Polychlorinated biphenyls in umbilical cord serum of newborns from Rio Grande do Sul state, Brazil

Clinica Chimica Acta 451 (2015) 323–328

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Polychlorinated biphenyls in umbilical cord serum of newborns from Rio Grande do Sul state, Brazil Susana Mohr a,1, Joice Sifuentes dos Santos b, Thiago Guilherme Schwanz a, Roger Wagner a, Joseane Oliveira Mozzaquatro a, Alessandra Scherer Lorenzoni c, Ijoni Hilda Costabeber d,⁎ a

Departamento de Tecnologia e Ciência dos Alimentos, Centro de Ciências Rurais, Universidade Federal de Santa Maria, CEP 97105-900 Santa Maria, RS, Brazil Mestrado em Ciência e Tecnologia de Leite e Derivados, Universidade Norte do Paraná, Rua Marselha, 591, CEP 86041-140 Londrina, PR, Brazil Mestrado em Ciências Farmacêuticas, Centro de Ciências da Saúde, Universidade Federal de Santa Maria, CEP 97105-900 Santa Maria, RS, Brazil d Departamento de Morfologia, Centro de Ciências da Saúde, Universidade Federal de Santa Maria, CEP 97105-900 Santa Maria, RS, Brazil b c

a r t i c l e

i n f o

Article history: Received 27 May 2015 Received in revised form 16 October 2015 Accepted 19 October 2015 Available online 22 October 2015 Keywords: PCBs POPs Prenatal exposure Human biomonitoring Food habits

a b s t r a c t Background: Polychlorinated biphenyls (PCBs) are food-chain contaminants that have been shown to contaminate foods worldwide. The newborn are exposed to these organochlorine compounds across the placenta and through breastfeeding. They are proven to be carcinogenic and may contribute to congenital malformation etiology. Methods: This study examined levels of five PCB congeners (28, 52, 138, 153 and 180) in umbilical cord serum samples from 148 newborns from Rio Grande do Sul state, Brazil. Serum concentrations of PCBs were analyzed by gas chromatography with electron capture detection and mass spectrometry. Results: Levels of ∑PCBs ranged from 0.35 to 55.17 ng/ml in umbilical cord serum positive samples, and PCB 138 was the most prevalent congener. Only 7.4% of samples presented no PCB congener. Conclusions: Some PCB congener cord serum levels were related to the locale of the mothers' residence, smoking and drinking habits, fruit consumption, and congenital malformation. © 2015 Elsevier B.V. All rights reserved.

1. Introduction Polychlorinated biphenyls (PCBs) were discovered before the turn of the 20th century and their usefulness for industry, because of their physical properties, was recognized early. They have become widely distributed in the environment throughout the world, and are persistent and accumulate in the food chain. PCBs bioaccumulate in the fatty tissues of humans and other animals and have caused toxic effects in both, particularly if repeated exposure occurs. The results of the studies on rodents suggest that some PCB congeners may be carcinogenic and that they can promote the carcinogenicity of other chemicals [1]. Previous studies suggested that human exposure to PCBs is associated with the occurrence of various types of cancer, such as breast cancer [2–4], colon cancer [5], liver, stomach, intestinal and prostate cancers[6,7] and a recent study showed that poisoning due to PCBs and dioxins can affect quality of life 30 y after the exposure [8]. PCBs were also associated with the occurrence of miscarriages and premature births [9], with damage to the ⁎ Corresponding author. E-mail addresses: [email protected] (S. Mohr), [email protected] (J. Sifuentes dos Santos), [email protected] (T.G. Schwanz), [email protected] (R. Wagner), [email protected] (J.O. Mozzaquatro), [email protected] (A.S. Lorenzoni), [email protected] (I.H. Costabeber). 1 Permanent address: Departamento de Defesa Agropecuária, Secretaria Estadual da Agricultura e Pecuária, Supervisão Regional de Ijuí, CEP 98700-000, Ijuí – RS, Brazil.

http://dx.doi.org/10.1016/j.cca.2015.10.019 0009-8981/© 2015 Elsevier B.V. All rights reserved.

