Chemosphere 119 (2015) 273–281

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

Arsenic pollution of agricultural soils by concentrated animal feeding

operations (CAFOs)
Xueping Liu a,b, Wenfeng Zhang a,b, Yuanan Hu a, Erdan Hu a,b, Xiande Xie a,b, Lingling Wang a,
Hefa Cheng a,⇑
a
State Key Laboratory of Organic Geochemistry, Guangzhou Institute of Geochemistry, Chinese Academy of Sciences, Guangzhou 510640, China
b
University of Chinese Academy of Sciences, Beijing 100049, China

h i g h l i g h t s

 Animal wastes bearing organoarsenic feed additives can cause soil As pollution.
 Soil As levels were elevated in the CAFO zone compared to other agricultural lands.
 p-Arsanilic acid was widely detected in the environmental matrices of the CAFO zone.
 ASA was a significant source of As in the surface soils of the CAFO zone.
 Release of As from swine farms is an environmental problem worthy of attention.

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

Article history: Animal wastes from concentrated animal feeding operations (CAFOs) can cause soil arsenic pollution due
Received 9 March 2014 to the widespread use of organoarsenic feed additives. This study investigated the arsenic pollution of
Received in revised form 20 June 2014 surface soils in a typical CAFO zone, in comparison with that of agricultural soils in the Pearl River Delta,
Accepted 22 June 2014
China. The mean soil arsenic contents in the CAFO zone were elevated compared to those in the local
background and agricultural soils of the Pearl River Delta region. Chemical speciation analysis showed
Handling Editor: Caroline Gaus that the soils in the CAFO zone were clearly contaminated by the organoarsenic feed additive, p-arsanilic
acid (ASA). Transformation of ASA to inorganic arsenic (arsenite and arsenate) in the surface soils was
Keywords: also observed. Although the potential ecological risk posed by the arsenic in the surface soils was
Soil pollution relatively low in the CAFO zone, continuous discharge of organoarsenic feed additives could cause
Animal wastes accumulation of arsenic and thus deserves significant attention.
Organoarsenic feed additive Ó 2014 Elsevier Ltd. All rights reserved.
Arsenate
Arsenite
Environmental fate

1. Introduction desiccants, and fertilizers are also important sources of arsenic in

Environmental pollution caused by arsenic compounds is an
important concern because of their toxicity and potential carcino-
genic properties (Han et al., 2004). Arsenic in surface soils is pri-
marily derived from lithogenic sources, followed by mining, and
emissions from industries such as ceramics and glass, electronics,
pigments and antifouling agents, cosmetics, and fireworks (Porter
and Peterson, 1977; Leonard, 1991;Yan-Chu, 1994). Pesticides
(mainly inorganic arsenic, such as sodium arsenite and calcium
arsenite) and herbicides (such as methylated arsenicals),
⇑ Corresponding author. Tel.: +86 20 8529 0175; fax: +86 20 8529 0706.
E-mail address: hefac@umich.edu (H. Cheng).
agricultural soils (Cullen and Reimer, 1989; Perkins and
Brushwood, 1991; Jiang and Singh, 1994; Marin et al., 1993).
Indiscriminate use of arsenic compounds in agricultural activities
during the 20th century has caused widespread soil pollution
(Mandal and Suzuki, 2002). Contamination of soils with arsenic
levels exceeding 1000 mg kg1 has been recorded at many sites
around the world (Smith et al., 1998).
Over the past two decades, phenylarsonic acid compounds have
been widely used as additives to animal feeds to control parasites,
promote growth, and increase feed utilization efficiency (Spiros
et al., 1997; Moore et al., 1998; Bednar et al., 2003; Jackson et al.,
2006). Roxarsone (3-nitro-4-hydroxyphenylarsonic acid, ROX) is
widely used in chicken feed, while p-arsanilic acid (ASA) is

