Ecotoxicology and Environmental Safety 208 (2021) 111642

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Ecotoxicology and Environmental Safety

journal homepage: www.elsevier.com/locate/ecoenv

Research Paper

Effects of low-level mercury exposure on brain-derived neurotrophic factor

in preschool children
Can-Can Zhou 1, Hui Fu 1, Guo-Yan Zhang , Jia-Wei Ma , Min Ni , Dong-Jie Li , Fu-Ming Shen *,
Fang Huang *
Department of Pharmacy, Shanghai Tenth People’s Hospital, Tongji University School of Medicine, Shanghai, China

A R T I C L E I N F O A B S T R A C T

Edited by: Professor Bing Yan Objectives: Mercury (Hg), a ubiquitous heavy metal, could affect the neurodevelopment of the children, however,
these associations are still equivocal. Brain derived neurotrophic factor (BDNF) plays an essential role in the
Keywords: central nervous system development in children. This study aimed to investigate the effects of low-level mercury
Mercury exposure on serum BDNF levels and the influence of sex and dietary intake on these relationships in children.
BDNF
Methods: In this cross-sectional study, a total of 541 pre-school children were recruited, the blood mercury
Sex
concentrations and serum BDNF levels were measured. The background information on demographic charac­
Children
teristics and dietary habits of the children was collected through questionnaires. Multivariable linear models
after adjustment for potential confounders were used to evaluate the associations between mercury exposure and
levels of BDNF in children.
Results: The GMs of blood mercury concentrations and serum BDNF levels were 1.06 μg/L, 20.4 ng/mL,
respectively. A significant positive association between blood mercury concentrations and serum BDNF levels
was found. After stratification by sex, the blood mercury concentrations in children were positive associated with
serum BDNF levels in girls but not in boys. However, these associations were attenuated when we further
adjusted the children’s dietary intake variables.
Conclusions: Our findings suggest that low-levels of mercury exposure may have sex-specific effects on BDNF
levels in young children and that dietary intake may be potential confounders in these relationships. However,
further studies are warranted to investigate the role of BDNF in the effects of mercury on neurodevelopment.

1. Introduction also observed a negative association between prenatal mercury exposure

Mercury as one of the environmental pollutants mainly comes from
atmospheric deposition and the primary organic form of mercury in the
food chain is methylmercury (MeHg), which is mainly transformed from
inorganic mercury by microorganism (Zupo et al., 2019). Exposure to
MeHg could result in learning and behavioral disorders, cardiovascular
diseases, and reproductive damages (Chen et al., 2020b; Santos-Lima
et al., 2020; Yao et al., 2020). Due to the higher absorption and lower
detoxification abilities for environmental toxicants as well as the sus­
ceptibility of the developing nervous system, children are more sensitive
to environmental MeHg exposure (Dufault et al., 2009). The conclusions
of the World Trade Center cohort study showed that increased cord
blood mercury was negatively associated with infant neuro­
developmental scores (Lederman et al., 2008). A study in Tohoku, Japan
and neonatal neurodevelopment (Suzuki et al., 2010). However, several
other researches about the influences of mercury exposure on childhood
neurodevelopment have found conflicting results. One study conducted
in a rural area of Laizhou cannot find the adverse effects of mercury
exposure on infant neurodevelopment (Hu et al., 2016). Another study
also showed that hair mercury levels were not related to neuropsycho­
logical functions (Surkan et al., 2009). Furthermore, a significantly
positive relationship between prenatal mercury exposure and infant
neurobehavioral development was also observed (Wu et al., 2014).
Therefore, additional evidences are still needed to clarify the influence
of mercury exposure on neurodevelopment in children.
Brain derived neurotrophic factor (BDNF), a member of the neuro­
trophin family, plays an important role in regulating neuronal survival,
differentiation, and synaptic plasticity. In the nervous system, BDNF can

* Corresponding authors.
E-mail addresses: fumingshen@tongji.edu.cn (F.-M. Shen), hazel13818753472@126.com (F. Huang).
1
These authors contributed equally to this work.

