Science of the Total Environment 706 (2020) 136086

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Science of the Total Environment

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

Biochar-induced migration of tetracycline and the alteration of microbial

community in agricultural soils
Hua-Yu Liu, Chao Song ⁎, Shan Zhao, Shu-Guang Wang ⁎
Shandong Key Laboratory of Water Pollution Control and Resource Reuse, School of Environmental Science and Engineering, Shandong University, Qingdao 266237, China

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

• Adsorption is the dominant dissipation

way for tetracycline in soils.
• Biochar promoted vertical migration of
tetracycline.
• Biochar and tetracycline greatly shifted
bacterial and fungal community compo-
sition in 90 days.

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

Article history: Recently, biochar is widely used as a soil amendment to improve soil properties, which might affect the fate and
Received 2 September 2019 behavior of contaminants in soil. In this study, we investigated the effect of biochar on the migration of tetracy-
Received in revised form 1 December 2019 cline (TC) in soil and their combined impacts on microbiome. Due to the strong interaction between soil and TC,
Accepted 10 December 2019
adsorption, rather than photolysis or biodegradation, was the dominating dissipation way of TC in soil. Moreover,
Available online 12 December 2019
biochar could promote the vertical migration of TC through the decreased soil bulk density and its lower adsorp-
Editor: Baoliang Chen tion capacity. After 90-day incubation, only slight impact of TC on soil bacterial community was observed due to
the rapid dissipation of TC in soil, whereas more available C supply induced by biochar significantly altered bac-
Keywords: terial community via the enhancement of copiotrophic bacteria. Besides, biochar could decrease the soil pH and
Tetracycline thus change the composition of fungal community. The effect of TC on fungal community was partially
Biochar counteracted by biochar, which could adsorb part of TC and thus decrease the contact of TC with microorganisms.
Soil This work will improve our understanding of the fate of organic pollutants and evolution of microbiome in soil
Microbiome where biochar servers as soil amendment.
Migration
© 2019 Elsevier B.V. All rights reserved.

1. Introduction

⁎ Corresponding authors. Nowadays, tetracycline antibiotics are widely used in the treatment
E-mail addresses: songchao@sdu.edu.cn (C. Song), wsg@sdu.edu.cn (S.-G. Wang). and prevention of infections for livestock (Zhou et al., 2013).

