Arch. Environ. Contam. Toxicol. 45, 30 –36 (2003)
DOI: 10.1007/s00244-002-0267-7
A R C H I V E S O F
Environmental
Contamination
a n d Toxicology
© 2003 Springer-Verlag New York Inc.
Trace Organic Contaminants, Including Toxaphene and Trifluralin, in Cotton
Field Soils from Georgia and South Carolina, USA
K. Kannan,1 S. Battula,2 B. G. Loganathan,3 C.-S. Hong,1 W. H. Lam,2 D. L. Villeneuve,2 K. Sajwan,4 J. P. Giesy,2
K. M. Aldous1
1
Wadsworth Center, New York State Department of Health and Department of Environmental Health and Toxicology, State University of New York
at Albany, Empire State Plaza, P.O. Box 509, Albany, New York 12201-0509, USA
2
National Food Safety and Toxicology Center, Michigan State University, East Lansing, Michigan 48824-1311, USA
3
Department of Chemistry and Center for Reservoir Research, Murray State University, Murray, Kentucky 42071, USA
4
Department of Biology and Life Sciences, Environmental Sciences Program, Savannah State University, Savannah, Georgia 31411, USA
Received: 23 November 2002 /Accepted: 13 January 2003
Abstract. Residues of organic contaminants—including toxaphene, DDT, trifluralin, hexachlorocyclohexanes, polychlorinated
biphenyls, polycyclic aromatic hydrocarbons (PAHs) and nonylphenol—were measured in 32 cotton field soils collected from
South Carolina and Georgia in 1999. Toxaphene, trifluralin, DDT
and PAHs were the major contaminants found in these soils. The
maximum concentration of toxaphene measured was 2,500 ng/g
dry weight. Trifluralin was detected in all the soils at concentrations ranging from 1 to 548 ng/g dry weight. Pesticide residues
were not proportional to soil organic carbon content, indicating
that their concentrations were a reflection of application history
and dissipation rates rather than air–soil equilibrium. Soil extracts
were also subjected to in vitro bioassays to assess dioxinlike,
estrogenic, and androgenic/glucocorticoid potencies. Relatively
more polar fractions of the soils elicited estrogenic and androgenic/glucocorticoid activities, but the magnitude of response was
much less than those found in coastal marine sediments from
industrialized locations.
The availability of inexpensive elemental chlorine together with
the development of industrial chlorination procedures in the twentieth century led to the production of a wide range of chlorinated
organic compounds for commercial applications. Because of their
persistence, bioaccumulation, and toxic potentials, production and
usage of several chlorinated compounds have been banned or
restricted in the United States. However, there has been an ongoing debate about the primary sources of emission of organochlorine compounds in recent years. Countries such as India and
Mexico continue to use DDT and hexachlorocyclohexanes
(HCHs) for agriculture and to combat malaria (Loganathan and
Kannan 1994; Kannan et al. 1997). This can contribute “fresh”
emissions into the environment. Another potential source is emis-
Correspondence to: K. Kannan; email: kkannan@wadsworth.org
sion from historically contaminated soil or sediments. Agricultural
soils in the United States are suspected to be an important source
of organochlorine pesticides that were used historically in large
quantities (Harris et al. 2000). Depending on the physicochemical
properties (Koc, half-life), soil characteristics (amounts and types
of clay materials, hydrous oxides and organic matter, redox status,
and pH), agricultural practices (e.g., tillage, irrigation), and meteorological conditions, organochlorines in soils can be released into
the atmosphere (Willis et al. 1983; Hitch and Day 1992; Boul et
al. 1994; Harris et al. 2000). For instance, concentrations of DDT
in ambient air above the soil of a California farm where DDT was
applied 23 years ago were two to three orders of magnitude greater
than those found in the Great Lakes region (Spencer et al. 1996).
It has been hypothesized that pesticides used historically in
agriculture in the Southern United States are being volatilized to
the atmosphere and likely contributing to the long-range transport
of these compounds (Spencer et al. 1996; Hoff et al. 1996; Harner
et al. 1999). Georgia and South Carolina are major cotton-growing regions in the United States. DDT and toxaphene were used
widely to control pests in cotton fields. Over 85% of toxaphene
use in the United States was in cotton-growing states from Texas
through Georgia. Only 1– 4% of its use occurred in the upper
Midwest, including the Great Lakes Basin (James and Hites
2002). DDT was used until 1972, and toxaphene was used until
1990. Relatively high concentrations of toxaphene found in the
Great Lakes water have been attributed to the atmospheric transport of toxaphene from the Southern United States (James and
Hites 2002).
