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Abstract
Total petroleum hydrocarbons (TPH) contaminated soil samples were collected from Shengli Oileld of China. Toxicity analysis was
carried out based on earthworm acute toxicity, plant growth experiment and luminescent bacteria test. The soil was contaminated bypetroleum hydrogcarbons with TPH concentration of 10.57%. With lethal and sub-lethal rate as endpoint, earthworm test showed that
the LD50 (lethal dose 50%) values in 4 and 7 days were 1.45% and 1.37% respectively, and the inhibition rate of earthworm body weight
increased with higher oil concentration. TPH pollution in the soil inhibited seed germination in both wheat and maize experiment when
the concentration of petroleum was higher than 0.1%. The EC50 (eective concentration 50%) for germination is 3.04% and 2.86%
in maize and wheat, respectively. While lower value of EC50 for root elongation was to be 1.11% and 1.64% in maize and wheat,
respectively, suggesting higher sensitivity of root elongation on petroleum contamination in the soil. The EC50 value in luminescent
bacteria test was 0.47% for petroleum in the contaminated soil. From the experiment result, it was concluded that TPH content of 1.5%
is considered to be a critical value for plant growth and living of earthworm and 0.5% will aect the activity of luminescent bacteria.
Key words: toxicity; total petroleum hydrocarbon; soil; earthworm; contamination
DOI: 10.1016/S1001-0742(10)60517-7
Citation: Tang J C, Wang M, Wang F, Sun Q, Zhou Q X, 2011. Eco-toxicity of petroleum hydrocarbon contaminated soil. Journal of
Environmental Sciences, 23(5): 845851
Introduction
Although pollutants and its concentration in soil can be
characterized rapidly by chemical method, the overall soil
quality can not be reected by only chemical analysis.
Concerning on the eco-toxicity of pollutants, biological
method is more suitable in determining possible hazard of
pollutants in soil on the ecology and environment. However, eorts to conduct site specic risk assessments using
ecotoxicity test and correlation to contaminant concentration is limited and relatively unsuccessful for hydrocarbon
contamination (Dorn and Salanitro, 2000).
Toxicity assessment of the contaminated soil with bioassays could provide meaningful information regarding a
characterization procedure in ecological risk assessment
(Al-Mutairi et al., 2008). The ecological toxicity diagnosis
includes dierent methods by using soil animal and plant
as testing approaches. Earthworm was used as test animal
in soil toxicity experiments and has been established as
standard method by International Organization for Standardization (ISO) in 1993 (ISO11268-1). Toxicity of heavy
metals on earthworm has been widely studied, and the
research has been conducted in subcellular level (Huang et
al., 2009; Klok and Thissen, 2009; Owojori et al., 2008).
* Corresponding author. E-mail: tangjch@nankai.edu.cn
846
The change of petroleum composition during bioremediation process will result in the change of biological toxicity
of the soil. Hydrocarbons inhibited microbial biomass, and
the greatest negative eect was observed in the gasolinepolluted sandy soil (Labud et al., 2007). Some of metabolic
compounds of TPH such as aromatic ketones, aldehydes,
carboxylic acids, fatty acids, esters, also contribute to
the biota toxicities of petroleum pollutants (Lee, 2003).
Although toxicities of crude oil in water and on aquatic
organisms have been studied (Couillard et al., 2005; Perkin
et al., 2005; Martinez-Jeronimo et al., 2005), the toxicity of
crude oil in soil has seldom been evaluated. In addition,
dierent test methods on toxicity reect toxicity from
only one aspect that may be not enough. Holistic toxicity
experiments were carried out in this study to test the TPH
toxicity in soil for a better evaluation of bioremediation
process and eco-safety of TPH.
Value
pH
TPH (%)
TN (g/kg)
TP (g/kg)
Cd (mg/kg)
Ni (mg/kg)
Cr (mg/kg)
Cu (mg/kg)
Pb (mg/kg)
7.9
10.57
2.75
0.11
ND
6.5
12
16.5
ND
/
/
/
/
1
200
300
400
500
ND: not detected under detection level of 1 mg/kg; /: items not regulated.