https://doi.org/10.1016/j.ecoenv.2020.111642
Received 21 September 2020; Received in revised form 4 November 2020; Accepted 8 November 2020
Available online 13 November 2020
0147-6513/© 2020 The Authors. Published by Elsevier Inc. This is an open access article under the CC BY-NC-ND license
(http://creativecommons.org/licenses/by-nc-nd/4.0/).
C.-C. Zhou et al.
be mediated by the activation of the tropomyosin receptor kinase B
(TrkB) pathway (Che et al., 2020). During development of the early
nervous system, increased BDNF could promote the survival, growth,
and differentiation of new neurons (Chen et al., 2020a). Previous studies
also suggested that the elevated peripheral BDNF levels are positively
associated with brain BDNF concentrations and neurocognitive func­
tions (Klein et al., 2011; Chen et al., 2020a). The decreased levels of
BDNF have been found significantly associated with neuro­
developmental disorders, symptoms of attention deficit hyperactivity
disorder (ADHD), and depression (Wang et al., 2019a, 2019b; Lee et al.,
2020). Moreover, it was also reported that serum BDNF level was a
significant factor related to decreased hippocampus volume in aging
(Erickson et al., 2010). Nevertheless, the influence of mercury exposure
on serum BDNF levels in children is still required to be investigated.
MeHg-contaminated fish is the most significant source of mercury
exposure for humans. Several studies have regarded fish consumption as
an important risk factor for children mercury exposure (Valent et al.,
2013; Basu et al., 2014; Burch et al., 2014). Food and Drug Adminis­
tration in the U.S. also proposed that pregnant women and young chil­
dren should limit their fish intake due to the accumulation of mercury
(Evans, 2002). However, the nutrients of long-chain polyunsaturated
fatty acids (LCPUFA), docosahexaenoic acid (DHA), and other essential
nutrients from fish were defined to be able to promote neuro­
development and improve memory functions (Das, 2000; Domingo
et al., 2007). Furthermore, childhood neurodevelopment might be
influenced by many other factors, among which dietary supplement of
cow-milk and multiple micronutrient supplementation (vitamins and
essential trace elements) are generally considered to be beneficial for
children’s cognitive development (Arellanes et al., 2020; D’Alonzo et al.,
2020; Li et al., 2020). It was also reported that nutritional status might
play a possible confounding role in the relationships between mercury
exposure and neurodevelopmental outcomes in children (Davidson
et al., 2008b). Therefore, these factors may be potential covariates and
may influence the conclusions of the relationships between mercury
exposure and BDNF in children. Nevertheless, limited studies considered
these factors and adjusted them during analysis.
Gender-related differences in the neuropsychological effects of
mercury exposure have been evaluated in several studies. However, the
results of sex interactions are still inconsistent. A cohort study conducted
in the Faroe Islands reported the negative associations between prenatal
mercury exposure and motor functions of the children aged 7 years in
males but not in females (Grandjean et al., 1998). Cord blood mercury
levels were also found to be inversely associated with the neonatal
behavioral ability only in boys (Gao et al., 2007). Furthermore, Myers
et al. observed that boys at age 9 years were more sensitive to
mercury-induced scholastic achievement decline than girls (Myers et al.,
2009). In contrast to these findings, higher susceptibility of female to
mercury exposure was also reported. Llop et al. (2012) demonstrated a
statistically significant inverse association between mercury exposure
and psychomotor development only among female infants. A negative
association between blood mercury levels and intelligence quotient (IQ)
scores in females was also observed (Myers et al., 2009). Therefore,
sex-specific differences may play an important role in childhood mer­
cury exposure and it is imperative to examine these factors.
Therefore, the objective of the current study was to investigate the
effects of mercury exposure on BDNF in children and to assess the sex-
specific differences in these associations after adjusting for potential
confounders.
2. Materials and methods
2.1. Study subjects and recruitment
This cross-sectional study was conducted in Wujiang city, near Taihu
Lake in Jiangsu China. Briefly, a total of 700 invitations were sent out to
the parents of the children in the kindergartens and were eligible to be
2
Ecotoxicology and Environmental Safety 208 (2021) 111642
recruited in our study if they had no disorders which might affect neu­
rodevelopment such as meningitis, traumatic brain injury, or epilepsy.
Meanwhile, further exclusion of the participants who did not finish the
questionnaires and whose parents refused to provide biological samples,
then, 541 children were finally included for this study. The study was
approved by the Ethics Committee of Shanghai Jiao Tong University
School of Medicine and all study participants (from the children’s par­
ents) provided written informed consent.
2.2. Data collection
The background information of the children was collected through
questionnaires. The parents completed the questionnaire and the nurses
retrieved them. Information on demographic characteristics and dietary
habits including children’s age, sex, BMI, maternal education, house­
hold monthly income, cow-milk intake, multivitamin supplements and
the frequency (vitamin intake), mineral supplements and the frequency
(mineral intake), fresh-water fish consumption, salt-water fish con­
sumption, shellfish consumption, and second-hand smoking were
collected.
2.3. Samples collection and blood mercury measurements
Venous blood was collected into EDTA-NA2-treated collection tubes
and the samples were stored at − 80 ℃ until analysis. The mercury
concentrations were detected using a Direct Mercury Analyzer 80 (DMA-
80, Milestone Inc., Bergamo, Italy). Detailed procedures and quality
control measures were conducted as we previously described (Wang
et al., 2019a, 2019b; Gao et al., 2018). Briefly, to confirm the accuracy
of the measurements, the procedural blanks were detected and standard
reference materials were used (heavy metal blood, Kaulson Labora­
tories, Inc., NJ, USA). The limit of detection (LOD) for mercury in blood
was 0.01 μg/L. The determination of mercury in all the samples were
performed in duplicate, and none of the samples in our study had
mercury levels less than the LOD.
2.4. Serum BDNF measurements
Venous blood was collected, and then the serum samples were
separated by centrifugation at 4 ℃ at 1,800 g for 10 min and serum
BDNF levels were detected by ELISA kits according to its instructions.
Briefly, the serum samples were diluted, added to a flat-bottom 96-well
plate (100 μl) coated with an anti-BDNF monoclonal antibody, and then
incubated for 90 min at 37 ℃. The 100 μl of HRP conjugate working
solution was added to each well and incubated for 1 h at 37 ℃ after
washing the plates with 1 × wash solution three times. One hour later,
the plate was washed with 1 × wash solution three times, and 90 μl of
substrate solution was added to each well. After the incubation for 15
min at 37 ℃, 50 ul of stop solution finally was added and the absorbance
value of each well was immediately detected at 450 nm. All samples and
standards were assayed in duplicate.
2.5. Statistical analysis
The children included in the study are stratified by sex and the dis­
tribution of the characteristics are presented as means ± SD or as a
number (n) with a percentage (%). The difference of the age and BMI
between the boys and girls were compared by Student’s t-test. The dif­
ference of the maternal education, household monthly income, cow-
milk intake, vitamin intake, mineral intake, fresh-water fish consump­
tion, salt-water fish consumption, shellfish consumption, and second-
hand smoking between the boys and girls were compared by Chi-
Square test. The distributions of the blood mercury concentration and
serum BDNF levels in all the children, boys, and girls were represented
by geometric means and 95% CI, arithmetic means, and SDs and quar­
tiles. Considering that dietary habits might be associated with both
C.-C. Zhou et al. Ecotoxicology and Environmental Safety 208 (2021) 111642