http://dx.doi.org/10.1016/j.chemosphere.2014.06.067
0045-6535/Ó 2014 Elsevier Ltd. All rights reserved.
274 X. Liu et al. / Chemosphere 119 (2015) 273–281
commonly used for swine production (Wang et al., 2010; Nachman
et al., 2013). They were approved as feed additives to animals in
the U.S. in 1958 (USDA, 1975), and in China in 1996. The sale of
ROX was voluntary suspended in the U.S. in 2011 in response to
the concern on elevated levels of inorganic arsenic in chicken liv-
ers. The use of three organoarsenic feed additives, including ROX
and ASA, in animal production was later banned in the U.S.
(USFDA, 2014). Meanwhile, ROX and ASA are still being widely
used in many countries, including China (Guo et al., 2013). The typ-
ical levels of ROX and ASA are 20–50 g t1 in poultry feeds and 45–
100 g t1 in swine feeds (Tian, 1996; Brown et al., 2005; Wang
et al., 2005, 2010).
Despite their clear veterinary benefits, the use of organoarsenic
feed additives in intensive animal agriculture is controversial
because they introduce soluble arsenic compounds into the envi-
ronment. Only very small fractions of these compounds can be
metabolized or uptaken in the animal bodies, while the rest pass
unchanged into the urine and manure (Aschbacher and Feil,
1991; Chiou et al., 1997; Christen, 2001). With the common prac-
tice of using poultry and swine wastes as fertilizers on agricultural
lands, potentially significant amount of arsenic can be released into
the soils (Mullins, 2000). In the U.S. alone, it has been estimated
that land application of poultry litter introduced approximately
1000 tonnes of organoarsenic feed additives and their degradation
products each year (Wershaw et al., 1999).
With the rising disposable incomes of over 1.3 billion people,
the demand for meat has been rising fast in China, and the per
capita consumption of meat has more than quadrupled since
1980. As the most popular meat in China, pork accounts for over
60% of the total domestic meat consumption in 2011 (NBS,
2012). Driven by the fast rising meat demands, concentrated ani-
mal feeding operations (CAFOs) have expanded rapidly, particu-
larly surrounding the urban centers in China. CAFOs were already
responsible for the production of approximately 80% chickens,
60% of pigs, and 40% cows in the country by 2012 (Liu et al.,
2013a). Because of the relatively high cost of transportation, the
bulk amounts of manure produced from the CAFOs are spread,
often repeatedly, on the local farm lands. Thus, the overwhelming
volume of animal wastes may result in significant accumulation of
pollutants, including organoarsenic feed additives, in the surface
soils of CAFO zones over time. The organoarsenic feed additives
can easily degrade into inorganic arsenic species (arsenate
[As(V)] and arsenite [As(III)]) through chemical and biological pro-
cesses in the surface soils (Wershaw et al., 1999; Bednar et al.,
2003). Meanwhile, arsenate and arsenite can be methylated to
monomethyl arsenic acid (MMA) and dimethyl arsenic acid
(DMA) by microorganisms under reducing conditions (Cullen and
Reimer, 1989; Tamaki and Frankenberger, 1992). The transforma-
tion of arsenic species has important environmental implications
as the inorganic arsenic compounds (arsenite and arsentate) are
more toxic the those in organic forms (WHO, 1981; Vega et al.,
2001).
This study was conducted to investigate the arsenic pollution
of surface soils in a typical CAFO zone, in comparison with the
general agricultural soils, in the Pearl River Delta, one of the
most densely urbanized regions in the world. The possible
sources of arsenic in the surface soils surrounding the CFAOs
was identified using correlation matrix and multivariate analy-
ses, including cluster analysis (CA) and principal component
analysis (PCA). Furthermore, the chemical speciation of the
arsenic compounds was analyzed to assess their sources and
the fate of the organoarsenic feed additives released by the ani-
mal wastes. Loading of arsenic in the surface soils and the con-
tribution from organoarsenic feed additives were analyzed, and
risk assessment was carried out to evaluate the potential ecolog-
ical risk of arsenic pollution.
2. Materials and methods
2.1. Field sample collection and preparation
To investigate the impact of organoarsenic feed additives used
in CAFOs on soil arsenic pollution, a typical CAFO zone in the Pearl
River Delta, south China was chosen in this study. Samples of farm
land soils and other environmental media were taken from a zone
with densely distributed swine farms in Boluo, Huizhou. Swine
farming industry has been established in the zone for more than
a decade, with the annual swine production capacity exceeding
3.8 million heads (Long et al., 2012). Fig. 1 shows the locations of
sampling sites in this study. A total of 72 samples of surface soils
(0–10 cm depth) were taken from plots of farm lands in the area
of approximately 180 km2 in two separate sampling campaigns
carried out in June and September of 2012. A total of 30 samples
of surface water and 15 samples of sediments were also taken from
nearby water bodies receiving discharges from the swine and poul-
try farms. A local background sample was collected from an undis-
turbed hill slope located far away from the CAFOs. Meanwhile, a
total of 65 surface soil samples, which were collected from agricul-
tural lands over the entire Pearl River Delta (approximately
7.2  104 km2) (Hu et al., 2013), were used as the regional back-
ground samples. Comparison was also made with soil samples
(n = 227) collected from all six major types of land uses (urban
area, forest land, agricultural land, industry area, waste disposal/
treatment site, and water source protection area) in the Pearl River
Delta (Hu and Cheng, 2013; Hu et al., 2013).
After removing gravel and plant debris, the soil and sediment
samples were freeze-dried and then grounded to pass 0.15-mm
Nylon sieves. The surface water samples were filtered with
0.45 lm cellulose acetate membranes, acidified with 5% nitric
acid, and stored at 4 °C for no more than 3 d before being
analyzed.
2.2. Chemical analyses
All chemicals and reagents used were checked for possible trace