peripheral nervous system [10], with respiratory infections in children from birth to 5 y [11], with reduced immunity in children [12], with diabetes [13] and with asymmetric hearing loss [14]. PCB concentration in umbilical cord plasma is a good indicator of prenatal exposure to PCBs [15]. Their levels detected in maternal and umbilical cord serum showed significant relationships, confirming their efficient transplacental transfer [16–18]. Entering the fetal blood system in this way, PCBs are a health risk to fetuses and newborns, which are much more vulnerable to environmental pollutants. The exposure to persistent organic pollutants in utero can have adverse effects on fetal growth (i.e. birth weight, length, head circumference) and health [19]. The largest number of studies on this subject is found in countries that produced PCBs, such as Italy, Germany, Japan, and the United States. Generally speaking, congeners 138, 153 and 180 are those that process the greatest concentrations of detectable residues [20– 23]. These three congeners, along with congeners 28, 52, 101 and 118 are denominated “the seven indicators” and are responsible for the greatest part of environmental contamination. This paper reports the residue concentrations of PCBs (28, 52, 138, 153, and 180) in umbilical cord serum of newborns from Rio Grande do Sul State, Brazil. The comparison among the PCB concentrations, sociodemographic characteristics and food habits of the mothers and the characteristics from the newborns, were also reported.

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2. Material and methods 2.1. Population and samples Arterial umbilical cord blood samples were collected from the babies of 148 mothers admitted to the Obstetrics Center of the University Hospital of Santa Maria, Rio Grande do Sul, Brazil, during 2006. Umbilical cord blood samples were taken immediately after the delivery, collected from the section of the umbilical cord next to the placenta, after clamping and cutting. To determine the PCB congeners, serum obtained from blood centrifugation was used. After centrifugation, the samples were stored at − 20 °C prior to analysis. The approximate volume of serum collected was 0.5 ml. Due to relatively low sample volumes, lipid analysis was not performed for this set of samples. All participants were asked to complete a questionnaire which detailed their weight, age, height, sociodemographic information as well as food habits. Details of the characteristics of the newborn infants were also documented. Not all participants responded to the questionnaires. The applied protocol was previously approved by the Research Ethical Committee of Federal University of Santa Maria through their Letter of Approval. 2.2. Standards, reagents and materials

70 eV. The temperature program was 60 °C (2 min), 5 °C/min to 220 °C (5 min), and 20 °C/min to 300 °C (2 min). The ions monitored were m/z 186, 256 and 258 (PCB 28); m/z 220, 222, 290 and 292 (PCB 52); m/z 290, 358, 360 and 362 (PCB 153); m/z 290, 235, 360 and 362 (PCB 138); m/z 324, 394, 396 and 398 (PCB 180). GC–MS analysis confirmed the identity of PCBs (Supplementary Material). 2.5. Quality control To evaluate the accuracy and reliability of the PCB analysis, the validation method procedure was previously performed [25]. Trueness was evaluated by spiked umbilical cord serum samples, with levels ranging from 5.0 to 20 ng/ml (n = 3) and related to recovery. Mean recoveries for the 5 PCBs ranged from 73 to 119% and the coefficient of variation was below 12.5%, indicating an adequate repeatability for the method. The limit of detection (LOD) was 0.1 ng/ml for all PCBs. The limit of quantification (LOQ) was 0.25 ng/ml for PCB 28 and 0.5 ng/ml for PCBs 52, 138, 153 and 180. The calibration curve (0.25 to 12 ng/ml of each analyte) was prepared in umbilical cord serum samples, linear and characterized by good correlation coefficients (N0.99) for all compounds studied. Intra- and inter-assay variations were calculated and the values were b18%. Blanks were made for each sample lot extracted, with the objective of eliminating possible interference.

A stock solution of PCBs containing 10 μg/ml of congeners 28 (2,4,4′-trichlorobiphenyl), 52 (2,2′,5,5′-tetrachlorobiphenyl), 138 (2,2′,3,4,4′,5′-hexachlorobiphenyl), 153 (2,2′,4,4′,5,5′hexachlorobiphenyl) and 180 (2,2′,3,4,4′,5,5′-heptachlorobiphenyl) was used for the identification method and the validation process. The standard solutions were obtained from SUPELCO Inc. n-Hexane for pesticide residue analysis was obtained from Mallinckrodt Baker, analytical grade sulfuric acid from Vetec Química Fina Ltda, and nitrogen 5.0 analytical grade from White Martins. Glassware used in the analyses and to store samples was previously washed following the method by Angulo et al. [24], with distilled water, rinsed with hexane and acetone alternately and dried at 150 °C, to assure chemical cleanliness.