https://doi.org/10.1016/j.scitotenv.2019.136086
0048-9697/© 2019 Elsevier B.V. All rights reserved.
2 H.-Y. Liu et al. / Science of the Total Environment 706 (2020) 136086
Unfortunately, 30%–90% of intake antibiotics cannot be absorbed by
body, and they are excreted as parent compounds or metabolites
through urine and feces into the soil (Erşan et al., 2013; Ji et al., 2012;
Zhao et al., 2010). Besides, the manure contaminated with antibiotics
is also applied as fertilizers onto arable lands, leading to the enrichment
of antibiotics in the soil (Duan et al., 2017). Residues of tetracycline have
already been detected at concentrations ranging from ng/g to μg/g in
soil (Song et al., 2017; Thiele-Bruhn and Beck, 2005). As a broad-
spectrum antibiotic, tetracycline could exhibit adverse influences on
soil ecosystem, such as inhibiting bacteria activity, disturbing microbial
metabolism and even changing soil microbiome (Jechalke et al., 2014;
Kong et al., 2006). The residues would also induce the evolution of anti-
biotic resistance genes, which are considered as a potential threat to
both ecosystem and human health (Hong et al., 2013). Therefore, un-
derstanding the fate and behavior of tetracycline in soil environment
becomes indispensable.
Biochar is a carbon-rich material which is produced by pyrolysis of
various forms of biomass (De la Rosa et al., 2019; Duku et al., 2011). In
recent years, biochar is commonly used as a soil amendment to improve
soil properties (Srinivasan et al., 2015). Moreover, biochar could in-
crease the microbial biomass and activity, and even reshape the struc-
ture of microbiome (Ahmad et al., 2014). It is worth noticing that
biochar is a porous material with high surface area and exhibits excel-
lent adsorption capacity on organic contaminants due to its surface aro-
maticity and functionality (Vithanage et al., 2016). Several studies have
reported the biochar based removal of antibiotics such as tetracycline,
ciprofloxacin and sulfonamides (Afzal et al., 2018; Ahmad et al., 2014;
Rajapaksha et al., 2016). Thus, biochar may exhibit a significant impact
on the migration of tetracycline in contaminated soils, which is highly
related to the behaviors of tetracycline in soils (Thiele-Bruhn, 2003).
However, the effects of biochar application on the migration and behav-
iors of tetracycline in soils, as well as its underlying mechanism, are still
unclear.
In this study, we investigated the impacts of biochar on the migra-
tion and behaviors of tetracycline in soils, and the objectives of this
study were as follows: (i) to quantify the temporal dissipation and ver-
tical migration of tetracycline in natural soils with/without biochar
amendment; (ii) to explore the roles of tetracycline and biochar on
soil microbial community; and (iii) to elucidate the combined effects
of tetracycline and biochar on soil microbiome. This work will extend
our understanding of the fate and behavior of antibiotics in soils,
which is vitally important to control the persistence and negative effects
of antibiotics in agricultural environment.
2. Materials and methods
2.1. Chemicals and reagents
Tetracycline hydrochloride (TC) was of USP grade and purchased
from Aladdin Industrial Corporation, USA. Acetonitrile, methanol and
other chemicals used for mobile phase were of HPLC grade and obtained
from Sinopharm Chemical Reagent Company, China. The biochar was
purchased from Huansheng Carbon Production Company, China,
which was produced from coconut shell at 300-500 °C in a vertical
kiln and its particle size was 1–3 mm. All other chemicals were of ana-
lytical reagent grade and used without further purification.
2.2. Soil samples and biochar characterization
Fresh soils were collected at a depth of 0–20 cm from Shandong Uni-
versity in Qingdao, China on June 2018. The samples were mixed and
sieved with a 2 mm-mesh screen to remove large particles. Meanwhile,
the initial soil was analyzed, and no tetracycline was detected.
The surface area and pore characteristics of biochar were measured
by N2 adsorption using a SSA-4300 surface area analyzer. The basic char-
acteristics of biochar were: special surface area (SSA) =1116.16 m2/g,
the total pore volume (TPV) =1.65 cm3/g, the average pore radius
(APR) = 4.17 nm. The isotherm curve (Fig. S1) showed hysteresis
loops in the relative pressure (p/p0) ranging from 0.2 to 1.0, suggesting
uniform mesopores in biochar structure.
2.3. Temporal dissipation of TC
The sieved soils were divided into four group: control group, dark
group, sterilized group, and dark-sterilized group. In each group, thirty
petri dishes were prepared with 10 g soil (dry weight) and TC at
50 μg/g soil. In control group, all dishes were placed on laboratory