metal contamination, and all tubes and vials were soaked in a 10%
(v/v) nitric acid solution for 24 h and rinsed with triple-distilled
water before use. Total digestion of the soil and sediment samples
was conducted following Method 3052 of the U.S. Environmental
Protection Agency (USEPA, 1996). The powdered samples (0.2 g)
were digested by trace metal grade acids (mixture of 9.0 mL con-
centrated HNO3 and 3.0 mL 40% HF (v/v)) using a Mars microwave
digestion system (CEM, USA). The recovery rates for the heavy met-
als reported in this study were reasonably good (77–119%) based
on digestion of four certified standard reference materials (SRMs),
including an estuarine sediment (SRM 1646A) and three soils
(GSS6, GSS7, and GSS16). Procedure blanks, SRMs, and duplicates
of 10% sample load were also processed along with the samples.
More details on the total digestion method and quality assur-
ance/quality control (QA/QC) can be found in our previous study
(Zhang et al., 2012a).
The arsenic compounds in the soil and sediment samples col-
lected in the CAFO zone were quantitatively extracted using the
method developed in our previous study (Liu et al., 2013b). In brief,
accurately weighted solid samples (0.5 g) were transferred into
10 mL centrifuge tubes and added with 5 mL of 0.5 M H3PO4 solu-
tion. The mixtures were shaken on a shaker for 16 h at room tem-
perature, followed by centrifugation at 5000 rpm for 20 min and
filtration with 0.45 lm cellulose acetate membranes to obtain
the final extracts. The recovery rates for As(V), MMA, DMA, ASA,
and ROX established from spiked samples were in the range of
67–102%, while that of As(III) was rather low (8.9%) because of
 X. Liu et al. / Chemosphere 119 (2015) 273–281
Fig. 1. Sampling locations in Boluo, Huizhou, a typical CAFO zone in the Pearl River Delta. Sampling time: June 2012 ( ) and September 2012 (4); background:
sites in close proximity are represented with a single marker on the map).
its oxidation during preparation of the spiked sample (Liu et al.,
2013b). Nonetheless, the true recovery rate of arsenite is expected
to be comparable to that of arsenate because of the similar physio-
chemical properties of the two inorganic arsenic species. More
details on the extraction method, recovery rates, and QA/QC can
be found in our previous study (Liu et al., 2013b).
The concentrations of total arsenic in aqueous solutions were
measured on an Agilent 7700X Inductively Coupled Plasma Mass
Spectrometer (ICP-MS) using 1 mg L1 Ge as the internal standard.
The potential interference from polyatomic species (e.g., 40Ar35Cl+,
35 40
Cl Ar+, 40Ca35Cl+, 39K36Ar+, 59Co16O+, 58Ni17O+, 74Ge1H+, and
74
Se1H+) on arsenic (m/z = 75) was eliminated by using helium
(4.5 mL min1) as a reaction gas. Other common heavy metals,
including Cr, Mn, Co, Ni, Cu, Zn, Cd, and Pb, were also detected
along with As in the ICP-MS analysis.
The concentrations of different arsenic species in aqueous solu-
tions were determined by the ICP-MS hyphenated with an Agilent
1200 high performance liquid chromatograph (HPLC) using the
method developed previously (Liu et al., 2013b). Separation of
the arsenic species, including ROX, ASA, As(III), As(V), MMA, and
DMA was achieved on a PRP-X100 anion exchange chromato-
graphic column (Hamilton, USA) by gradient elution using water
and 50 mM (NH4)2HPO4 solution (pH = 6) as the mobile phase
(Liu et al., 2013b). The signal of m/z 75 was recorded while that
of m/z 35 was monitored simultaneously to track the potential
inference from 35K+ and 35Cl+, which could form interference ions
of 40Ar35Cl+, 35Cl40Ar+, and 35K40Ar+. The concentrations of the ana-
lytes were calculated from the areas of the corresponding chro-
matogram peaks based on calibration curves established using
standard solutions.
For total organic carbon (TOC) analysis, the soil samples (ca.
1.0 g) were weighted into ceramic crucibles, and then added with
10 mL 5% (v/v) HCl. The crucibles were then heated in a 60 °C water
bath for 1 h to remove the carbonate minerals (He et al., 2009). The
residue was washed by triple-distilled water until the pH of the
solution reached neutral. The TOC contents of the residues were
analyzed with a Leco C230 elemental analyzer after being oven
dried at 60 °C for 24 h.
275
(sampling
2.3. Data analyses
Statistical analysis of the data was conducted using the SAS
software package. The potential ecological risk posed by the soil
arsenic in the CAFO zone was evaluated with the ecological risk
index (Er) proposed by Hakanson (1980):
C
Er ¼ T r  ð1Þ
C0
where C and C0 are the concentration of arsenic measured in the
sample and the reference value, respectively, and the toxicity factor
(Tr) for arsenic is 10 (Xu et al., 2008; Wang et al., 2011).
3. Results and discussion
3.1. Status of soil pollution by arsenic
Fig. 2a shows the distribution histograms for the levels of
arsenic in the surface soils from farm lands in the CAFO zone, in
the surface soils from the agricultural lands of the Pearl River Delta,
and in the surface soils from lands of all use types in the region.
Obviously, the arsenic contents in the surface soils from the CAFO
zone shifted towards the higher level compared to those in the sur-
face soils of the region. The mean concentrations of total arsenic in
the surface soil samples were 16.0 and 12.2 mg kg1 in the CAFO
zone and the agricultural soils of the Pearl River Delta, respectively
(Table 1). The mean background levels of arsenic in the surface
soils are 11.2 mg kg1 in China (SEPA, 1990; Wei et al., 1991),
and 10.8 mg kg1 in the Guangdong province (Zhang et al.,
2012b). The mean arsenic content in surface soils of the CAFO zone
(16.0 mg kg1) was higher than the local background value
(14.8 mg kg1) and that of agricultural soils (12.2 mg kg1) in the
Pearl River Delta, suggesting the surface soils of the farm lands in
the CAFO zone had been polluted by anthropogenic arsenic
sources. It has been reported the arithmetic mean for the soil
arsenic background level in Guangdong Province had increased
from 8.9 mg kg1 in the late 1980s to 10.8 mg kg1 in the late
276 X. Liu et al. / Chemosphere 119 (2015) 273–281