Statistical analyses were performed using the Statistica® 7.0 software package. PCBs measured in the umbilical cord serum samples that did not fit normal distribution, were analyzed using nonparametric tests. The Mann–Whitney test was used to compare the means of PCBs between groups of variables. A value equal to half of the LOD was given to each PCB not detectable in a sample. Positive samples were calculated for samples with levels above the LOD. ∑ PCB was calculated as the sum of PCB congeners. All statistical significance was set at p b 0.05 and p b 0.10.

2.3. Serum extraction and clean-up

3. Results and discussion

Details of the procedure have been described previously [25]. Briefly, arterial umbilical cord serum samples were placed in centrifuge tubes. Then, n-hexane and sulfuric acid were added and the mixture was stirred in a vortex mixer. The supernatant n-hexane layer was separated by centrifugation and transferred to a second centrifuge tube using a Pasteur pipette. This n-hexane extraction step of sulfuric acid digested sample was repeated twice. The combination of all n-hexane extracts provided a total volume of 7 ml. Additional sulfuric acid was added to this volume and the mixture was vortex stirred and centrifuged as before. The supernatant was transferred to a conical bottomed, graduated tube and the volume was reduced to near dryness under a gentle stream of nitrogen. After this stage, the sample was reconstituted in 0.5 ml n-hexane.

A description of the main characteristics of mothers and newborns is shown in Table 1. The mean age of pregnant women was 25.5 y, with mean weight gain during the pregnancy of 14.1 kg. The mean weight of the newborns was 3.2 kg, and half of them were boys. Among these pregnant women, an estimated 74% were multiparas. Although PCBs were banned more than three decades ago, in the present study, we found PCB congeners in umbilical cord serum samples from newborns. Recent studies also have noted the presence of these substances in biological fluids and human tissues worldwide [16,26– 28]. Levels of ∑PCBs ranged from 0.35 to 55.17 ng/ml in umbilical cord serum positive samples, with a mean of 7.04 ng/ml in positive samples and of 6.65 ng/ml in all samples (Table 2). PCB 138 was the most prevalent congener (63.5% of positive samples), while PCB 28 was the least prevalent (18.9%). This pattern was observed due to the biodegradability of PCBs, that is diminished considerably when halogenation degree increases [29]. PCB 138 is hexachlorinated, while PCB 28 is three chlorinated. PCB 138 was also the main contributor to ∑PCB (36.8%), followed by PCB 52 (25.7%) and PCB 180 (22.4%). A different result was observed by Lopes et al. [18], where the major congener was PCB 153. Umbilical cord samples presented at least one PCB congener in a frequency of 92.6%. The majority of samples had two (29.1%) or three (25.7%) PCB congeners. One PCB congener was detected in 16.2% samples, four in 16.9% and five in 4.7% of the samples. The PCB congeners found were ranked as follows: 138 N 52 N 180 N 153 N 28 (Table 2). The levels of PCBs found in umbilical cord serum samples for this study were greater than those reported by studies in other countries [18,22,30–32] (Table 3). In the present work, levels of PCBs 52 and 28

2.4. Instrumental analysis Identification and quantification analyses were performed in an Agilent 6890 Plus gas chromatograph (Agilent Technologiesprovided with a μECD and an Agilent HP-5 (crosslinked 5% phenyl methyl siloxane) capillary column (30 m long × 0.32 mm i.d. × 0.25 μm film thickness). Nitrogen was the carrier gas (1.5 ml/min). The injector and detector temperatures were 280 and 320 °C, respectively. The temperature program was 60 °C (2 min), 30 °C/min to 190 °C (5 min), 5 °C/min to 220 °C (5 min), and 20 °C/min to 300 °C (1 min). Selected samples were analyzed using a Hewlett-Packard GC–MS HP 6890-5973 to confirm the qualitative results with the same column used in the GC–μECD analysis. The carrier gas was helium (1.5 ml/min). The MS detector was used in selected ion monitoring mode (SIM), with electron energy of

2.6. Statistical analysis

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Table 1 Main characteristics of mothers and newborns participants of the study (n = 148). Characteristics

n (%)

Maternal Age (y) Body mass index before pregnancy (kg/m2) Weight gain during pregnancy (kg) Parity 1st child 2nd child N2 child Newborn Weight (kg) Length (cm) Head circumference (cm) Number of boys/girls (%) Type of birth (natural/cesarean) (%)

148 (100) 117 (79.1) 102 (68.9)

Mean ± S.D.

