Spectrochimica Acta Part A: Molecular and Biomolecular Spectroscopy 217 (2019) 263–270

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Spectrochimica Acta Part A: Molecular and Biomolecular

Spectroscopy
journal homepage: www.elsevier.com/locate/saa

Investigation of mineralogical and bacteria diversity in Nanxi River

affected by acid mine drainage from the closed coal mine: Implications
for characterizing natural attenuation process
Jianwen He a, Wenxu Li a, Jing Liu b,⁎, Shu Chen a, Ray L. Frost c,⁎
a
The Key Laboratory of Solid Waste Treatment and Resource Recycle, Ministry of Education, Southwest University of Science and Technology, Mianyang, Sichuan 621010, PR China
b
The School of Resource and Environment, Southwest University, Chongqing 400715, PR China
c
School of Chemistry, Physics and Mechanical Engineering, Science and Engineering Faculty, Queensland University of Technology, 2 George Street, GPO Box 2434, Brisbane, Queensland 4001,
Australia

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

Article history: Due to the supply-side reform and environmental protection in China, many small coal mines have been closed
Received 18 November 2018 since 2015. However, acid mine drainage from these coal mines are continuously discharging into many rural
Received in revised form 16 March 2019 creeks, which requires the systematical investigation on the variations of geochemical and environmental biolog-
Accepted 18 March 2019
ical aspects in these water systems. In this study, from a classic acid mine drainage (AMD) from a closed coal mine
Available online 21 March 2019
of Hunan, China, various sediments and water samples in different sections were collected and analyzed. Accord-
Keywords:
ing to the corresponding Mineralogical and simple bacterial characteristics analysis (16S rRNA gene sequencing),
Closed coal mine the main findings were: 1) Secondary iron-containing minerals gradually transited from Gr(CO2− 3 ) (green rust),

Mineral-water interface Sh (schwertmannite) to Akg (Akaganeite) and more stable Gt (Goethite); 2) compared to the pristine sediment,
Mineralogy these minerals decreased the acid-neutralizing capacity and cation exchange capacity (CEC) of sediments;
16S rRNA 3) Proteobacteria and Firmicutes were the dominant phyla and the obvious variation of Firmicutes species was ob-
Natural attenuation served in the creek affected by AMD, which probably could been a biological index to diagnose the natural atten-
uation of AMD. These results could be greatly significant to understand typical variations of creek attenuation and
bacterial community in the presence of high metal and sulfate concentration.
© 2019 Published by Elsevier B.V.

1. Introduction
Acid mine drainage (AMD) is environmental problem in many coun-
tries due to the potential heavy metals and corrosive acidity. Pyrite is a
common mineral associated with coal and is a major contributor to
AMD. Coal often contains high concentrations of Fe (0.1–300 mg/L), Al
(0.1–60 mg/L), Mn (0.1–100 mg/L) and SO2− 4 oxyanions
(200–2000 mg/L) [1], which readily affect local creaks, rivers and oceans.
In AMD, based on their self-physical and chemical factors, including
saturate index, water chemistry and transport velocity [2,3], various
metals finally form complex secondary minerals, such as labyrinthian
mineral combination with different deposit types. Additionally, due to
the geochemical variation along the river, amorphous iron hydroxides
and aluminum-containing minerals, such as goethite (α-FeOOH), Jarosite
(KFe3(SO4)2(OH)6) [4], aluminum-flocs (Al13O4(OH)24(H2O)7+ 12 ), alum
(KAl3(SO4)2(OH)6) and gibbsite (Al(OH)3), are formed. More than forty
iron, aluminum and manganese-containing secondary minerals have
been found in AMD [5].
⁎ Corresponding authors.
E-mail addresses: Liujing-vip@163.com (J. Liu), r.frost@qut.edu.au (R.L. Frost).
These minerals have different surface areas thereby having varying
potentials to remove. For example, schwertmannite is an iron-sulfate
hydroxide with a specific area of 200 m2/g compared to goethite with
40 m2/g. The ability of these secondary minerals to adsorb heavy and
toxic metals results in the natural attenuation of AMD within creeks
[6]. However, the understanding on this field is still limited probably
due to the emphasis on active remediation on AMD [7].
Due to the supply-side reform and environmental protection, the
Chinese office of the state council promulgated a file in 2013 to
strengthen the closure of N2000 small-scale coal mines (annual output
is b6 × 105 tons of standard coal) since 2015, especially in Hunan prov-
ince with abundant coal resources and complete coal types in China [8].
Additionally, the exposed pyrite-containing strata and excavation tun-
nel in these mines are continuously exposed to oxidative wreathing.
These AMD-affected creeks and rivers often are in rural area; thus, the
contemporaneous and effective treatment cannot be done. Therefore,
natural attenuation comes from the secondary minerals, primary prop-
erty of strata along watershed could be a main remediation mechanism.
Due to bacteria diversity physiological habits, the distinct zoning
maybe occurs in the runoff of AMD. For example, various acidophilic
bacteria (YARDs1-4), acidithiobacilus ferroxidase and sulfur oxidizing