Trifluralin (␣-,␣-,␣-trifluoro-2,6-dinitro-N,N-dipropyl-p-toluidine) is a selective, preemergence dinitroaniline herbicide most
commonly used in cotton cultivation. Half-lives of trifluralin in
soils vary from 45 days to 8 months (Wheeler et al. 1979;
Environment Canada 1987). Very few studies have reported the
occurrence of trifluralin in soils. In this study, we determined the
concentrations of organochlorine pesticides, including DDT and
its derivatives (p,p⬘-DDE, p,p⬘-DDD, and p,p⬘-DDT), HCHs,
hexachlorobenzene (HCB), toxaphene, and organofluorine herbicide trifluralin in soils collected from cotton fields in Georgia and
Trace Organic Contaminants in Cotton Field Soils
South Carolina. Furthermore, soils were also tested for the presence of polychlorinated biphenyls (PCBs), polycyclic aromatic
hydrocarbons (PAHs), and nonylphenol (NP).
In addition to instrumental analysis, extracts of a few soil
samples were analyzed for dioxinlike, estrogenic, and combined androgen receptor (AR)– and glucocorticoid receptor
(GR)–mediated activities using in vitro bioassays. In vitro
bioassays provide information on the potency of all compounds
present in soil extracts that act through a known mechanism of
action. This approach has been used to characterize contaminants in sediments or biota (Khim et al. 1999; Kannan et al.
2000; Hilcherova et al. 2000). In vitro luciferase assay with
recombinant rat hepatoma cells (H4IIE-luc) was used to screen
for compounds capable of modulating aryl hydrocarbon receptor (AhR)–mediated gene expression (Sanderson et al. 1996).
In vitro luciferase assay with recombinant MCF-7 human
breast carcinoma cells (MVLN) was used to screen extracts for
compounds that can modulate gene expression through an
estrogen receptor (ER)–mediated mechanism (Pons et al.
1990). MDA-kbs cells are MDA-MB-453 breast cancer cells
that were transformed with a MMTV.lucifierase.neo reporter
gene construct that is under control of both the AR and GR
(Wilson et al. 2002; Hartig et al. 2002). Compounds that act
through either the AR or GR can activate the MMTV luciferase
reporter gene, resulting in increased luciferase expression and
therefore greater light production in the bioassay.
31
Fig. 1. Map of the United States showing soil sampling locations in
South Carolina and Georgia
concentrated to a final volume of 1 ml prior to injection. In terms of
polarity, F1 was nonpolar, F2 was moderately polar, and F3 was polar.
Chemical Analysis
Materials and Methods
Sample Collection
Surface soils (0 –10 cm) were collected from 32 cotton fields: 16 from
South Carolina on 14 November 1999 and 16 from Georgia on 4
December 1999 (Figure 1, Table 1). Cotton was the standing crop
when samples were collected. Samples from several locations in a
given field were pooled to obtain a representative sample. Samples
were transferred to precleaned I-Chem glass jars and stored at ⫺20°C
until extraction.
Sample Preparation and Cleanup
Soil moisture content was determined by weighing soils before and
after drying for ⬃20 h at 100°C. Total organic content (TOC) of the
soil samples was analyzed at the Soil and Plant Nutrient Laboratory at
Michigan State University. Pesticides, PCBs, PAHs, and NP were
analyzed following methods described elsewhere (Kannan et al. 2001).
Briefly, 35 g of soil (wet weight) were weighed and mixed with
anhydrous sodium sulfate to remove the moisture content until a fine,
grainy consistency was achieved. PCB-30 (50 ng) was added as an
internal standard before extraction. Samples were extracted with 400
ml dichloromethane (DCM)/hexane (3:1) for ⬃ 18 h. Extracts were
then concentrated to 10 ml and treated with acid-activated copper for
the removal of sulfur. Extracts were passed through 10 g of activated
florisil column (10 mm ID). The first fraction (F1), eluted with 100 ml
hexane, contained PCBs, HCB, and p,p⬘-DDE. The second fraction
(F2), eluted with 100 ml 20% DCM in hexane, contained PAHs,
p,p⬘-DDT, p,p⬘-DDD, HCHs (␣-, ␤-, and ␥-isomers) and toxaphene.
NP was eluted in the third fraction (F3) using 100 ml 50% DCM in
methanol. Trifluralin eluted in both F2 and F3. The fractions were
Organochlorines and trifluralin were quantified using a gas chromatograph (GC) equipped with a 63Ni electron capture detector, and PAHs
were quantified using GC interfaced with a mass spectrometer (Kannan et
al. 2001). The mass spectrometer was operated under selected ion monitoring mode using the molecular ions selective for individual PAHs
(Kannan et al. 2001; Khim et al. 1999). NP was analyzed using a
reverse-phase, high-pressure liquid chromatograph with fluorescence detection (Kannan et al. 2001). A solution containing 100 individual PCB
congeners with known composition and content was used as a standard,
and concentrations of 100 individually resolved peaks were summed to
obtain total PCB concentration (Khim et al. 2000). The PAH standard
consisted of 16 priority pollutant PAHs as identified by the U.S. Environmental Protection Agency (Method 8310). Further details of the instrumental analysis have been described elsewhere (Kannan et al. 2001).