Median

Range

25.5 ± 7.1 24.6 ± 5.9 14.1 ± 6.2

24.0 23.2 13.0

14.0–44.0 16.9–55.2 3.0–36.0

3.2 ± 0.5 48.5 ± 3.1 34.8 ± 2.6

3.2 49.0 34.5

1.1–4.7 34.0–56.0 29.0–51.5

48 (32.4) 42 (28.4) 32 (21.6) 148 (100) 130 (87.8) 131 (88.5) 74/71 (50/48) 45/103 (30.4/69.6)

S.D. = Standard deviation. Sum of % is different of 100 when missing data.

were 154 and 75-fold higher than those detected in Spain [32]. In a previous study conducted by members of our research group, Lucena et al. [4] verified that these congeners were significantly related to breast cancer in Spain. Thus, is important to highlight that even in low concentrations, these substances are of high toxicological risk. Levels of PCBs 153 and 138 (p b 0.05) and PCB 180 (p b 0.10) were lower among Santa Maria residents than among residents from other towns (Table 4). The study was conducted in the Hospital of UFSM, which receives pregnant women from Santa Maria (urban area) and nearby towns (urban and rural areas). This region is not heavily industrialized, and agriculture is characterized by rice, corn, soy, tobacco and bean cultivation, as well as dairy and beef cattle. No significant difference (p N 0.05) was observed between housewives and mothers that work in other activities. Umbilical cord serum samples from smoker (p b 0.05) and drinker (p b 0.10) mothers presented higher PCB 28 levels than mothers who do not smoke or drink (Table 4). Lopes et al. [18] also observed that smoking habits were related to PCB concentrations in human serum. Mainstream tobacco smoke contains over 3500 different volatile and nonvolatile compounds, many of them not yet identified [33]. Among these compounds, PCBs can be identified [34]. Little is known about the relation between alcohol consumption and PCB levels. Alcohol affects the xenobiotic metabolism, and can alter its biotransformation, distribution and elimination. Although PCBs have a long half-life and tendency to bioaccumulation, in the present work a positive correlation between the age of the mothers and levels of PCBs in umbilical cord blood (p N 0.05) was not observed. No significant difference (p N 0.05) was noted among age groups (b20 y — 43 mothers, ∑PCB = 7.03 ng/ml; from 21 to 24 y — 38, ∑PCB = 6.03 ng/ml; from 25 to 31 y — 33, ∑PCB = 6.12 ng/ml; and N32 y — 34 mothers, ∑ PCB = 7.37 ng/ml). Tsang et al. [35]

Table 2 Levels of PCBs in umbilical cord serum (ng/ml) (n = 148). Congener

PCB 28 PCB 52 PCB 153 PCB 138 PCB 180 ∑PCB

All samplesa

Positive samplesb

Mean ± S.D.

Md

n (%)

Mean ± S.D.

Md

Range

0.18 ± 0.40 1.71 ± 4.49 0.82 ± 1.04 2.45 ± 3.07 1.49 ± 1.77 6.65 ± 7.45

0.05 0.59 0.55 0.94 0.73 4.84

28 (18.9) 81 (54.7) 76 (51.4) 94 (63.5) 82 (55.4) 137 (92.6)

0.75 ± 0.69 3.08 ± 5.73 1.55 ± 1.01 3.83 ± 3.11 2.64 ± 1.63 7.04 ± 7.54

0.47 1.11 1.18 3.03 2.17 4.95

0.26–3.40 0.50–35.15 0.52–5.39 0.50–16.62 0.53–8.04 0.35–55.17

∑PCB was calculated as the sum of PCB congeners. S.D. = Standard deviation. Md = Median. a Samples with non-detectable levels of PCB congeners were considered as half limit of detection (1/2 LOD) on statistical analysis. b Positive samples were calculated for samples with levels above limit of detection.