blood mercury concentration and serum BDNF levels, the relationships mean BMI was 15.2. More than 60% of the participating families the
between dietary habits and blood mercury concentration or serum BDNF monthly household income was > 15,000. The proportion of higher
levels were investigated by linear regression analyses. The associations family income (> 15,000) among the boys tended to higher than that
between mercury concentrations in the blood and serum BDNF levels in among the girls (64.1% vs. 56.1%, P = 0.06). The frequency of vitamin
preschool children were analyzed by multivariable linear regression consumption and sell-fish consumption of the girls also tended to be
models. The mercury concentrations were also categorized into tertiles higher than these of the boys (43.0% vs. 34.0%, P = 0.01; 47.1% vs.
in the linear models and the lowest tertile was used as the reference 38.7%, P = 0.01). However, no differences in age, BMI, maternal edu­
group. The dietary habits items of the children associated with both cation, cow-milk intake, trace elements consumption, water-fish con­
blood mercury concentrations and serum BDNF levels were selected as sumption, sea-fish consumption, and passive smoking were found
potential confounders and those only associated with serum BDNF levels between the boys and girls.
as covariates. For the analyses, three different models were used: Model The GMs of blood mercury concentrations in all the children, boys
1, adjusted for age, sex, maternal education, household monthly income; and girls were 1.06 μg/L (range: 1.00, 1.12 μg/L), 1.08 μg/L (range:
Model 2, adjusted for Model 1 and the fish intake items associated with 1.00, 1.16 μg/L) and 1.03 μg/L (range: 0.94, 1.13 μg/L), respectively
serum BDNF levels; and Model 3, adjusted for Model 2 and the dietary (Table 2). Approximately 50% of the blood samples from the children
intake items associated with serum BDNF levels. All the statistical ana­ had mercury levels higher than 1.00 μg/L. The GMs of serum BDNF
lyses were performed using SAS 9.4 software package (SAS Institute, levels in all the children, boys and girls were 20.4 ng/mL (range: 19.4,
Cary, NC). The significance level was set at P < 0.05, whereas 0.05 < P 21.4 ng/mL), 19.7 ng/mL (range: 18.4, 21.0 ng/mL) and 21.3 ng/mL
< 0.1 were considered borderline statistically significant. (range: 19.8, 22.9 ng/mL), respectively.
Table 3 presented the dietary supplement of nutrients frequency
3. Results items that were significantly associated with blood mercury concen­
trations or the serum BDNF levels in the total children, boys, and girls.
The main characteristics of the children included in our study are The dietary items associated with BDNF (P < 0.1) were included as
shown in Table 1. The study population included 541 pre-school chil­ covariates in the multiple linear regression.
dren with the mean age of the children was 69.3 months. The numbers of The associations of childhood mercury exposure with BDNF after
boys and girls were 318 (58.8%) and 223 (41.2%), respectively. The adjusting for potential covariates were presented in Table 4. Through
multivariable linear regression analysis, we found a significantly posi­
tive association between mercury concentrations and serum BDNF levels
Table 1