Identity of trifluralin was confirmed by injecting the extracts into a
Finnigan Thermoquest Trace 2000 Series GC interfaced with a GCQ plus
ion trap mass spectrometer. Selected ion chromatogram and total ion
spectrum of trifluralin standard and a sample are shown in Figure 2.
Toxaphene was quantified by integrating all of the toxaphene peaks.
p,p⬘-DDT and p,p⬘-DDD interfered with toxaphene peaks and were isolated for quantifying total toxaphene concentrations.
Bioassay Analysis
Selected soil extracts from the florisil column fractions were screened for
their ability to induce AhR-, ER-, and AR/GR-mediated activity using in
vitro bioassays. The procedures applied to conduct the bioassays have
been described in detail previously (Villeneuve et al. 2000a, 2000b;
Hilcherova et al. 2001; Wilson et al. 2002). Luciferase assay was conducted after 72 h of exposure of H4IIE-luc and MVLN cells and 48 h of
exposure of MDA cells to soil extracts. Responses of cells, determined as
mean relative luminescence units (RLUs) over three replicate wells, were
expressed as a percentage of the maximum response observed for 2,3,7,8tetrachlorodibenzo-p-dioxin (TCDD; %-TCDD-max.) or 17-␤ estradiol or
32
K. Kannan et al.
Table 1. Concentrations of total organic carbon (%), DDTs, HCHs, and PAHs (ng/g dry weight) in cotton field soils from Georgia and South
Carolina
Sample ID
Location
South Carolina
SC soil #1
SC soil #2
SC soil #3
SC soil #4
SC soil #5
SC soil #6
SC soil #7
SC soil #8
SC
SC
SC
SC
soil
soil
soil
soil
#9
#10
#11
#12
SC soil #13
SC soil #14
SC soil #15
SC soil #16
Georgia
GA soil #1
GA soil #2
GA soil #3
GA soil #4
GA soil #5
GA
GA
GA
GA
GA
GA
GA
GA
GA
GA
GA
soil
soil
soil
soil
soil
soil
soil
soil
soil
soil
soil
#6
#7
#8
#9
#10
#11
#12
#13
#14
#15
#16
Hampton County, Nixville Road, 2 miles from Estill
Hampton County, Nixville Road, 2 miles from Estill
Hampton County, Hwy 3, 3 miles from Estill
Hampton County, Hwy 3N, Gravel Road, 6 miles east of
Estill
Hampton County, Estill
Hampton, Hwy 321, 14 miles from Estill
Hampton County, Hwy 321, 18 miles west of Estill,
Columbia Road, Gifford
Hampton County, Columbia Road, Gifford, 18 miles north
of Estill
Hampton County, Columbia Road, Gifford
Hampton County, Columbia Road, Gifford
Annandale County, Hwy 321, Buford’s Bridge Hwy
Annandale County, Hwy 321 (right), Pond Town Rd,
Scycamore Town
Annandale County, Hwy 321 (left), Pond Town Rd,
Scycamore Town
Border of Annandale and Orangeburg County, Hwy 301
Border of Annandale and Orangeburg County, Hwy 301
Orangeburg County, Hwy 301
Bulloch County, Statesboro, Hwy 67
Bulloch County, Statesboro, 1 mile from #1, Hwy 67
Bulloch County, Statesboro, 3 miles from #1, Hwy 46E
Bulloch County, Statesboro, 4 miles from #1, Hwy 46E
Bulloch County, Statesboro, Red Hill Church Road, 8 miles
from #1
Bulloch County, Statesboro, 9 miles from #1, Hwy 46E
Bulloch County, Statesboro, mile mark 20, Hwy 46E
Bulloch County, Statesboro, 46W
Bulloch County, 46W, mile 13
Bulloch County, Hwy 301 (harvested cotton field)
Statesboro city, Hwy 67
Statesboro city, Hwy 67
Statesboro, south Wynn Road
Statesboro, Rushing Road
Georgia Hwy 67S
Georgia Hwy 67S
TOC
(%)
p,p⬘DDT
p,p⬘DDD
p,p⬘DDE
0.67
0.57
0.69
1.67
⬍0.1
⬍0.1
⬍0.1
⬍0.1
⬍0.1
2.0
0.1
0.3
3.62
0.11
0.31
⬍0.1
⬍0.1
⬍0.1
8.2
6.4
13.9
1.06
0.68
1.48
1.25
1.13
X
0.53
⬍0.1
X
1.9
1.8
14.3
3.68
2.93
14.3
⬍0.1
⬍0.1
0.39
27.1
⬍5
138
1.54
0.50
0.10
0.9
1.50
0.19
1.59
1.65
1.21
