Chemosphere 151 (2016) 171e177

Contents lists available at ScienceDirect

Chemosphere
journal homepage: www.elsevier.com/locate/chemosphere

Effects of plastic film residues on occurrence of phthalates and

microbial activity in soils
Jun Wang a, b, **, Shenghong Lv a, Manyun Zhang b, Gangcai Chen a, Tongbin Zhu c,
Shen Zhang a, Ying Teng b, *, Peter Christie b, Yongming Luo b
a
Chongqing Research Academy of Environmental Sciences, Chongqing 401147, China
b
Key Laboratory of Soil Environment and Pollution Remediation, Institute of Soil Science, Chinese Academy of Sciences, Nanjing 210008, China
c
Karst Dynamics Laboratory, Institute of Karst Geology, Chinese Academy of Geological Sciences, Guilin 541004, 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

Relationships among plastic residues,

PAE occurrence and soil microbial
activity were examined.

PAEs in soil were correlated with
PAEs released from plastic film
residues.

Soil microbial activities declined
partly due to PAEs derived from
plastic film residues.

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

Article history: Plastic film mulching has played an important role in Chinese agriculture, especially in vegetable pro-
Received 10 December 2015 duction, but large amounts of film residues can accumulate in the soil. The present study investigated the
Received in revised form effects of plastic film residues on the occurrence of soil PAEs and microbial activities using a batch pot
16 February 2016
experiment. PAE concentrations increased with increasing plastic film residues but the soil microbial
Accepted 16 February 2016
Available online 15 March 2016
carbon and nitrogen, enzyme activities and microbial diversity decreased significantly. At the end of the
experiment the PAE concentrations were 0e2.02 mg kg1 in the different treatments. Soil microbial C
Handling Editor: Tamara S. Galloway and N, enzyme activities, AWCD value, and Shannon-Weaver and Simpson indices declined by about 28.9
e76.2%, 14.9e59.0%, 4.9e22.7%, 23.0e42.0% and 1.8e18.7%, respectively. Soil microbial activity was
Keywords: positively correlated with soil PAE concentration, and soil PAE concentrations were impacted by plastic
Phthalates color and residue volume. Correlations among, and molecular mechanisms of, plastic film residues, PAE
Plastic film mulching occurrence and microbial activity require further study.
Soil microorganisms © 2016 Elsevier Ltd. All rights reserved.
Enzyme activities

1. Introduction

Plastic film mulch technology was introduced to China in the

late 1970s and has played an important role in Chinese agriculture
due to soil warming, moisture conservation effects, increased crop
* Corresponding author.
production and control of weeds (Yan et al., 2010; Wang et al.,
** Corresponding author. Chongqing Research Academy of Environmental Sci-
ences, Chongqing 401147, China. 2013). The use of plastic film mulching has helped to increase the
E-mail address: yteng@issas.ac.cn (Y. Teng). yields of cash crops, especially vegetables, by 20e60% (Liu et al.,