table at room temperature without further operation. In dark group,
aluminum foil was used to cover the dishes, which were then kept in
dark place. For sterilized group, the dishes with 10 g soil were
autoclaved at 121 °C for 30 min before adding TC, and then they were
placed in the same condition with control group. In dark-sterilized
group, all dishes were covered with aluminum foil and placed in dark
after autoclaved. At predetermined time intervals, three soil samples
in one dish of each group were taken for further analysis to determine
the amount of TC. Each dish was used for one-time sample collection.
2.4. Vertical transport of TC
The sieved soils (60 g) were filled into a tube (3.5 cm inner diame-
ter), where they were designated as Layer 1 to Layer 6 from top to bot-
tom equally based on their depth.
TC was spiked to Layer 1 at the dosage of 100 μg TC/g soil. In addition,
biochar was mixed in Layer 2 at 2% w/w, which was considered as the
most suitable proportion (Huang et al., 2018). The tube without biochar
was also prepared as control. All the tubes were covered with aluminum
foil and kept in dark. Deionized water was sprayed to soils as fine drop-
lets using a plastic nebulizer. The spray was very slow with suitable time
interval to make the water equally distributed. The total volume of
sprayed water was 2.4 mL, which was amount to the rainfall of a
heavy rain in China (Day et al., 2018). Then, amounts of TC in each
layer were measured to investigate its acute vertical migration in soils.
All experiments were repeated in triplicate, and the error bars represent
the standard deviations calculated for each independent experiment.
2.5. Amount of TC in soil
TC in soils was extracted using the method described in previous
study (Pan and Chu, 2016) with minor modifications. Briefly, soil
sample (1 g) was transferred to a 50 mL centrifuge tubes with
10 mL of 1/1(v/v) EDTA-Mcllvaine buffer (pH = 4.0). Then, the sam-
ple was vortexed for 1 min, ultrasonically extracted for 30 min, and
centrifuged at 4500 min for 5 min. The supernatants were all
decanted into a 50 mL centrifuge tube, and the sediment was treated
twice again with the same procedure to achieve a high extraction ef-
ficiency. All supernatants were collected together, filtered with
0.45 μm fiber filter (purchased from Shanghai Xinya purification de-
vice factory), and then pumped into an Oasis HLB cartridge (6 mL,
500 mg, Waters) at a flow of 3–5 mL/min. Then, the cartridge was
rinsed with 15 mL ultrapure water and eluted with 8 mL methanol
(containing 0.1% formic acid, v/v), followed by collecting the eluent
for HPLC analysis.
The quantification of TC was carried out using a high-performance
liquid chromatography (HPLC) system equipped with an ODS C18 col-
umn (4.6 × 150 mm, 5-μm). The wavelength of UV detector was set at
360 nm. The mobile phase (v/v) consisted of 0.01 M anhydrous oxalic
acid (80%), acetonitrile (16%) and methanol (4%) at a flow rate of
1.0 mL/min. Each sample was filtered through a 0.22 μm membrane fil-
ter before analysis.
 H.-Y. Liu et al. / Science of the Total Environment 706 (2020) 136086
2.6. Soil microbiome analysis
Batch experiments were designed to evaluate the alteration of
soil microbiome. As shown in Fig. 1, glass column reactors (internal
diameter 7.5 cm, height 13 cm) were designed with porous PVC
plates and 1 cm-diameter pipes at the bottom. A piece of filter
paper covered on the PVC plate was arranged to prevent the leak-
age of soil. The sieved soils (480 g) were added into reactors and
designated as Layer 1 to Layer 6 (2 cm for every Layer) from top
to bottom. Based on the amounts of TC and biochar, all reactors
were divided into four groups: control group with only sieved
soils (CK group), control group with 2% (w/w) biochar in Layer 2
(BC group), experimental group with 50 μg/g TC in Layer 1 (TC
group), experimental group with both TC (50 μg/g in Layer 1) and
biochar (2% in Layer 2) (BT group). Artificial rain (11.55 mL, equiv-
alent to 2.5 mm) was sprayed to the soils by a plastic nebulizer
within 1 h to simulate rainfall and maintain moisture. The rain
was conducted every 3 days and the entire process lasted for
90 days. Three parallel reactors were set in identical operation for
each group. Soil samples were collected from each reactor after
90-day operation for further analysis.
DNA was extracted from 500 mg of soil using PowerSoil DNA
Isolation Kit (MoBio Laboratories, Carlsbad, CA) according to the
manufacturer's instructions. The extracted DNA was amplified via
PCR on a Mastercycler Gradient (Eppendorf, Germany), and the
products were purified using a QIA quick Gel Extraction Kit
(QIAGEN, Germany). Then, the DNA samples were analyzed with
