Environmental Pollution 248 (2019) 18e27

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Environmental Pollution
journal homepage: www.elsevier.com/locate/envpol

Effect of calcium peroxide on the water quality and bacterium

community of sediment in black-odor water*
Wen-Huai Wang a, Yi Wang a, *, Pan Fan a, Lin-Feng Chen a, Bao-Hua Chai a,
Jing-Chan Zhao b, Lu-Qin Sun c
a
School of Environmental and Municipal Engineering, Xi'an University of Architecture and Technology, China
b
College of Chemistry & Materials Science, Northwest University, Xi'an, 710069, China
c
Environmental Science Department, University of San Francisco, California, CA, 94117, USA

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

Article history: This study investigated how efficiently CaO2 could treat black-odor landscape water caused by low
Received 10 September 2018 dissolved oxygen (DO) in a field experiment of 600 m2. The study demonstrated that CaO2 could
Received in revised form significantly elevate the DO concentration in waters and the oxidationereduction potential (ORP) level in
5 November 2018
sediments (p ¼ 0.003 and p ¼ 0), which is conducive to improving the anoxic environment of landscape
Accepted 22 November 2018
Available online 23 November 2018
water. The concentrations of total chemical oxygen demand (TCOD) and S2 in overlying and interstitial
waters were considerably decreased. The average concentrations of TCOD in the overlying and interstitial
waters of the test zone (TZ) were 52.98% and 66.05% of those of the control zone (CZ), and the average
Keywords:
Calcium peroxide
concentrations of S2 in the overlying and interstitial waters of TZ were 29.63% and 39.79% of those of CZ.
Landscape water Meanwhile, CaO2 could obviously reduce turbidity but increase the transparency in the overlying water.
Sediment release The mean value of turbidity in the overlying water of TZ was 39.46% of that of CZ, whereas the trans-
Black-odor governance parency in the overlying water of TZ was 2.07 times that of CZ. Furthermore, CaO2 changed the microbial
Microbial flora community structure in the sediments, where the relative abundance of anaerobic bacteria was
decreased but that of the aerobic bacteria was increased with some functional bacteria. In summary,
CaO2 could significantly increase the DO and ORP in black-odor landscape water, obviously inhibit the
release of pollutants from sediment, and increase the diversity of microbial strains. Consequently, the
black-odor phenomenon of landscape water could be alleviated effectively by adding CaO2.
© 2018 Published by Elsevier Ltd.

1. Introduction to anoxic or anaerobic conditions due to the rapid consumption of

Urban landscape water has an important ecological value in
improving local climate and beautifying urban environment
(Mcgoff et al., 2017). Nevertheless, this type of water is seriously
polluted by area-source pollution because it is commonly located in
urban districts with large population density (Wang et al., 2018).
Meanwhile, a large amount of pollutants are gradually accumulated
in the sediments of rivers and lakes in the form of endogenous
pollutant due to the defects of landscape water, such as shallow
water, poor fluidity, and weak self-purification capacity (Ao et al.,
2018). In summer, higher temperature contributes to the higher
microbial metabolisms which will convert the water environment
*
This paper has been recommended for acceptance by Dr. Sarah Harmon.
* Corresponding author.
E-mail address: wangyi1003@xauat.edu.cn (Y. Wang).
dissolved oxygen (DO) in the water. The proliferation, growth, and
metabolism of anaerobic bacteria are accompanied with the cor-
ruption, decomposition, fermentation, and transformation of
organic compounds in sediments (Li et al., 2016), which strengthen
the production of substances, such as ammonia nitrogen (NHþ 4 eN),
humus, hydrogen sulfide, methane, and mercaptan. Among these
substances, ammonia (NH3), hydrogen sulfide, and mercaptan
inevitably diffuse and spread out into the surroundings, which are
probably the main reasons turn the water into smelly or olfactory
water. Meanwhile, some metals, such as iron and manganese, are
reduced in the anoxic environment and further combined with S2
to form ferrous sulfide and other compounds (Ololade et al., 2016),
which usually cause the water turning into black color in the
landscape water due to a large quantity of negative colloids (FeS
and MnS) being adsorbed by suspended particles and floated in the
overlying water. The formation of black and smelly water body will