http://dx.doi.org/10.1016/j.chemosphere.2016.02.076
0045-6535/© 2016 Elsevier Ltd. All rights reserved.
172 J. Wang et al. / Chemosphere 151 (2016) 171e177
2014;Teng et al., 2014). The use of agricultural plastic mulch films in
China increased nearly four times from 0.32 million to 1.25 million
tonnes from 1991 to 2011 (China Agricultural Statistical Yearbook,
2012; Wang et al., 2013). Over the past 20 years all of the prov-
inces and regions of China have shown increasing trends in plastic
film mulch use intensity, especially northern regions which have
exhibited higher absolute intensity (Liu et al., 2014), for example
from 7.0 kg ha1 in 1991e34.8 kg ha1 in 2011 in Xinjiang (China
Agricultural Statistical Yearbook, 2012). Moreover, the use of
mulching film is expected to increase at a rate of 7% from the cur-
rent 1.25 million tonnes to over two million tonnes in the next ten
years (Wang et al., 2013; Liu et al., 2014).
The large amounts of mulching film residue resulting from
widespread use of plastic film in China represent a series of
pollution hazards (Jambeck et al., 2015). Yank et al. (2010) reported
that film residue decreases soil porosity and air circulation, changes
microbial communities and may lead to low soil fertility, with
consequent effects on crop seed germination and seedling growth
(Subrahmaniyan and Mathieu, 2012). Cuello et al. (2015) found that
plastic film mulching significantly decreased soil organic matter
content and greatly increased greenhouse gas emissions. Further,
several studies show that crop yields decreased when plastic film
residues were present in the soil at rates of up to 58.5 kg ha1 (Li
et al., 2014; Dong et al., 2015). Fragments of plastic film have
been found to accumulate pesticides and other toxins that exist in
the soil (Ramos et al., 2015). Moreover, plastic film residue may
decrease the soil microbial biomass and microbial activity with the
degree of influence affected by the type and color of the mulching
film (Moreno and Moreno, 2008). Plastic film residues in soil may
also release phthalate acid esters (PAEs) which are potentially
carcinogenic and mutagenic (Fu and Du, 2011; Wang et al., 2015a).
The plastic mulching film used in China is mainly polyvinyl
chloride that is not readily degradable and it gradually becomes
thin and fragile as it degrades physically with time (Liu et al., 2014;
Jambeck et al., 2015). Investigations indicate that the volume of soil
residue was 50e260 kg ha1 in arable areas with plastic film mulch
cover over a period of ten years (Yan et al., 2010). Plastic film
contains about 20e60% phthalates that will be gradually released
into the soil and potentially affect the quality of crops such as
vegetables and thus pose a human health risk via the food chain (Fu
and Du, 2011; Wang et al., 2015a). PAEs have strong hydrophobicity
and are readily adsorbed by soil organic matter and their residues
may persist in the soil for long time periods (Chen et al., 2013). They
may affect the soil microbial population and enzyme activities
(Zhou et al., 2005; Chen et al., 2013). However, few studies have
focused on the relationships among plastic film residues, phthalate
occurrence and soil microbial activities. A detailed study of the
effects of plastic film residue on soil PAE occurrence and microbial
ecology will therefore be of considerable interest in relation to
maintaining greenhouse soil quality and minimizing environ-
mental risk.
In the present study the effects of different plastic film residues
on soil PAEs and soil microbial activity were studied in a pot
experiment. The main objectives were to formulate the character-
istics of plastic film phthalate release and soil phthalate dynamics,
to analyze soil microbial biomass C and N, soil enzyme activities
and soil microbial diversity in the presence of different plastic film
residues. Correlations were examined among plastic film residues,
PAEs and microbial activities in the soil. The results may provide
more direct evidence of the role of plastic film residues in declining
soil environmental quality.
2. Methods and materials
2.1. Soil and plastic film
The study soil was collected from the top 50 cm of an unpolluted
shrub field after discarding the litter in suburban Nanjing, China, in
October 2011. The soil collected was air-dried in the laboratory and
sieved (2 mm). Before the pot experiment the dried soil was
moistened to 50% of water holding capacity and pre-conditioned by
incubation for 1 week at 25  C. The soil is classified as a Luvisols and
the main properties are as follows: pH 7.40, organic carbon 0.85%,
cation exchange capacity (CEC) 17.03 cmol kg1 soil, clay, 15.2% (v/
v), and the concentration of PAEs was below the detection limit.
Two colors (black and white) of plastic film without cadmium and
lead were purchased from an agricultural supply store. The poly-
