hormone metabolism. 2. Hepatic effects. 3. Hematopoietic system effects. 4. Lung function. Effects on IQ and behaviour have been documented in children on both sides of the Atlantic Ocean. Non-dioxin-like PCBs, measured in maternal and cord blood and current plasma samples have been implicated. Interference with thyroid hormone metabolism in the mother, in the foetus and in the newborn baby could be responsible for these effects on brain development. During early gestation the foetus is completely dependent on maternal thyroxine (T4). Lower T4 levels in the mother, caused by dioxins and PCBs, might negatively influence (early) brain development. It is plausible that the intrauterine dependency on maternal T4 and the high T4 need shortly after birth makes both these periods vulnerable for environmental influences. Effects of dioxin exposure on thyroid hormone metabolism have been described in the period shortly after birth. These effects are no longer found after two years of age indicating a transient effect. In animal studies, in utero exposure has led to effects on brain development due to abnormal induction of liver enzymes. This induction resulted in lower testosterone and estrogen levels, interfering with brain development in the vulnerable period of language development and the development of visuo-spatial abilities. In humans this developmental period occurs around the thirtieth week of pregnancy. Follow-up studies in puberty and adolescence of the different cohorts studied is necessary to evaluate these negative influences. Damaging effects on the liver found shortly after birth have proven to be transient. Effects on the haematopoietic system are clear immediately after birth, for instance on white blood cells and thrombocytes. An increase in middle ear infections (otitis media) in relation to current levels of PCBs at the age of 4 years was described in the Rotterdam study. Negative effects on lung function in the sense of increased obstruction was found after 8 years in relation to perinatal exposure to dioxins in the Zaandam study. This rather new finding might explain the sharp increase in lung problems in children in the Western world.     

Cytochrome p450 induction and gonadal status alteration in common carp (Cyprinus carpio) associated with the discharge of dioxin contaminated effluent to the Hikiji River,Kanagawa Prefecture,Japan

Accumulations of polychlorinated dibenzo-p-dioxins, dibenzofurans and coplanar polychlorinated biphenyls were analyzed in common carp (Cyprinus carpio) collected in the Hikiji River, Kanagawa Prefecture, Japan in which dioxin contaminated effluent was released during the period starting from November 1992 to March 2000. Higher levels of 2,3,7,8-tetrachlorinated dibenzo-p-dioxin toxic equivalents were observed in carps collected downstream to the dioxin release site (contaminated site) than the reference site. Modulations of cytochrome p450 (CYP) enzyme in liver, serum estrogen concentration and gonadal somatic index (GSI) were also measured as biomarkers for the contaminants. Total CYP content in livers was markedly higher in male and female carps from the contaminated site relative to the reference site fish. The expression level of the cytochrome p450 1A and Ethoxyresorufin O-deethylase activity were significantly higher in female carps from the contaminated site than from the reference site. A lower level of plasma estrogen was observed in carps from the contaminated site. The GSI in female carps from the contaminated site was smaller than that recorded at the reference site. The present study indicates that dioxins released to the Hikiji River might induce the CYP enzyme and inhibit the reproductive functions in common carps dwelling downstream from the release site.     

Assessment of current serum levels of PCDD/Fs, dl-PCBs and PBDEs in a Dutch cohort with known perinatal PCDD/F exposure

As part of a longitudinal cohort study, now in its second decade, we determined PCDDs/Fs dl-PCBs and PBDEs in serum of adolescents with known perinatal PCDD/F exposure. Of the original cohort, 33 adolescents aged 14-19 years, who had been studied previously during their neonatal (n=60), toddler and pre-pubertal period (n=41) agreed to participate in the current follow-up. PCDD/F-, dl-PCB- and PBDE congeners were measured using GC/MS. Current serum levels of PCDD/Fs determined in our cohort were relatively low (mean of 2.2pg/g) compared to the perinatal exposure. No correlation between perinatal exposure and current serum PCDD/F was found. Planar PCB TEQ levels were 2.2pg/g. Current summation operatorPBDE levels were 8.7ng/g lipid. There was one outlier with a summation operatorPBDE of 74ng/g lipid. The presence of this high value indicates that the exposure pathway is different from PCDD/F and PCB, most likely by dust and food contaminated with dust. Concluding we can say that current PCDD/F levels are quite low compared to the perinatal PCDD/F exposure of the cohort. PBDE levels however are relatively high compared to other European countries, more research on possible health effects of these levels, especially for subjects with outlier concentrations, should be performed.     