16S rRNA and 18S rRNA high-throughput pyrosequencing. The
raw data and sequences were first quality-filtered and clustered
into operational taxonomic units (OTUs) at a similarity level of
97% using Illumina Analysis Pipeline Version 2.6 and QIIME
(Edgar, 2013). And then the sequences were classified into differ-
ent taxonomic groups by Ribosomal Database Project (RDP) Classi-
fier tool (Cole et al., 2009). For bacterial analysis, a total of 18,649
reads belonging to 2733, 2564, 2664, 2651 operational taxonomic
units (OTUs) of CK, BC, TC and BT treatment were found, respec-
tively. For fungal analysis, 17,560 reads belonging to 681, 850,
585, 836 operational taxonomic units (OTUs) of CK, BC, TC and BT
treatment were found, respectively. The richness and diversity in-
dices were calculated and PCA were used using R (Wang et al.,
2012). And an Unweighted Pair Group Method with Arithmetic
Mean (UPGMA) was used to describe the dissimilarity between
multiple samples (Jiang et al., 2013).
Fig. 1. Schematic design of the soil column experiments among different groups. CK: control group; BC: biochar group; TC: tetracycline group; BT: biochar and tetracycline group.
3
3. Results and discussion
3.1. The short-term fate of TC in soil
The temporal dissipation of TC was first investigated in natural soils.
As shown in Fig. 2, the concentration of TC decreased from 50 μg/g to
9.98 ± 0.89 μg/g after 30 days in control group, indicating that most
TC (80.0% ± 1.77%) could be removed under natural conditions. Similar
results were also obtained in the dark group with final concentration of
9.54 ± 2.01 μg/g, suggesting that little TC was eliminated via
photodegradation. Previous study monitored dissipation of antibiotics
in manure piles and found no apparent effects of light on tetracycline
(Storteboom et al., 2007). Similarly, photodegradation of antibiotics is
assumed to be limited due to poor light penetration in realistic soils
(Ozaki et al., 2011). The role of hydrolysis was also evaluated in this
study, and there was no obvious change in Fig. S2, indicating the little
role of hydrolysis. Meanwhile, more TC residuals were detected in ster-
ilized group, resulting from the role of microorganism. The loss via bio-
degradation was calculated to be 9.34% ± 0.15% in this study. Compared
to 30% oxytetracycline loss due to biodegradation in soil under aerobic
conditions (Yang et al., 2009), the contribution of biodegradation is
quite small in our study. This could be explained by two reasons: first,
microorganisms need time to be screened out to degrade TC which
didn't exist in soil before. Second, the stronger adsorption property of
TC (kd: 1093 L/kg) makes it more difficult to desorb from soil than oxy-
tetracycline (kd: 1026 L/kg) (Wszolek and Alexander, 1979). Further-
more, there was no significant difference (RMSE = 1.69) between the
TC amounts on the 30th day in sterilized group and those in dark-
sterilized group, which confirmed the limited contribution of photoly-
sis. After 30-day operation, 64.01% ± 6.63% TC was removed from ster-
ilized soil in dark condition, which was attributed to the strong
adsorption of TC by soils.
The adsorption of antibiotics to soil include reversible equilibrium
process, sequestration and non-extractable residues (Jechalke et al.,
2014). TC dissipation in soils could be divided into two phases: the
faster phase before the 8th day and the slower phase after (Fig. 2). It is
consistent with the two dynamic processes proposed to characterize
TC adsorption to soils: a rapid initial adsorption on the outer surface
and then the slower intercalation to microscopes (Chang et al., 2009).
Most TC was sequestered into micro- and nanopores or formed non-
extractable residues with soil via complexation or other mechanisms,
decreasing its bioavailability and making it difficult to be extracted or
detected by the method used here. Moreover, pH is an important factor
4 H.-Y. Liu et al. / Science of the Total Environment 706 (2020) 136086
Fig. 2. Dynamic dissipation and removal rate of TC under different conditions during
30 days incubation. Error bars represent the mean ± standard deviations (n = 3).
that affects complexation. TC mainly exists as zwitterion in common soil
with pH 3.3–8.0 (pH 6.7 in this study), where surface complexation be-
tween TC and soil is the dominant mechanism (Sassman and Lee, 2005).
However, this part of TC will be released with slow rates and subse-
quently degraded by microorganisms. The incubation period (30 days)
was too short to reveal this process. Moreover, the removal data was
fitted with first-order kinetic model, and the estimates and 95% confi-
dence intervals of TC half-life were listed in Table 1. The half-life of TC
