Science of the Total Environment 671 (2019) 1017–1025

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Science of the Total Environment

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Polybrominated diphenyl ethers in human serum, semen and indoor

dust: Effects on hormones balance and semen quality
Yun-jiang Yu a, Bi-gui Lin a, Xi-chao Chen a, Jing Qiao b, Liang-zhong Li a, Yi Liang c, Guo-zhi Zhang b, Yan Jia a,
Xiu-qin Zhou b, Cai-rong Chen b,⁎, Hai-dong Kan d,⁎
a
State Environmental Protection Key Laboratory of Environmental Pollution Health Risk Assessment, South China Institute of Environmental Sciences, Ministry of Ecology and Environment, Guang-
zhou 510655, China
b
Reproductive Medicine Center, People’s Hospital of Qingyuan, The Sixth Affiliated Hospital of Guangzhou Medical University, Qingyuan 511518, China
c
State Key Laboratory of Organic Geochemistry, Guangzhou Institute of Geochemistry, Chinese Academy of Sciences, Guangzhou 510640, China
d
School of Public Health, Key Lab of Public Health Safety of the Ministry of Education and NHC Key Laboratory of Health Technology Assessment, Fudan University, Shanghai 200032, China

H I G H L I G H T S G R A P H I C A L A B S T R A C T

• There are significant correlations among

PBDE congeners in dust, serum, and
semen.
• PBDEs in serum, semen, and dust all can
decrease the semen quality.
• PBDEs also disrupt the balance of pro-
duction and thyroid hormone.
• PBDEs in semen were more responsive
to the balance of hormones than that
in serum.
• PBDEs might affect the main compo-
nents of male reproductive system.

a r t i c l e i n f o a b s t r a c t

Article history: Exposure to polybrominated diphenyl ethers (PBDEs), especially their intake from indoor dust, may result in the
Received 28 December 2018 dysregulation of human thyroid hormones, endocrine disruption, and in poor semen quality. We used multiple
Received in revised form 20 March 2019 linear regression (MLR) models to determine the influence of the levels of PBDEs present in indoor dust and in
Accepted 21 March 2019
the serum and semen of male individuals on the quality of human semen, as well as on the levels of reproductive
Available online 26 March 2019
and thyroid hormones. The results revealed significant interactions between PBDEs (such as BDE47, BDE153, and
Editor: Wei Huang BDE154) in these media and the male reproductive system after adjusting for covariates. The levels of PBDE con-
geners in semen had pronounced negative association with semen volume, sperm count, and motility, and also
Keywords: had positive or negative association with reproductive hormones (including testosterone, luteinizing hormone,
PBDEs and estradiol), aromatase activity, and Leydig cell function, and thyroid hormone (including sensitivity to
indoor environment thyroid-stimulating hormone). The results were the same for the levels of PBDE congeners in serum and house
sperm quality dust, which had similar negative or positive correlations with semen quality, sex hormones, and thyroid hormone
production hormone factors. Taken together, our findings suggest that exposure to PBDEs might decrease the semen quality and dis-
thyroid hormone
rupt the balance of reproductive and thyroid hormones in the human body.
© 2019 Elsevier B.V. All rights reserved.

⁎ Corresponding authors.
E-mail addresses: cairong1222@163.com (C. Chen), kanh@fudan.edu.cn (H. Kan).