among children (β = 4.91, 95% CI = 1.20–8.63, P = 0.01) (Table 4-
Main characteristics in all the children, boys and girls.
model 1). When the variables of sea-fish and shell-fish consumption
Characteristic Total Boys (318) Girls (223) P- were included in the models (Table 4-Model 2), the coefficients
(541) value
remained virtually the same. However, when vitamins, mineral intake,
Age 69.3 ± 69.5 ± 69.1 ± 0.46 and passive smoking were further included in the models (Model 3), we
6.56 6.52 6.63
observed that the positive association between blood mercury levels and
BMI 15.2 ± 15.3 ± 15.1 ± 0.23
1.65 1.72 1.56
serum BDNF levels became borderline significant (β = 3.65, 95% CI = −
Maternal education 0.07 to 7.37, P = 0.05). After stratifying by sex, among girls, mercury
Lower than high school 238 151 87 (39.01) 0.15 concentrations were found to be significantly positively associated with
(43.99) (47.48) serum BDNF levels (β = 7.37, 95% CI = 1.46 – 13.27, P = 0.02). These
High school and higher 286 158 128
relationships remained significant after additionally adjusting for sea-
(52.87) (49.69) (57.40)
Household monthly income fish and shell-fish consumption in girls (Table 4-model 2). Addition­
(CNY) ally, when the trace elements consumption was further adjusted, the
< 10,000 212 (39.2) 114 (35.9) 98 (43.9) 0.06 associations between blood mercury concentrations and serum BDNF
> 15,000 329 (60.8) 204 (64.1) 125 (56.1)
levels became borderline significant (β = 5.75, 95% CI = − 0.16 to
Cow-milk intake
NO 216 (39.9) 125 (39.3) 91 (40.8) 0.73
Yes 325 (60.1) 193 (60.7) 132 (59.2)
Table 2
Vitamins intake 0.10
Geometric mean (GM) and percentiles of the blood mercury levels and the serum
≦ 1 times per month 337 (62.3) 210 (66.0) 127 (57.0)
≦ 3 times per week 177 (32.7) 93 (29.3) 84 (37.7) BDNF concentrations in the participants .
> 3 times per week 27 (5.0) 15 (4.7) 12 (5.3) GM Mean Percentiles
Mineral intake 0.51 (Range) (SD)
≦ 1 times per month 197 (36.4) 122 (38.4) 75 (33.6) 10 25 50 75 90
≦ 3 times per week 317 (58.6) 180 (56.6) 137 (61.4) Mercury
> 3 times per week 27 (5.0) 16 (5.0) 11 (5.0) Total 1.06 1.28 0.51 0.82 1.16 1.67 2.21
Water-fish consumption 0.40 (1.00, (0.77)
≦ 1 times per month 147 (27.2) 93 (29.3) 54 (24.2) 1.12)
≦ 1 times per week 203 (37.5) 118 (37.1) 85 (38.1) Boys 1.08 1.24 0.51 0.85 1.16 1.53 1.90
> 1 times per week 191 (35.3) 107 (33.6) 84 (37.7) (1.00, (0.69)
Sea-fish consumption 0.55 1.16)
≦ 1 times per month 191 (35.3) 118 (37.1) 73 (32.8) Girls 1.03 1.31 0.51 0.78 1.14 1.70 2.33
≦ 1 times per week 153 (28.3) 86 (27.0) 67 (30.0) (0.94, (0.83)
> 1 times per week 197 (36.4) 114 (35.9) 83 (37.2) 1.13)
Sell-fish consumption 0.09 BDNF
≦ 1 times per month 313 (57.8) 195 (61.4) 118 (52.9) Total 20.4 23.7 9.9 15.0 21.2 28.8 39.0
≦ 1 times per week 140 (25.9) 79 (24.8) 61 (27.4) (19.4, (13.40)
> 1 times per week 88 (16.3) 44 (13.8) 44 (19.7) 21.4)
Second-hand smoking 0.74 Boys 19.7 23.0 9.3 14.6 20.9 28.2 37.9
≦ 1 times per month 211 (39.0) 121 (38.0) 90 (40.3) (18.4, (12.93)
≦ 3 times per week 224 (41.4) 136 (42.8) 88 (39.5) 21.0)
> 3 times per week 106 (19.6) 61 (19.2) 45 (20.2) Girls 21.3 24.7 10.1 15.3 21.8 30.7 40.5
(19.8, (14.0)
P value was obtained from comparing the difference of characteristics between
22.9)
boys and girls