Fig. 2. Arsenic contents and pollution levels in the surface soils from the CAFO zone and in the surface soils from the agricultural lands in the Pearl River Delta: (a) distribution
histograms of soil arsenic contents (data outliers, i.e., values smaller or larger than 1.5 times the interquartile range, have been removed), and (b) distribution of soil pollution
level. Agriculture soils are classified into four categories based on the arsenic contents: Class I (<15 mg kg1), Class II (15–30 mg kg1), Class III (30–40 mg kg1), and Heavily
polluted (>40 mg kg1) based on the Environmental Quality Standard for Soils in China (SEPA, 1995). Class I soils are regarded as unpolluted.

2000s (Zhang et al., 2012b). Metals are mobile in the soil environ-
ment, and the background levels might have been altered over the
past two decades by natural weathering processes, anthropogenic
inputs, and movement of metals among surface water, groundwa-
ter, and the atmosphere. A few soils samples in the Pearl River
Delta contain arsenic at levels exceeding the range of arsenic con-
tent in the CAFO zone, which might be attributed to land pollution
by mining and metal smelting activities in the west and north parts
of the region (Hu et al., 2013).
Fig. 2b shows the levels of arsenic pollution for the agricultural
soils in the CAFO zone and the Pearl River Delta. Based on the clas-
sification criteria in the Environmental Quality Standard for Soils in
China (SEPA, 1995), the fraction of polluted (i.e., Class II, Class III,
and Heavily polluted) soils accounted for 50.0% (n = 72) of the agri-
cultural soil samples in the CAFO zone. In contrast, 35.4% (n = 65)
of the agricultural soils in the Pearl River Delta were polluted by
arsenic, which is comparable to that of all types of soils (34.8%,
n = 227) in the region. These results indicate that arsenic pollution
of agricultural soils was more prevalent in the CAFO zone com-
pared to the other soils in the Pearl River Delta.
The mean arsenic concentrations in the agricultural soils from
the CAFO zone and the Pearl River Delta were higher than the back-
ground concentrations in Guangdong Province (10.8 mg kg1) and
China (11.2 mg kg1), suggesting the surface soils from the agricul-
tural lands in the Pearl River Delta had been polluted by anthropo-
genic sources of arsenic. The higher concentrations of arsenic in the
agricultural soils from the CAFO zone compared to those in the
agricultural soils from the Pearl River Delta indicate that animal
wastes from the CAFO had probably contributed to the elevated
levels of arsenic in local soils. Another study has reported that
 X. Liu et al. / Chemosphere 119 (2015) 273–281 277

Table 1
Basic summary statistics of the contents of heavy metals in the surface soils from farm lands in the CAFO zone and from agricultural lands in the Pearl River Delta (data outliers,
i.e., values smaller or larger than 1.5 times the interquartile range, have been removed).