https://doi.org/10.1016/j.scitotenv.2019.03.319
0048-9697/© 2019 Elsevier B.V. All rights reserved.
1018 Y. Yu et al. / Science of the Total Environment 671 (2019) 1017–1025
1. Introduction
Semen quality is a vital indicator of male reproductive capability.
Low quality of semen is one of the important reasons for infertility
(Cooper et al., 2010). In United States, Europe, Australia, and India, the
quality of semen has decreased significantly in the past decades
(Swan et al., 2000; Adiga et al., 2008; Burton, 2013; Rolland et al.,
2013). A decline in most of the semen parameters has been reported
in recent studies on sperm samples donated by healthy reproductive
aged Chinese adults, which indicates that there might be a severe and
generalized decrease in semen quality among males in China (Huang
et al., 2017; Wang et al., 2017; Yuan et al., 2018). The decline in
semen quality has, therefore, been an important issue in the health
community in recent decades.
Environmental factors are considered to pose threat to the male re-
productive function, especially to the semen quality (Oliva et al., 2001;
Stefankiewicz et al., 2006). Environmental endocrine disruptors
(EDCs), which can simulate, increase, or inhibit hormones, may also af-
fect the human reproductive development, even at very low concentra-
tions (Hauser et al., 2002; Hauser et al., 2003; Hauser et al., 2005).
Therefore, EDCs deserve most attention worldwide. Polybrominated
diphenyl ethers (PBDEs) are persistent flame-retardants that were pre-
viously used in consumer products, such as plastics, electronics, con-
struction, and textile (Hites, 2004; Zhu et al., 2015). PBDE congeners
are also a group of ubiquitous EDCs, which have been linked to adverse
health outcomes, including dysregulation of thyroid hormone, endo-
crine disruption, and poor semen quality (Legler, 2008; Abdelouahab
et al., 2011; Den Hond et al., 2015).
People usually spend N80% of their time indoors. The indoor environ-
ment is one of the main sources for exposure of humans to PBDEs
(Jones-Otazo et al., 2005; Harrad et al., 2010; de Boer et al., 2016). Pre-
vious studies found that the PBDE content in dust was positively corre-
lated with that in male serum (Karlsson et al., 2007), and was negatively
correlated with the levels of luteinizing hormone (LH) and follicle stim-
ulating hormone (FSH) in serum (Meeker et al., 2009). Furthermore, in
many studies, it was reported that BDE153 levels in serum were in-
versely correlated with sperm concentration and testicular size
(Akutsu et al., 2008), and BDE47, BDE100, and ∑PBDEs levels in adult
serum were negatively correlated with sperm motility and the levels
of thyroid hormones (Abdelouahab et al., 2011). Our previous pilot
study revealed that exposure to PBDEs from house dust might have ad-
verse effects on the quality parameters of human semen, including
sperm count, motility, and viability (Yu et al., 2018).
The effects of PBDEs on male reproductive health have been evalu-
ated in only a few studies, and much less is known about the mecha-
nisms underlying such effects. The present study aimed to
comprehensively analyze the different factors that may affect the repro-
ductive health of male individuals in electronic waste areas of South
China where PBDE levels are particularly high (Wang et al., 2010;
Zheng et al., 2015), including the levels of PBDE congeners in indoor
dust and in male serum and semen. The association between internal
and external exposure to PBDE congeners on the levels of reproductive
and thyroid hormones was also analyzed, including the correlation be-
tween PBDE congeners levels in serum and in semen.
2. Methods
2.1. Study participants and sampling
We recruited 76 adult male volunteers aged ~20–50 years during the
period from October 2016 to September 2017. Thirty-eight of these vol-
unteers were from an e-waste area and lived near an e-waste disman-
tling plant, and the other 38 were lived in the vicinity of the e-waste
dismantling area at Qingyuan city in South China. Individuals with re-
productive health problems, such as epididymitis, vasectomy, orchiditis,
and vesiculitis, and those taking any medication were excluded from
the study. Prior to signing the informed consent, all the eligible subjects
were informed of the study procedures and their possible conse-
quences. They all completed the questionnaire under the guidance of
the staff at Qingyuan People's Hospital, China. This study proposal was
reviewed and approved by the medical ethics committee of Qingyuan
People's Hospital.
Semen samples from eligible participants were collected by mastur-
bation after 2–7 days of abstinence. In addition, semen samples were re-
collected for nine participants whose semen volume was b1.5 mL within
2–3 days, and the two samples were pooled for detection of PBDEs,
which was to increase the sample volume to facilitate the detection of
samples. While just the parameters of semen samples collected for the
first time were used for statistical analysis. The serum samples of volun-
teers were collected on the same day. Indoor dust samples were col-
lected from the residences (n = 76) of the participants within
2 weeks before or after the collection of semen and serum samples.
Dust samples were sieved through a stainless-steel sieve (150 μm) to re-
move large debris and were stored at −20 °C until further analysis. The
details of dust sampling method have been described in one of our pre-
vious studies (Yu et al., 2018).
2.2. Analysis of semen quality and serum hormone levels
For assessing the semen quality, sperm counts and motility were de-
termined by computer assisted semen analysis (CASA, version WLJY-
9000, Beijing Weili Co. Ltd., Beijing, China), sperm morphology was
scored by rapid Diff-Quick staining according to the manual of World
Health Organization (WHO, 2010), and sperm viability was assessed
by eosin staining and visualization under a microscope at high magnifi-
cation. The levels of reproductive and thyroid hormones were deter-
mined by automated chemiluminescence immunoassay system
(DXI800, Beckman Coulter, Inc., CA) and microparticle chemilumines-
cence immunoanalyzer (Coulter ACCESS2, Beckman Coulter, Inc., CA),
respectively. The remaining semen and serum samples were kept in liq-
uid nitrogen (−196 °C) for the analysis of PBDE congeners.
2.3. Determination of the levels of PBDEs in serum, semen, and dust samples
2.3.1. PBDEs in serum and semen samples
The details of the analytical procedure used for determining the
levels of PBDEs in serum have been described previously (Sjodin et al.,
2004), and the extraction and analysis procedures of PBDE congeners
and PBB153 in semen samples were the same as those for the serum.
Serum and semen samples, spiked with surrogate standards (BDE77,
BDE181, and 13C-BDE209), were subjected to solid-phase dispersion
on diatomaceous earth, and were then dried by pressurized nitrogen.
The samples were transferred into a soxhlet and extracted with a mix-
ture of hexane/acetone (1:1, v/v) for 48 h with a soxhlet extractor.
After evaporation of the solvent, the extracts were transferred to pre-
weighed glass tubes and evaporated under a gentle stream of ultra-
high purity nitrogen, and then the lipid content was of serum and
semen determined gravimetrically using a semimicro balance (Sarto-
rius RC210P, Goettingen, Germany). Concentrated extracts were re-
solved with n-hexane and purified by gel permeation chromatography
(GPC), and were further cleaned and fractionated on a multi-layer col-
umn (10 mm i.d.) packed, from the bottom to the top, with neutral alu-
mina (6 cm), neutral silica gel (2 cm), alkaline acid silica (5 cm), neutral
silica gel (2 cm), 44% sulfuric acid silica (8 cm), and anhydrous sodium
sulfate (2 cm). After sample loading, the columns were eluted with
70 mL of dichloromethane/hexane (1:1, v/v). The eluates were con-
densed until dryness under a gentle flow of nitrogen spiked with inter-
nal standards (BDE118 and 128), and were finally concentrated to 50 μL.
2.3.2. PBDEs in dust samples
The analytical approach for determining the levels of PBDEs in in-
door dust samples has been described in detail elsewhere (Van den
 Y. Yu et al. / Science of the Total Environment 671 (2019) 1017–1025 1019