vinyl chloride film width  thickness was 2000  0.004 mm. The
films were cut with scissors into 20 mm  20 mm pieces and then
mixed with the test soil.
2.2. Experimental design and sampling
The experimental work was carried out in a glass greenhouse
and a laboratory. Pre-incubated soil was added to 1.5-kg ceramic
pots (20 cm height  18 cm diameter) and amended with different
plastic film residues. There were three plastic film residue treat-
ments comprising zero plastic film residue, conventional plastic
film residue (67.5 kg ha1) and five times the conventional plastic
film residue (337.5 kg ha1) with two colors. The treatments are
described as control without film residue (CK), black film residue 1
(BR1), black film residue 3 (BR3), white film residue 1 (WR1) and
white film residue 3 (WR3). The water holding capacity of the soil
was adjusted every other day to maintain it constant (50%) during
the experiment and there were three replicates of each treatment.
The pot experiment was completed within 60 days and the soil
and residual plastic film were sampled as follows. Plastic film
samples were collected 0 and 60 d and the soil samples were
collected 30 and 60 d from the start of the experiment. Plastic film
samples were rinsed clean with deionized water and air-dried for
extraction. Soil samples were collected with a clean stainless steel
soil auger from the pots and small pieces of residual plastic films
were removed. The soil samples were divided into two parts, one of
which was freeze-dried, ground and homogenized by sieving
through a stainless steel sieve (60-mesh) for PAE extraction and the
other directly ground and passed through a 15 mm sieve to
determine soil microbial activity. Soil samples collected at 60 d
were used to determine soil microbial activity. Precautions were
taken during sampling and sample processing to avoid PAE
contamination.
2.3. Materials and reagents
Mixed standard solutions containing DMP, DEP, DBP, BBP, DEHP
and DOP (1 mg ml1) and the isotope surrogate standard di-n-butyl
phthalate-d4 (100 mg ml1) were supplied by Dr. Ehrenstorfer
(Augsburg, Germany). The stock solutions of six mixed phthalates
were prepared in n-hexane at a concentration of 1 mg L1. Neutral
silica gel and alumina (100- to 200-mesh) and anhydrous sodium
sulfate were activated at 400 ± 1  C for 6 h. The acetone and n-
hexane used were HPLC grade and were purchased from Tedia
Company Inc., Fairfield, Ohio. Before use, all glassware was washed
in a detergent solution in a laboratory ultrasonic washer (KQ-
600DB, Kun Shan Ultrasonic Instruments Co., Ltd., Jiangsu Province,
China), air dried and immersed in sulphuric acid (guaranteed re-
agent) and washed with tap water and ultrapure water before
oven-drying and rinsing with acetone and n-hexane. No plastic
 J. Wang et al. / Chemosphere 151 (2016) 171e177
vessels were used in the experimental procedures.
2.4. Extraction and measurement of PAEs
Extraction and analysis of PAEs in soil and plastic film were
performed according to Wang et al. (2013). Briefly, soil (5.0 g) and
plastic film (1.0 g) samples were weighed into glass centrifuge
bottles and mixed with 20 mL acetone: hexane (1:1 v/v), left
overnight and extracted for 30 min and the procedure was repeated
a further two times. All of the filtrates were concentrated with the
isotope surrogate standard, adjusted to 1 mL and transferred
through a 0.25 mm membrane filter to a brown sample bottle for
sample injection prior to instrumental analysis. The PAEs were
determined with an Agilent 7890 GC-5975MSD gas chromatog-
raphy mass spectrometry system (GCeMS) (Agilent Technologies,
Avondale, PA) operating in electron impact and selective ion
monitoring mode and with a DB-5 (30 m  0.25 mm  0.25 mm)
fused-silica capillary column for chromatographic separation. The
temperature program column oven was set to 50  C for 1 min and
raised to 200  C at 15  C min1, maintained for 1 min, then up to
280  C at 8  C min1 and held for 3 min and injected into the
GCeMS system in non-pulse and splitless mode with an injector
temperature of 250  C. The GCeMS transfer line was set at 280  C
and the post run temperature was 285  C for 2 min.
2.5. Microbial biomass C and N
Soil microbial biomass carbon and nitrogen (biomass C and
biomass N) were determined by the fumigation extraction method
with some modification (Brookes et al., 1985; Vance et al., 1987).
Briefly, fresh soil (equivalent to 10 g oven-dry weight) was weighed
into 6-cm-diameter glass culture dishes (three per treatment) and
fumigated with alcohol-free chloroform for 24 h at 25  C. Excess
chloroform was removed by repeated evacuation and then the
samples were extracted with 50 ml 0.5 M K2SO4 on a rotary shaker
at 220 rpm for 30 min and then filtered with a qualitative filter
paper. The remaining three unfumigated samples from each
treatment were immediately extracted and filtered as described