observed no correlation between total PCBs in human milk collected from Hong Kong and maternal age. Among primiparous (∑ PCB = 5.39 ng/ml) and multiparous (∑PCB = 5.71 ng/ml) no significant difference (p N 0.05) was observed. The major source of PCB human contamination is food. Table 5 presents the levels of PCBs in umbilical cord serum of newborns and the food habits of the mothers. PCB levels were not different between consumers and non-consumers of meat, milk, eggs, salad, fruits, rice and beans. The sole exception was PCB 153, which presented higher levels in umbilical cord of newborns whose mothers consumed fruits (p b 0.05). This result was not expected, because PCBs are lipophilic compounds, presenting higher levels in food from animal origin. To the best of our knowledge, no study was conducted in Brazil to evaluate PCB levels in fruits. Lopes et al. [18] observed higher levels of PCB 153 in serum of women that consumed more portions of fresh vegetables and fat rich foods. Similar to our result, Mariscal-Arcas et al. [36] found no significant association between Mediterranean Diet adherence and the presence of organochlorine residues in umbilical cord blood serum from Southern Spain. The food frequency questionnaire focused on the dietary intake of the year before pregnancy, which is a relatively short period compared to the age of the participants. No data was found in the literature consulted in respect to contamination of biological samples from other Brazilian type groups. Investigative studies on contaminated food products from the same Brazilian State were previously performed, which noted the presence of contamination in cheese, milk, meat products, corn, rice and beans. The mean sum of the PCBs 28, 52, 138, 153, and 180 was 24.85 ng/g fat for cheese [37], 15.5 ng/g fat for pasteurized milk [38], and 31.19 ng/g fat for hotdog sausage [39]. Honey is considered as a biomarker of environmental contamination. PCBs 52 and 28 were the most frequent in honey, being detected in 41 and 39% of the samples, respectively, and followed by PCB 180 (11%) [40]. In fish, PCB 153 was observed in 98.3% of samples, followed by PCBs 138 and 180, detected in 95% of samples [41]. Among foodstuffs of vegetable origin, PCBs 28, 52, 138, 153 and 180 were detected at mean concentrations of 0.39 ng/g in corn [42], and PCBs 28, 52, 101, 118, 138, 153 and 180 at mean concentrations of

Table 3 Concentrations (ng/ml) of PCBS in umbilical cord serum samples from different countries. Country Germany Germany Portugal Slovakia Spain

PCB 28

PCB 52

PCB 153

PCB 138

PCB 180

Reference

0.21

0.17

0.18

0.97 0.38 0.21

0.11 0.28 0.17

0.36 0.33 0.20

Bucholski et al., 1996 [30] Lackmann et al., 1996 [31] Lopes et al., 2014 [18] Park et al., 2008 [22] Grimalt et al., 2010 [32]

0.47

0.01

0.02

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Table 4 Levels of PCBs in umbilical cord serum (ng/ml) of newborns and socio-demographic characteristics of the mothers (n = 148). Variable

Residence Santa Maria Out Santa Maria Main occupation Housewife Others Smoke habits Smoker Non-smoker Alcohol habits Drinker Non-drinker

N (%)

PCB 28

PCB 52

PCB 153

PCB 138

∑PCB

PCB 180

M

Md

M

Md

M

Md

M

Md

M

Md

M

Md

106 (71.6) 41 (27.7)

0.19 0.16

0.05 0.05

1.90 1.25

0.62 0.51

0.63b 1.30a

0.05 1.06

2.26b 2.98a

0.75 1.95

1.27B 2.03A

0.59 1.88

6.26 7.72

4.06 7.32

88 (59.4) 59 (39.8)

0.21 0.15

0.05 0.05

2.00 1.30

0.66 0.50

0.82 0.81

0.55 0.52

2.32 2.68

0.93 1.45

1.56 1.37

1.04 0.05

6.91 6.31

5.09 3.65

36 (24.3) 111 (75.0)

0.28a 0.15b

0.05 0.05

0.95 1.97

0.05 0.60

0.68 0.87

0.05 0.55

2.16 2.56

0.96 0.98

1.18 1.58

0.65 0.81

5.24 7.13

4.67 4.97

21 (14.2) 123 (83.1)