in soils varied from 13 to 16 days, which were consistent with that at
6 to 15 days in other study (Storteboom et al., 2007).
3.2. Vertical migration of TC in soil
To evaluate the effect of biochar on vertical migration of TC in soil,
we analyzed TC concentration in different depth of soil after TC spiking
on soils with and without biochar, respectively. As shown in Fig. 3, TC
was only found in Layer 1 in the control group and undetectable in
deeper soils, resulting from the strong adsorption of soil on TC (Pan
and Chu, 2016). However, TC was detected in all layers with the addi-
tion of biochar. Most TC (81.68 ± 3.72 μg/g) was retained in Layer 1
while 7.56 ± 1.25 μg/g in Layer 2 and 11.32 ± 12.94 μg/g in other layers.
These results suggest that biochar amendment promotes the migration
of TC to deeper soils. The adsorption isotherms for tetracycline on bio-
char and soil also indicated the slightly lower adsorption capacity of bio-
char (Fig. S3). Similarly, the adsorption affinity of TC on biochar was
lower than that on soil, largely due to the strong interaction between
soil clay and TC (Zhang et al., 2013). Meanwhile, biochar can decrease
the bulk density of soil through creating large void spaces (Jones et al.,
2011), making it easier for TC to spread. However, some studies found
contrary result between antibiotics and biochar/soil. For instance, the
application of burcucumber-derived biochar showed an enhancement
on the adsorption of sulfamethoxazole in soil (Vithanage et al., 2014).
This discrepancy mainly results from the different properties of both
biochar and molecular structures of pollutants. The larger molecular
size of TC (MW = 444.43) causes stronger steric hindrance when ap-
proaching biochar, which weaken its adsorption on biochar (Zhang
et al., 2013).
Table 1
First-order kinetic parameters for the dissipation of tetracycline in soils among control,
dark, sterilized and dark-sterilized groups.
Treatments k (mg kg−1d−1)a R2
Control 0.05484 0.992
Dark 0.05064 0.991
Sterilized 0.04318 0.986
Dark-sterilized 0.04594 0.993
a
k, rate constant of the first-order reaction kinetics; R2, coefficient of determination.
b
t1/2, half-life of dissipation (ln 2/k).
Fig. 3. Acute vertical migration of TC from Layer 1 to Layer 6 in soils with/without biochar.
Error bars represent the mean ± standard deviations (n = 3). CK: control group; BC:
biochar group.
3.3. Evolution of bacterial community in soils
Soil samples in the four groups were collected after 90-day incuba-
tion, and both bacterial and fungal community were investigated
using high-throughput pyrosequencing. The differences in bacterial
communities were analyzed with Partial least squares Discriminant
(PLS-DA) plot. As shown in Fig. 4, there was significant difference be-
tween samples in CK, BC, and BT group whereas slight difference was
observed between samples in CK and TC group. It has been reported
that biochar could increase bacterial diversity and richness in rice
paddy soil from south China (Chen et al., 2015). In contrast, TC exhibited
limited impacts on bacterial community, as no significant effect was
found on dominant microbial species in soil during 120-day incubation
(Ma et al., 2016). Therefore, biochar exhibited more profound shaping
influence on soil microbial community than that of TC, which might
be attributed to the rapid dissipation of TC in soil.
The phylogenetic classification at phylum and genus level was sum-
marized in Fig. 5. After 90-day incubation, the most dominant phyla
were Acidobacteria and Proteobacteria in all samples with relative abun-
dance at 32.2%~36.5% and 28.4–36.4%, which is similar to other studies
on bacterial communities with carbon materials (Xu et al., 2014).
Other prevalent phyla included Nitrospirae, Bacteroidetes,
Gemmatimonadetes, and Verrucomicrobia with relative abundance
above 1%. At the genus level, RB41 was found to be the dominant
genus (relative abundance = 10%). Other highly abundant genus in-
cluding Sphingomonas, Nitrospira, 11–24, and H16 were also detected
with relative abundance above 2% of the total community.
Furthermore, Bacterial community in biochar-amended soil was dif-
ferent from that in CK. The addition of biochar increased the relative
abundance of Actinobacteria and decreased that of Acidobacteria and
Nitrospirae. Biochar addition to soil commonly increases soil C/N ratio
t1/2 (days)b
13
14
16
15
Fig. 4. Bacterial community changes: Partial least squares Discriminant (PLS-DA) analysis
of bacterial community at OTUs level. CK: control group; BC: biochar group; TC:
tetracycline group; BT: biochar and tetracycline group.
 H.-Y. Liu et al. / Science of the Total Environment 706 (2020) 136086 5