above. Soil filtrates were stored at 20  C prior to analysis for soil
microbial biomass C and N. Organic C in the filtrate was determined
with a total organic C analyzer (Phoenix-8000, Teledyne Tekmar,
Mason, OH). Total soluble N in the filtrate was followed by alkaline
persulfate oxidation and measured by dual-wavelength ultraviolet
spectrophotometry (Cabrera and Beare, 1993). Soil biomass C was
calculated as biomass C ¼ (Cfumigated  Cunfumigated)/KC, where Cfu-
migated and Cunfumigated are the extracted organic C contents of
fumigated and unfumigated samples and KC is 0.45 according to
Vance et al. (1987). Soil biomass N was calculated as biomass
N ¼ (Nfumigated  Nunfumigated)/KN, where Nfumigated and Nunfumigated
are the extracted N contents of fumigated and unfumigated sam-
ples and KN is 0.54 according to Brookes et al. (1985).
2.6. Enzyme activity and biodiversity
Enzyme activity and functional diversity were analyzed ac-
cording to the methods of Sofo et al. (2012) and Wang et al. (2015b)
with some modifications. Briefly, soil fluorescein diacetate hydro-
lysis (FDA) induces fluorescein diacetate to generate yellow fluo-
rescein by hydrolysis and dehydrogenation. Soil dehydrogenase
(DHO) induces 2,3,5-triphenyltetrazolium chloride to generate red
triphenyl formazance. The concentrations of FDA and DHO were
measured with a spectrophotometer at 490 nm and 485 nm,
respectively.
Functional diversity of soil microbial communities was tested
with Biolog Eco Plates (Biolog, Hayward, CA). The plates were
173
incubated at 25  C and the color development in each well was
recorded as optical density (OD) at 590 nm at 12-h intervals. Mi-
crobial activity in each microplate, expressed as average well-color
development (AWCD), was determined by: AWCD ¼ SODi/31,
where ODi is the optical density value from the well i, corrected by
subtracting the blank well (inoculated but without a carbon source)
values from plate well i. The Shannon-Weaver index (H) and
Simpson index (l) were calculated by: H ¼ -Spi ln pi and l ¼ Sp2i ,
where pi is the ratio of ODi to the sum of OD values on all wells
(SODi).
2.7. Statistical analysis
Statistical analysis was conducted using the SPSS software
package version 20.0. All the data were subjected to one-way
analysis of variance and pairs of mean values were compared by
Duncan's multiple range test at the 5% level.
3. Results
3.1. Dynamics of soil phthalate with plastic film residues
The concentrations of PAEs in different plastic films are shown
in Fig. 1. The total concentrations of PAEs in black and white plastic
films were 157.0 and 233.0 mg kg1 in new plastic films and 57.2
and 79.7 mg kg1 60 days after their application. It is clear that
DnOP was the predominant PAE used in the different plastic films
with S6PAE contributions ranging from 36.0 to 61.1%. The second
dominant PAE was DEHP, ranging from 24.3 to 50.6%, followed by
DnBP with a contribution of 7.2e11.7% and the total contribution of
DMP, DEP and BBP was about 3e5.4%. All of the six PAE in white
plastic films were significantly (p < 0.05) higher than those in the
black plastic films. The concentrations of the six priority control
PAEs in soil with different plastic film residues ae shown in Fig. 2.
PAE concentrations in the soil increased with increasing weight of
plastic film residue. PAEs decreased significantly (p < 0.05) with
increasing time of incorporation of plastic film residues. The total
concentration of the six PAE ranged from 0.05 to 5.72 mg kg1 after
30 days but only 0.00e2.02 mg kg1 after 60 days. The components
of individual PAEs in soil were similar to those in the plastic films.
DnOP and DEHP were also the predominant phthalates in soils.
Plastic film residues may have been the main sources of PAEs in the
soil.
3.2. Effects of plastic film residues on soil microbial biomass
nitrogen and carbon
Variation in soil biomass C and N with different plastic film
residues are shown in Fig. 3. Biomass C or N decreased with
increasing weight of plastic film residues. Biomass C in BR1, BR3,
WR1, and WR3 declined by 28.9, 73.5, 29.5 and 57.1% while biomass
N declined by 41.4, 74.3, 38.2 and 76.2%, respectively, compared
with the control values of 740 and 139 mg kg1, respectively. Plastic
film residues significantly decreased the soil biomass C and N but
there were no statistically significant differences between black
and white plastic films.
3.3. Effects of plastic film residues on soil enzyme activities
Fluorescein diacetate hydrolysis and dehydrogenase activities
with different plastic film residues are shown in Table 1. Plastic film
residues inhibited soil enzyme activities clearly with increasing
weight of plastic film residue. Compared with the control, fluo-
rescein diacetate hydrolysis activities declined by about 1.6, 15.6,
10.2 and 30.7% in BR1, BR3, WR1 and WR3 treatments, respectively.
174 J. Wang et al. / Chemosphere 151 (2016) 171e177