0.28A 0.17B

0.05 0.05

0.91 1.88

0.05 0.60

1.09 0.79

0.76 0.52

2.17 2.54

1.03 0.98

1.70 1.46

1.70 0.65

6.16 6.85

6.02 4.88

∑PCB was calculated as the sum of PCB congeners. Results are presented as Mean (M) and Median (Md). Different lower case superscript letters in the same column indicate statistical difference by Mann–Whitney test (p b 0.05). Different capital superscript letters in the same column indicate statistical difference by Mann–Whitney test (p b 0.10). Sum of % is different of 100 when missing data.

that PCBs can have an influence when present in the congenitally malformed, an example of which has already been shown in other studies on carcinogenicity in humans [2,3,5,6,46]. The types of malformations observed were hydrocephalus (2 babies), auricular malformations (2), penile malformation (1), renal malformation (1), hollow chest (1), anal malformation (1), brachial palsy (1), cardiac and osteoarticular malformation (1), Crouzon's Syndrome, brachycephaly and ogival palate (1). The largest value for ∑ PCBs was detected in the newborn with penile malformation, followed by those with auricular malformation. The types of congenital malformations identified in the present study are of diverse etiologies. Nonetheless, comparing PCB levels in both groups, we found higher levels in the congenital malformation group. The incidence of congenital malformation in the present study (7.4%) was higher than those observed in Brazil (2 to 3% [47]), India (2.4% [48]), Europe (2.4% [49]) and United States (2.9% [50]). This may be attributed to the status of important center to risk pregnancy, receiving difficult cases of the region. Studies involving investigations into the influence of PCBs in human congenital malformations are very rare. No previous study has been performed to investigate the levels of PCBs in

4.39 ng/g for rice and 4.17 ng/g for beans [43]. In accordance with Brazilian legislation [44], these values are below the established maximum limits, however, due to the high frequency of determination and the bioaccumulative characteristic of PCBs, along the time these values have contributed to body accumulation. Drinking and agriculture water and soil are other sources of contamination. Additionally, no work was conducted to evaluate the disposal of electrical devices in Rio Grande do Sul state, responsible for the major use of PCBs. Nevertheless, food contamination is present and is also detected in the umbilical cord serum taken from subjects in the same region. Table 6 presents some characteristics of newborns and levels of PCBs in umbilical cord serum. No difference (p N 0.05) was observed between PCBs and gender, gestational time (preterm was considered when babies were born before 37 weeks [45]) and birth weight (normal birth weight was considered when babies had ≥ 2.5 kg [45]). When levels of PCBs were compared between normal and malformed newborns, PCB 52 and PCB 180 presented higher levels (p b 0.10) in malformed babies (5.75 and 2.59 ng/ml, respectively), when compared to normal babies (1.39 and 1.41 ng/ml, respectively). These results suggest

Table 5 Levels of PCBs in umbilical cord serum (ng/ml) of newborns and food habits of the mothers (n = 148). Variables

Meat Consume No consume Milk Consume No consume Eggs Consume No consume Salad Consume No consume Fruits Consume No consume Rice Consume No consume Beans Consume No consume

n (%)

PCB 28

PCB 52

PCB 153

PCB 138

∑PCB

PCB 180

M

Md

M

Md

M

Md

M

Md

M

Md

M

Md

136 (91.9) 8 (5.4)

0.19 0.20

0.05 0.05

1.82 0.44

0.61 0.05

0.84 0.68

0.58 0.05

2.41 4.17

0.93 4.47

1.50 1.24

0.75 0.43

6.76 6.72

4.90 6.36

102 (68.9) 42 (28.4)

0.17 0.22

0.05 0.05

1.93 1.28

0.62 0.05

0.81 0.90

0.63 0.05

2.49 2.54

0.88 1.89

1.54 1.37

0.87 0.65

6.93 6.32

4.90 5.31

58 (39.2) 86 (58.1)

0.20 0.18

0.05 0.05

2.58 1.17

0.60 0.57

0.90 0.79

0.65 0.05

2.28 2.66

0.74 1.67

1.58 1.43

0.93 0.65

7.53 6.23

4.63 5.07

104 (70.3) 40 (27.0)

0.15 0.27

0.05 0.05

1.79 1.61

0.57 0.67

0.89 0.69

0.29 0.62

2.42 2.71

0.85 1.73

1.53 1.38

0.75 0.73

6.79 6.66

4.76 5.14

46 (31.1) 98 (66.2)