3
C.-C. Zhou et al. Ecotoxicology and Environmental Safety 208 (2021) 111642

Table 3
Dietary intake is associated with blood mercury levels or serum BDNF levels in children, boys, and girls.
Total Boys Girls

Items β (95% CI) P Items β (95% CI) P Items β (95% CI) P

Mercury Mercury Mercury

Sea-fish consumption Second-hand Sea-fish
smoking consumption
≦ 1 times per month Ref Never ref ≦ 1 times per month ref
≦ 1 times per week 0.15 (− 0.01, 0.32) 0.07 Sometimes 0.11 (− 0.09, 0.31) 0.30 ≦ 1 times per week 0.13 (− 0.10, 0.36) 0.27
≧ 1 times per week 0.18 (0.03, 0.34) 0.02 Always 0.38 (0.12, 0.63) 0.004 ≧ 1 times per week 0.21 (− 0.01, 0.42) 0.06
Cow-milk intake BDNF Cow-milk intake
NO Ref Vitamins intake NO ref
Yes − 0.14 (− 0.27, − 0.04 ≦ 1 times per month ref Yes − 0.18 (− 0.36, 0.06
0.01) 0.00)
Second-hand ≦ 3 times per week 4.05 (0.91, 7.18) 0.01 BDNF
smoking
≦ 1 times per month Ref > 3 times per week 6.35 (− 0.38, 0.06 Mineral intake
13.08)
≦ 3 times per week 0.059 (− 0.09, 0.20) 0.43 Mineral intake ≦ 1 times per month ref
> 3 times per week 0.24 (0.06, 0.42) 0.01 ≦ 1 times per month ref ≦ 3 times per week 5.00 (1.08, 8.92) 0.01
BDNF ≦ 3 times per week 1.97 (− 0.10, 4.95) 0.19 > 3 times per week 7.51 (− 1.30, 16.32) 0.09
Vitamins intake > 3 times per week 6.45 (− 0.29, 0.06 Sea-fish
13.19) consumption
≦ 1 times per month ref Sea-fish consumption ≦ 1 times per month ref
≦ 3 times per week 2.93 (0.50, 5.36) 0.02 ≦ 1 times per month ref ≦ 1 times per week 3.70 (− 0.87, 8.27) 0.11
> 3 times per week 4.24 (− 0.10, 9.48) 0.11 ≦ 1 times per week 2.05 (− 1.54, 5.63) 0.26 > 1 times per week 7.61 (3.28, 11.95) 0.001
Mineral intake > 1 times per week 4.04 (0.72, 7.36) 0.02 Sell-fish
consumption
≦ 1 times per month ref Sell-fish ≦ 1 times per month ref
consumption
≦ 3 times per week 3.26 (0.90, 5.63) 0.01 ≦ 1 times per month ref ≦ 1 times per week 3.73 (− 0.62, 8.07) 0.09
> 3 times per week 6.87 (1.52, 12.23) 0.01 ≦ 1 times per week 4.29 (0.93, 7.65) 0.01 > 1 times per week 2.29 (− 2.58, 7.15) 0.36
Sea-fish consumption > 1 times per week 4.19 (− 0.02, 8.39) 0.05
≦ 1 times per month ref
≦ 1 times per week 2.75 (− 0.06, 5.57) 0.06
> 1 times per week 5.53 (2.90, 8.17) < 0.0001
Sell-fish
consumption
≦ 1 times per month ref
≦ 1 times per week 4.15 (1.50, 6.81) 0.002
> 1 times per week 3.46 (0.32, 6.61) 0.03
Second-hand
smoking
≦ 1 times per month ref
≦ 3 times per week − 0.65 (− 3.17, 1.86) 0.61
> 3 times per week 2.90 (− 0.22, 6.02) 0.07