Effects of dietary habits and CYP1A1 polymorphisms on blood dioxin concentrations in Japanese men

The major source of dioxin impurities in Japan in the past was agrochemical formulations; more recently, it has been exhaust from waste incinerators. To examine the environmental and genetic factors that influence blood dioxin concentration, we investigated the relationship among dioxin concentrations, dietary habits and cytochorome P450 1A1 (CYP1A1) polymorphisms (MspI type and Ile-Val type) in Japanese fishermen and farmers, including also a group of office workers as controls. The mean dioxin concentrations in the fishermen, the farmers and the controls were 161369, 79079 and 100500 pg/g fat, respectively. The elevated dioxin concentration with polychlorinated dibenzo-p-dioxins, polychlorinated dibenzofurans and coplanar-PCBs found in the fishermen may be due to the frequent consumption of fish; no such relationship was found both in the farmers and the controls. We found that the concentrations of congeners found as impurities in certain chemicals such as those previously used in agriculture showed no significant differences among the three groups; we consider it unlikely that the farmers would be directly exposed to dioxins from such chemicals. Thus, it is probable that the primary route of dioxin exposure in the Japanese population is through the food chain via fish consumption, regardless of occupation. No meaningful relationship between blood dioxin concentration and CYP1A1 polymorphisms was found in this study, although there was a significant difference between the concentration of total non-ortho-PCBs in genotypes A and B. Further studies on more subjects, including those of genotype C, are needed to confirm the relationship between blood dioxin concentrations and MspI polymorphisms.     

Increased thyroid volume and frequency of thyroid disorders signs in schoolchildren from nitrate polluted area

Thyroid volume (ThV) and echogenicity by ultrasound were estimated in 324 schoolchildren (aged between 10 and 13-years) from high nitrate area (HNA) located in agricultural lowland with high nitrate drinking water supply (51-274 mg/l) from shallow wells. The data were compared to children of the same age from low nitrate area (LNA) consisting of 168 children from the neighboring area with very low nitrate (< 2 mg/l) drinking water and of 596 children from the city of Kosice located in a vicinity of LNA and also supplied by low nitrate water. Blood samples were obtained from 315 willing children from HNA and 109 children from LNA and the levels of thyrotropin (TSH), total thyroxine (TT4), free triiodothyronine (FT3) and thyroperoxidase antibodies (anti-TPO) in serum were determined. ThV (mean +/- SE) in 10-year (5.10 +/- 0.14 ml) and 13-year (5.97 +/- 0.11 ml) old children from HNA was significantly higher than that in two groups of respective age from LNA, 4.58 +/- 0.17 (p < 0.02) and 5.23 +/- 0.15 ml (p < 0.05), and from the city of Kosice, 4.77 +/- 0.10 ml (p < 0.05) and 4.87 +/- 0.1 0ml (p < 0.0001). The frequency of hypoechogenicity in HNA was also significantly higher than that in pooled LNA plus Kosice, 13.7% vs. 4.7% (p < 0.01) in 10-year and 10.6% vs. 5.7% (p < 0.03) in 13-year, respectively. The frequency of TSH level in the range of subclinical hypothyroidism (> 4.0 mU/l) in pooled age groups from HNA was 13/324 (4.0%) and that of positive anti-TPO was 8/324 (2.5%), while no case of either increased TSH or positive anti-TPO was found in 109 children from LNA. Finally, no differences in the levels of TT4 and FT3 were found between HNA and LNA. It was concluded that long-term exposure to high nitrate intake by drinking water and home made meals from local products results in increased thyroid volume and increased frequency of signs of subclinical thyroid disorders (thyroid hypoechogenicity by ultrasound, increased TSH level and positive anti-TPO).     