Vol. 23
No. 5
847
2 Results
50
40
30
20
10
0
0.5
1
1.5
TPH content (%)
Fig. 2 Inhibition rate of earthworm body weight for 7 days in dierent
TPH contaminated soils.
4
3
2
-0.2
Fig. 1
4
3
y = 5.1456x + 4.3009
0.2
y = 5.1931x + 4.1636
R2 = 0.8449
-0.4
Probability unit
Probability unit
R2 = 0.8708
1
0.4
-0.4
-0.2
0.2
0.4
logD
logD
Regression analysis of LD50 for 4 (a) and 7 days (b) in dierent TPH contaminated soils. D: soil TPH content.
0.6
848
4 days
germination
4 days
control
10 days
germination
10 days
control
Wheat
Maize
Cotton
Mexican corn grass
Tall fescue
20
10
0
0
0
95
90
0
30
95
60
45
0
0
10
100
100
10
75
100
100
100
a
b
80
Root length inhibition (%)
80
Germination inhibition (%)
Vol. 23
60
40
20
60
40
20
0
0.1
0
0.1
1
2
3
TPH content (%)
TPH content (%)
Fig. 3 Average inhibition rate of maize germination (a) and root elongation (b) in 72 hr by dierent TPH contaminated soils.
100
80
100
0.5
0.5
-20
60
40
20
0
80
60
40
20
0
0.1
0.5
1
2
3
1
2
3
TPH content (%)
TPH content (%)
Fig. 4 Average inhibition rate of wheat germination (a) and root elongation (b) in 72 hr by dierent TPH contaminated soils.
0.1
0.5
No. 5
120
100
80
60
40
y = 105.86x - 0.03
R2 = 0.91
20
0
0
Fig. 5
method.
0.4
0.8
1.2
3 Discussion
The mortality of E. fetida increased in soils contaminated with 2% petroleum (Geissen et al., 2008). The
earthworm toxicity test also demonstrated that diesel concentrations in soil exceeding 1% caused a dose-dependent
weight loss in earthworms and increased mortality and oil
concentration of 1.5% has reduced survival to less than
40% (Hanna and Weaver, 2002; Shin et al., 2005). While
another report showed that 0.1% TPH content did not
show any lethal eects (Schaefer, 2003). TPH may cause
death, swelling, body lesions, stiening, coiling and low
reproduction of earthworm (Oboh et al., 2007). The above
result is in accordance with our result with LD50 of 1.45%
for 4 days experiment in earthworm. The TPH analysis by
GC method suggests that C6C25 hydrocarbons are the
more volatile, soluble, and biodegradable constituents in
crude oil and may provide an indicator of acute toxicity
to worms. Bioavailability and sequestration of TPH as
aected by soil properties were considered to be related
to the toxicity of petroleum hydrocarbons in the soil
(Dorn and Salanitro, 2000). In addition, earthworm is a
good option for bioremediation of TPH contaminated soil
849
4 Conclusions
This study demonstrated the ecotoxicity of petroleum
pollutants in soil by using earthworm, plant and luminescent bacteria toxicity experiments. The LD50 of petroleum
contaminated soil for earthworm is 1.45% in 4 days, while
850
EC50 for root elongation was 1.64% and 1.11% for wheat
and maize, respectively, and EC50 in luminescent bacteria
test is about 0.5%. The result suggests much higher toxicity
of petroleum on bacteria activity, while some plant species
such as wheat is tolerant to petroleum contamination than
earthworm and bacteria.
Acknowledgments
This work was supported by the National High Technology Research and Development Program (863) of China
(No. 2007AA061201), the Special Fund Project for Technology Innovation of Tianjin City (No. 08FDZDSF03402),
and the Tianjin Natural Science Foundation (No. 09JCYBJC08800).
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