11.65, P = 0.06). However, no significant associations between blood
mercury concentrations and serum BDNF levels were observed in boys.
4. Discussion
In this cross-sectional study, we observed that developmental low-
levels mercury exposure was significantly positively associated with
serum BDNF levels. When we stratified by sex, we only observed these
associations in girls. Moreover, when dietary habits and fish intake were
further adjusted, the associations between blood mercury and serum
BDNF became borderline significant.
Mercury exposure in children could damage various organs in the
body, especially the developing central nervous system. In this study,
the mean of the blood mercury concentrations in preschool children was
1.28 μg/L, which was lower than those observed in southeastern of
China, Guangdong (1.98 μg/L) and a national study in Chinese children
aged 0–6 years (1.39 μg/L), but still higher than that reported in
northeastern of China, Qinghai (0.37 μg/L) (Gao et al., 2018). Once
comparing with international studies, the levels of mercury in this study
were also significantly lower than the blood mercury concentrations of
children in Korea (2.07 μg/L). However, the blood mercury in our study
was still higher than that reported in European countries (0.12–0.94
μg/L) (Hruba et al., 2012; Lim et al., 2015). The relative lower levels of
mercury observed in our study might due to that only one-third of the
children included in our study consumed fish more than once per week.
Therefore, the blood mercury levels of the children in our study were
relatively low.
Numerous studies have indicated that fish consumption is an
important risk factor related to childhood mercury exposure (Hertz-­
Picciotto et al., 2010; Zhang and Zhang, 2015). In our study, we also
observed that salt-water fish intake, but not water-fish and shell-fish
consumption, was significantly positively associated with increased
blood mercury levels in children. This phenomenon may be resulted
from the higher size of marine fish and subsequent higher accumulation
of mercury (Muir et al., 2005; Zupo et al., 2019). In addition, the
second-hand smoking was also regarded as one of the sources of expo­
sure to heavy metals for children. A previous study had shown that
smoking could induce increased levels of mercury concentration in
smokers (Kim et al., 2019). Karatela et al. (2019) verified that maternal
smoking may be another pathway for mercury exposure in children. In
line with these observations, in this study, we also observed that the
blood mercury concentrations in children were positively associated
with the frequency of passive smoking. Though the contribution of
passive smoking for the mercury exposure seems low, it is still worthy of
attention considering mercury’s accumulation and adverse effects.
Intriguingly, a negative association between the frequency of cow-milk
intake and the levels of mercury in blood was also found. However, more
evidences are needed to verify the relationships of cow-milk intake on
mercury effects.
The relationships between developmental mercury exposure and