Eede et al., 2012). Briefly, an aliquot (0.5 g) of the sample was accurately variables were used to adjust all the MLR models. All statistical analyses
weighed and spiked with surrogate standards (BDE77, BDE 181, and were conducted using SPSS software, and statistical significance was de-
13
C-BDE209). Samples were extracted using 3 mL of hexane/acetone noted as p ≤ .05.
(3:1, v/v) by a combination of vortex and ultrasonic extraction (1 min
vortex and 10 min ultrasonic extraction), which was repeated three 3. Results
times. The extracts were centrifuged at 4000 rpm for 5 min. The super-
natants were transferred into clean glass tubes and evaporated to 1 mL, 3.1. Participants
and were subsequently purified through a Florisil cartridge eluted with
10 mL of hexane and an acidified silica 44% cartridge eluted with 10 mL The characteristics of the study participants are described in Table 1.
of hexane/dichloromethane (1,1, v/v). The purified extracts were con- The age of participants was between 20 and 50 years, and the mean age
densed until dryness under gentle nitrogen flow spiked with internal was 37.5 years. Those who lived near the e-waste dismantling park (n
standards (BDE118 and 128), and finally concentrated to 100 μL. = 38) were prone to exposure to home environments with higher con-
centrations of PBDEs and their mean age was 40.5 years. The BMI of
2.3.3. Instrumental analysis 73.7% of the participants was b25 kg/m2. Their residence time in the
The levels of eight PBDE congeners (BDE28, 99, 100, 153, 154, 183, study area ranged from 3 to 42 years, with an average time of
and 209) and one polybrominated biphenyl (PBB) congener (PBB153) 17 years. More than half (56.6%) of the participants used alcohol, but
were measured in the purified extracts of serum, semen, and dust sam- less than half (44.7%) used tobacco.
ples. These congeners were analyzed by gas chromatograph with a mass
spectrometer (GC-MS, 7890A-5975C, Agilent, Inc., Palo Alto, CA, USA) in
electron capture negative ionization mode (NCI). The mass spectrome- 3.2. Concentrations of PBDE congeners in dust, serum, and semen samples
ter was employed in selected ion monitoring (SIM) mode. Congeners
were separated with a DB-5HT (15 m × 0.25 mm i.d., 0.10 μm film thick- 3.2.1. PBDE congeners in dust samples
ness, J&W Scientific, Folsom, CA) capillary column. The concentrations The detection rate of PBDE congeners in dust samples was 100%,
of serum PBDE congeners were adjusted for lipids and were expressed whereas that of PBB153 in dust samples was 94.7% (Table S1). Concen-
in units of nanograms per gram total lipids, whereas those of semen trations of ∑PBDEs (sum of eight congeners) ranged from 102 to
were adjusted for wet weight (ww). 26,098 ng/g dust. The geometric mean of ∑PBDEs was 1876 ±
3673 ng/g. BDE209 was the dominant PBDE congener, accounting for
2.4. Quality assurance and quality control on average 94.6% of the total PBDEs by weight, and was followed by
BDE183 and BDE99.
The quality assurance and quality control samples (QA/QC samples)
were simultaneously analyzed for each batch (10) of samples, including 3.2.2. PBDE congeners in serum and semen samples
procedural blanks, solvent blanks, and parallel samples. All analytic runs The detection rates of PBDE congeners in serum and semen samples
included a blank, and the amount of each PBDE congener was corrected were 53.9%–97.4% and 46.1%–96.1% (Table S1), respectively. The detec-
by its amount in the procedural blank. tion rates of BDE153 in both the serum and semen samples were the
The limit of detection (LOD) was defined as three times the noise highest (97.4% and 96.1%, respectively). On the contrary, rates of detec-
level. The LOD for individual PBDE congeners in semen ranged from tion of BDE100 in serum and semen samples were the lowest (53.9%
0.94 to 4.76 pg/g ww (median, 1.74 pg/g ww), and in serum it ranged and 46.1%, respectively). The mean concentrations of ∑PBDEs in
from 0.104 to 0.529 ng/g lipid (median, 0.193 ng/g lipid) (Table S1). serum and semen were 17.8 ± 11.9 ng/g lipid and 49.5 ± 36.5 pg/g
The average contents of lipids in the semen and serum samples were ww, respectively (Table 2). The concentration of serum BDE47 and
0.315 ± 0.045% and 0.545 ± 0.140%, respectively. The recoveries of sur- semen BDE153 were both the highest (except BDE209), being 3.32 ±
rogate standards in semen and in serum were 88.2 ± 9.3% for BDE77, 4.93 ng/g lipid and 4.17 ± 3.98 pg/g ww, respectively. The median con-
80.7 ± 8.5% for BDE181, and 70.8 ± 9.6% for 13C-BDE209. The LOD of in- centrations of BDE28, 47, 153, and PBB153 in semen were 2.88, 4.47,
dividual congeners in dust samples ranged from 0.005 ng/g to 4.63, and 1.75 pg/g ww, respectively (Table S1). The levels of individual
0.407 ng/g (median, 0.010 ng/g), and average recoveries for the surro- PBDEs (BDE28, 47, 153) and PBB153 determined in this study are com-
gates spiked in samples were 97.2 ± 8.9% for BDE77, 81.3 ± 9.2% for parable with those reported previously by us from this study area (Yu
BDE181, and 72.3 ± 10.5 for 13C-BDE209. et al., 2018) as well as with those reported previously from East China
(Liu et al., 2012).
2.5. Statistical analysis
Table 1
PBDE levels in different media b LOD, including serum, semen, and Demographic characteristics of the study participants (n = 76).a
dust, were substituted with a value of 1/2 LOD for calculating the total Covariate Mean ± SD Range
concentrations of ∑PBDEs and for statistical analysis. The
Age(years) 37.5 ± 8.6 20–50
Kolmogorov–Smirnova (K–S) and Shapiro–Wilk (S–W) methods were No.%:
used to test the normal distribution. Spearman's correlation coefficients 20–39 42(55.3%)
(rs) assessed the relationship among the concentrations of PBDE conge- 40–50 34(44.7%)
ners in the serum, semen, and dust samples. BMI 23.74 ± 2.81 18–32.3
No.%:
We used multiple linear regression (MLR) models to explore the sta-
b25 56(73.7%)
tistical interactions, in terms of the comprehensive effects of all the fac- 25- b 30 19(25.0%)
tors on male reproductive health, especially on semen quality. Before ≥30 1(1.3%)
MLR analysis, all data that did not conform to the Gaussian distribution Abstinence time (days) 4.66 ± 1.57 2–7
Smoking Yes 34 (44.7%)
and constant variance were transformed by Box–Cox transformation
No 42 (55.3%)
using Minitab 17 Mathematical software (State College, PA, USA) Alcohol use Yes 43 (56.6%)
(Louis et al., 2015; Vitku et al., 2016), and were subsequently standard- No 33 (43.4%)
ized with SPSS software (version 20 for Windows, SPSS Inc., Chicago, IL, Residence time (years) 12.3 ± 9.8 3–42
USA). The list of potential confounders included age, abstinence time, a
Abbreviations: SD, standard deviation; No.%, Number of subjects; BMI, body mass
body mass index (BMI), smoking, and alcohol consumption, and these index.
1020 Y. Yu et al. / Science of the Total Environment 671 (2019) 1017–1025