0.77
0.84
11.8
3.38
1.80
0.17
0.44
1.88
0.39
3.0
14.5
6.2
9.1
4.03
26.7
11.5
11.3
⬍0.1
0.25
0.1
⬍0.1
0.86
10.5
1.28
14.7
26.5
0.97
1.27
0.71
1.44
18.1
3.74
1.77
0.20
3.70
1.28
0.33
⬍0.1
23.0
10.8
8.9
0.5
1.12
1.03
0.76
0.69
4.19
1.75
6.73
1.39
0.21
⬍0.1
1.03
⬍0.1
0.57
1.26
0.92
0.56
0.75
1.01
0.68
1.35
0.8
0.96
1.07
1.43
6.6
14
0.36
3.69
4.96
0.36
9.68
0.56
1.25
0.23
1.94
0.32
0.54
2.23
⬍0.1
0.35
0.40
0.24
2.79
⬍0.1
⬍0.1
0.11
0.26
⬍0.1
⌺DDT
⌺HCH
PAHs
34
8.3
78.8
7.9
⬍5
0.16
241
44.8
15.9
11
0.75
0.54
⬍0.1
⬍0.1
⬍0.1
⬍5
⬍5
14
26
14.5
6.9
14.0
11.1
18.9
8.64
21.8
12.5
⬍0.1
⬍0.1
⬍0.1
⬍0.1
16.5
49.3
19.6
⬍5
18.3
17.3
0.8
5.3
14.6
0.3
10.3
⬍0.1
4.9
⬍0.1
16.7
0.6
25.4
33.6
1.16
9.36
19.9
0.88
22.8
0.56
6.15
0.34
18.9
0.90
⬍0.1
0.16
⬍0.1
⬍0.1
⬍0.1
⬍0.1
0.49
⬍0.1
⬍0.1
⬍0.1
⬍0.1
⬍0.1
⬍5
13.3
23.8
13.3
9.3
18.3
11
189
6.7
6.9
7.7
24.1
Concentrations of PCBs, HCB, and NP were ⬍5, ⬍1, and ⬍5 ng/g dry weight, respectively.
X: interference due to high toxaphene concentrations.
testosterone standard curves generated on the same day. The MDA cells
measure both AR- and GR-mediated responses. The two responses can be
distinguished by addition of an AR inhibitor, but this has not been done in
this study. As a result, any responses should be considered combined
AR/GR activity. Testosterone was used as a standard for calibrating MDA
cell responses. Further details of the bioassay procedures are presented
elsewhere (Villeneuve et al. 2000a, 2000b; Khim et al. 1999; Kannan et
al. 2000; Hilcherova et al. 2001; Wilson et al. 2002).
Results and Discussion
Chemical Analysis
Toxaphene and trifluralin were the most commonly detected
pesticides in soils. Concentrations of toxaphene in cotton soils
from South Carolina ranged from 3.3 to 2,500 ng/g dry weight
(mean: 277), whereas those from Georgia ranged from 9.5 to
269 ng/g dry weight (mean: 83) (Table 2). The highest toxaphene concentration, 2,500 ng/g dry weight, was found in a soil
sample collected from Hampton County in South Carolina.
This value was similar to the highest concentration of 2,420
ng/g dry weight, reported for a farm soil collected from southwestern Alabama in the late 1990s (Harner et al. 1999). In
general, the range of toxaphene concentrations found in soils
from South Carolina are similar to those observed in farm soils
in Alabama (⬍3–2,420 ng/g dry weight) (Harner et al. 1999).
Measured concentrations of toxaphene in soils were 10- to
100-fold greater than those observed in surface sediments from
Grand Traverse Bay, Lake Michigan (Schneider et al. 2001). A
regression of toxaphene concentrations versus percent organic
Trace Organic Contaminants in Cotton Field Soils
33
Fig. 2. Selected ion and total ion spectrum of trifluralin standard and a soil sample using ion trap gas chromatograph–mass spectrometer
carbon showed a lack of significant relationship (r2 ⫽ 0.002;
outlier removed).
Occurrence of notable concentrations of toxaphene in soils
has implications for atmospheric contamination by this compound by soil–air exchange. For a soil–air system in equilibrium, the concentration of a hydrophobic chemical is expected
to be proportional to the organic carbon content of the soil. The
wide variability in soil concentrations of toxaphene and the
poor correlation with organic carbon suggest that the soil
burdens are largely dependent on the application history of
toxaphene and the soil–air concentrations are not in equilibrium.