4
C.-C. Zhou et al. Ecotoxicology and Environmental Safety 208 (2021) 111642

Table 4 supplement also provided protection effects from mercury neurotoxicity

Associations between the blood levels of mercury and serum BDNF concentra­ via anti-apoptosis and autophagy pathway (El Asar et al., 2019). Sup­
tions in the participants, boys, and girls. porting our study, a multicentre birth cohort study carried out in Spain
Model 1 Model 2 Model 3 reported the positive associations between postnatal mercury exposure
β (95% CI) P β (95% CI) P β (95% CI) P
and the MSCA scores, however, these relationships were significantly
attenuated when the variables of children’s fish intake were further
Total
adjusted (Llop et al., 2020). Intriguingly, it is generally considered that
Continuous 4.91 (1.20, 0.01 4.10 (0.44, 0.03 3.65 (− 0.05
log 8.63)a 7.76)b 0.07, BDNF is a protein majorly secreted by neurons and plays a key role in
7.37)c synaptic plasticity, synaptogenesis, learning, and memory. However,
Tertile 1 Ref Ref Ref BDNF exacerbated methylmercury-induced cell death in rat cerebellar
Tertile 2 1.89 (− 0.37 1.25 (− 0.38 2.17 (− 0.13 granule cells were also reported (Sakaue et al., 2009). Therefore,
0.99, 4.76) 1.52, 4.02) 0.63, 4.97)
Tertile 3 3.00 (0.20, 0.09 2.31 (− 0.15 1.76 (− 0.28
although we have observed that increased mercury levels were associ­
5.79) 0.87, 5.48) 1.44, 4.97) ated with elevated serum BDNF concentrations, the role of BDNF in the
Boys effects of mercury on neurodevelopment are still needed to be
Continuous 2.93 (− 0.23 2.55 (− 0.30 2.89 (− 0.24 investigated.
log 1.89, 2.25, 1.93,
When we stratified by sex, a positive association between postnatal
7.75)a 7.34)d 7.71)e
Tertile 1 Ref Ref Ref exposure to mercury and serum BDNF levels was only observed in girls.
Tertile 2 − 0.95 (− 0.60 − 1.14 (− 0.53 − 0.33 (− 0.85 Although its potential molecular mechanisms of this difference were still
4.52, 2.63) 4.70, 2.41) 3.86, 3.20) unknown, many epidemiological studies have regarded gender as a
Tertile 3 0.66 (− 0.74 0.63 (− 0.75 0.74 (− 0.71 potential modifier. For example, a negative association between pre­
3.33, 4.66) 3.34, 4.60) 3.23, 4.71)
Girls
natal mercury exposure and psychomotor development among females
Continuous 7.37 (1.46, 0.02 5.97 (0.19, 0.04 5.75 (− 0.06 was detected in Spanish children (Llop et al., 2012). Postnatal mercury
log 13.27)a 11.76)f 0.16, exposure was also observed negatively associated with intelligence
11.65)g quotient (IQ) and visuospatial abilities among girls (Davidson et al.,
Tertile 1 Ref Ref Ref
2008a; Myers et al., 2009). Moreover, an experimental animal study also
Tertile 2 4.36 (− 0.06 4.38 (− 0.06 5.81 (1.17, 0.01
0.24, 8.97) 0.11, 8.86) 10.44) found that compared with the male mice, the female mice take up more
Tertile 3 6.10 (0.64, 0.03 4.50 (− 0.10 4.18 (− 0.13 mercury into their motor neurons in the same dose exposure (Pamphlett
11.56) 0.85, 9.86) 1.19, 9.55) et al., 1997). However, a positive relationship between postnatal mer­
a
Adjusted for age, sex, maternal education, monthly household income per cury exposure and scholastic achievement in girls was also documented
capita. (Davidson et al., 2010). These results suggested that sex differences
b
Adjusted for the covariates listed in a and for sea-fish consumption, shell-fish played an important role in the effects of mercury exposure. Therefore,
consumption. sex might play an important role in the effects of mercury on children’s
c
Adjusted for the covariates listed in b and for vitamin intake, mineral intake, neurodevelopment.
passive smoking. Our study still has some limitations. First, in addition to the analysis
d
Adjusted for the covariates listed in a and for sea-fish consumption, shell-fish of the relationships between blood mercury levels and serum BDNF
consumption. levels, children’s cognitive tests which may affect children’s intelligence
e
Adjusted for the covariates listed in d and for vitamin consumption, mineral
quotient directly were not performed and analyzed. Second, the dietary
intake.
f habits in the questionnaire were somewhat rough, for example, although
Adjusted for the covariates listed in a and for sea-fish consumption, shell-fish
consumption. the frequency of fish consumption was collected, the intake of fish
g
Adjusted for the covariates listed in f and for mineral intake. species was not distinguished, which may affect dietary mercury expo­
sure. Third, the levels of fatty acid such as n-3 LC-PUFA, and DHA in the
serum of the children were not further determined and analyzed.
BDNF in children were also assessed. We observed that increasing blood
In conclusion, our study suggested that developmental low-level
mercury concentrations were positively associated with higher serum
mercury exposure positively was associated with serum BDNF levels
BDNF levels, which were unexpected and conflicted with other studies.
and that these effects might differ by sex. Furthermore, when dietary
Previously, we observed that there was no association between blood
intake items were further adjusted, these associations became borderline
mercury concentrations and serum BDNF levels in a cross-sectional
significant. The second-hand smoking and sea-fish consumption might
study conducted in Taizhou city (Zhou et al., 2019). Another study
be confounders in the relationships between mercury exposure and
also reported that they did not find any adverse effects of mercury
BDNF. Further studies are warranted to confirm these findings and
exposure on BDNF during early pregnancy (Zaw and Taneepanichskul,
reveal the possible neurotoxic effects of developmental mercury
2019). The heterogeneity of these conclusions may be attributed to
exposure.
different populations and methodologies included in the analysis.
However, an experimental study performed in Japan found that meth­
ylmercury treatment significantly increased BDNF expression in the CRediT authorship contribution statement
cerebral cortex of the mice (Fujimura and Usuki, 2017). Furthermore,
when we included fish consumption in the models, the coefficients for Can-Can Zhou: Formal analysis, Data curation, Methodology,
this association remained positive and statistically significant. However, Writing - original draft, Writing - review & editing. Hui Fu: Writing-
when dietary intake items were further adjusted, these associations revised draft, Writing - review & editing. Guo-Yan Zhang: Formal
between blood mercury concentrations and serum BDNF levels became analysis, Data curation. Jia-Wei Ma: Formal analysis, Data curation.
borderline significant. It is well known that apart from mercury and Min Ni: Methodology. Dong-Jie Li: Data curation. Fu-Ming Shen:
other toxicants, fish and dietary intake also contain numerous beneficial Writing - review & editing. Fang Huang: Writing-revised draft, Data
nutrients including vitamins, essential trace elements, and unsaturated curation.
fatty acids, which are necessary for brain development. Recently, several
studies also verified its antagonism to the toxicity of mercury. For Acknowledgments
example, vitamin E protected the rats’ brains against mercury-induced
toxicity through antioxidant way (Agarwal et al., 2010). Selenium This study was supported by the Fundamental Research Funds for the
Central Universities (No. 22120200244); the Shanghai Sailing Program