Table 2 The correlations among the levels of PBDE congeners in semen,

Descriptive statistics of analytes of PBDE congeners in indoor dust, serum and semen.a serum, and indoor dust samples (semen vs. serum, semen vs. dust,
Compound Dust Serum (n = 76) Semen (n = 76) serum vs. dust) are shown in Table 3. The concentrations of BDE99,
(ng/g) (ng/g lipid) (pg/g ww)
BDE154, BDE153, BDE183, ∑PBDEs, and PBB153 were significantly cor-
related in serum and semen samples (rs = 0.28–0.685, p b .05). Signifi-
BDE28 2.05 (6.05) 0.53 (0.68) 3.14 (4.62)
cant correlations were also observed for concentrations of individual
BDE47 6.82 (32.22) 0.95 (2.30) 4.17 (3.98)
BDE100 2.68 (9.47) 0.28 (0.62) 1.76 (2.74) PBDEs (BDE47, BDE99, BDE154, BDE153, BDE183) in semen and dust
BDE99 10.6 (62.5) 0.55 (1.64) 2.87 (4.78) samples and ∑PBDEs (rs = 0.279–0.652, p b .05), and between concen-
BDE154 3.08 (8.23) 0.42 (1.29) 1.79 (4.14) trations of individual PBDEs (BDE47, BDE100, BDE99, BDE154, BDE153)
BDE153 7.37 (24.3) 3.32 (4.93) 3.54 (9.32) in serum and dust samples and ∑PBDEs (rs = 0.454–0.712, p b .05).
BDE183 22.4 (123) 0.90 (3.58) 1.22 (2.87)
BDE209 1767 (3535) 6.12 (5.99) 17.6 (28.7)
The three-dimensional distribution of the levels of PBDE congeners in
∑PBDEs 1876 (3673) 17.8 (11.9) 49.5 (36.5) these three matrices are shown in Fig. S1. Spearman correlation coeffi-
PBB153 2.05 (6.05) 0.27 (0.91) 1.41 (2.79) cients suggested that BDE99 concentrations in dust, serum, and semen
a
Abbreviations: GM (GSD), GM = geometric mean; GSD = geometric standard devi- samples strongly correlated with one another, and were the same for
ation; ww, wet weight. BDE154, BDE153, and ∑PBDEs.

3.3. Semen quality, and levels of reproductive and thyroid hormones

Table 3 The semen parameters of participants and levels of reproductive and

Correlations between PBDEs concentration in human semen, serum and indoor dust.a
thyroid hormones in serum are summarized in Table 4. The median
Congeners Semen VS. serum Semen VS. dust Serum VS. dust values for semen volume, sperm concentration, total sperm count,
rs p-Valueb rs p-Valueb rs p-Valueb sperm progressive motility, total motile spermatozoa motility, mor-
phology, and sperm viability were 2.9 mL, 27.8 million sperms/mL,
BDE 28 −0.074 0.527 0.060 0.607 0.182 0.116
BDE 47 0.185 0.109 0.288* 0.012 0.712** 0.000
66.4 million, 35.6% motile sperm, 57.1% motile sperm, 4.0% normal
BDE 100 0.008 0.948 0.080 0.494 0.454** 0.000 sperm, and 75.0% survival, respectively. The qualified rates of these
BDE 99 0.363** 0.001 0.522** 0.000 0.650** 0.000 semen parameters were only 52.60%–84.20% based on the World Health
BDE 154 0.469** 0.000 0.492** 0.000 0.362** 0.001 Organization reference values (WHO, 2010) (Table S2). Except for
BDE 153 0.450** 0.000 0.652** 0.000 0.496** 0.000
semen volume, the median values of all these semen parameters were
BDE183 0.327** 0.004 0.451** 0.000 0.112 0.336
BDE 209 0.095 0.415 0.094 0.417 0.190 0.100 lower than those for fertile men (20–45 years, n = 1213) in Guangdong
∑PBDEs 0.286* 0.012 0.279* 0.015 0.502** 0.000 Province, and it was the same for the qualified rate according to the
PBB153 0.685** 0.000 −0.047 0.684 −0.150 0.195 WHO criteria. The average sperm concentration of healthy men aged
Bold values statistically significant at p b 0.05. 20 to 60 in China ranged from 66.8 to 73.3 million sperms/mL (n =
a
p-Values were derived from Spearman test 998) (Zhu et al., 2011). The sperm concentrations in this study area
b
*p b .05 and **p b .01 indicates significant correlation among semen, serum and indoor were, therefore, also lower than those in healthy Chinese men. More-
dust.
over, as shown in Table S3, the levels of thyroid and reproductive

Table 4
Summary statistics of semen quality, reproductive and thyroid hormone (n = 76).a