Fig. 1. PAE residues in different plastic films (B and W indicate black and white plastic films and 0 and 60 denote time of plastic film application to the soil).

Fig. 2. PAE concentrations in soils with different plastic residues.

However, dehydrogenase activity changed more with changing
weight of plastic film. Dehydrogenase activity in BR1, BR3, WR1 and
WR3 declined by about 14.9, 58.9, 19.8 and 59.0% compared with
the control. Both fluorescein diacetate hydrolysis and dehydroge-
nase activities in soil with black plastic film residues were signifi-
cantly higher than in the corresponding white plastic film
treatments. The inhibitory effect of plastic film residue on dehy-
drogenase activity was stronger than that on fluorescein diacetate
hydrolysis activity.
value of AWCD decreased markedly with increasing weight of
plastic film residues and the response of the AWCD value was more
sensitive to white plastic film residues than to black plastic film.
The value of AWCD increased slowly with increasing incubation
time and tended to stablilize after 156 h of incubation. From 156 h
the AWCD value and Shannon-Weaver and Simpson indices
declined by about 4.9e22.7%, 23.0e42.0% and 1.8e18.7%, respec-
tively, with different plastic film residues (Table). There was a sta-
tistically significant difference (p < 0.05) in soil functional diversity
with different plastic film residue treatments.

3.4. Effects of plastic film residues on soil microbial diversity

The diversity of cultivable microorganisms and the change in

ecological function index of the soil with different plastic film types
and residues after incubation for 60 days are shown in Fig. 4. The
 J. Wang et al. / Chemosphere 151 (2016) 171e177 175