5
C.-C. Zhou et al. Ecotoxicology and Environmental Safety 208 (2021) 111642

(No. 20YF1437600); the National Natural Science Foundation of China Hertz-Picciotto, I., Green, P.G., Delwiche, L., Hansen, R., Walker, C., Pessah, I.N., 2010.
Blood mercury concentrations in CHARGE Study children with and without autism.
(No. 81703590); Shanghai Key Clinical Specialist Construction Pro­
Environ. Health Perspect. 118, 161–166.
grams (to Fu-ming Shen); Shanghai Science and Technology Commis­ Hruba, F., Stromberg, U., Cerna, M., Chen, C., Harari, F., Harari, R., Horvat, M.,
sion Experimental Animal Grants (19140904900). Koppova, K., Kos, A., Krskova, A., Krsnik, M., Laamech, J., Li, Y.F., Lofmark, L.,
Lundh, T., Lundstrom, N.G., Lyoussi, B., Mazej, D., Osredkar, J., Pawlas, K.,
Pawlas, N., Prokopowicz, A., Rentschler, G., Spevackova, V., Spiric, Z., Tratnik, J.,
Skerfving, S., Bergdahl, I.A., 2012. Blood cadmium, mercury, and lead in children:
Conflict of interest an international comparison of cities in six European countries, and China, Ecuador,
and Morocco. Environ. Int. 41, 29–34.
None. Hu, Y., Chen, L., Wang, C., Zhou, Y., Zhang, Y., Wang, Y., Shi, R., Gao, Y., Tian, Y., 2016.
Prenatal low-level mercury exposure and infant neurodevelopment at 12 months in
rural northern China. Environ. Sci. Pollut. Res. Int. 23, 12050–12059.
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