Of the various contaminants measured, trifluralin, a preemergence organofluorine herbicide, was the second abundant compound found in soils (Table 2). Concentrations of trifluralin in
cotton field soils from South Carolina ranged from 3.3 to 574
ng/g dry weight (mean: 115), whereas those from Georgia
ranged from 1 to 548 ng/g dry weight (mean: 150) (Table 2).
There was no significant difference in the concentrations of
trifluralin in soils from Georgia and South Carolina (p ⬎ 0.05).
Trifluralin is applied as a preemergence herbicide during
March–May. Occurrence of this herbicide in soils collected in
November–December suggests its persistence in soils. Occur-
rence of trifluralin at concentrations of 50 to 130 ng/g dry
weight has been reported in soils 30 months after the last
application (Corbin et al. 1994). Concentrations of trifluralin in
soils collected from vegetable farms in British Columbia varied
from 55 to 310 ng/g dry weight (Szeto and Price 1991). These
values are similar those observed in this study. Trifluralin is
strongly adsorbed to soil organic matter (Wheeler et al. 1979),
and the adsorbed herbicide becomes inactive. Therefore, higher
application rates are needed for soils rich in organic matter.
Nevertheless, concentrations of trifluralin were not proportional to soil organic carbon. This suggests that in addition to
application rates, farm management practices, such as tillage
and irrigation, and soil temperature play an important role in
determining persistence of trifluralin in soils.
Residues of DDT were found in almost all of the soils (Table
1). Total concentrations of DDT (including DDE and DDD)
ranged from 0.11 to 45 ng/g dry weight (mean: 11) in soils
from South Carolina and from 0.34 to 34 ng/g dry weight
(mean: 13) in soils from Georgia (Table 1). p,p⬘-DDE was the
predominant metabolite of DDT, accounting for, on average,
69% of the total DDT concentrations. p,p⬘-DDT accounted for
33% of the total DDT concentrations. DDT concentrations
were similar to those observed in soils from Alabama (mean:
34
K. Kannan et al.
Table 2. Concentrations of toxaphene and trifluralin (ng/g dry
weight) in cotton field soils from Georgia and South Carolina
Location
South Carolina
SC soil #1
SC soil #2
SC soil #3
SC soil #4
SC soil #5
SC soil #6
SC soil #7
SC soil #8
SC soil #9
SC soil #10
SC soil #11
SC soil #12
SC soil #13
SC soil #14
SC soil #15
SC soil #16
Georgia
GA soil #1
GA soil #2
GA soil #3
GA soil #4
GA soil #5
GA soil #6
GA soil #7
GA soil #8
GA soil #9
GA soil #10
GA soil #11
GA soil #12
GA soil #13
GA soil #14
GA soil #15
GA soil #16
TOC (%)
Trifluralin
Toxaphene
0.67
0.57
0.69
1.06
0.68
1.48
1.54
1.59
1.65
1.21
0.77
0.86
0.97
1.27
0.71
1.44
41.0
35.8
50.1
17.8
3.3
67.7
212
109
104
176
176
5.7
16.6
574
178
69.5
26.6
3.3
63.2
75.8
10.4
2500
48.5
26.2
116
99.0
100
275
698
286
94.8
5.7
1.12
1.03
0.76
0.69
0.57
1.26
0.92
0.56
0.75
1.01
0.68
1.35
0.8
0.96
1.07
1.43
22.3
16.6
122
548
1.0
297
197
1.0
323
269
7.1
127
181
105
58.7
132
43.3
96.9
166
14.8
138
130
12.1
76.2
112
56.6
269
101
21.3
21.7
58.3
9.5
50 ng/g dry weight) (Harner et al. 1999). The mean concentrations were 5- to 20-fold less than those observed in cotton
field soils in India (mean: 1,000; median: 280 ng/g dry weight),
where DDT was used until the late 1990s (Kawano et al. 1992).
There was no significant relationship between the concentrations of total DDT and soil organic matter content. DDT was
extensively used as an agricultural and vector control pesticide
in the United States with peak production occurring in 1963.
Usage of DDT was banned in the United States in 1972.
However, DDT persists in soil for several years with reported
half-lives ranging from 20 –30 years (Dimond and Owen
1996).