The constitutively active Ah receptor (CA-AhR) mouse as a model for dioxin exposure – Effects in reproductive organs

The dioxin/aryl hydrocarbon receptor (AhR) mediates most toxic effects of dioxins. In utero/lactational exposure to 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) impairs fetal/neonatal development and the developing male reproductive tract are among the most sensitive tissues. TCDD causes antiestrogenic responses in rodent mammary gland and uterus and in human breast cancer cell lines in the presence of estrogen. Also, more recently an estrogen-like effect of TCDD/AhR has been suggested in the absence of estrogen. A transgenic mouse expressing a constitutively active AhR (CA-AhR) was developed as a model mimicking a situation of constant exposure to AhR agonists. Male and female reproductive tissues of CA-AhR mice were characterized for some of the effects commonly seen after dioxin exposure. Sexually mature CA-AhR female mice showed decreased uterus weight, while an uterotrophic assay in immature CA-AhR mice resulted in increased uterus weight. In immature mice, both TCDD-exposure and CA-AhR increased the expression of the estrogen receptor target gene Cathepsin D. When co-treated with 17β-estradiol no increase in Cathepsin D levels occurred in either TCDD-exposed or CA-AhR mice. In sexually mature male CA-AhR mice the weights of testis and ventral prostate were decreased and the epididymal sperm reserve was reduced. The results of the present study are in accordance with previous studies on dioxin-exposed rodents in that an activated AhR (here CA-AhR) leads to antiestrogenic effects in the presence of estrogen, but to estrogenic effects in the absence of estrogen. These results suggest the CA-AhR mouse model as a useful tool for studies of continuous low activity of the AhR from early development, resembling the human exposure situation.     

Levels of PCDDs, PCDFs and Co-PCBs in human milk in Saitama, Japan, and epidemiological research

From 1998 to 2000, the dioxin levels were measured in the milk of 299 mothers who lived in Saitama prefecture, Japan. Factors that influenced the dioxin levels were investigated based on a questionnaire given to the milk donors at that time. It was found that the dioxin levels in the milk of the mothers who smoked were lower than those of non-smokers, and the dioxin levels were generally low in the mothers who were heavy smokers. The average dioxin levels in the milk of mothers who had been breast-fed were higher than those given formula, and there was a significant difference in the dioxin congeners. It was also found that the dioxin levels in milk of the women who regularly consumed fish and shellfish were generally higher. There was a strong correlation between summation operator TEQ and PCB126. The data suggested that PCB126 could be a useful indicator for the simplified analysis of dioxin in human milk.     

Maternal-fetal distribution and transfer of dioxins in pregnant women in Japan, and attempts to reduce maternal transfer with Chlorella (Chlorella pyrenoidosa) supplements

Dioxins can be transferred from mother to fetus via the placenta, or to nursing infants via breast milk, potentially causing developmental health problems in children. To assess pediatric health risks from dioxins, exposure of mothers and children to dioxins must be clarified. Methods of reducing maternal transfer of dioxins should also be investigated. Concentrations of 28 dioxin (polychlorinated dibenzo-p-dioxins, polychlorinated dibenzofurans, and co-planar polychlorinated biphenyls) congeners in blood, adipose tissue, breast milk, cord blood and placenta collected from 44 pregnant Japanese women were measured. In addition, to investigate potential reductions in maternal transfer of dioxins, 23 pregnant women were instructed to take Chlorella pyrenoidosa supplements during pregnancy. Correlations were observed between dioxin total toxic equivalents (total TEQ) in blood and total TEQ in adipose tissue (r=0.913, P<0.0001), breast milk (r=0.695, P=0.0007), and cord blood (r=0.759, P<0.0001). Dioxin levels transferred to fetuses and nursing infants reflect cumulative maternal concentrations of dioxins. A linear regression equation was introduced to predict total TEQ in breast milk and cord blood from dioxin levels in maternal blood, which should prove useful in evaluating fetal and infant risk of dioxin exposure. Total TEQ in cord blood were approximately 26% lower than in maternal blood (P<0.0001). The results of this study suggest that transplacental transfer differs depending on the dioxin congener. Total TEQ in breast milk were approximately 30% lower in the Chlorella group than in controls (P=0.0113). This finding suggests that maternal transfer of dioxins can be reduced using dietary measures such as Chlorella supplements.     