Heavy metals Surface soils from farm lands in the CAFO zone Surface soils from agricultural lands in the Pearl River Delta
(mg kg1)
n Mean Std Min Q1 Median Q3 Max n Mean Std Min Q1 Median Q3 Max
As 70 15.95 8.49 3.44 8.57 14.36 21.20 36.41 59 12.23 8.73 1.92 5.86 10.04 16.88 38.39
Cd 69 0.15 0.07 0.06 0.10 0.13 0.21 0.37 63 0.14 0.08 0.02 0.07 0.11 0.18 0.35
Co 68 4.16 2.95 0.57 1.87 3.09 5.79 12.01 61 6.38 3.41 1.98 3.45 5.57 8.47 14.18
Cr 71 45.42 23.92 8.56 25.06 40.26 58.12 104.05 64 48.10 29.66 5.07 24.76 41.27 69.94 130.93
Cu 69 24.20 14.27 6.77 12.79 19.00 31.12 62.43 62 16.87 10.01 3.69 8.20 14.75 22.31 41.16
Mn 63 138.54 121.91 22.67 51.77 96.49 189.25 538.19 59 230.04 141.54 42.24 137.36 188.38 294.59 608.26
Ni 68 14.53 9.26 1.50 6.23 13.14 20.64 43.42 65 12.61 9.36 0.00 4.66 10.63 20.53 33.45
Pb 65 24.30 13.82 6.90 14.83 19.67 28.43 64.35 65 39.10 19.23 7.67 21.77 37.68 55.32 87.81
Zn 70 48.23 36.10 8.34 17.65 40.20 72.30 146.64 62 57.42 30.41 9.79 38.01 51.51 79.56 128.16

the arsenic concentrations in the surface soils of farm lands sur-
rounding 15 swine farms using ASA as feed additives in Guangdong
province were in the range of 26–56 mg kg1, far exceeding the
background levels of arsenic in surface soils (Wang et al., 2006).
These results consistently indicate that the widespread use of
organoarsenic feed additives in CAFO zones could cause arsenic
pollution of local agricultural soils, which deserves significant
attention.
3.2. Identification of the sources of soil arsenic
The heavy metals in surface soils of the Pearl River Delta could
come from both natural sources, such as erosion of rocks and soil
parent materials, and anthropogenic sources, including industrial
and municipal wastewaters, pesticide and fertilizer applications,
and industrial and traffic emissions (Cheng et al., 2009; Cheng
and Hu, 2010; Hu and Cheng, 2013; Hu et al., 2013). The possible
anthropogenic sources of arsenic in the surface soils from the farm
lands in the CAFO zone include applications of agrochemicals, fer-
tilization by animal wastes, and deposition of aerosol particles
from industrial sources and traffic emissions in the surrounding
areas. Due to the relatively large numbers of surface soil samples
collected in the CAFO zone and from the agricultural lands in the
Pearl River Delta, and the number of heavy metals measured for
each sample (Table 1), there is a need to reduce the multidimen-
sional data sets to lower dimensions (Barona and Romero, 1996).
PCA and CA analyses have been successfully employed to identify
the possible sources of the heavy metals based on their associa-
tions in the soil samples (Li et al., 2004; Rodríguez Martín et al.,
2006; Aelion et al., 2008; Zhang et al., 2012a; Hu et al., 2013).
Therefore, correlation matrix and multivariate analyses (PCA and
CA) were used here to help identify the possible sources of the
arsenic pollution in the surface soils of the CAFO zone based on
the associations between the heavy metals in the soils.
Table 2 shows the Pearson’s correlation matrix for arsenic and
other common heavy metals found in the agricultural soils from
the CAFO zone and the Pearl River Delta. Arsenic exhibited strong
positive correlations with the major heavy metals (Cr, Mn, Co, Ni,
Zn, Cd, and Pb) with the exception of Cu in the CAFO zone. In con-
trast, it showed strong positive correlations with Cr, Mn, Co, Ni, Cu,
and Zn, but no significant correlations with Cd and Pb in the agri-
cultural soils in the Pearl River Delta. Although the presence of sig-
nificant linear correlations among arsenic and the other common
heavy metals makes it difficult to identify the sources of arsenic,
the different distribution patterns suggests that the sources of
arsenic in the CAFO zone were not identical to those in the agricul-
tural soils of the Pearl River Delta in general.
The dendogram in Fig. 3a shows that Cr and As formed a cluster
in the surface soils in the CAFO zone, while Fig. 3b indicates that
arsenic was isolated from the other heavy metals in the agricul-
tural soils of the Pearl River Delta. The associations between the
arsenic and other common heavy metals in the agricultural soils
from the CAFO zone and the Pearl River Delta are shown in
Fig. 3c and d. As expected, the factor loadings of arsenic in the soils

Table 2
Pearson’s correlation matrix for arsenic and other common heavy metals in the agricultural soils of the CAFO zone and the Pearl River Delta.