Concentrations of total HCHs were less than 1 ng/g in all the
soils. Among HCH isomers, ␤- and ␥-HCH isomers were
found in some soils. HCB, PCBs, and NP were not detected at
the detection limits of 1, 5, and 5 ng/g dry weight, respectively
(Table 1). The application of sludge in agriculture can be a
potential source for these compounds in soils. In addition,
presence of these compounds as impurities in other chlorinated
pesticide formulations and atmospheric deposition can contribute to their occurrence in soils. Occurrence of NP in soils
treated with sewage sludge has been reported (Vikelsoe et al.
2002). The results our study suggest that contamination by
HCB, PCBs, and NP in cotton soils is not a significant concern.
PAHs are a group of common environmental contaminants
that originate from anthropogenic sources such as waste incineration, coal gasification and accidental oil spills as well as
natural processes like fossil fuel and wood combustion. The
total PAH concentrations ranged over two orders of magnitude
from ⬍5 to 241 ng/g dry weight (mean: 38 ng/g) (Table 1).
Three of the 32 soils analyzed contained PAH concentrations
greater than 100 ng/g dry weight. Fluoranthene was the most
commonly detected PAH in soils. In soils that contained PAH
concentrations greater than 100 ng/g, pyrene, chrysene, and
benzo[b]fluoranthene were also found at notable levels. The
concentrations of total PAHs in the soil samples are in excess
of the reported natural concentrations in soils of 1–10 ng/g
(Edward 1987) but less than those in rural soils from the United
Kingdom (median: 190 ng/g) (Wild and Jones 1995).
Bioassay Analysis
Dioxinlike (AhR), estrogenic (ER), and androgenic/glucocorticoid (AR/GR)–like activities of 5 of the 32 soils analyzed in
this study are shown in Figure 3. The soil samples tested had
low to insignificant AhR-, ER-, and AR/GR-mediated activity.
None of the F1 samples elicited significant dioxinlike activity,
whereas one of the F2 samples and four of the F3 samples
elicited significant dioxinlike activity, although the maximum
response was only 10% of that elicited by 1,500 pM TCDD.
Low activity in F1 extracts is consistent with the lack of
coplanar PCBs, but greater activities in F2 and F3 extracts
suggest the presence of dioxinlike compounds in these fractions. Polychlorinated dibenzo-p-dioxins (PCDDs) and polychlorinated dibenzofurans (PCDFs) are expected to be eluted in
these fractions. However, these compounds were not quantified
in soils. PAHs that elute in F2 can act through AhR-mediated
mechanism of action (Villeneuve et al. 2002). Concentrations
of PAHs in GS (Georgia Soil) 1, GS2, GS3, GS4, and GS5
were 8.2, 6.4, 14, 27, and ⬍5 ng/g dry weight, respectively.
Relatively high concentration of PAHs in GS4 corresponds
with the greater dioxinlike activity observed in this sample.
Occurrence of dioxinlike activities in F2 and F3 soil extracts
was similar to those observed in coastal sediments (Khim et al.
1999; Kannan et al. 2000), but the magnitude of response in
soils extracts was severalfold lower than those in coastal sediments from industrialized areas.
None of the F1 extracts of soils were estrogenic. One of the
five F2 extract was estrogenic. Some of the PAHs that elute in
F2 are weakly estrogenic (Villeneuve et al. 2002). Lack of
estrogenic activity in most of the F2 samples suggests that the
concentrations of PAHs measured were not high enough to
elicit a significant response. Three of the five F3 soil extracts
elicited estrogenic activity at ⬃10% of that elicited by 100 pM
17-␤ estradiol. Estrogenic activity associated with F3 suggests
the presence of unknown/unidentified estrogenic compounds in
this polar fraction of soils. This is similar to that reported for
marine sediments (Khim et al. 1999).
p,p⬘-DDE, which appears in F1 of the florisil column fractionation procedure, has been reported as an androgenic compound (Kelce et al. 1995). To date, potential xenobiotic GR
Trace Organic Contaminants in Cotton Field Soils
35
Fig. 3. Dioxinlike, estrogenic, and androgenic/glucocorticoid-like activities of florisil fractions 1, 2, and 3 of Georgia cotton soils (GS). GS1 ⫽
Georgia soil #1, GS2 ⫽ Georgia soil 2, and so on. Response magnitude presented as a percentage of the maximum response observed for a 1,500
pM TCDD, 1,000 pM 17-␤ estradiol, and 3,170 pM testosterone standards (100% max), respectively, for dioxinlike, estrogenic, and androgenic
activities. Error bars ⫽ SD; sig ⫽ magnitude of response equal to 3 SD above the mean solvent control response (0% max)
agonists have not been well characterized. None of the five F1
extracts elicited significant AR/GR-mediated activity, which
suggests that the concentrations of p,p⬘-DDE measured in these
soils were not adequate enough to elicit significant androgenic
or glucocorticoid-like response. Three of the F3 extracts elicited AR/GR-mediated activity, suggesting the presence of AR
and/or GR agonists in this fraction. Additional assays conducted in the presence of an AR inhibitor (such as hydroxyflutamide) would be needed to distinguish from glucocorticoidlike contributions to the response observed (Wilson et al.