The constitutively active Ah receptor (CA-AhR) mouse as a model for dioxin exposure - effects in reproductive organs

The dioxin/aryl hydrocarbon receptor (AhR) mediates most toxic effects of dioxins. In utero/lactational exposure to 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) impairs fetal/neonatal development and the developing male reproductive tract are among the most sensitive tissues. TCDD causes antiestrogenic responses in rodent mammary gland and uterus and in human breast cancer cell lines in the presence of estrogen. Also, more recently an estrogen-like effect of TCDD/AhR has been suggested in the absence of estrogen. A transgenic mouse expressing a constitutively active AhR (CA-AhR) was developed as a model mimicking a situation of constant exposure to AhR agonists. Male and female reproductive tissues of CA-AhR mice were characterized for some of the effects commonly seen after dioxin exposure. Sexually mature CA-AhR female mice showed decreased uterus weight, while an uterotrophic assay in immature CA-AhR mice resulted in increased uterus weight. In immature mice, both TCDD-exposure and CA-AhR increased the expression of the estrogen receptor target gene Cathepsin D. When co-treated with 17β-estradiol no increase in Cathepsin D levels occurred in either TCDD-exposed or CA-AhR mice. In sexually mature male CA-AhR mice the weights of testis and ventral prostate were decreased and the epididymal sperm reserve was reduced. The results of the present study are in accordance with previous studies on dioxin-exposed rodents in that an activated AhR (here CA-AhR) leads to antiestrogenic effects in the presence of estrogen, but to estrogenic effects in the absence of estrogen. These results suggest the CA-AhR mouse model as a useful tool for studies of continuous low activity of the AhR from early development, resembling the human exposure situation.     

Characterization of dioxin exposure in firefighters, residents, and chemical workers in the Irkutsk Region of Russian Siberia

The goal of this study was to characterize body burdens of polychlorinated dioxins (PCDDs), polychlorinated dibenzofurans (PCDFs), and polychlorinated biphenyls (PCBs) in three groups of Siberians living in the Irkutsk Region of Russia. These groups included firefighters exposed to a mixture of toxic substances extinguishing a large fire at the Shelekhovo Cable Factory in 1992, chemical workers from the Khimprom chemical plant, and residents living in proximity to large chemical factories in Sayansk and Angarsk. Blood samples from all groups were obtained in the fall of 1998. Dioxin analyses of samples were performed in Germany, Canada, and in a Russian dioxin laboratory in Ufa, Bashkortostan Republic. The average levels of dioxin toxic equivalents (TEQs) are 23.6 parts per trillion (ppt) total TEQ (PCDD/F only) in the disabled firefighters, 25.0 in the non-disabled firefighters, 28.7 in residents, and 45.6 in the Khimprom workers blood. Two workers did have elevated total TEQs of 91.4 and 102.2 ppt. Dibenzofurans and coplanar PCBs substantially contribute to the total elevated TEQ seen here. The average TEQs suggest levels of dioxin exposure in this part of the former Soviet Union not dissimilar to levels measured in industrialized countries of Europe and Northern America.     

Exposure of arc-furnace-plant workers to polychlorinated dibenzo-p-dioxins and dibenzofurans (PCDD/Fs)

This study aimed to evaluate serum polychlorinated dibenzo-p-dioxins and dibenzofurans (PCDD/Fs) levels in electric-arc-furnace workers according to their corresponding occupational exposure. In addition, the ambient PCDD/Fs of the work environment were measured to provide additional support for the exposure and accumulation inside the electric arc furnace. The ambient PCDD/F concentrations inside the electric arc furnace were 1.557-1.917 pg-TEQ/Nm(3), 5-24 folds higher than those outside (0.080-0.385 pg-TEQ/N m(3)). In addition, higher average serum levels were measured in the workers with high occupational dioxin exposure (24.0 pg WHO-TEQ/g lipid) than in those with lower occupational dioxin exposure (13.8 pg WHO-TEQ/g lipid). Higher PCDFs/PCDDs ratios were found in serum samples from high-exposure groups than in low-exposure groups. The higher ratio of PCDFs/PCDDs was also found in ambient samples consistent with other metallurgical processes reported previously. Our results suggest that PCDD/Fs exposure exits in the smelting process of electric arc furnace, and the occupational hygiene should be taken more seriously concern in that workplace.     