https://doi.org/10.1016/j.envpol.2018.11.069
0269-7491/© 2018 Published by Elsevier Ltd.
 W.-H. Wang et al. / Environmental Pollution 248 (2019) 18e27
not only reduces the aesthetic value of landscape water but also
harms the ecological balance and introduces many hygienic prob-
lems (Andreasen et al., 2013; Shen et al., 2014). A total of 1861 water
bodies (85.7% of rivers and 14.3% of lakes) in China were labeled as
black-odor water by the end of February 2017. Thus, how to deal
and treat black-odor landscape water is a common and serious
issue existing in China.
The popular treatments used today to restore black-odor land-
scape water are physical methods (e.g., sediments dredging, sedi-
ments coverage, and artificial aeration) (Richardson et al., 2011),
chemical methods (e.g., addition of aluminum salt, sodium nitrate,
calcium nitrate, and hydrogen peroxide) (Ronen et al., 2010), and
biological methods (e.g., purification of aquatic plants, microor-
ganism strengthening, and exogenous microorganism delivery)
(Faccioabbba, 2012). These techniques can accordingly improve the
quality of black-odor landscape water but have some obvious de-
fects in their applications. Physical methods are often companied
with the issues of high cost, low efficiency, and secondary pollution.
For example, the sediments after dredging out from the bottom
need to be properly further treated, and more pollutants could be
released into the water from sediments by artificial aeration (Wang
et al., 2017b; Uggetti et al., 2016). While using chemical methods
dealing with the black-odor water have many problems too, such
as, applying aluminum salt is commonly known to be toxic to
aquatic organisms (Freitas et al., 2016), using nitrate would increase
the concentration of nitrate nitrogen in the overlying water (Liu
et al., 2016; Bindhu et al., 2016), Na2CO4 has a short timeliness,
therefore is easily soluble in water and moist atmosphere (Sindelar
et al., 2014), and the use of hydrogen peroxide is dangerous and
unstable (Zingaretti et al., 2016). Biological methods are usually
commented as low efficiency, poor stability, and high susceptibility
to environmental conditions. Therefore, a safer, greener, more
economical and efficient technology is absolutely needed to solve
the black-odor problem of landscape water caused by low DO.
Calcium peroxide (CaO2) can release oxygen molecules slowly in
moist air or water and it is safe to aquatic ecosystem as a common
food additive; thus, CaO2 can be used in the in-situ treatment of
black-odor landscape water (Lu et al., 2017). Currently, there are
considerable literature study the effects of CaO2, which mainly
concentrates on the removal of trichloroethylene in water, degra-
dation of endocrine-disrupting compounds in waste activated
sludge (Zhang et al., 2015a; Zhang et al., 2015b), and reduction in
triamcinolone acetonide, benzene, and persistent organic pollut-
ants in the environment (Wang et al., 2017a; Zhang et al., 2018).
However, the effects of CaO2 on black-odor landscape water and the
microbial community structure in sediments are seldom studied. In
this study, CaO2 was applied into the severely black-odor landscape
water of 600 m2, as a slow-releasing peroxide. The feasibility and
stability of CaO2 for emergency treatment of black-odor landscape
water were explored and evaluated and the improvement of anoxic
environment in water and the change of microbial community
structures in sediments were recorded and studied. This study is
important in managing black-odor landscape water and provides a
valuable field case for reference in the future.
2. Materials and methods
2.1. General situation and design of the experiment
Xi'an moat surrounds the ancient city wall of Xi'an, and its
original function was to prevent military attacks. At present, Xi'an
moat serves as urban landscape and storm-water storage. Xi'an
moat has a circumference of 14.8 km, a width of 15e20 m, and a
depth of 1e8 m. The water velocity in Xi'an moat is extremely small,
which allows its sediment to reach the depth of 3 m high.
19
The test site of this study is located in a relatively closed zone of
the northeast corner of Xi'an moat, where the overlying water is
17 m in width and 1.6 m in depth and the sediments are 0.9 m in
depth. The control zone (CZ) and test zone (TZ) were set along the
direction of water flow in the test site, which were both 30 m in
length and 10 m in width. The two zones were naturally separated
by a platform used as a tourists resting place, which is 15 m in
length and 10 m in width. At the beginning of the experiment, 75 kg
of CaO2 with a purity of 76.3% was sprinkled evenly on the surface
of sediment in TZ, whereas CZ was in the original condition. In
addition, five sampling points were selected for both zones (i.e., in
the center and four corners), and these sampling points in the same
zone were studied as a test areas.
The experiment lasted for 2 months in the summer of 2017. The
average water temperature of the sampling days was
30.2  C ± 0.7  C during the experiment. At the 48th and 61st days of
the experiment, reservoir water was replenished to the Xi'an moat
at the upstream 2.5 km of the test site to improve the water quality
by the Management Committee of Xi'an moat.
2.2. Sampling
The overlying water was collected individually using a sampler
at 0.5 m below the surface water at five sampling points both in CZ
and TZ. The sediment samples were grabbed below the surface
sediment phase approximately 0.10 m by a gravity sediment col-
lector from five sampling points in the CZ and TZ respectively. The
supernatant of the sediments sample centrifuged by a refrigeration
centrifuge (5804R) was obtained as a representative sample of the
interstitial water. Water and sediment samples were collected at
9:00 a.m. every 7 days. The sediment samples from five sampling
points in CZ and TZ were collected and mixed respectively as
representative samples of each experimental zone to determine the
microbial community structure for the study site.
2.3. Analysis of samples
2.3.1. Analysis of water quality and physicochemical index of
sediment