https://doi.org/10.1016/j.saa.2019.03.069
1386-1425/© 2019 Published by Elsevier B.V.
264 J. He et al. / Spectrochimica Acta Part A: Molecular and Biomolecular Spectroscopy 217 (2019) 263–270
bacteria were identified according to the 16S rRNA sequence. Robbins
investigated microbiology in AMD with pH b 4.5 and reported that
there were 86 genera or species in it. Gónzalez-Toril [9] collected AMD
from the LaZarza-Perrunal mine in the Iberian pyrite belt and also
found sulfate reducers in the initial site (pH = 3.1) and ion-oxidizers oc-
curred with the downstream distance.
To investigate the potential natural attenuation in AMD-affected
water systems, the AMD-affected stream water is an ideal site. The min-
eralogy of sediments in the creek was characterized and bacteria diver-
sity also was investigated with 16S RNA [10,11]. The potential
neutralization and re-adsorption ability of sediments from downstream
was also evaluated. This study provides a better understanding for the
potential of rehabilitating AMD-affected creeks and rivers in Hunan.
2. Studied area
The AMD-affected creek (Nanxi River) in Shimen, Changde, Hunan,
China was chosen. The creek flows through Baiyun Town, where the
state has compulsorily closed a coal mine in October 2015, and finally
joins into Shishui River. The creek is in suburb without influence from
any industrial and mining enterprise and other creeks, the creek should
be a typical one to study the natural attenuation affected by AMD from
Fig. 1. Map of the Nanxi River showing eight sampling points; the sampling position in the figure was S5 (bottom right corner).
closed coal mine. The creek is approximately 14 km (from 29°38′00″N
to 29°40′00″N and from 111°08′00″E to 111°11′00″E) (Fig. 1).
3. Materials and methods
3.1. Collection of sediment and aqueous samples
Sediment and water samples were collected from eight locations
along the Nanxi River where changes in pH of the water were markedly
different. A sample (S0) from clean creek before AMD input into the
river was collected for the necessary comparative sample. The sample
(S1) was located at the entrance of the closed mine, not the tributary
of the Nanxi River. The longitude and latitude of these sites was deter-
mined with a portable GPS receiver (Trimble). Positions of sampling
sites were listed in Fig. 1. The pH values of water samples were mea-
sured with calibrated Portable Hach HQ40D immediately. Samples
were transferred to the laboratory where sediments were kept at 4 °C
for b2 days until the geochemical analysis were performed.
All water samples were filtered using a 0.45 μm membrane and an-
alyzed for total calcium concentration (Ca(T) detection limit was
0.002 mg/L), total iron concentration (Fe(T) detection limit was
0.002 mg/L) and total magnesium concentration (Mg(T) detection
 J. He et al. / Spectrochimica Acta Part A: Molecular and Biomolecular Spectroscopy 217 (2019) 263–270 265

limit was 0.001 mg/L) using an Inductively Coupled Plasma Emission immediately were sent to Changsha of Hunan, and send to Tianjin
Spectrometer (ICP-AES, iCPA6500 Thermo Fisher) after samples passed Nuohe Zhiyuan biological analytical center with refrigerated transport
through 0.45 μm membrane. Sulfate concentrations of aqueous samples within 2 days.
were determined with Ion chromatographic method (IC, Metrohm, 881
Compact pro) [12,13]. 4. Results and discussion