Fig. 5. Distribution of partial sequences of bacterial 16S rRNA genes from different groups
at the phylum (a) and genus (b) level on day 90. Phylogenetic groups accounting for 1% or
less of the classified sequences in all samples were summarized in the group “others”. CK:
control group; BC: biochar group; TC: tetracycline group; BT: biochar and tetracycline
group.
Fig. 6. Distribution of partial sequences of fungal 18S rRNA genes from different groups at
the phylum (a) and genus (b) level on day 90. Phylogenetic groups accounting for 1% or
less of the classified sequences in all samples were summarized in the group “others”.
CK: control group; BC: biochar group; TC: tetracycline group; BT: biochar and
tetracycline group.

(Chan et al., 2007). Meanwhile, micro- and macropores in biochar hold biochar (Table 2), which was due to the acidic property of biochar
more oxygen and air content which has been reported to stimulate (Lehmann et al., 2011). Then, the pH increased to 5.58 ± 0.04 in BC
more soil organic matter (SOM) decomposition (Parsons and Smith, group and 5.47 ± 0.16 in BT group after 30-day incubation, which was
1989). The resulting available C leads to the promotion of copiotrophic similar to another study (Xu et al., 2014). It has been reported that
taxa (i.e. Actinobacteria and Proteobacteria), which was consistent with higher pH (7.29 ± 0.02) was beneficial to the growth of Ascomycota
other studies (Jiang et al., 2016; Xu et al., 2016). In this study, biochar (Li et al., 2019). The genus Staurothele from Ascomycota was also
might be utilized by microorganisms such as Actinobacteria, which adapted to neutral to alkaline environments (Gueidan et al., 2014).
could degrade recalcitrant polymers (Jones et al., 2011; Kirby, 2006), These could explain the decreased relative abundance of Ascomycota
causing the shift of community composition. At genus level, biochar in BC and BT group. Moreover, the relative abundance of Basidiomycota
also enhanced the growth of Sphingomonas, which is classified in slightly increased from 6.5% in CK group to 8.0% in BC group and 7.5% in
alphaproteobacteria class and able to degrade refractory contaminants BT group. Similarly, biochar also slightly increased the relative abun-
(Leys et al., 2005). In contrast, no obvious change was observed on the dance of Basidiomycota in a forest soil conducted for 96 days (Hu et al.,
relative abundance of Acidobacteria, implying little impact of biochar 2014). Basidiomycota could degrade organic materials such as lignin,
on these bacteria. Acidobacteria prefers oligotrophic soils with low car- hemicellulose and cellulose (Nie et al., 2018), and thus the carbon-rich
bon availability (Mao et al., 2012). Moreover, the relative abundance conditions induced by biochar promoted its growth in BC and BT
of Nitrospira at genus level decreased slightly with biochar amendment, group in this study. At genus level, Trinema in groups with biochar
which is because that Nitrospira prefers to live under oligotrophic condi- was detected three times more than that in CK group. The amount of
tions (Shen et al., 2013). Trinema is generally dependent on the moisture content of the soil
(Wilkinson and Mitchell, 2010), and the presence of biochar could pro-
3.4. Evolution of fungal community in soils mote the migration of rainwater to deeper soil in this work. Tausonia, a
genus of Basidiomycota, was also observed at higher abundance in BC
All samples were roughly clustered into four regions in the PLS-DA
plot (Fig. S4), suggesting the alteration of fungal community induced Table 2
by biochar or tetracycline. It is in agreement with other studies pH values of soils in different groups during 30 days incubation.
(Jechalke et al., 2014; Yao et al., 2017). The effect of biochar on fungal
Day 1 Day 10 Day 20 Day 30
community was first evaluated at the phylum level. As shown in
CK 6.72 ± 0.05a 6.12 ± 0.21a 6.90 ± 0.03a 7.11 ± 0.20a
Fig. 6a, the addition of biochar decreased the relative abundance of As-
BC 4.22 ± 0.51b 4.88 ± 0.25b 5.08 ± 0.05b 5.58 ± 0.04b
comycota from 40.4% in CK group to 26.6% in BC group, which was at- TC 6.57 ± 0.08a 6.54 ± 0.05a 6.80 ± 0.04a 7.09 ± 0.04a
tributed to the low pH in soil with biochar. Soil pH has been BT 4.38 ± 0.07b 5.21 ± 0.21b 5.04 ± 0.17b 5.47 ± 0.16b