Temperature (T), transparency, DO, and oxidationereduction
potential (ORP) were monitored at the field site, whereas other
parameters (e.g., turbidity, S2, SO2
4 , and total chemical oxygen
demand (TCOD)) were tested in the Key Laboratory of the Ministry
of Education. T, DO, and ORP were measured with a HACH portable
multifunctional water quality meter (HQ-30d), transparency was
metered with a Secchi disk, turbidity was analyzed with a spec-
trophotometer (XINMAO752N), and TCOD was detected by potas-
sium dichromate method. S2 was measured by dimethyl-aniline
spectrophotometry after pretreatment by NaOHezinc acetate,
whereas SO24 was analyzed by anion chromatography (ICS 1100).
2.3.2. Identification of microbial strains in sediment
High-throughput sequencing of the sediments in CZ and TZ
were conducted with the Illumina MiSeq system, which contrib-
uted to understanding the difference in bacterial community
structure in both sediments. The specific measurement steps are
discussed as follows:
The total DNA was extracted from sediment using the OMEGA
kit (Life, USA) while its completeness was tested by agarose gel
electrophoresis. Genomic DNA was accurately quantified using the
Qubit 2.0 DNA Assay Kit (Life, USA) to determine the amount of DNA
that should be added to the PCR reaction. PCR was amplified with
the primers of 341F (50 eCCCTACACGACGCTCTTCCGATCTG-30 ) and
805R (50 -GACTGGAGTTCCTTGGCACCCGAGAATTCCA-30 ).
The PCR amplification procedure was conducted as follows: pre-
20 W.-H. Wang et al. / Environmental Pollution 248 (2019) 18e27
denaturation at 94  C for 3 min, denaturation at 94  C for 30 s,
annealing at 45  C for 20 s, extension at 65  C for 30 s, repeating 5
cycles; denaturation at 94  C for 20 s, annealing at 55  C for 20 s,
extension at 72  C for 30 s, repeating 20 cycles; introduction of
Illumina bridge PCR compatible primers, pre-denaturation at 95  C
for 30 s, denaturation at 95  C for 15 s, annealing at 55  C for 15 s,
extension at 72  C for 30 s, repeating 5 cycles.
2.4. Statistical analysis
The experimental data were statistically analyzed, calculated,
and plotted using Origin 9.0 and Excel software. In the strain
identification phase, the base sequence obtained from high-
throughput sequencing was first decontaminated using Trimmo-
matic, and high-quality double-end sequences were connected by
the flash software to obtain optimized sequences. Sequences were
clustered into operational taxonomic unit (OTU) at a 97% identity
threshold with the using UPARSE software, then the OTU repre-
sentative sequence was compared with the bacterial RDP database
by using the RDP classifier software to obtain OTU species infor-
mation. The Mothur, R language barplot, and R language pheatmap
software packages were used to analyze the Alpha diversity of the
strains, species richness, and differences in species abundance
distribution.
The average value, standard deviation, and variance of the data
were analyzed using SPSS software (PASW Statistics 20.0). The
homogeneity of variance among samples was tested using f-test
method. The means were tested using t-test method with a sig-
nificance level of p < 0.05.
3. Results
3.1. Effect of CaO2 on quality of landscape water
3.1.1. Effect of CaO2 on the overlying water
1) Time course of turbidity and transparency in the overlying
water
Fig. 1 shows the variations in turbidity and transparency in the
overlying water of different zones. As shown in the figure, the
turbidity in the overlying water of CZ increased gradually compared
Fig. 1. Time course of turbidity and transparency in the overlying water of different zones: (a) turbidity, (b) transparency.
to the initial value, but that of TZ decreased. Unsurprisingly, the
variation trends of transparency in both zones were opposite to
those of turbidity.
Fig. 1(a) shows that the initial values of turbidity in the overlying
water of CZ and TZ were 15.26 ± 0.87 and 16.12 ± 0.93 NTU,
respectively. The turbidity in the overlying water of CZ increased
gradually compared to the initial value, and it reached the
maximum value (23.16 ± 1.07 NTU) at the 43rd day. However, the
turbidity in the overlying water of TZ decreased stably compared to
the initial value, and it reached the minimum value (4.08 ± 0.36
NTU) at the end of the experiment. As shown in Fig. 1(b), the
transparency in the overlying water of CZ decreased gradually
compared to the initial value, and it reached the minimum value
(22.2 ± 1.3 cm) at the 36th day. However, the transparency in the
overlying water of TZ increased stably, and it reached the maximum
value (69.8 ± 1.1 cm) at the 50th day. The average values of turbidity
in the overlying water of CZ and TZ were 19.92 ± 0.99 and
7.86 ± 0.87 NTU (p ¼ 0), respectively, throughout the test. By
contrast, those of transparency were 29.2 ± 1.9 and 60.5 ± 2.1 cm
(p ¼ 0). Thus, the mean value of turbidity in the overlying water of
TZ was 39.46% of that of CZ, whereas the mean value of trans-
parency in the overlying water of TZ was 2.07 times that of CZ.
Therefore, CaO2 could significantly increase the transparency in
the overlying water of black-odor landscape water but decrease the
turbidity.
2) Time course of DO and ORP in the overlying water
Fig. 2 shows the variations in DO and ORP in the overlying water
of both areas. As shown in Fig. 2, the DO concentration and ORP
level in the overlying water of TZ were constantly higher than those
of CZ during the experiment.
As shown in Fig. 2, the initial values of DO concentration and
ORP level in the overlying water of CZ were similar to those of TZ.
The initial DO (2.08 ± 0.05 mg L1) and initial ORP (23.9 ± 2.3 mv)
were relatively low in the overlying water of CZ; both decreased
stably compared to the initial value and reached the minimum
value (0.13 ± 0.04 mg L1 and 178.9 ± 7.3 mv) at the 36th and 57th
day, respectively. However, the variation trends of DO and ORP in
the overlying water of TZ increased gradually first and then reached
the maximum value (7.26 ± 0.11 mg L1 and 83.7 ± 3.6 mv) at the
8th and 36th days, respectively. The DO concentration and ORP
 W.-H. Wang et al. / Environmental Pollution 248 (2019) 18e27 21