(Yan et al., 2010; Fu and Du, 2011; Dong et al., 2015). Mulching film
residues release multiple heavy metals and organic pollutants in
soils, especially phthalates which represent potential human health
risks (Teng et al., 2014). Several studies indicate that plastic films
contain about 20e60% phthalates are these are the commonest
source of PAEs in soils (Wang et al., 2013, 2015). The present study
shows that PAEs in the residual plastic films and soils decreased
simultaneously, likely due to the combined effects of photochem-
ical and soil microbial degradation of PAEs. However, residual PAE
concentrations in the soil remained at up to 2.02 mg kg1 and PAE
accumulation in the soil will continue to increase with repeated
annual application of mulching films (Fu and Du et al., 2011; Wang
et al., 2013). Based on (eco)toxicology and environmental chemistry
data, the environmental risk limits for DnBP and DEHP in fresh soils
and sediments with 10% organic matter are 0.7 and 1.0 mg kg1
(van Wezel et al., 2000) so that the concentrations of DnBP and
DEHP in some treatments have exceeded the risk limits in the
present study. In addition, Ramos et al. (2015) reported that the
Fig. 3. Soil microbial biomass C and N contents in different treatments. presence of plastic residues may lead to pesticide migration to the
inside of the plastic matrix. Film residues may act as potential
pesticide collectors in soils. Therefore, recycling measures for re-
Table 1 sidual films need to be developed to avoid PAE accumulation in the
Soil enzyme activities in different plastic film residues. soil and increase crop quality and health.
Treatment Fluorescein diacetate hydrolysis Dehydrogenase activity Soil biomass C and N provide an important index of soil fertility
mg fluorescein g1 dwt soil 20 min1 mg TPF g1 dwt soil h1
and the most active part of soil organic components mainly refer to
*
the biomass with volume <5000 mm in the soil (Brookes et al., 1985;
Control 170.8 ± 4.84 a 93.6 ± 0.78 a
Garland, 1996; Li et al., 2004). Plastic film mulching with no
BR1 168.1 ± 0.18 a 79.7 ± 0.91 b
BR3 144.2 ± 0.30 b 38.5 ± 0.26 c flooding reduced microbial biomass C and N markedly (Liu et al.,
WR1 153.4 ± 4.09 b 75.2 ± 0.52 b 2010). In the present study soil biomass C and N were significantly
WR3 118.4 ± 0.60 c 38.4 ± 1.04 c (p < 0.05) reduced with increasing mulching film residue and
*
Different letters within a row indicate significant difference (p < 0.05). showed little difference between the two plastic film colors. These
results are in agreement with previous studies. For example,
Moreno and Moreno (2008) found that the use of plastic films
resulted in the lowest values of soil microbial biomass C and soil
organic matter mineralization. However, in other studies Li et al.
(2004) found that the highest soil microbial biomass C contents
were found in the mulched treatments and this might be related to
the original soil organic matter content (Moreno and Moreno,
2008). Our results indicate that PAE concentrations and biomass
C and N were weakly and positively correlated with the plastic film
residue rate in the soil. The declines in soil biomass C and N might
be due to the combined effects of film residues and PAEs.