Metal Cr Mn Co Ni Cu Zn As Cd Pb
CAFO zone
Cr 1.00
Mn 0.52** 1.00
Co 0.59** 0.80** 1.00
Ni 0.94** 0.68** 0.74** 1.00
Cu 0.76** 0.63** 0.58** 0.80** 1.00
Zn 0.33** 0.67** 0.56** 0.46** 0.43** 1.00
As 0.39** 0.31** 0.47** 0.34** 0.17 0.39** 1.00
Cd 0.37** 0.60** 0.58** 0.47** 0.51** 0.44** 0.40** 1.00
Pb 0.27* 0.67** 0.77** 0.41** 0.28* 0.70** 0.63** 0.55** 1.00
Pearl river delta
Cr 1.00
Mn 0.11 1.00
Co 0.03 0.75** 1.00
Ni 0.80** 0.16 0.34** 1.00
Cu 0.53** 0.15 0.26* 0.48** 1.00
Zn 0.22 0.64** 0.70** 0.15 0.24 1.00
As 0.45** 0.35** 0.32** 0.44** 0.48** 0.28* 1.00
Cd 0.13 0.43** 0.42** 0.10 0.22 0.64** 0.09 1.00
Pb 0.26* 0.38** 0.48** 0.00 0.26* 0.57** 0.02 0.59** 1.00
*
Level of significance: p < 0.05.
**
Level of significance: p < 0.01.
278 X. Liu et al. / Chemosphere 119 (2015) 273–281

Fig. 3. Association of arsenic and other common heavy metals in the agricultural soils from the CAFO zone and the Pearl River Delta revealed by multivariate analyses: (a)
hierarchical dendogram for heavy metals in the CAFO zone; (b) hierarchical dendogram for heavy metals in the Pearl River Delta; (c) 3-D plots for PCA loadings of heavy
metals in the CAFO zone; and (d) 3-D plots for PCA loadings of heavy metals in the Pearl River Delta.

from the CAFO zone and the Pearl River Delta were significantly
different. Overall, both CA and PCA results consistently indicate
that arsenic had distinctly different sources in the soils of the CAFO
zone compared to the agricultural soils of the Pearl River Delta.
Fig. 4 shows the correlations between the contents of total
arsenic and TOC in the agricultural soils from the CAFO zone and
the Pearl River Delta. Animal manure is composed primarily of
organic matter, and contributes to soil fertility by increasing the
contents of organic matter and nutrients (e.g., nitrogen). Animal
wastes are disposed off on local farm lands as an organic fertilizer
in the CAFO zone, thus the surface soils receiving more animal
wastes should have higher levels of TOC and arsenic contributed
by the animal wastes. The content of arsenic in the surface soils
from the CAFO zone was weakly correlated to TOC
(r2 = 5.7  102, p = 0.047), while no significant linear correlation
(r2 = 2.7  105, p = 0.97) was observed between arsenic and TOC
contents in the agricultural soils from the Pearl River Delta. Chem-
ical fertilizers are overwhelmingly used on agricultural lands in the
Pearl River Delta. In contrast, animal wastes are the predominant
form of fertilizers used in the CAFO zone due to their ready avail-
ability and the short transportation distance required. The correla-
tion between the contents of arsenic and organic carbon in the
surface soils from the CAFO zone suggests that the fertilization of
agricultural lands with animal wastes was a likely source of arsenic
in the surface soils. The r2 value of the linear model in the CAFO
zone is small because lithogenic materials remain the predominant
source of arsenic in the soils. As discussed earlier, the mean con-
tent of soil arsenic (16.0 mg kg1) was not too much higher than
that (12.2 mg kg1) in the agricultural soils in the Pearl River Delta,
thus TOC in the surface soils could not be the only predictor for soil
arsenic content, even if the soil organic matter came completely
from the animal wastes.
As the arsenic in the soils of the CAFO zone is contributed by
both natural and anthropogenic sources, identifying the exact
sources and their contributions can be quite challenging. The rela-
tively fast environmental transformation of organoarsenic feed Fig. 4. The correlation between the levels of total arsenic and TOC contents of the
additives in the environment further adds to the challenge as their agricultural soils in (a) the CAFO zone and (b) the Pearl River Delta.
 X. Liu et al. / Chemosphere 119 (2015) 273–281 279