Fig. 2. Time course of DO and ORP in the overlying water of different zones: (a) DO, (b) ORP.

level in the overlying water of TZ were constantly higher than those
of CZ during the experiment although the numbers went down a
little bit as time passed. The average DO concentrations in the
overlying water of TZ and CZ were 4.50 ± 1.12 and 1.19 ± 1.10 mg L1
(p ¼ 0.003), respectively; the former was 3.78 times the latter. The
average ORP levels in the overlying water of TZ and CZ were
62.3 ± 3.2 and 73.5 ± 6.2 mv (p ¼ 0), respectively; the former was
significantly higher than the latter.
Therefore, CaO2 could significantly increase the DO concentra-
tion and ORP level of the overlying water in the black-odor water
body.
3) Time course of TCOD and sulfur concentration in the overlying
water
Fig. 3 shows the variations of TCOD, S2, and SO2
4 in the over-
lying water of both zones. As shown in the figure, the concentra-
tions of TCOD and S2 increased first and then decreased in the
overlying water of CZ, while the concentration of SO24 decreased
stably compared with the initial value. Nevertheless, the variation
trends of TCOD, S2, and SO2 4 in the overlying water of TZ were
contrary to those of CZ.
Fig. 3(a) shows the variation trends of TCOD concentrations in
the overlying water for both zones. As shown in the figure, the
initial TCOD concentrations in the overlying water of CZ and TZ
were 68.33 ± 3.52 and 70.05 ± 3.37 mg L1, respectively. These
values considerably exceeded the concentration limit of Chinese
Surface Water Quality Standards for Class V (COD  40 mg L1,
GB3838-2002). However, the TCOD concentration in the overlying
water of CZ was constantly higher than that of TZ during the entire
experiment. TCOD concentration increased gradually and reached
the maximum value (105.32 ± 4.18 mg L1) at the 36th day in the
overlying water of CZ, whereas the concentration decreased grad-
ually and reached the minimum value (29.06 ± 2.28 mg L1) at the
36th day in the overlying water of TZ. The average TCOD concen-
trations in the overlying water of TZ and CZ were 43.78 ± 3.47 and
82.63 ± 4.32 mg L1 (p ¼ 0), respectively, which indicated that CaO2
could reduce 47.02% TCOD in the overlying water of TZ compared