Unit Mean ± SD Range Median Reference values

Semen quality WHO reference

Semen volume mL 2.98 ± 1.5 0.55–7.03 2.9 ≥1.5
Sperm concentration 106 sperm/mL 39.94 ± 43.56 0–255.8 27.8 ≥15
Total sperm count 106 sperm 119.83 ± 145.41 0–652.16 66.4 ≥39
Sperm motion WHO reference
Sperm progressive motility % 36.27 ± 19.06 0–80 35.6 ≥32
Total motile spermatozoa % 57.44 ± 23.11 0–97.1 57.1 ≥40
LIN % 42.93 ± 13.86 0–67.5 43.8 /
STR % 68.47 ± 16.81 0–91.3 70.7 /
WOB % 60.49 ± 14.71 0–79 62.9 /
ALH μm 1.92 ± 0.61 0–3.5 2.0 /
BCF Hz 7.37 ± 2.56 0–10.6 8.4 /
Sperm morphology WHO reference
Rate of normal morphology % 3.57 ± 2.41 0–10 4.0 ≥4
Sperm viability (%) % 71.01 ± 22.12 0–99 75.0 ≥58
Reproductive hormone Reference range
T nmol/L 18.26 ± 9.38 3.58–49.13 15.6 8.64–29.0
LH mIU/mL 5.42 ± 3.3 1.23–22.6 4.5 1.70–8.60
FSH mIU/mL 5.9 ± 3.66 0.51–20.56 5.0 1.5–12.4
E2 pg/mL 24.13 ± 12.62 2.5–65 20.5 25.8–60.7
Thyroid hormone Reference range
fT3 pmol/L 5.15 ± 1.25 1.76–14.04 5.1 3.10–6.80
fT4 pmol/L 17.09 ± 3.33 5.81–32.89 16.8 12.0–22.0
TT3 nmol/L 1.73 ± 0.37 0.98–4.02 1.7 1.30–3.10
TT4 nmol/L 90.63 ± 22.01 1.28–165.9 92.5 66.0–181.0
TSH μIU/mL 1.59 ± 0.88 0.01–4.45 1.4 0.27–4.20
a
Abbreviations: WHO, World Health Organization; LIN, Linearity of sperm; STR, Straightness of sperm; WOB, Wobble of sperm; ALH, Amplitude of sperm lateral head displacement;
BCF, Beat-cross frequency of sperm; T, Testosterone; LH, Luteinizing hormone; FSH, Follicle-stimulating hormone; E2, Estradiol; fT3, Free triiodothyronine; fT4, Free thyroxine; TT3, Total
triiodothyronine; TT4, Total thyroxine; TSH, Sensitivity thyroid-stimulating hormone.
 Y. Yu et al. / Science of the Total Environment 671 (2019) 1017–1025 1021

Table 5
Results of multiple regression analyses of associations among semen quality, hormone levels and PBDE congeners level in semen.

Parameters β 95%CI p-Value PBDE congeners level

Semen quality
Semen volume −0.381 (−0.563, −0.199) 0.000 BDE47 in semen
Semen volume −0.381 (−0.583, −0.178) 0.000 BDE153 in semen
Sperm concentration −0.331 (−0.552, −0.110) 0.004 PBB 153 in semen
Total sperm count −0.362 (−0.627, −0.096) 0.008 BDE153 in semen
Rate of normal morphology −0.255 (−0.498, −0.011) 0.041 BDE 99 in semen
LIN −0.386 (−0.637, −0.136) 0.003 BDE153 in semen
STR −0.432 (−0.678, −0.187) 0.001 BDE153 in semen
BCF −0.385 (−0.636, −0.134) 0.003 BDE153 in semen

Reproductive hormone
T 0.278 (0.054, 0.501) 0.016 BDE183 in semen
T 0.238 (0.017, 0.458) 0.035 PBB153 in semen
LH 0.306 (0.074, 0.538) 0.010 PBB153 in semen
E2 0.337 (0.124, 0.550) 0.002 PBB153 in semen
E2 −0.330 (−0.562, −0.098) 0.006 BDE154 in semen
T:LH ratio 1.216 (0.761, 1.656) 0.000 BDE209 in semen
T:LH ratio −1.122 (−1.572, −0.658) 0.000 ∑PBDEs in semen
T:LH ratio 0.343 (0.105, 0.584) 0.005 BDE154 in semen
T:LH ratio −0.267 (−0.500, −0.031) 0.027 BDE28 in semen
E2:T ratio 0.251 (0.053, 0.450) 0.014 BDE28 in semen
E2:T ratio −0.276 (−0.487, −0.066) 0.011 BDE 47 in semen
E2:T ratio −0.204 (−0.395, −0.013) 0.037 BDE154 in semen

Thyroid hormone
TSH −0.304 (−0.524, −0.083) 0.008 PBDEs in semen
TSH −0.285 (−0.517, −0.052) 0.017 BDE154 in semen

hormones in the serum of 90.8%–96.1% and 90.8%–96.1% of the partici- serum BDE47 levels. The levels of BDE47 in the serum also had a signif-
pants, respectively, were within the recommended reference range pro- icant negative correlation with the aromatase activity (E2:T ratios).
vided by the hospital. Among the thyroid hormones, fT3 levels negatively correlated with