Possible effects of persistent organochlorinated pollutants cocktail on thyroid hormone levels and pituitary-thyroid interrelations

In polluted district of Michalovce in East Slovakia (POLL) and two districts with background pollution (BCGR) 2046 adults (834 males and 1212 females aged 20-75 years) were examined. Serum levels of thyrotropin (TSH), free thyroxine (FT4), total triiodothyronine (TT3) and antithyroperoxidase antibodies (TPOab) were estimated by electrochemiluminiscent assay and also these of 15 polychlorinated biphenyl congeners (PCBs), p,p'-DDE, p,p'-DDT, hexachlorobenzene (HCB) and hexachlorocyclohexane were measured by high resolution gas chromatography/mass spectrometry. In addition, also dioxins, furans, coplanar- and mono-ortho-PCBs as well as selected hydroxylated and methylsulphonated PCBs and DDE metabolites were measured by appropriate methods based on gas chromatography/mass spectrometry principle. In POLL significantly higher levels of all organochlorines were found than these in BCGR. When pooled values from both areas were stratified in terms of PCBs level and treated as continuous variables, positive association of PCBs with FT4 and TT3 was found, the latter two being also mutually associated. However, within the category of PCBs level <530 ng/glipid (n=232) the association between PCBs and both the FT4 (p<0.09) and TT3 (p<0.03) was negative and any association of these was not found within the category of PCBs level of 531-1000 ng/g (n=691). In contrast, in the category of 531-2000 ng/g (n=1307) positive association appeared between PCBs and FT4 (p<0.001) as well as TT3 (p<0.05). Highly significant association of PCBs with FT4 (p<0.001) was further found in the categories with PCBs level of 1001-101414 ng/g (n=1307) and 2001-101414 (n=1123), while significant association with TT3 was observed only in the category of 531-2000 ng/g. Such findings suggest possible threshold level in positive effect of PCBs on FT4 and TT3 level which seems to be individual and located somewhere around the PCBs level of 1000 ng/g. However, highly significant negative association of both FT4 and TT3 with TSH was found in each of above indicated PCBs categories. Considerable difference in FT4 and TT3 level between large groups of subjects with the same range of PCBs level was also found suggesting different individual susceptibility to the effects of organochlorines. Among a total of 26 cases from POLL with very low TSH level (<0.5 mU l(-1)) 13 cases showed very high level of PCBs, FT4 and TT3, thus supporting a hypothesis on a novel sporadic form of high PCBs related peripheral subclinical hyperthyroidism possibly resulting from the long-term disruption of equilibrium between bound and free thyroxine in plasma by high PCBs level followed by a striking inhibition of TSH release from the pituitary.     

Dioxins and related compounds: Status and regulatory aspects

Several countries have put much effort in dioxin research programmes with the aim to assess the risks of dioxin exposure, to identify dioxin sources and to quantify their contribution to environmental emissions and human exposure. In this review, the accent has been put on: sources of dioxins and their contribution to the total dioxin emissions, sources of human exposure and their contribution to exposure and measures to reduce emission and exposure. The information from countries with existing dioxin regulations is reviewed and compared. Uncertainties which hamper comparability of data are indicated. Results of measures and expectations for the near future are presented.     

Concentrations of organochlorine pollutants in mothers who gave birth to neonates with congenital hypothyroidism

We have investigated the effects of prenatal exposure to dioxin-like compounds (PCDDs, PCDFs and dioxin-like PCBs), PCBs and organochlorine pesticides (DDT, HCH, chlordane, HCB and their metabolites) on the incidence of congenital hypothyroidism and/or cretinism in Fukuoka, Japan from 2001 to 2004. Thirty-four positive neonates of the mass-screening for cretinism were classified into three groups by the pediatrician after the reevaluation of the serum TSH level, that is, negative in reevaluation group, hyper thyroid stimulating hormone (TSH) emia group and cretinism group. One-hundred and two negative neonates of the mass-screening were classified into the normal group. Concentrations of these organochlorine compounds in the breast milk of mothers, which were considered as the indicator of prenatal exposures to them, were gradually increased from the normal group to the cretinism group in the four groups and were around two times higher in the cretinism group than in the normal group. According to the case-control study adjusted for the parity and mother's age, odds ratios of PCBs, DDT and HCB were 10 (p=0.003), 10 (p=0.003) and 22 (p=0.004), respectively and in dioxin-like compounds, PCDFs showed the highest odds ratio, 9.8 (p=0.005). Based upon those findings, these compounds seemed play an important role in the incidence and/or causation of the cretinism.     

Status of centers for disease control dioxin studies of U.S. population

Although the toxic effects of dioxin have been extensively studied in animals and documented in cases of accidental high-level exposure in humans, little is known of the health effects to humans produced by long-term exposure to relatively low levels of dioxin. The Centers for Disease Control (CDC) has been involved in several studies designed to assess these health risks