Fig. 3. Time course of TCOD and S concentrations in the overlying water of different zones: (a) TCOD, (b) S2, and SO2
4 .
22 W.-H. Wang et al. / Environmental Pollution 248 (2019) 18e27

with that of CZ. period of 50 days.

Fig. 3(b) shows the variation trends of S2 and SO2 4 concen-
trations in the overlying water of different zones. The initial S2
concentrations in the overlying water of CZ and TZ were the same
(0.13 ± 0.02 mg L1). The S2 concentration increased first and
reached the maximum value (0.42 ± 0.03 mg L1) at the 43rd day in
the overlying water of CZ, but the concentration decreased steadily
(except for the end of the experiment) and reached the minimum
value (0.03 ± 0.01 mg L1) at the 50th day in the overlying water of
TZ. During the entire experiment, the average S2 concentrations
were 0.27 ± 0.02 and 0.08 ± 0.01 mg L1 (p ¼ 0) in the overlying
water of CZ and TZ, respectively, which indicated CaO2 could
decrease 70.37% S2 in the overlying water. The variation trends of
SO2
4 in the overlying water of different zones were contrary to
those of S2. The initial SO2
4 concentrations in the overlying water
of CZ and TZ were similar, which were 32.26 ± 2.16 and
31.68 ± 1.87 mg L1, respectively. The SO2
4 concentration decreased
uninterruptedly and reached the minimum value
(23.36 ± 1.72 mg L1) at the 36th day in the overlying water of CZ,
but the concentration increased first and then reached the
maximum value (92.68 ± 2.82 mg L1) at the 43rd day in the over-
lying water of TZ. The average SO2 4 concentrations were
28.06 ± 1.32 and 70.75 ± 1.87 mg L1 (p ¼ 0) in the overlying water
of CZ and TZ, respectively; the latter was 2.52 times the former.
In summary, CaO2 could greatly reduce the concentrations of
TCOD and S2 in the overlying water of the black-odor landscape
water, but it is possible to increase the concentration of SO2 4 .
Therefore, CaO2 could reduce sulfur effectively but promote sulfur
oxidation to a high valence (SO24 ).
3.1.2. Effect of CaO2 on DO concentration of the sedimentewater
interface
Table 1 shows the variations in DO concentration at the
sedimentewater interface of different zones. As shown in the table,
the initial DO concentrations in CZ and TZ were the same. The DO
concentration at the sedimentewater interface of TZ increased first,
reaching the maximum value (4.02 ± 0.18 mg L1) at the 22nd day
and then dropped gradually. The concentration was higher than
1.60 mg L1 at the sedimentewater interface of TZ throughout the
test (except the starting point). However, the DO concentration at
the sedimentewater interface of CZ was low lesser than
0.20 mg L1 for the entire experiment. Thus, the average DO con-
centrations were 2.56 ± 0.11 and 0.08 ± 0.04 mg L1 (p ¼ 0) at the
sedimentewater interface of CZ and TZ, respectively; the former
was 32 times the latter.
Therefore, CaO2 could significantly elevate the DO concentration
at the sedimentewater interface of the landscape water. Moreover,
CaO2 could help maintain the DO concentration at a high level for a
3.1.3. Effect of CaO2 on TCOD and sulfur concentrations in the
interstitial water
Fig. 4 shows the changes in TCOD, S2, and SO2 4 concentrations
in the interstitial water of different zones. As shown in the figure,
the variation trends of these indices were the same as those in the
overlying water of different zones. The concentrations of TCOD and
S2 in the interstitial water of CZ increased gradually compared to
the initial value, but the concentration of SO2 4 decreased stably.
However, the concentrations of TCOD, S2, and SO2 4 in the inter-
stitial water of TZ were contrary to those of CZ.
Fig. 4(a) shows the variation trends of TCOD concentration in the
interstitial water of different zones. As shown in the figure, the
initial TCOD concentrations were 131.74 ± 5.28 and
136.42 ± 6.53 mg L1 in the interstitial water of CZ and TZ, respec-
tively; the latter was slightly higher than the former. During the
experiment, TCOD concentration increased first and reached the
maximum value (198.33 ± 7.25 mg L1) at the 43rd day in the
interstitial water of CZ. On the contrary, the TCOD concentration
decreased stably and reached the minimum value
(86.06 ± 5.27 mg L1) at the 50th day in the interstitial water of TZ.
Therefore, the average TCOD concentrations in the interstitial water
of CZ and TZ were 163.28 ± 5.76 and 107.84 ± 5.32 mg L1 (p ¼ 0),
respectively; the latter was 66.05% of the former.
Fig. 4(b) shows the changes of S2 and SO2 4 concentrations in
the interstitial water of both areas. The initial S2 concentrations in
the interstitial water of different zones were the same at approxi-
mately 0.31 mg L1. S2 concentration increased continually and
reached the maximum value (0.92 ± 0.13 mg L1) at the 50th day in
the interstitial water of CZ, whereas the S2 concentration
decreased gradually and reached the minimum value
(0.13 ± 0.02 mg L1) at the 57th day in the interstitial water of TZ.
During the entire experiment, the average S2 concentrations were
0.61 ± 0.04 and 0.20 ± 0.03 mg L1 (p ¼ 0) in the interstitial water of
CZ and TZ, respectively, which indicated CaO2 could reduce 67.21%
S2 in the interstitial water. The initial SO2
4 concentrations in the
interstitial water of CZ and TZ were similar, which were 20.05 ± 2.11
and 21.18 ± 1.73 mg L1, respectively. SO24 concentration decreased
continuously in the interstitial water of CZ, whereas the concen-
tration increased stably and reached the maximum value
(52.37 ± 2.09 mg L1) at the 50th day in the interstitial water of TZ.
The average SO2 4 concentrations were 15.51 ± 0.98 and
45.27 ± 1.62 mg L1 (p ¼ 0) in the interstitial water of CZ and TZ,
respectively; the latter was 2.92 times the former.
In summary, CaO2 could significantly decrease the concentra-
tions of TCOD and S2 in the interstitial water but could increase
the concentration of SO2 4 , which could provide a good reference in
the purification and improvement of black-odor landscape water.