3.2. Mineralogical characterization of sediments 4.1. Aqueous chemical characteristics

Mineral composition of the sediment sample was determined using
X-ray diffraction analysis. The minerals were identified from the XRD
patterns (diffract grams) according to position, intensity, shape and
breadth of peaks [14]. Most of the important peaks were identified
within the 2θ range from 8° to 43°. After sediments were dried at 80
°C for 24 h, they were passed through 100 mesh sieves for the XRD.
And in the attenuated total reflectance Fourier transform infrared spec-
troscopy (ATR-FTIR) analysis, ATR mode was done directly on the sieved
powder with Perkin Elmer Spectrum (4000–400 cm−1, a spectral reso-
lution of 4 cm−1).
3.3. Cation exchange capacity (CEC)
After the creek goes through clay-rich barriers, the migration of cat-
ions and molecules might be influenced, which can be characterized by
CEC. Adsorbed cations and anions are in equilibrium with similar cat-
ions and anions in solution. When a particular ion is adsorbed, it is
largely replaced by some previously-adsorbed on soil particles. Accord-
ing to the cation exchange ability of sediment, the adsorption perfor-
mance of sediment can be determined to a certain extent. Compared
with the common Ca, Mg, Na and K in the soil [11,15], under the influ-
ence of AMD, the sediment cations at the bottom of Nanxi River were
mainly Fe, Ca and Mg. Therefore, in this study, only these three cations
were determined. The potential purification capacity of the bottom sed-
iment of the river could be measured according to the changes of CEC.
The cation exchange capacity of sediment sample was measured ac-
cording to the NaOAc (pH 8.2) method. The sum of adsorbed positive
charge per kg is the cation exchange capacity (CEC, units of cmolc/kg).
The pH values and chemical analysis of filtered samples (Table 1) re-
vealed the presence of sulfate, Ca(T), Fe(T) and Mg(T)at concentrations
much greater than of comparative point (S0), which represents a tribu-
tary to the Nanxi River that is not contaminated. Due to Hunan province
stratum is the product of pre-Cambrian boundary event of abrupt
change in paleoenvironment [17], the geological layer contains a lot of
calcium and sulfate. The concentrations of sulfate and calcium compar-
ative also higher than others elements in natural creek (S0). As showed
in Table 1, all pH values were from 4.77 to 8.10, closing to that of the
comparative sample S0. The Fe(T) in these samples were decreased
from 46.27 to 0.05 mg/L, Ca(T) from 26.79 to 5.46 mg/L, and Mg(T)
from 14.76 to 1.12 mg/L. And, synchronously, the sulfate concentrations
were decreased from 2509.36 to 74.04 mg/L. Compared with the com-
parative sample S0, the AMD-contaminated Nanxi River water quality
seemed to recover to its initial level after about 2 km from the source
through self-purification.
4.2. Sediment of FTIR and XRD analysis
The representative XRD and FTIR of sediments are shown in Figs. 2
and 3. The XRD and FTIR shows GrCO2– 3 (green rust) was found in S1
and Gt (goethite) was found in S2, while Sh (schwertmannite) in both
S1 and S2. Green rust compounds can be represented with the general
formula [FeII(6−x) FeIIIx (OH)12] x+[Ax/n− ∗ yH2O] x−, where A is an n−-
valent anion (mainly Cl−, CO2− 2−
3 and SO4 ) (Eq. (1)). In groundwater,
carbonate forms of Gr precipitated preferentially over its sulfate form
[18], these form can exchange due to sulfate was the dominant anion
in the Nanxi River.

3.4. Neutralization ability of sediment Fe6 ðOHÞ12 SO4 þ CO2− 2−

3 →Fe6 ðOH Þ12 CO3 þ SO4 ð1Þ
h i
The collected sediments were air-dried, passed through 100 mesh 2−  
K 1 ¼ SO2−
4 =CO3 Fe6 ðOH Þ12 CO3 =Fe6 ðOH Þ12 SO4
sieve and stored in a plastic bag. Approximately 1.0 g of each sample
was weighed and put into glass bottle (6 glass bottles in total), and
100 mL working solution with initial pH as 3.8 were added into each Gr (SO2−
4 ) consists of alternating positively charged brucite-like iron

bottle, which were agitated (150 r/min) at 30 °C. The pH values of solu- hydroxide layers of [Fe2+ 3+
4 Fe2 (OH)12]
2+
and negatively charged inter-
2−
tions were adjusted with H2SO4 solution. The pH of solution was mea- layers of anions [SO4·8H2O] , through which the electroneutrality is
sured at reaction time of 10, 20, 30, 40, 60, 80, 110, 140, 200, 320, 440, maintained [19]. Due to the oxidation of intermediate amorphous
1040 and 1760 min. According to the pH changes of solution, the neu- metal sulfides, such as amorphous FeS (asFeS), the unstable Gr(SO2− 4 )