3.4. Association among semen quality, hormones, and exposure to PBDEs Table 6
Results of multiple regression analyses of associations among semen quality, hormone
3.4.1. PBDE congeners in semen levels and PBDE congeners level in serum.
We observed a negative association of semen volume with levels of Parameters β 95%CI p-Value PBDE
BDE47 (β = −0.381, 95% CI: −0.563, −0.199, p b .001) and BDE153 (β congeners
= −0.381, 95% CI: −0.583 to −0.178, p b .001) in the semen (Table 5). level
Furthermore, negative association was observed among a) semen Semen quality
PBB153 and sperm concentration; b) semen BDE99 and rate of normal (−0.493, BDE 153 in
Total sperm count −0.247 0.048
morphology; c) semen BDE153 and total sperm count, linearity (LIN), −0.002) serum
straightness (STR), and beat-cross frequency (BCF). Rate of normal (−0.220, BDE183 in
−0.121 0.017
morphology −0.022) serum
The levels of PBB153 in semen had significant positive correlation BDE 153 in
with the levels of reproductive hormones (including T, LH, E2), and it LIN −0.364 (−0.589,-0.139) 0.002
serum
was the same for BDE183 and T. However, significant inverse associa- (−0.581, BDE154 in
tion was observed between the levels of BDE154 and E2. The function STR −0.358 −0.134) 0.002 serum
(−0.570, BDE153 in
of Leydig cells, which was measured as the ratio of T:LH (Meeker
WOB −0.341 −0.113) 0.004 serum
et al., 2011), was positively correlated with the levels of BDE209 and (−0.485, BDE153 in
BDE154, but was negatively correlated with the levels of ∑PBDEs and BCF −0.250 −0.016) 0.037 serum
BDE28 in semen. The aromatase activity measured as the ratio of E2:T
Reproductive hormone
was positively correlated with the level of BDE28 and negatively corre- BDE153 in
lated with those of BDE47 and BDE 154 in semen. Among the thyroid T 0.285 (0.058, 0.513) 0.015 serum
hormones, only the TSH levels had a significant negative correlation PBB153 in
with the levels of ∑PBDEs and BDE154 in semen. LH 0.240 (0.007, 0.472) 0.043 serum
PBB153 in
E2 0.331 (0.121, 0.542) 0.002 serum
3.4.2. PBDE congeners in serum (−0.532, BDE47 in
Similar to PBDE congeners in semen, inverse associations between E2 −0.285 −0.038) 0.024 serum
BDE154 and BDE153 in serum and semen quality were observed in (−0.554, BDE47 in
E2:T ratio −0.319 −0.085) 0.008 serum
these participants (Table 6). We found inverse associations between
BDE153 and total sperm count, as well as between BDE183 and normal Thyroid hormone
morphology. In addition, serum BDE154 and BDE153 levels were nega- (−0.659, ∑PBDEs in
fT3 −0.413 −0.167) 0.001 serum
tively correlated with sperm motility parameters (including LIN, STR,
BDE47 in
WOB, and BCF). fT3 0.360 (0.087, 0.632) 0.010 serum
Among the reproductive hormones, significant positive associations BDE183 in
were observed between testosterone (T) and BDE153 levels, as well as TT3 0.235 (0.012, 0.457) 0.039 serum
between LH and PBB153 levels. Furthermore, the E2 levels positively (−0.478, BDE183 in
TT4 −0.256 −0.033) 0.025 serum
correlated with those of PBB153, but inversely correlated with the
1022 Y. Yu et al. / Science of the Total Environment 671 (2019) 1017–1025
Table 7
Results of multiple regression analyses of associations among semen quality, hormone levels and PBDE congeners level in dust.
Parameters β 95%CI
Semen quality
Semen volume −0.369
Sperm concentration −0.315
Total sperm count −0.397
sperm progressive motility −0.311
Total motile spermatozoa −0.306
Sperm viability (%) −0.328
LIN −0.714
STR −0.404
WOB −0.457
BCF −0.428
Reproductive hormone
LH −0.251
Thyroid hormone
TSH −0.296
the levels of ∑PBDEs in the serum, but were positively correlated with
serum BDE47 levels, and serum BDE183 levels were positively corre-
lated with those of TT3 but negatively correlated with fT3 levels.
3.4.3. PBDE congeners in dust
Significant negative correlation was observed between the levels of
BDE183 in dust and semen volume, sperm concentration, and total
sperm count (Table 7). The levels of ∑PBDEs in the dust samples
were also inversely related with the progressive motility [(A + B)%] of
sperms, total motile spermatozoa [(A + B + C)%], and sperm viability
(%). Moreover, the levels of BDE153, BDE209, and ∑PBDEs in dust sam-
ples were negatively correlated with sperm motility parameters (in-
cluding LIN, STR, WOB, and BCF).
Among the serum hormones, LH and BDE154 showed significant
negative correlation, and it was the same for TSH and BDE99. However,
there was no significant statistical correlation between the PBDE conge-
ners in dust and other reproductive hormones or thyroid hormones.
Our results showed that the levels of BDE153 in semen and serum
were negatively correlated with the total sperm count. Only the levels
of PBDE congeners in the dust samples were significantly correlated
with sperm progressive motility, total motile spermatozoa, and sperm
viability, but no significant correlation was found between such param-
eters and the levels of PBDE congeners in serum and semen.
Most of the sperm motility parameters were inversely correlated
with the levels of PBDE congeners (mainly BDE153 and BDE154) in
serum, semen, and dust samples, suggesting that PBDE congeners had
a significant effect on sperm motility. Especially, BDE153 levels in
serum and semen had inverse correlation with LIN, STR, and BCF.
The levels of thyroid hormones were closely associated with those of
PBDE congeners in the serum, whereas the levels of PBDE congeners in
semen and dust were negatively associated with the TSH levels. The
levels of reproductive hormones were associated with those of PBDE
congeners in serum and semen samples, but not with the levels of
PBDE congeners in dust samples. The aromatase activity (E2:T ratio)
might be affected by BDE47 levels in semen and serum. Only the PBDE
congeners in semen were significantly correlated with the function of
Leydig cells, whereas the levels of PBDE congeners in serum and dust
samples were not significantly correlated with it. Therefore, this dem-
onstrates that PBDE congeners in semen might affect the male repro-
ductive function by changing the T:LH and E2:T ratios. Structure
diagrams linking reproductive hormones and semen quality parameters
with PBDE levels in semen and in serum were shown in Fig. S2.
4. Discussion
PBDEs are a kind of endocrine disruptors that can interfere with thy-
roid and reproductive hormones (Legler and Brouwer, 2003). The
p-Value PBDE congeners level
(−0.592, −0.145) 0.002 BDE 183 in dust
(−0.573, −0.057) 0.017 BDE183 in dust
(−0.651, −0.144) 0.003 BDE183 in dust
(−0.552, −0.070) 0.012 ∑PBDEs in dust
(−0.552, −0.060) 0.015 ∑PBDEs in dust
(−0.571, −0.084) −0.009 ∑PBDEs in dust
(−1.100, −0.329) 0.000 BDE 153 in dust
(−0.637, −0.171) 0.001 BDE 153 in dust
(−0.686, −0.228) 0.000 BDE 153 in dust
(−0.660, −0.196) 0.000 ∑PBDEs in dust
(−0.501, −0.001) 0.049 BDE154 in dust
(−0.538, −0.054) 0.017 BDE99 in dust