0.20 0.18

0.05 0.05

1.69 1.76

0.64 0.56

1.15a 0.69b

0.80 0.05

2.69 2.42

1.37 0.95

1.70 1.39

0.93 0.64

7.43 6.44

5.82 4.34

130 (87.8) 14 (9.4)

0.17 0.34

0.05 0.05

1.72 1.90

0.56 0.95

0.84 0.83

0.57 0.30

2.46 2.89

0.88 3.23

1.45 1.90

0.66 1.41

6.64 7.86

4.90 6.38

124 (83.8) 20 (13.5)

0.17 0.29

0.05 0.05

1.79 1.41

0.58 0.62

0.82 0.90

0.58 0.05

2.42 3.03

0.85 2.61

1.45 1.72

0.66 0.92

6.66 7.35

4.90 6.38

∑PCB was calculated as the sum of PCB congeners. Results are presented as Mean (M) and Median (Md). Different lower case superscript letters in the same column indicate statistical difference by Mann–Whitney test (p b 0.05). Sum of % is different of 100 when missing data.

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Table 6 Levels of PCBs in umbilical cord serum (ng/ml) of newborns and its characteristics (n = 148). Variable

Gender Male Female Gestational time Preterm Full term Birth weight Normal Low birth weight Malformation Normal Malformed

n (%)

PCB 28

PCB 52

PCB 153

PCB 138

∑PCB

PCB 180

M

Md

M

Md

M

Md

M

Md

M

Md

M

Md

74 (50.0) 71 (48.0)

0.13 0.24

0.05 0.05

1.65 1.68

0.50 0.62

0.83 0.85

0.29 0.65

2.16 2.63

0.89 1.03

1.37 1.64

0.64 0.86

6.13 7.04

4.27 4.99

18 (12.2) 129 (87.2)

0.29 0.17

0.05 0.05

3.44 1.47

0.30 0.60

0.99 0.79

0.29 0.55

2.94 2.38

0.88 0.90

1.25 1.53

0.05 0.88

8.92 6.34

2.43 4.92

138 (93.2) 10 (6.8)

0.19 0.05

0.05 0.05

1.35 6.66

0.60 0.05

0.81 1.02

0.57 0.05

2.50 1.74

1.12 0.05

1.48 1.52

0.85 0.05

6.33 10.99

4.84 3.31

136 (91.9) 11 (7.4)

0.15 0.58

0.05 0.05

1.39B 5.75A

0.57 1.73

0.77 1.26

0.29 1.12

2.33 3.90

0.94 0.64

1.41B 2.59A

0.65 2.30

6.06b 14.08a

4.34 7.71

∑PCB was calculated as the sum of PCB congeners. Results are presented as Mean (M) and Median (Md). Different lower case superscript letters in the same column indicate statistical difference by Mann–Whitney test (p b 0.05). Different capital superscript letters in the same column indicate statistical difference by Mann–Whitney test (p b 0.10). Sum of % is different of 100 when missing data.

umbilical cord serum from malformed newborns in Brazil. In the literature consulted, only two recent studies were encountered, which investigated the relationship of maternal pregnancy serum levels of PCBs with the occurrence of hypospadias and cryptorchidism in male offspring. However, in these two studies the researchers did not encounter any statistically significant difference between concentrations in the control group and the malformed group [51,52]. 4. Conclusions Levels of PCBs found in the samples were greater than those reported by other studies, which cause concern about environmental and human contamination. PCB levels in umbilical cord serum are affected by food and the life habits of the mothers. Acknowledgments This work was supported by CNPq (475787/2007-2). S. Mohr was the recipient of CAPES Master degree fellowship. J.O. Mozzaquatro and A. Lorenzoni were the recipients of scientific initiation fellowships of CNPq and FAPERGS, respectively. I. Costabeber was the recipient of CNPq research fellowship. The authors also thank the Special Program of Incentives from International Publications/PRPGP/UFSM, the University Hospital of Santa Maria (HUSM), all the volunteers that accepted to participate in this study and Prof. Dr. Hugo Aurélio Becker Amaral, for the assistance given to us throughput this project.

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Appendix A. Supplementary data Supplementary data to this article can be found online at http://dx. doi.org/10.1016/j.cca.2015.10.019.

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