Table 1 3.2. Effect of CaO2 on the sediment of landscape water

DO concentration at the sedimentewater interface of different zones during the
experiment.
3.2.1. Effect of CaO2 on sediment ORP
Time (d) DO (mg L1) Table 2 shows the variations of ORP levels in the sediment of
CZ TZ different zones. As shown in the table, the initial values of ORP level
0 0.03 ± 0.02 0.03 ± 0.02
in different zones were similar, as low as 342.9 ± 15.6
4 0.06 ± 0.04 2.88 ± 0.15 and 341.8 ± 13.5 mv in the sediments of CZ and TZ respectively.
8 0.12 ± 0.04 3.15 ± 0.14 During the experiment, the ORP level in the sediment of CZ
15 0.16 ± 0.05 3.96 ± 0.17 decreased steadily and reached the minimum value (426.4 ± 14.7
22 0.11 ± 0.07 4.02 ± 0.18
mv) at the 57th day. However, the ORP level in the sediment of TZ
29 0.07 ± 0.04 3.27 ± 0.18
36 0.02 ± 0.01 3.11 ± 0.18 increased first and reached the maximum value (160.4 ± 13.4 mv)
43 0.03 ± 0.02 2.26 ± 0.12 at the 29th day. Although the ORP levels in the sediment of TZ
50 0.05 ± 0.04 2.08 ± 0.16 declined later, it was still higher than that of CZ in the corre-
57 0.08 ± 0.04 1.78 ± 0.18 sponding time. The average values of ORP were 373.28 ± 11.1
64 0.16 ± 0.06 1.62 ± 0.18
and 223.3 ± 9.8 mv (p ¼ 0) in the sediments of CZ and TZ,
 W.-H. Wang et al. / Environmental Pollution 248 (2019) 18e27 23

Fig. 4. Time course of TCOD and sulfur in the interstitial water of different zones: (a) TCOD, (b) S2 and SO2
4 .

Table 2 the sequencing depth was sufficient. The values of OTU, Chao, and
ORP levels in the sediments of different zones during the experiment. ACE indices in the sediment of TZ were higher than those of CZ,
Time (d) ORP (mv) whereas the Shannon, Simpson, and Shannoneven indices of mi-
CZ TZ
crobial species in the sediment of TZ were slightly smaller than
those of CZ. Therefore, CaO2 could increase the diversity and
0 342.9 ± 15.6 341.8 ± 13.5
abundance of microbial community in the sediment of landscape
4 312.6 ± 13.7 317.3 ± 14.6
8 313.7 ± 12.8 276.4 ± 14.2 water but could decrease the uniformity of microbial community
15 339.6 ± 15.4 233.2 ± 11.3 structure in the sediment.
22 378.6 ± 14.5 182.2 ± 13.8
29 392.7 ± 16.7 160.4 ± 13.4
2) Analysis of microorganism community structure of phylum level
36 394.2 ± 17.8 162.5 ± 12.5
43 378.7 ± 15.1 177.8 ± 11.7
50 410.3 ± 16.2 189.2 ± 11.9 Fig. 5 shows the species abundances of phylum level in the
57 426.4 ± 14.7 200.3 ± 12.5 sediment of different zones, obtained by the RDP classifier soft-
64 416.8 ± 13.7 213.2 ± 12.1 ware. Bacteria having less than 0.5% abundance, together with
undetermined class were uniformly named as “Others.”
Among different species in the sediment of CZ shown in Fig. 5,
respectively. Therefore, CaO2 could significantly elevate the ORP Proteobacteria was the dominant species (31.94%), followed by
level in the sediment of landscape water. Chloroflexi (17.26%), Firmicutes (10.48%), and Bacteroidetes (10.31%).
Although the relative abundances of the species varied greatly, the
dominant bacteria in the sediment of TZ were similar to those of CZ.
3.2.2. Effect of CaO2 on microorganisms in sediment Proteobacteria was still the dominant species (27.82%), followed by
Bacteroidetes (12.80%), Firmicutes (12.76%), and Chloroflexi (5.33%).
1) Analysis of microbial diversity CaO2 could also affect the relative abundance of other bacteria in
the sediment of TZ to a certain extent. The relative abundance of
Table 3 shows the microbial diversity indices in the sediments of Caldiserica in the sediment of TZ was 2.39%, which was larger than
different areas. The Chao and ACE indices were used to reflect the that of CZ (1.55%). Meanwhile, the relative abundances of Actino-
richness of the species without considering the uniformity, which bacteria, Cyanobacteria, Spirochaetes, and Synergistetes in the sedi-
were positively correlated with the species. The Shannon and ment of TZ were 1.26%, 0.63%, 1.27%, and 0.38%, respectively, which
Simpson indices were used to evaluate the species richness under were lower than those of CZ (2.38%, 1.02%, 2.33%, and 2.05%). Be-
the premise of considering species uniformity. The Shannoneven sides those species being found in CZ, some unique bacteria, such as
index was used to assess the uniformity of the species distribution Aminicenantes (1.06%), Nitrospirae (1.22%), and Thermodesulfoba
in the samples. The Goods coverage index was used to obtain the (1.11%) were also detected in the sediment of TZ in phylum level.
probability of the species being detected in the samples. Consequently, CaO2 not only could change the relative abun-
As shown in Table 3, the Goods coverage indices in the sedi- dance of microbial species in the sediment but also could derive a
ments of different zones were larger than 99%, which indicated that