results of our previous pilot study suggested that exposure to PBDEs
from house dust might have adverse effects on the quality of semen
(Yu et al., 2018). The present study describes a comprehensive assess-
ment of the effects of PBDEs on male reproductive health, wherein we
measured the levels of PBDE congeners in house dust, serum, and
semen, as well as the levels of thyroid and reproductive hormones in
serum. Moreover, one of our most important purposes was to explore
whether PBDE congeners levels in semen and serum have different ef-
fects on semen parameters, as well as their possible influence mecha-
nism. These results may offer insights into the potential mechanism
underlying the effects of PBDEs.
4.1. Association among the concentrations of PBDEs in serum, semen, and
indoor dust
For BDE99, BDE154, BDE153, and ∑PBDEs, Spearman correlation
coefficients (rs) suggested significant association between the concen-
trations of PBDEs in any two matrices of indoor dust, serum, and
semen (p b .05), indicating close correlation between the levels of
PBDE congeners in the human body and indoor dust accumulation. A
significant correlation between dust and serum concentrations of pre-
dominant PBDE congeners (namely BDE47, 99, and 100) has been re-
ported in previous studies (Karlsson et al., 2007; Johnson et al., 2010).
Bramwell et al. (2016) pointed out that the levels of penta-BDE conge-
ners in human generally correlated strongly with those in dust, based
on studies published over the years (up to 2015) on the relationship be-
tween internal dose of PBDE congeners and indoor dust and dietary in-
take. However, there have been few studies on the correlation among
the concentrations of PBDEs in semen, serum, and indoor dust.
4.2. Effect of exposure to PBDEs on semen quality
In this study, serum BDE153 and semen BDE153 were observed to
have a significant negative association with total sperm count, and
there was a significant correlation between the BDE153 content in
serum and semen (rs = 0.450, p b .001), suggesting that the internal
dose of BDE153 might affect the total sperm count. The presence of
BDE153 in serum and in semen both had significant negative associa-
tion with sperm motility. Previous studies have shown that serum
PBDEs may interfere with the sperm quality in the general population.
Akutsu et al. (2008) reported that the level of BDE153 in human
serum was negatively correlated with sperm concentration and testicu-
lar size. Abdelouahab et al. (2011) found that the levels of BDE47,
BDE100, and ∑PBDE in adult serum were negatively correlated with
sperm motility, but were not significantly correlated with other semen
parameters. Moreover, our results demonstrate that predominant
PBDE congeners and ∑PBDE in indoor dust might have a significant
 Y. Yu et al. / Science of the Total Environment 671 (2019) 1017–1025
inhibitory effect on sperm count and motility. However, to the best of
our knowledge, the effect of dust PBDEs on male semen quality has
not been previously reported.
Although experiments on animals revealed that environmentally-
relevant perinatal exposure to PBDEs can result in long-lasting changes
in DNA methylation in sperms (Suvorov et al., 2018), no in-depth study
on the effect of PBDEs on human reproductive function have been con-
ducted. Our results show that PBDE congeners might play a significant
role in determining the quality of semen, but the mechanism underlying
the effects need further investigations.
4.3. Effect of exposure to PBDEs on reproductive hormones
We found a positive association between the levels of BDE153 and T
in serum, which is consistent with the results of another study wherein
it was reported that serum BDE47 levels in adult males in USA were pos-
itively associated with the T levels (Turyk et al., 2008). Our results also
demonstrate that the presence of BDE154 in dust was negatively corre-
lated with the serum luteinizing hormone (LH) level, which is consis-
tent with the results of a previous study in which inverse correlations
between PBDE congeners levels in dust and LH and FSH were observed
(Meeker et al., 2009). In addition, levels of PBB153 and T, LH, and E2 in
semen were significantly correlated, indicating that the presence of
PBB153 in semen affects the levels of reproductive hormones. More-
over, there was a significant correlation between the levels of BDE183
and T, as well as between the levels of BDE154 and E2 in semen.
There are only a few reports on the effects of PBDE congeners on
human reproductive hormones, especially with regard to the mecha-
nism underlying such effects. Nevertheless, animal experiments have
shown that PBDE congeners have certain estrogenic effects. Mercado-
Feliciano and Bigsby (2008a, 2008b) found that DE71, a mixture of
PBDEs (including BDE47, 49, 100, and 154), did not bind to estrogen re-
ceptor in rats, but had a weak effect on the induction of estrogen, and
their hydroxyl metabolites exhibited good binding with estrogen
receptor-α (ER-α), showing an estrogen induction effect. In vivo exper-
imental studies also showed that PBDEs and their metabolites interfered
more significantly with androgen than with estrogen (Hamers et al.,
2006), and might have endocrine-disrupting effects through nuclear
hormone receptors (Kojima et al., 2009). Pubertal exposure to DE-71
delayed the puberty and prefabricated androgen-dependent organ de-
velopment in male Wistar rats, suggesting that it induced the androgen
metabolism or it acts as an androgen receptor (AR) antagonist (Stoker
et al., 2005).
Leydig cells are the primary source of androgen, and can synthesize
and secrete T (Eladak et al., 2018). Furthermore, this process is regu-
lated by hypothalamus and pituitary, and is easily affected by some
EDCs (Zhao et al., 2011). Our results show that the levels of PBDEs in
semen and serum were positively correlated with T and LH, but only
the level of PBDEs in semen was significantly correlated with the func-
tion of Leydig cells. This suggests that semen PBDEs might directly affect
the function of Leydig cells following the secretion of testosterone,
thereby, altering the level of T in the serum.
Aromatase (CYP19) can convert T to E2 in the gonads and in other
tissues; therefore, the balance of estrogen and androgen in the body
can be altered by promoting or inhibiting the activity of aromatase
(Eladak et al., 2018). Our results demonstrated that the presence of
BDE47 in serum and in semen both was negatively correlated with the
activities of the aromatase, as was the case for semen BDE154. In con-
trast, the presence of semen BDE28 was positively correlated with aro-
matase activity. These results are consistent with those of in vitro
studies on the inhibition and induction of aromatase activity by PBDEs
and their hydroxylated metabolites. Canton et al. (2005) reported that
the presence of BDE28 and BDE38 in serum caused a minor induction