degradation products (arsenate and arsenite) cannot be differenti-
ated from the inorganic arsenic species commonly present in sur-
face soils. Another important fact is that unlike the case of heavy
industrial pollution, the arsenic input with animal wastes is rela-
tively small compared to the arsenic already present in the soils.
As a result, several statistical tools are used to help identify the
potential sources of the soil arsenic in the CAFO zone. Even though
the correlations between animal wastes and soil arsenic pollution
in the CAFO zone are rather weak, the results do consistently indi-
cate that the arsenic in the surface soils in the CAFO has different
input sources, and animal wastes serve as a potential source.
3.3. Arsenic species in environmental matrices of the CAFO zone
The chemical forms of the arsenic compounds in the surface
soils, and in the samples of surface water and sediment as well,
were analyzed to further assess the contributions from the organ-
oarsenic feed additives. Table 3 summarizes the concentrations of
six arsenic species, including inorganic arsenic [As(III) and As(V)],
common organoarsenic compounds found in the environment
(MMA and DMA), and organoarsenic feed additives (ASA and
ROX) in the environmental matrices of the CAFO zone. It should
be noted that the total amount of arsenic species extracted by
the 0.5 M H3PO4 is much less than the total arsenic in the soil
and sediment samples determined by total digestion. This is not
surprising, as inorganic arsenic is primarily present within the
matrices of soil and sediment minerals and this fraction is not
extractable by weak acids, while the results of total digestion
include this fraction (Zhang et al., 2012a). On the other hand, the
fractions of arsenic species extracted by 0.5 M H3PO4 could closely
represent those that are potentially bioavailable and pose environ-
mental risk. Therefore, the results for soil and sediment samples
summarized in Table 3 could reflect more accurately the fractions
introduced by external sources and are bioavailable compared to
total arsenic contents. Arsenate was found to be the major inor-
ganic arsenic species, while ASA was the major organoarsenic spe-
cies in the surface soils and sediments. The concentrations of
arsenite in the sediments were relatively high compared to surface
soils, which was probably caused by the more reducing environ-
ment. The fact that arsenic is first methylated to MMA, and then
to DMA explains the higher concentrations of MMA relative to
those of DMA in the soils and sediments (Tamaki and
Frankenberger, 1992; Tlustoš et al., 2004). With dilution by stream
flow and sorption on the sediments, the arsenic compounds were
found in the surface water samples at rather low concentrations.
Nonetheless, ASA was consistently detected in the surface water
samples, suggesting continuous input of ASA from the CAFOs,
and possibly from leaching from the surrounding farm lands as
well. The detection of ASA and the absence of ROX, which is more
stable than ASA, in the environmental matrices is consistent with
the fact that ASA had long been used as the feed additive in
large-scale swine farms in Guangdong province (Wang et al.,
2006). The widespread occurrence of ASA in the environmental
matrices surrounding the CAFOs proves that animal wastes from
the swine farms are an important source of arsenic pollution in
the CAFO zone.
Fig. 5 shows the concentrations of the arsenic species in the sur-
face soils in the CAFO zone during the two sampling campaigns.
Overall, the concentrations of ASA decreased while those of arse-
nate and arsenite increased in the surface soils over the three
month period. As the first sampling campaign (June 2012) was car-
ried out just before crop planting while the second one (September
2012) was carried out prior to crop harvesting, no additional ani-
mal wastes would have been applied in between based on the
common agricultural practices. The weather in the Pearl River
Delta from July to September is hot and humid with frequent
typhoons. Such weather conditions favor biotic and abiotic degra-
dation of ASA, and leaching of the arsenic species from surface soils
as well. Meanwhile, leaching of ASA from the surface soils is
expected to be much less compared to arsenate and arsenite, as
it is relatively hydrophobic, less water soluble, and adsorbs more
strongly on the soil components compared to the inorganic arsenic
species. Consequently, the increase in the contents of inorganic
arsenic species could qualitatively support the reduction in ASA
contents resulted largely from transformation of ASA in the surface
soils between the two sampling campaigns. Overall, the mean total
arsenic concentration did not change significantly, while that of
ASA decreased (from 0.52 to 0.12 mg kg1) and those of inorganic
arsenic species increased in the surface soils over the three month
period. These results show that ASA could degrade relatively
quickly into inorganic arsenic species in the surface soils, which
is consistent with the findings of previous reports on the environ-
mental fate of organoarsenic feed additives (Bednar et al., 2003;
Garbarino et al., 2003; Jackson et al., 2006). As the inorganic
arsenic species are more toxic than the organoarsenic compounds
(WHO, 1981; Vega et al., 2001), the environmental degradation of
organoarsenic feed additives leads to an increase in the overall tox-
icity of the arsenic compounds discharged with the animal wastes.
3.4. Soil loading and potential ecological risk of arsenic pollution in the
CAFO zone
Accumulation of arsenic in agricultural soils can potentially
impact crop production, thus a quantitative understanding on the

Table 3
Summary of the concentrations of six arsenic species detected in the environmental matrices of the CAFO zone.

Samples As(III) As(V) MMA DMA ASA ROX

Recovery ratea (%) 8.9 ± 0.53 102 ± 16 96 ± 3.6 91 ± 6.1 67 ± 21 91 ± 2.9
Surface soils (mg kg1)
Concentration 0.11 ± 0.24 4.6 ± 3.9 0.015 ± 0.024 nd 0.24 ± 0.33 ndb
Range nd  1.6 nd  22 nd  0.16 nd  0.11 nd  1.2 ndb
Local background 0.037 1.9 nd nd nd nd
Sediments (mg kg1)
Concentration 0.28 ± 0.22 4.1 ± 2.7 0.047 ± 0.069 nd 0.37 ± 0.43 ndb
Range 0.039  0.91 0.85  10 nd  0.27 nd  0.026 nd  1.5 ndb
Surface waters (lg L)
Concentration 0.45 ± 0.74 0.70 ± 3.16 nd nd 0.29 ± 0.40 ndb
Range nd  2.8 nd  9.4 nd  0.47 nd  0.98 nd  1.5 ndb
Local background nd nd nd nd nd nd