tralization ability of sediment was determined. tends to convert into Gt(α-FeOOH), which is the most stable iron
oxide mineral [20] (Eq. (2)). While Sh is commonly found in near-
3.5. 16S rRNA analysis on sediments from different sections surface environments and in unstable compounds. The dissolution reac-
tion of Sh was (Eq. (3)).
According to previous studies, microbial diversity was affected by
pH, ion concentration [16]. The natural attenuation trend of AMD pollu- 4Fe2þ þ HS− þ 7OH − →FeSðamÞ þ 3FeðOHÞ2 ðamÞ þ H 2 O ð2Þ
tion had an impact on the distribution of sediment biodiversity at the
bottom of the river. 16S rRNA sequence similarity is now accepted as a Table 1
method for judging the diversity of microbial communities. Therefore, The pH values and elemental compositions of water samples along the Nanxi River.
pelagic and particle-associated organisms from sediments were used Samples pH Element content
for 16S rRNA gene cloning and sequencing, including amplicon genera-
Fe(mg/L) Ca(mg/L) Mg(mg/L) SO4(mg/L)
tion, PCR products quantification and qualification, library preparation
and sequencing. The rarefaction curves, species richness estimators S0 8.50 0.03 3.84 0.56 281.84
S1 4.77 46.27 26.79 14.76 2509.36
and community diversity indices were obtained based on Qiime soft-
S2 3.83 23.09 25.20 14.78 2458.72
ware (Version 1.7.0). S3 5.66 2.26 10.94 4.62 796.61
In order to determine the bacterial diversity, five samples were S4 5.56 0.92 10.57 3.97 651.28
taken along line of creek. They were stored in centrifugal tubes and S5 7.68 0.03 7.67 1.83 224.81
kept in a 5 °C box with ice bags in order to decrease their bacteria vari- S6 7.61 0.06 6.26 1.39 111.27
S7 8.10 0.05 5.46 1.12 74.04
ation. Due to the sampling were done in summer, these samples
266 J. He et al. / Spectrochimica Acta Part A: Molecular and Biomolecular Spectroscopy 217 (2019) 263–270

Fig. 3. ATR-FTIR spectra of sediment samples.

Fig. 2. XRD patterns of sediments from AMD-polluted Creek (GrCO2−

3 : Green Rust; Gp:
Gypsum; Sh: Schwertmannite; Gt: Goethite; Akg: Akaganeite; Kln: Kaolinite; Qz:
Quartz; Ill:illite).
The peaks from 900 to 1100 cm−1 of S5 and S6, are different from S1-
S4, could be the characteristic bending of Al-OH and stretching of Si\\O
vibrations, principally illite and kaolinite [25]. The peaks at 1163, 1088,
778, 693, and 438–475 could be the presence of quartz mineral [26–28].
Fe8 O8 ðOHÞ8−2x ðSO4 Þx þ ð24−2xÞHþ →8Fe3þ þ xSO2−
4
The XRD and ATR-FTIR comparing the sediments of each samples
þ ð16−2xÞH2 O ð3Þ showed that amorphous particles likely formed first, then aged to be-
come more iron oxyhydroxides, such as, Akg and Gt.