2002). No known AR or GR agonists were quantified in this
fraction.
These results suggest that the residues of organochlorine
pesticides in cotton field soils in Georgia and Alabama varied
two to three orders of magnitude. Toxaphene, trifluralin, and
DDT were the predominant pesticides detected in these soils.
Concentrations of these pesticides were not proportional to
organic carbon content, which suggests that the history of the
pesticide usage and farm management practices play a major
role in influencing residue levels. It is hypothesized that toxaphene and trifluralin residues in soils in Georgia and South
Carolina are continually being volatilized to the atmosphere
and likely to contribute to long-range transport as suggested
earlier (Spencer et al. 1996; Harner et al. 1999). Bioassay
analysis of selected soils suggests the presence of estrogenic
and androgenic and/or glucocorticoid-like compounds in polar
fractions. Further studies are needed to identify and characterize the estrogenic and androgenic compounds present in these
fractions.
Acknowledgments. We thank Steven Connor, Wadsworth Center, for
assistance with ion trap mass spectrometer analysis of trifluralin. The
U.S. Department of Energy and the U.S. Environmental Protection
Agency funded this study through Savannah State University.
References
Boul ML, Garnham ML, Hucker D, Baird D, Aislabie J (1994)
Influence of agricultural practices on the levels of DDT and its
residues in soil. Environ Sci Technol 28:1397–1402
Corbin BR, McClelland M, Frans RE, Talbert RE, Horton D (1994)
Dissipation of fluometuron and trifluralin residues after long-term
use. Weed Sci 42:438 – 445
Dimond JB, Owen RB (1996) Long-term residue of DDT compounds
in forest soils in Maine. Environ Pollut 92:227–230
Edward NTJ (1987) Polycyclic aromatic hydrocarbons (PAHs) in the
terrestrial environment—a review. J Environ Qual 12:427– 441
Environment Canada/Agriculture Canada (1987) Pesticide registrant
survey, 1986. Commercial Chemicals Branch, Environment Canada, Ottawa
36
Harner T, Wideman JL, Jantunen LMM, Bidleman TF, Parkhurst WJ
(1999) Residues of organochlorine pesticides in Alabama soils.
Environ Pollut 106:323–332
Harris ML, Wilson LK, Elliott JE, Bishop CA, Tomlin AD, Henning
KV (2000) Transfer of DDT and metabolites from fruit orchard
soils to American robins (Turdus migratorius) twenty years after
agricultural use of DDT in Canada. Arch Environ Contam Toxicol
39:205–220
Hartig PC, Bobseine KL, Britt BH, Cardon MC, Lambright CR,
Wilson VS, Gray LE (2002) Development of two androgen receptor assays using adenoviral transduction of MMTV-Luc reporter and/or hAR for endocrine screening. Toxicol Sci 66:82–90
Hilcherova K, Machala M, Kannan K, Blankenship AL, Giesy JP
(2000) Cell bioassays for detection of dioxin-like and estrogen
receptor mediated activity in environmental samples. Environ Sci
Pollut Res 7:159 –171
Hilcherova K, Kannan K, Kang Y-S, Holoubek I, Machala M, Masunaga S, Nakanishi J, Giesy JP (2001) Characterization of dioxinlike activity of riverine sediments from the Czech Republic.