4.2. Enzyme activities and microbial functional diversity in soil with

plastic residues

In general, the biological and biochemical properties of soils

Fig. 4. Changes in the per hole absorbance values of soil samples with plastic film
residues.
4. Discussion
4.1. Phthalate occurrence and soil biomass C and N in soil with
plastic film residues
Wide usage of mulching technology has weakened the agro-
nomic limitations of season in agricultural production, increased
yields and controlled weeds but adverse impacts on crop yields and
soil environmental quality have also gradually appeared with
plastic film residue volume and the number of years of application
play a very important role in soil pollution and can indicate the
influence of pollutants on the soil ecosystem (Wang et al., 2009).
Soil enzyme activity can indicate the general condition of soil mi-
crobial populations (Wang et al., 2015b). Fluorescein diacetate hy-
drolysis has been used as an indicator of soil metabolic activity
(Sofo et al., 2012) and dehydrogenase has been used as an impor-
tant index of soil microbial activity and functional diversity
(Sebiomo et al., 2011). In the present study plastic film residues
significantly reduced soil FDA and DHO activities compared with
control soil receiving no plastic film residues (Table 1). This might
be due to the release of PAEs by film residues in the soil which
might impact enzyme activity (He et al., 2015). Wang et al. (2015b)
found that FDA and DHO activities increased significantly (P < 0.05)
with decreasing PAE concentrations in soil. Some other studies
have shown that PAEs can enhance dehydrogenase, catalase, pro-
teinase and phosphatase activities but inhibit urease, cell enzymes
and beta glucose activities in soil (Zhou et al., 2005; Wang et al.,
2009; Xie et al., 2010). The differences in the results from these
176 J. Wang et al. / Chemosphere 151 (2016) 171e177
Table 2
The functional diversity of microbial communities in soil with different plastic film
residues.
Treatment AWCD(156h) Shannon-Weaver index Simpson index
Control 1.738 ± 0.075 a* 5.38 ± 0.531 a 43.49 ± 0.320
BR1 1.653 ± 0.040 b 4.137 ± 0.827 b 42.70 ± 0.471
BR3 1.545 ± 0.023 c 3.544 ± 0.329 b 35.37 ± 0.626
WR1 1.486 ± 0.232 d 3.946 ± 0.287 b 40.27 ± 0.348
WR3 1.344 ± 0.112 e 3.122 ± 0.779 c 35.61 ± 0.162
*
Different letters in a row indicate significant difference (p < 0.05).
studies might be attributable to different environmental conditions
and types and concentrations of phthalates in addition to plastic
film residues.
The soil microbial community plays a crucial role in nutrient
cycling, maintenance of soil structure and detoxification of noxious
chemicals and its diversity is a sensitive indicator of soil quality that
can reflect subtle changes and provide information for evaluation of
soil function (Sebiomo et al., 2011; He et al., 2015). The AWCD and
the Shannon-Weaver and Simpson indices reflect the sole carbon
source utilization ability of the soil bacterial community and soil
bacterial activity and the actual richness and evenness of the mi-
crobial community in the soil (Simpson, 1949; Garland, 1996). In
the present study the plastic film residues significantly influenced
the soil AWCD and the Shannon-Weaver and Simpson indices,
indicating that the soil microbial community and its diversity
declined sharply (Table 2). Plastic films contain lipophilic PAEs
which affect soil microbial activity and diversity, community
structure and metabolic activity by destroying the fluidity of the
cell membrane (Zhou et al., 2005; Xie et al., 2010). DEP can reduce
bacterial numbers and the activity of pseudomonads by 47 and 62%,
respectively, in one day when present at a concentration of
>1 mg g1 (Cartwright et al., 2000) and PAEs significantly inhibit
the respiration of the microbial community (Larsson et al., 1986).
Wang et al. (2016) demonstrated that the operational taxonomic
unit (OTU) richness and the Shannon index of the bacteria were
reduced by DBP contamination in black soils. PAE contamination
inhibited soil microbial activity (Chen et al., 2013). The results
indicate that the decline in the soil community might be due to
plastic film residues as a result of PAE pollution. Plastic films of
different colors have different PAE concentrations and also exert
different effects on soil enzyme activity and microbial community
diversity. The toxicity of PAEs on soil microorganisms may there-
fore impact soil quality and affect the yield and quality of crops
grown for human consumption.
5. Conclusions
Using two types of plastic film with three residue gradients the
present study demonstrates that, overall, soil PAEs, biomass C and
N, enzyme activities and microbial community diversity were
clearly related to mulch film residue levels in the soil. In addition,
soil microbial activity was significantly and positively related to the
volumes of the residues, with white plastic film exerting a greater
residual effect than black film. Soil microbial biomass C and N,
enzyme activity and microbial community diversity show positive
relationships with plastic film residues. The decline in soil micro-
bial activity might be due to the release of PAEs from plastic film
residues. Further studies are required to elucidate the molecular
mechanisms of changes in soil microbial community structure due
to plastic film residues so that appropriate measures can be intro-
duced to reduce plastic film residues and protect soil quality.
Acknowledgments
This study was funded by the Key Laboratory of Soil Environ-
ment and Pollution Remediation (SEPR2014-08), Institute of Soil
a Science, Chinese Academy of Sciences, by the Project of Basic Sci-
a entific Research Business Expenses Research (2014cstc-jbky-
c 01602), and by Chongqing Research Program of Basic Research and
b
Frontier Technology (No. cstc2015jcyjA20002).
c
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