of the steroidogenic enzyme, aromatase (CYP19), whereas BDE206
and 209 could inhibit the activity of this enzyme. Moreover,
1023
hydroxylated BDE47 metabolites could stimulate the aromatase activity
(Karpeta et al., 2013).
Our results demonstrate that the presence of PBDE congeners in
semen strongly correlates with aromatase activity and Leydig cell func-
tion, whereas only the serum BDE154 correlates with aromatase activ-
ity, suggesting that PBDE congeners in semen are more responsive to
the balance of reproduction hormones than that in serum.
4.4. Effect of PBDE exposure on thyroid function
In this study, we observed a significant correlation between fT3 and
serum BDE47 and ∑PBDEs levels, as was the case for relationship be-
tween serum BDE183 and fT3 and TT4. Our results are consistent with
those of other studies wherein TT4 was shown to be inversely associated
with the levels of PBDE congeners (namely BDE47, BDE99, and
∑PBDEs) in serum; they, however, differ from other results which
showed that serum concentrations of PBDEs were not strongly associ-
ated with those of other thyroid hormones (Abdelouahab et al., 2011;
Makey et al., 2016). However, Turyk et al. (2008) found that associa-
tions of PBDE congeners (namely BDE47, BDE99, BDE100, and
BDE153) with thyroid hormones varied; for instance, PBDEs were posi-
tively related to T4 whereas they were inversely related to TT3 and TSH.
Additionally, we found that the levels of BDE99 in dust were inversely
associated with serum TSH in our study population, but this result dif-
fered from those of another study where a positive relation was ob-
served between the levels of PBDEs in dust and the level of fT4 in the
serum of male individuals whereas no effect was observed on TSH and
TT3 (Meeker et al., 2009). Moreover, our results show that the levels
of both BDE154 and ∑PBDEs in semen were inversely correlated with
those of TSH, suggesting that the presence of PBDEs in semen may
also interfere with thyroid hormones.
As the molecular structures of PBDE congeners are very similar to
those of thyroid hormones T3 and T4 (Meerts et al., 2000; Birnbaum
and Staskal, 2004), they can stimulate, inhibit, or mimic the biological
effects of thyroid hormones, and can, thereby, disrupt the function of
thyroid hormones even at low doses (Jacobson et al., 2016; Zheng
et al., 2017). Results of animal and in vitro experiments have shown
that PBDEs may indirectly disrupt the endocrine system by interacting
with nuclear receptors other than hormone receptors. It has, therefore,
been hypothesized that PBDEs can interfere with the hypothalamic–
pituitary–thyroid axis (Lema et al., 2008; Kojima et al., 2009).
It has been reported that the level of PBDEs in serum of residents in
high-exposure areas was higher than that in low-exposure areas. Bi
et al. (2007) found that the median concentration of PBDE in serum
from residents of an e-waste dismantling region (Guiyu, South China)
was three times higher than that in control areas. Sjodin et al. (1999) re-
ported that the serum PBDE levels in a group of computer dismantlers
were about eight times higher than those in a reference group of hospi-
tal cleaners. Studies have also shown that there was a tendency toward
lower TT4 and higher fT3 with increasing PBDE in young children
(Jacobson et al., 2016). Our study also indicated that PBDE levels in
serum and semen were significantly correlated with semen quality
and hormone levels. Therefore, we should pay close attention to the re-
productive health of resident from high exposure areas such as e-waste
recycling area.
To our knowledge, this is the first study to explore the correlation
among the levels of PBDE congeners in semen and reproductive and
thyroid hormones. These results indicated that PBDE congeners in
serum, semen, and dust all were significantly correlated with reproduc-
tive hormones, thyroid hormones, and semen quality in the study area.
Serum PBDE congeners might bind to reproductive hormone receptors
or thyroid hormone receptors and interfere with the regulation of
these hormones, resulting in disordering of the hypothalamus–
pituitary–testicular axis, which affects spermatogenesis and sperm via-
bility. Moreover, PBDE congeners could also enter semen, directly af-
fecting the motility of sperms. However, there are certain limitations
1024 Y. Yu et al. / Science of the Total Environment 671 (2019) 1017–1025
of the present study. The exposure levels to PBDE congeners were rela-
tively high in the e- waste dismantling area and in its vicinity; the effects
of exposure to low concentrations of PBDE congeners remain unknown.
Moreover, the sample size was also relatively small in the present study.
In the literature on andrology, semen quality and blood hormone levels
have been discussed extensively with regard to seasons (Chen et al.,
2003; De Giorgi et al., 2015); as such, one limitation of our work is
that we did not include season as a covariate for semen quality and hor-
monal levels. The mechanisms underlying the observed effects need
further experimental verification, which should include experimental
studies on the operative mechanisms at molecular and genetic levels,
and further large-scale epidemiological investigations.
5. Conclusions
In the present study, we systematically studied the effects of PBDE
congeners, ingested through indoor dust, on male reproductive health
in an e-waste dismantling area in South China. Regression analyses re-
vealed that the concentrations of PBDE congeners in serum and
semen, which were strongly associated with the concentrations in in-
door dust, were significantly correlated with the semen quality, and
the levels of reproductive and thyroid hormones. Our results suggest
that PBDEs might affect the main components of male reproductive sys-
tem, including testis, thyroid and adrenal glands, indicating that the ef-
fect of PBDEs on reproductive health should not be neglected. It is
necessary to perform further experimental studies on deciphering the
mechanisms operative in animals at molecular and genetic levels, and
to explore the mechanisms underlying the effects of PBDE congeners
on hypothalamus–pituitary–testicular axis dysfunction, which can pro-
vide deeper insights into the mechanisms responsible for the effects of
PBDE congeners on the male reproductive health.
Acknowledgments
This project was sponsored by the Joint fund of the National Natural
Science Foundation of China (NSFC) and People's Government of
Guangdong Province (Grant No. U1401233), National Key Research
and Development Program (Grant No. 2018YFC1801105), and
Qingyuan Science and Technology Planning Project (Grant No.
170807091720894).
Appendix A. Supplementary data
Supplementary data to this article can be found online at https://doi.
org/10.1016/j.scitotenv.2019.03.319.
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