Notes:
nd – not detected.
a
Recovery rates for arsenic species in soil and sediment samples established in Liu et al. (2013b).
b
Not detected for all samples.
280 X. Liu et al. / Chemosphere 119 (2015) 273–281
Fig. 5. Box and whisker plots for the concentrations of the arsenic species in the
surface soils in (a) June 2012 (n = 25) and (b) September 2012 (n = 47) in the CAFO
zone.
soil arsenic loading and the corresponding potential ecological risk
in the CAFO zone is essential. According to the numbers of pigs
raised in the CAFO zone, which are 0.5, 1, and 3.8 million heads
in 2008, 2009, and 2011, respectively (Township of Shiba, 2009;
Long et al., 2012), the annual growth rate of swine farming is esti-
mated to be about 100% in recent years. Based on the historic data
and the rough annual growth rate of 100%, it is estimated that
approximately 15 million pigs had been raised in the CAFO zone
by the end of 2012. According to the typical swine feed formula,
each tonne of feed contains 45–100 g of ASA (mean 80 g, or 28 g
As), while each pig consumes approximately 0.3 tonne of feed over
its life span. As organoarsenic feed additives are essentially not
uptaken by the animal bodies, almost all the ASA added to the
swine feed is discharged into the environments (in the forms of
ASA and its organic and inorganic metabolites). As a result, the
use of ASA in the farming of 15 million pigs (8.4 g of As each) is
estimated to result in the release of about 126 tonnes of arsenic
into the environmental matrices in the CAFO zone by 2012. With
a land area of 180 km2, the loading of arsenic in the surface soils
of the CAFO zone is estimated to be approximately 461.2 tonnes
based on the mean content of arsenic in the surface soils (with
6.7 tonnes in the form of ASA from the mean content of soil ASA,
as most of the ASA released already degraded). It is further esti-
mated that natural background and normal use of agrochemicals
on the farm lands accounted for approximately 351.6 tonnes of
the soil arsenic based on the mean content of arsenic in the agricul-
tural soils (12.2 mg kg1) of the Pearl River Delta. Consequently,
the difference (109.5 tonnes) is believed to be contributed by the
organoarsenic feed additives used in the swine farming in the
CAFO zone. The net accumulation of arsenic in soil (109.5 tonnes)
is less than the estimated input (126 tonnes), probably because
significant fractions of the arsenic species (primarily in the forms
of arsenate and arsenite) had been leached out of the surface soils
by rainwater and irrigation water, which subsequently impacted
local surface water and groundwater. Therefore, the extensive
use of organoarsenic feed additives makes animal wastes a signif-
icant anthropogenic source of arsenic pollution in the CAFO zone.
The ecological risk index (Er) values calculated for arsenic in the
surface soils of the CAFO zone are all lower than 40 (data not
shown), indicating low to moderate potential ecological risk
(Hakanson, 1980). Nonetheless, with the continuous expansion of
swine farming industry in the CAFO zone, fast accumulation of
arsenic from the animal wastes can significantly raise the potential
ecological risk of arsenic pollution in the near future if no action is
taken to limit the use of organoarsenic feed additives in intensive
animal farming.
4. Conclusion
Surface soils in the CAFO zone showed higher levels of arsenic
compared to the agricultural soils in the Pearl River Delta.
Correlation matrix, CA, and PCA results indicate that the sources
of the soil arsenic were significantly different between the CAFO
zone and the agricultural lands in the Pearl River Delta. These
observations, together with the correlation between soil arsenic
and TOC contents in the surface soils of the CAFO zone, suggest
that organoarsenic feed additives in the animal wastes are an
important source of arsenic pollution in the CAFO zone. Analysis
of the arsenic species showed the presence of ASA at significant
levels in the environmental matrices of the CAFO zone, while
ROX was absent. Degradation of ASA in the surface soils was also
observed between the two sampling campaigns (about 3 months).
Even though the potential ecological risk posed by arsenic in the
surface soils of the CAFO zone was relatively low, continuous dis-
charges of arsenic-bearing animal wastes from CAFOs might ele-
vate the arsenic risk. The findings of this study shows that soil
arsenic pollution caused by the extensive use of organoarsenic feed
additives in areas concentrated with swine CAFOs is worthy of sig-
nificant attention.
Acknowledgements
The authors gratefully acknowledge the anonymous reviewers
for valuable comments and suggestions. This work was supported
in parts by the Natural Science Foundation of China (Grant Nos.
41202251, 41322024, and 41121063), the Chinese Academy of
Sciences (Y234081A07 and ‘‘Interdisciplinary Collaboration Team’’
programs), and the National Program for Support of Top-notch
Young Professionals. H.C. also acknowledges the support of K.C.
Wong Education Foundation.
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