Table 3
Microbial diversity indices in the sediments of different areas.

Sample Observed OTU Chao ACE Shannon Simpson Goods Coverage Shannoneven

Control 1006 1081 1072 5.916 0.9934 0.9923 0.8438

Test 1129 1254 1221 5.853 0.9923 0.9919 0.8226
24 W.-H. Wang et al. / Environmental Pollution 248 (2019) 18e27

Fig. 5. Relative abundances of phylum level in sediment.

certain number of new bacteria.
3) Analysis of microorganism community structure in genera level
Bacteria having less than 0.3% abundance in genera level,
together with undetermined sequence were uniformly named as
“Others.” Fig. 6 shows the species abundances of genera level in the
sediment of different areas.
In this experiment, there are 243 and 258 different microbial
species in genera level that were detected in the sediments of CZ
and TZ, respectively. The number of microbial genus found in the
sediments of CZ and TZ were 21 and 22, respectively, in addition to
the bacteria of relative abundance less than 0.3% with undeter-
mined sequence. As shown in Fig. 6, the relative abundance of
microbial genus in the sediments of different zones was relatively
different. Among the dominant species (the relative abundance

Fig. 6. Relative abundances of genera level in sediment.

 W.-H. Wang et al. / Environmental Pollution 248 (2019) 18e27
higher than 2.0%) in the sediment of CZ, Petrimonas was ranked first
(10.83%), followed by Smithella (9.51%), Syntrophorhabdus (3.09%),
Proteus Hauser (2.38%), and Levilinea (2.13%). Correspondingly,
Petrimonas was the most dominant species (8.04%) in the sediment
of TZ, followed by Smithella (7.28%), Proteus Hauser (6.79%), Planc-
tomyces (2.91%), Desulfomicrobium (2.58%), and Verrucomicrobium
(2.36%). Besides the same species composition in the sediments of
both zones, the unique bacteria of Enterobacteriaceae (1.89%),
Gemella (1.67%), Haemophilus (1.06%), and Streptococcus (1.95%)
were detected in the sediment of CZ in genera level, whereas the
unique bacteria of Desulfomicrobium (2.58%), Nitrobacter (1.13%),
Nitrosomonas (0.52%), Syntrophomonas (1.82%), and Thiobacillus
(1.62%) were also found in the sediment of TZ. Meanwhile, the
relative abundances of specific or facultative anaerobes in genera
level were decreased obviously in the sediment of TZ. For example,
the relative abundances of Bellilinea, Caldisericum, Clostridium,
Parabacteroides, and Desulfobacterium in the sediment of TZ were
0.77%, 0.54%, 0.66%, 0.42%, and 0.37%, respectively, which were
lower than those in the sediment of CZ (1.09%, 1.39%, 1.59%, 1.58%,
and 0.96%).
In summary, CaO2 could obviously reduce the relative abun-
dance of anaerobic bacteria but maybe increase that of aerobic
bacteria. CaO2 could also derive some new bacteria which were
well adaptive to the environment.
4. Discussion
4.1. Improving water quality by adding CaO2 to black-odor
landscape water
CaO2, an odorless and tasteless alkaline earth metal peroxide,
has long persistence and good thermal stability, and is not easily
affected by air humidity and CO2 contents. Meanwhile, CaO2 can
release oxygen slowly in water, which can improve the hypoxic
environment of river and lake theoretically (Chen et al., 2016).
According to the stoichiometric ratio, the oxygen release rate of
CaO2 is as high as 0.222 g-O2/g-CaO2. CaO2, as one of the common
food additives, which is safe to the environment and has a high
industrial purity of 60%e80%. (Nelson et al., 2015). Therefore, CaO2
has been widely used for oxygen generation and deodorization in
environmental engineering.
CaO2 þ H2O / 0.5O2[þ Ca(OH)2 (1)
CaO2 þ 2H2O / H2O2 þ Ca(OH)2 (2)
H2O2 / 2H2O þ O2[ (3)
CaO2 / 0.5O2[þ CaO (4)
H2O2 þ activator / OH þ HO (5)
Ca(OH)2 þ H2O2 / CaO2 þ 2H2O (6)
Formulas (1)e(6) show the reactions between CaO2 and water.
Wang et al. demonstrated in their study that CaO2 was slightly
soluble in water, where it continued to release O2 and H2O2 in the
process of dissolution, and H2O2 was further reacted with active
substances to produce O2 and HO (Wang et al., 2016). Due to the
strong oxidizing properties of O2 and HO, CaO2 could increase the
DO concentration and improve the quality of black-odor landscape
water and had good removal effects on other contaminants (such
as: 2,4-dichlorophenol, arsenic, benzene and heavy metals) (Qian
et al., 2016; Olyaie et al., 2012; Xue et al., 2016; Chen et al., 2016).
The reaction of CaO2 and H2O to produce H2O2 and O2 is
25
accompanied with the formation of Ca(OH)2, which may cause a
large increase in pH value of the trial water theoretically. Never-
theless, the mean values of pH in CZ and TZ were 7.19 ± 0.02 and
7.26 ± 0.02 (p ¼ 0.568) for overlying water; 7.05 ± 0.02 and
7.32 ± 0.03 (p ¼ 0) for interstitial water; 6.85 ± 0.03 and 7.23 ± 0.04
(p ¼ 0) for sediment (Table A in the supplementary material) in our
study. Thus CaO2 had no significant impact on the pH value of the
overlying water. Although the interstitial water and sediment in TZ
was in a weak-alkaline environment, the pH values were slightly
higher than those of CZ. This phenomenon was mainly attributed to
the actual river had a strong buffer capacity for the variances of pH
caused by CaO2 owing to the existence of sediment and the flowing
and open of water area (Nyka €nen et al., 2012).
In summary, CaO2 could not only improve the black-odor phe-
nomenon of landscape water but also remove other pollutants
effectively from water and sediments. Therefore, CaO2 is a preferred
medicament for emergency treatment of black-odor landscape
water.
4.2. Mechanism of improving water quality in black-odor landscape
water by CaO2
4.2.1. Effect of CaO2 on basic environment of landscape water and
microorganisms in sediment
As described in Section 4.1, CaO2 could produce O2, H2O2, O2,
and HO during its dissolution procedure in water, which would
effectively increase the DO concentration of water and the ORP
level of sediments. In our experiment, the DO concentrations in the
overlying water and the sedimentewater interface of TZ were 3.78
and 32 times those of CZ, whereas the ORP levels were also
significantly elevated (p ¼ 0) in the overlying water and the sedi-
ments of TZ.
CaO2 not only could improve the basic environment of the
overlying water and sediment but also could influence the micro-
bial community structure of sediment to a certain extent. The high-
throughput sequencing results showed that the relative abun-
dances of Bacteroidetes in the sediments of CZ and TZ were 10.31%
and 12.80%, respectively. Bacteroidetes, as a common aerobic bac-
teria, could promote the degradation of organic matter in sedi-
ments (Oun et al., 2017). The relative abundances of Firmicutes in
the sediments of CZ and TZ were 10.48% and 12.76%, respectively,
and its abundance was positively correlated with DO concentration
but negatively correlated with N content (Marra et al., 2012). The
relative abundances of Proteobacteria in the sediments of CZ and TZ
were 31.94% and 27.81%, respectively; the abundance of this com-
mon specific or facultative anaerobes in water was negatively
correlated with DO concentration (Lim et al., 2018). The relative
abundances of Chloroflexi, one of facultative anaerobes, in the
sediments of CZ and TZ were 17.26% and 15.33% respectively. Sor-
okin et al. indicated that Chloroflexi lacked the functions of pro-
ducing O2 and fixing nitrogen, whereas it could accelerate the
hydrolysis process of macromolecular substances in sediments
(Sorokin et al., 2012).
Therefore, CaO2 could effectively improve the anoxic condition
of landscape water in summer, reduce the abundance of specific or
facultative anaerobes, and increase the relative abundance of aer-
obic bacteria. Moreover, a certain number of functional bacteria
would be generated, which were contributed to the water quality
purification of black-odor landscape water.
4.2.2. Effect of CaO2 on the concentrations of TCOD and S in water
Our experiment exhibited that DO concentration of water and
ORP level of sediments in TZ were elevated effectively, which could
prevent pollutants releasing from sediment. Meanwhile, the TCOD
concentrations in the overlying and interstitial waters of TZ
26 W.-H. Wang et al. / Environmental Pollution 248 (2019) 18e27
reduced obviously and some pollutants are removed from the
landscape water, which were due to the enhanced biochemical
reaction and oxidation effects of CaO2. The S2 concentrations in