Environ Toxicol Chem 20:2768 –2777
Hitch RK, Day HR (1992) Unusual persistence of DDT in some
western USA soils. Bull Environ Contam Toxicol 48:259 –264
Hoff RM, Strachan WMJ, Sweet CW, Chan CH, Shackleton M,
Bidleman TF, Brice KA, Burniston DA, Cussion S, Gatz DF,
Harlin K, Schroeder WH (1996) Atmospheric deposition of chemicals to the Great Lakes: a review of data through 1994. Atmos
Environ 30:3505–3527
James RR, Hites RA (2002) Atmospheric transport of toxaphene from
the southern United States to the Great Lakes region. Environ Sci
Technol 36:3474 –3481
Kannan K, Kober JL, Kang Y-S, Masunaga S, Nakanishi J, Ostaszewski A, Giesy JP (2001) Polychlorinated—naphthalenes, -biphenyls, -dibenzo-p-dioxins, -dibenzofurans, polycyclic aromatic hydrocarbons and alkylphenols in sediment from the Detroit and
Rouge Rivers, Michigan, USA. Environ Toxicol Chem 20:1878 –
1889
Kannan K, Tanabe S, Giesy JP, Tatsukawa R (1997) Organochlorine
pesticides and polychlorinated biphenyls in foodstuffs from Asian
and Oceanic countries. Rev Environ Contam Toxicol 152:1–55
Kannan K, Villeneuve DL, Yamashita N, Imagawa T, Hashimoto S,
Miyazaki A, Giesy JP (2000) Vertical profiles of dioxin-like and
estrogenic activities associated with a sediment core from Tokyo
Bay, Japan. Environ Sci Technol 34:3560 –3567
Kawano M, Ramesh A, Thao VD, Tatsukawa R, Subramanian AN
(1992) Persistent organochlorine insecticide residues in some
paddy, upland and urban soils of India. Intern J Environ Anal
Chem 48:163–174
Kelce WR, Christy RS, Laws SC, Gray LE, Kemppainen JA, Wilson
EM (1995) Persistent DDT metabolite p,p⬘-DDE is a potent androgen receptor. Nature 375:581–585
Khim JS, Kannan K, Villeneuve DL, Koh CH, Giesy JP (1999)
Characterization and distribution of trace organic contaminants in
sediment from Masan Bay, Korea. 2. Instrumental analysis. Environ Sci Technol 33:4199 – 4205
K. Kannan et al.
Khim JS, Villeneuve DL, Kannan K, Hu WY, Giesy JP, Kang SG,
Song KJ, Koh CH (2000) Instrumental and bioanalytical measures
of persistent organochlorines in blue mussel (Mytilus edulis) from
Korean coastal waters. Arch Environ Contam Toxicol 39:360 –
368
Loganathan BG, Kannan K (1994) Global organochlorine contamination trends: an overview. Ambio 23:187–191
Pons M, Gagne D, Nicolas JC, Mehtali M (1990) A new cellular model
of response to estrogens: a bioluminescent test to characterize
(anti)estrogen molecules. Biotechniques 9:450 – 459
Sanderson JT, Aarts JMMJG, Brouwer A, Froese KL, Denison MS,
Giesy JP (1996) Comparison of Ah receptor–mediated luciferase
and ethoxyresorufin-O-deethylase induction in H4IIE cells: implications for their use as bioanalytical tools for the detection of
polyhalogenated aromatic hydrocarbons. Toxicol Appl Pharmacol
137:316 –325
Schneider AR, Stapleton HM, Cornwell J, Baker JE (2001) Recent
declines in PAH, PCB, and toxaphene levels in the northern Great
Lakes as determined from high resolution sediment cores. Environ
Sci Technol 35:3809 –3815
Spencer WF, Singh G, Taylor CD, LeMart RA, Cliath MM, Farmer
WJ (1996) DDT persistence and volatility as affected by management practices after 23 years. J Environ Qual 25:815– 821
Szeto SY, Price PM (1991) Persistence of pesticide residues in mineral
and organic soils in the Fraser valley of British Columbia. J Agric
Food Chem 39:1679 –1684
Vikelsoe J, Thomsen M, Carlsen L (2002) Phthalates and nonylphenols in profiles of differently dressed soils. Sci Total Environ
296:105–116
Villeneuve DL, Blankenship AL, Giesy JP (2000a) Derivation and
application of relative potency estimates based on in vitro bioassay results. Environ Toxicol Chem 19:2835–2843
Villeneuve DL, Kannan K, Khim JS, Falandysz J, Nikiforov VA,
Blankenship AL, Giesy JP (2000b) Relative potencies of individual polychlorinated naphthalenes to induce dioxinlike responses in
fish and mammalian in vitro bioassays. Arch Environ Contam
Toxicol 39:273–281
Villeneuve DL, Khim JS, Kannan K, Giesy JP (2002) Relative potencies of individual polycyclic aromatic hydrocarbons to induce
dioxinlike and estrogenic responses in three cell lines. Environ
Toxicol 17:128 –137
Wheeler W, Stratton G, Twilley R, Ou L-T, Carlson D, Davidson J
(1979) Trifluralin degradation and binding in soil. J Agric Food
Chem 27:702–706
Wild SR, Jones KC (1995) Polynuclear aromatic hydrocarbons in the
United Kingdom environment: a preliminary source inventory and
budget. Environ Pollut 88:91–108
Willis GH, McDowell LL, Harper LA, Southwick LM, Smith S (1983)
Seasonal disappearance and volatilization of toxaphene and DDT
from a cotton field. J Environ Qual 12:80 – 85
Wilson VS, Bobseine K, Lambright CR, Gray LE (2002) A novel cell
line, MDA-kb2, that stably expresses an androgen- and glucocorticoid-responsive reporter for the detection of hormone receptor
agonists and antagonists. Toxicol Sci 66:69 – 81