This might also explain Sh was not detected by XRD in S3–S6. An in-
tense band at 1124 cm−1 reflect a strong splitting of V3(SO4) due to the 4.3. Cation exchange capacity (CEC) of sediments
formation of a bidentate bridging complex between SO4 and Fe, while
the bend at 976 cm−1 was due to the presence of structural SO4 in VI As shown in Table 2, the CEC was decreased at the beginning and
(SO4) (Fig. 3) [21]. then increased within a broad range, indicating that the AMD had a sig-
Akg(akaganeite), occurs rarely in soils and is more commonly asso- nificant effect on the CEC of river sediments, probably including parent
ciated with chloride rich environments such as hot brines, has been material, age of weathering surface, climatic factors, degree of wreath-
found in marine environments, carbonate concretions and sulfide-rich ing, completeness of chemical and physical dispersion [29]. With the
sediments [22]. According some previous study [22,23], sulfate- discharge of AMD, the content of Fe and other cations in the river was
containing akaganeite was an intermediate phase between Akg and Sh gradually increased, causing the change in the form of ferriferous min-
[(Fe8O8(OH)−x 6 (SO4)x·nH2O), a structural analog of akaganeite, con- erals with a clear color change of river bottom sediment, from tan to
taining sulfate instead of chloride in tunnels]. In this creek, with the reddish brown. Thus, the change trend of CEC decreases. After the
high concentration of sulfate and the dissolution of Sh, sulfate- river self-purification and mineral natural attenuation, the CEC was
containing was formed in sample S4. Due to Gt(α-FeOOH) and Akg(β- gradually recovered to its initial natural level (approximately 45.21
FeOOH) have simile bands, the XRD results of S3-S4 had no significant cmolc/kg).
difference vibration peak. While the SO4 bands of S4 in 976 and
1174 cm−1 weaker compared with S1–S3, there are possible that chlo- Table 2
ride replace sulfate in Akg's tunnels. An intense bands at 615, 798, The cation exchange capacity (CEC cmolc/kg) of sediments.
905 cm−1 and 693, 844 cm−1 were due to the presence of Gt and Akg Samples S0 S1 S3 S4 S5 S6 S7
[24] (Fig. 3), which was consistent with XRD results, showing that Akg
CEC (cmolc/kg) 45.21 20.37 8.08 12.72 9.12 44.07 43.34
and Gt were mainly Fe-mineral forms in this acidic creek.
 J. He et al. / Spectrochimica Acta Part A: Molecular and Biomolecular Spectroscopy 217 (2019) 263–270
4.4. Neutralization ability
As shown in Fig. 4, all sediments had a different respond to acidic so-
lution (pH = 3.8). Sh has two dissolution reaction Eqs. (3) and (4),
Fe8 O8 ðOHÞ8−2x ðSO4 Þx ¼ 4Fe2 O3 þ 2xH þ þ xSO2−
4 þ ð4−2xÞH 2 O
In S1 and S2, pH value was slight decreased to 3.5 and 2.4 with extra
H+ added. In S3, pH value was gradually increased due to the neutrali-
zation reaction between H+ and the stable Fe-containing minerals. In
S4, because Gt (α-FeOOH) was more stable than Akg (β-FeOOH) [30].
Additionally, because OH– and H2O could form a complex with Fe3+
or Fe2+, the formation of a complex in an aqueous solution was actually
a reaction in which a ligand replaced H2O in a hydrated ion to form a
new complex. Furthermore, H+ with high concentration in the reaction
mixture competed with positive ions for the adsorption sites [31], and
the pH value of S4 solution was decreased to 3.3.
Self-purification of the stream is attributed to the mineral that com-
pose fraction of the sediments, these minerals include Qz, Kln and Ill.
The neutralization capacity of the downtown sediments (S5 and S6)
Fig. 4. The pH changes with the reaction time after an acidic solution was added.
267
has returned to its natural level. Compared to the initial sediment (S0)
and the lowest downstream sediments, the pH value of sediment from
AMD-affected creek was quickly decreased to pH = 3–4, indicating
the formation of secondary iron-containing minerals might result in
the enhance of an acidity in the creek.
ð4Þ
4.5. 16S rRNA diversity analysis
According to the results of species annotation, 10 species with the
highest abundance of each sample at phylum classification level were
selected to generate a column cumulative graph of relative abundance
to visually characterize the relative abundance of species and their pro-
portions at phylum levels for each sample. The result (Fig. 5) showed
the bacterial composition and the similarity of the all eight Nanxi
River sediment samples, indicating that Proteobacteria and Firmicutes
were the dominant phyla, accounting for 39.2%–57.2% and 0.5%–25.6%
relative abundance in phylum, respectively. This is consistent with pre-
vious studies [32,33]. In addition to Proteobacteria and Firmicutes, the
other two most AMD-affected microbial phyla are Acidobacteria
(3.1–10.7%), and Bacteroidetes (4.4–20.9%). Bacteroidetes was one of
268 J. He et al. / Spectrochimica Acta Part A: Molecular and Biomolecular Spectroscopy 217 (2019) 263–270

Fig. 5. UniFrac UPGMA cluster of microbial communities in different sediment samples, which was constructed on the basis of Illumina sequencing data (left); relative abundance of
bacteria in sediments as term of phylum (right).