the overlying and interstitial waters were decreased obviously,
whereas the corresponding SO2 4 concentrations were increased
significantly. Accordingly, the water quality of black-odor landscape
water could be improved greatly.
During the experiment, the mean TCOD concentrations in the
overlying and interstitial waters of TZ were 52.98% and 66.05% of
those of CZ, which indicated that the addition of CaO2 could
effectively reduce the TCOD concentration of landscape water.
The relative abundances of Proteus Hauser in the sediments of CZ
and TZ were 2.38% and 6.79%, respectively, whereas those of
Planctomyces were 0.82% and 2.91%. Chen et al. claimed that
Proteus Hauser was a common microbial strain in the nitrogen
and phosphorus removal system, and it had a good degradation
ability to organic matter in sewage wastewater (Chen et al., 2010).
Liu et al. indicated that Planctomyces could increase the degra-
dation efficiency of TN, NHþ 4 ‒N, and COD in leachate when they
were used a two-phase anoxic/aerobic combined membrane
bioreactor system to dispose landfill leachate (Liu et al., 2017).
Therefore, CaO2 could obviously reduce the concentration of
TCOD in the landscape water through increasing the microbial
diversity, oxidation reaction and functional bacteria abundance in
the sediments of TZ.
The average S2 concentrations in the overlying and interstitial
waters of TZ were 29.63% and 32.79% of those of CZ throughout the
test. On the contrary, the corresponding values of SO2 4 were 2.52
and 2.92 times those of CZ, which was mainly caused by the
changes of microbial community structure in the sediments of TZ.
In the experiment, the removal rates of S2 and TCOD by adding
calcium peroxide were better than that of sediments directly
dredging in a typical eutrophic shallow lake (Liu et al., 2015).
Caldisericum could reduce sulfate and thiosulfate to H2S under
anaerobic or anoxic conditions when it used lactic or propionic
acid as an electron donor in the reduction reaction (Shivaji et al.,
2011). Smithella could degrade alkanes and had mutual promo-
tion effect on metabolization with methanogenic archaea (Li et al.,
2017). Syntrophorhabdus could produce methane, acetic acid, and
butyric acid through anaerobic fermentation (Li et al., 2014). In this
experiment, the above-mentioned genera were all detected both
in the sediments of CZ and TZ. However, the relative abundances of
these genera in the sediment of CZ were much higher than those of
TZ. Meanwhile, the unique genera of specific or facultative an-
aerobes, such as Enterobacteriaceae, Gemella, Haemophilus, and
Streptococcus, were also detected in the sediment of CZ in genera
level besides the same composition of species in TZ. These genera,
commonly generated during the anaerobic digestion process,
could promote the anaerobic fermentation process, thereby
increasing the concentrations of S2 in the overlying and inter-
stitial waters of CZ. The unique genera in the sediments of TZ
included Desulfomicrobium, Nitrobacter, Nitrosomonas, and Thio-
bacillus, which could enhance the removal of nitrogen and sulfide
in water by promoting nitrification and biodesulfurization (Gu
et al., 2018; Han et al., 2017). NO 3 could oxidize H2S, sulfide,
thiosulfate, and tetrathionate to sulfate but decrease the S2 con-
centrations in the overlying and interstitial waters of TZ when it
acted as an electron acceptor in those reactions. Furthermore, the
products (such as O2, H2O2, HO and O2) from the reaction of CaO2
with H2O could reduce the pollutants concentrations by the
oxidation effect.
Therefore, CaO2 could significantly reduce the concentrations of
TCOD and S2 in the overlying and interstitial water but increase
the concentrations of SO24 . Accordingly, the black-odor phenom-
enon of landscape water could be alleviated.
4.2.3. Effect of CaO2 on turbidity and transparency of landscape
water
As previously described, CaO2 could effectively inhibit the pro-
cess of anaerobic hydrolysis, reduce the release of endogenous
pollutants from sediments, and improve the quality of black-odor
landscape water by improving the basic environment of land-
scape water and microbial community structure of sediment.
Meanwhile, CaO2 could produce O2 and HO during its dissolution
procedure in the water, which had strong oxidizing and bleaching
properties for oxidizing Fe2þ and Mn2þ to a high valence. Then, the
high-valence metal ions could produce colloid with OH, thereby
decolorizing in the black-odor landscape water. Moreover, CaO2
could promote the function of microorganism to assimilate and
degrade pollutants in water body and enhance the effect of strong
oxidative free radicals on decolorization and deodorization. As a
result, the turbidity of landscape water could be decreased,
whereas the transparency could be increased. The average values of
turbidity in the overlying water of TZ was significantly lower than
those of CZ (p ¼ 0), whereas those of transparency were obviously
higher than those of CZ. During the entire experiment, the mean
value of turbidity in the overlying water of TZ was 39.46% of that of
CZ, whereas the mean value of transparency in the overlying water
of TZ was 2.07 times that of CZ. The comprehensive purification
effect of black-odor water by adding CaO2 was superior to using
chitosan-modified clays (Huang et al., 2015).
In summary, the addition of CaO2 could effectively inhibit the
release of pollutants from sediment and hinder anaerobic hydro-
lysis by improving the anoxic condition of landscape water and the
microbial community structure in sediment. Simultaneously, CaO2
could obviously decrease the TCOD concentration, S2 concentra-
tion, and turbidity in the overlying and interstitial waters. On the
contrary, CaO2 could increase the transparency and restore the
ornamental value of the black-odor landscape water.
5. Conclusions
In the field experiment, the purification effect of CaO2 on black-
odor landscape water was systematically studied, and the purifi-
cation mechanism was explored. The main conclusions were ob-
tained as follows:
1) CaO2 could effectively alleviate the black-odor phenomenon of
landscape water by improving the anoxic condition of water
body. Meanwhile, its attributes of releasing oxygen slowly pro-
vides a time effect of CaO2 on the treatment of black-odor
landscape water, and the DO concentration in the overlying
water of TZ was maintained at a high level (DO > 3 mg L1) in
the entire experiment of 2 months.
2) CaO2 could increase the diversity of the microbial flora in sedi-
ments, reduce the relative abundance of the specific or facul-
tative anaerobes in sediments, and derive some aerobic and
functional bacteria, such as Nitrobacter, Nitrosomonas, Desulfo-
microbium, and Thiobacillus.
3) CaO2 could obviously reduce the concentrations of TCOD and
S2 in the overlying and interstitial waters of landscape water
but increase those of SO24 . Thus, the average concentrations of
TCOD in the overlying and interstitial waters of TZ were 52.98%
and 66.05% of those of CZ. The average concentrations of S2
were 29.63% and 32.79%, which were 2.52 and 2.92 times those
of SO2
4 . As a result, the formation of odorant substances and
black matters were inhibited effectively.
4) CaO2 could effectively improve the basic environment of land-
scape water, inhibit the release of pollutants from sediment, and
promote microbial assimilation and degradation rate to
 W.-H. Wang et al. / Environmental Pollution 248 (2019) 18e27
pollutants. Consequently, the turbidity in the overlying water
could be decreased, whereas the transparency could be
increased.
In summary, CaO2 could effectively alleviate the black-odor
phenomenon and excellently restore the ornamental value of
landscape water. Thus, for practical applications, addition of CaO2
to the black-odor landscape water has a positive effect on water
ecological environment and opens new avenues for the use of CaO2
in the future.
Acknowledgments
This work was funded by the National Natural Science Foun-
dation of China (Project No. 21677115) and the Shaanxi Provincial
Natural Science Foundation Research Key Project (Project No.
2016JZ019).
Appendix A. Supplementary data
Supplementary data to this article can be found online at
https://doi.org/10.1016/j.envpol.2018.11.069.
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