considered as a key factor on soil fungal community (Zhang et al., CK: control group, BC: biochar group, TC: tetracycline group, BT: biochar and tetracycline
2016). In this study, soil pH decreased from 6.72 ± 0.05 to 4.22 ± group. Different letters in a row demonstrate significant difference among the treatments
0.51 in BC group and 4.38 ± 0.07 in BT group with the addition of at p b 0.05.
6 H.-Y. Liu et al. / Science of the Total Environment 706 (2020) 136086
and BT group due to the carbon-rich conditions induced by biochar (Nie
et al., 2018). Moreover, the genus of Mortierella, saprotrophs in soil, was
also enhanced by biochar amendment, implying that biochar could pro-
vide a better survival habitat for Mortierella (Liu et al., 2015; Zheng et al.,
2016).
Tetracycline exhibited no significant effect on fungal diversity
(Fig. S5) but it obviously changed the composition of fungal community
in soil (Fig. 6). The most frequent fungal phyla such as Ascomycota, Nem-
atoda and Basidiomycota were found less abundance in TC group. Sev-
eral studies have also highlighted the adverse impact of veterinary
antibiotics on soil microbial biomass (Jechalke et al., 2014; Kong et al.,
2006). However, there were some fungal genus with higher abundance
in TC group than others, such as Thermothelomyces, a kind of fungi able
to degrade aromatic compounds (Périgon et al., 2019). Hence, the pres-
ence of TC could also enrich the microorganism which can degrade an-
tibiotics. Compared to TC group, fungal community was less impacted in
BT group, and the abundance of some dominant phyla recovered to nor-
mal levels with Basidiomycota at 7.5% and Nematoda at 6.3%, which was
attributed to the combined effect of TC and biochar. Liu et al. (2014) also
found that the diversity of soil microbial community stimulated by
chlortetracycline could be partially counteracted by organic matter. Be-
sides, TC would be partly adsorbed by biochar due to its large surface
area (Quilliam et al., 2013). Therefore, biochar could adsorb some quan-
tities of tetracycline, which limited the contact between TC and micro-
organisms, decreasing its bioavailability and mitigating its effect on
fungal community in soil.
4. Conclusion
This study evaluated the effect of biochar on the migration of tet-
racycline in soil and their combined impacts on microbiome. TC was
mainly dissipated in soil via adsorption with little fraction being
photolyzed or biodegraded. Moreover, the vertical migration of TC
was accelerated in the presence of biochar, mainly due to the de-
creased soil bulk density and lower adsorption capacity of TC on bio-
char. Biochar could stimulate available C to promote the growth of
copiotrophic bacteria and thus significantly change the composition
of bacterial community in soil, whereas TC played a minor role due to
its rapid dissipation in soil. Besides, biochar changed the composi-
tion of fungal community due to its acidic impact on soil and partially
counteracted the role of TC through limiting its contact with
microbiome.
CRediT authorship contribution statement
Hua-Yu Liu: Conceptualization, Methodology, Validation, Investiga-
tion, Formal analysis, Writing - original draft, Writing - review & editing.
Chao Song: Conceptualization, Investigation, Writing - original draft,
Writing - review & editing. Shan Zhao: Investigation, Formal analysis,
Writing - review & editing. Shu-Guang Wang: Conceptualization, Su-
pervision, Project administration, Writing - review & editing.
Declaration of competing interest
The authors declare that they have no known competing financial
interests or personal relationships that could have appeared to influ-
ence the work reported in this paper.
Acknowledgement
The research was supported by the China Major Science and Tech-
nology Program for Water Pollution Control and Treatment
(2017ZX07101003) and National Natural Science Foundation of China
(21676161 and 21808127). The authors wish to thank Project funded
by China Postdoctoral Science Foundation (2019M652389) and Young
Scholars Program of Shandong University.
Appendix A. Supplementary data
Supplementary data to this article can be found online at https://doi.
org/10.1016/j.scitotenv.2019.136086.
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