the most abundant phyla in mining area sediment, up to 17.2% [34]. In
this study, the abundance was gradually increased from 7.4% to 20.9%
relative abundance in phylum with the AMD influence, and then de-
crease to 4.4%.
Further analysis of microbial composition of Nanxi River sediments
showed a community dominated by microorganisms commonly found
in AMD environments such as iron-, sulfur-, sulfide- and thiosulfate-
oxidizing bacteria (Fig. 5). And the tendency of Acidobacteria abundance
was gradually decreased from 7.0% to 2.6% when the pH value was de-
creased from 8.5 to 5.6. When pH was increased from 5.6 to 8.1, the
abundance was increase from 2.6% to 10.7%. This correlation between
Acidobacteria and pH was consistent with a series of studies on soil
and sediment [35]. However, a negative correlation between
Acidobacteria and pH was also observed in the presence of some
material stream [36,37] and some metal sediments. In this acidic
creek, may due to the influence of AMD and other bacterial, with the
pH was higher than 5.6, the Acidobacteria abundance was increased.
The similarity among the microbial communities in the eight sedi-
ment samples was evaluated with cluster analysis. As shown in Fig. 5,
bacterial communities could be clustered into two groups. Group I con-
tain samples S1 to S4. Groups II contain S0, S5-S7, respectively. These re-
sults indicated that microbial community structure was probably
correlated directly with the AMD, which could be also demonstrated
in the canonical correspondence analysis (CCA) [38]. As shown in
Fig. 6, the microbial communities of S0, S6, S5 and S7 were positively
correlated with pH, while those of S3, S2, S1 and S4 were positively cor-
related with sulfate and metal ion (Fe, Mg and Ca). These results con-
firmed the effect of AMD on bacterial diversity in sediments. The

Fig. 6. The canonical correspondence analysis (CCA) between 16S rRNA gene date and environmental parameters. The arrows show the direction and size of environmental parameters
associated with bacterial community structure.
 J. He et al. / Spectrochimica Acta Part A: Molecular and Biomolecular Spectroscopy 217 (2019) 263–270
stream self-purification and natural attenuation, the downstream (S5
and S6) bacterial diversity had returned to a level similar to that of S0,
before the Nanxi River passes through the Baiyun Town.
In conclusion, we present a possibility on the potential for iron
change process in this acid creek affected by indigenous bacterial com-
munities. Amorphous particles likely formed first, then aged to become
more crystalline iron oxyhydroxides, such as Akg and Gt in this acid
creek, containing high concentrations of sulfate and iron. Some of ini-
tially smooth, spherical particle in S1 may formed Fe-bearing flocculent
substance, which could potential provide high reactive surface area for
chemical and biological reactions in the aging process. The tendency
of metal concentrations in creek water revealed that elements such as
Ca and Mg also could be adsorbed and transformed at iron change
process (Table, 1). Pelagic and particle-associated organisms of
sediments showed that members of the Proteobacteria (mainly
Betaproteobacteria, Alphaproteobacteria and Gammaproteobacteria)
dominated bacterial communities, while the Firmicutes changed sig-
nificantly bacterial communities. Firmicutes increased from 3.1% to
25.6% relative abundance in phylum suddenly under the influence
of high concentration of metal ion and sulfate [39]. There is active
microbial iron cycling in heavy metal contaminated creeks. Based
on the above phenomenon, we guess the biomass of Firmicutes
could be used as a biological index to diagnose the natural attenua-
tion of Nanxi River containing high concentration of sulfate and
metal ions (Fe, Ca and Mg). As for the functioning of Firmicutes
within this environment need further research.
5. Conclusions
In this study, the natural attenuation of Nanxi River sediments and
the changes of microbial communities under the influence of AMD
were investigated. The main results are:
(1) With the high concentration of metal ions and sulfate in Nanxi
River, Fe-containing minerals like amorphous particles likely
formed first, then aged to Sh (schwertmannite), last become
more crystalline iron oxyhydroxides, such as Akg (Akaganeite)
and Gt (Goethite).
(2) Due to the variation of mineral species in sediment, the
sediment's adsorption ability and CEC were changed.
(3) Through CCA and UniFrac UPGMA cluster analysis, the relation-
ship between the Firmicutes abundance and environmental con-
ditions was interpreted. The biomass of Firmicutes response
significantly to sediment natural attenuation in the presence of
high concentration of metal and sulfate.
Acknowledgments
This study was supported by the National Natural Science Founda-
tion of China (41772367).
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