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Environment International 26 (2001) 409 ± 411

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Bioremediation of oil sludge-contaminated soil


N. Vasudevan*, P. Rajaram
Centre for Environmental Studies, Anna University, Chennai 600 025, India

Abstract

Bioremediation has become an important method for the restoration of oil-polluted environments by the use of indigenous or selected
microbial flora. Several factors such as aeration, use of inorganic nutrients or fertilizers and the type of microbial species play a major role in
the remediation of oil-contaminated sites. Experiments were undertaken for bioremediation of oil sludge-contaminated soil in the presence of
a bacterial consortium, inorganic nutrients, compost and a bulking agent (wheat bran). Experiments were conducted in glass troughs for the
90-day period. Bulked soil showed more rapid degradation of oil compared to all other amendments. During the experimental period, wheat
bran-amended soil showed 76% hydrocarbon removal compared to 66% in the case of inorganic nutrients-amended soil. A corresponding
increase in the number of bacterial populations was also noticed. Addition of the bacterial consortium in different amendments significantly
enhanced the removal of oil from the petroleum sludge from different treatment units. D 2001 Elsevier Science Ltd. All rights reserved.

Keywords: Bioremediation; Contaminated soil; Oil sludge; Degradation

1. Introduction was below 10% (Freijer, 1986). Tillage is a mechanical


manipulation of soil to improve soil conditions (Hillel,
Oil production and shipping operations result in acciden- 1980). It alters physical and chemical properties of soil in
tal contamination of soil with petroleum hydrocarbons. such a way that it stimulates microbial activity (Melope et
Petroleum refining also results in the generation of large al., 1987). Tillage redistributes carbon, nitrogen and water
quantities of oil sludge consisting of hydrophobic substan- and reduces spatial distribution within the soil (Rhykerd et
ces and substances resistant to biodegradation. Clean-up al., 1999).
technologies such as incineration and burial of sludge in Bulking agents are materials of low density that lower
secure landfills are expensive. Land treatment disposal of oil soil bulk density, increase porosity and oxygen diffusion,
refinery sludge generally gives good results (Bartha, 1986). and can help to form water-stable aggregates. These
Controlled land treatment, i.e., land farming, is cheaper and activities increase aeration and microbial activity (Hillel,
also environmentally safe (Bonnier et al., 1980; El-Nawawy 1980). The aim of this study was to enhance the remedia-
et al., 1987). tion of soil contaminated with oil sludge by the use of a
Aerobic conditions and appropriate microorganisms are bacterial consortium, inorganic supplements, bulking
necessary for an optimal rate of bioremediation of soils agents and compost.
contaminated with petroleum hydrocarbons. In soils, the
oxygen content depends on microbial activity, soil texture,
water content and depth. A low oxygen content in soils has 2. Materials and methods
been shown to limit bioremediation of soils contaminated
with petroleum hydrocarbons (von Wedel et al., 1988) and 2.1. Soil preparation
in a laboratory experiment, mineralization of hydrocarbons
from soil was severely limited when the oxygen content The soil used in the study was collected from open fields
near the petroleum refinery site in Chennai city. Surface
litter was removed and soil was collected to a depth of 25
* Corresponding author. Tel.: +91-44-2354-296; fax: +91-44-2354-
cm and sieved to remove large roots, macrofauna and
717. stones. For physico-chemical properties, the soil was air-
E-mail address: nvasu30@yahoo.com (N. Vasudevan). dried and passed through a sieve (2 mm); soil pH was

0160-4120/01/$ ± see front matter D 2001 Elsevier Science Ltd. All rights reserved.
PII: S 0 1 6 0 - 4 1 2 0 ( 0 1 ) 0 0 0 2 0 - 4
410 N. Vasudevan, P. Rajaram / Environment International 26 (2001) 409±411

measured in water using a 1:5 soil/water ratio. Soil texture, 53% ether-extractable hydrocarbons. The troughs were
pH and organic matter content were determined according to covered with sterile aluminium foil and incubated at
Parmer and Schmidt (1964). 30°C for 90 days. To achieve sufficient aeration, the
contents of the troughs were mixed thoroughly every
2.2. Sludge alternative day. After the start of the experiment and at
intervals of 15 days, ether extractable hydrocarbons and
Petroleum refinery sludge was analyzed gravimetrically bacterial population in soil were determined. The ether
as described by Dibble and Bartha (1994). The following extractable hydrocarbons were determined by extracting
results were obtained after Soxhlet extraction. It con- 25 g of soil using diethyl ether. The bacterial counts in
tained 24% ether-extractable hydrocarbons, 5% water and different treatment units were determined by plating on
71% ash. nutrient agar medium and the colony forming units were
counted after 24 h of incubation at 30°C. All determina-
tions were carried out in duplicate.
2.3. Biodegradation experiments

Experiments were carried out in glass troughs of


23  12.5 cm containing 5 kg soil and 5% w/w oil sludge. 3. Results and discussion
Experiments were conducted with the following treatment
combinations: The soil from site near industrial area had a loamy
texture (34% sand, 40% silt and 26% clay). The soil
Soil + oil sludge (abiotic control) organic matter content was 2.5%. Gas chromatographic
Soil + oil sludge analysis revealed that the oil sludge contained 53% satu-
Soil + oil sludge + compost rated hydrocarbons, 24% aromatic hydrocarbons and 12%
Soil + oil sludge + bacterial consortium asphaltic hydrocarbons.
Soil + oil sludge + inorganic nutrients + bacterial consor-
tium
Soil + oil sludge + wheat bran + bacterial consortium 3.1. Biodegradation of oil sludge

The abiotic control containing soil, oil sludge and 0.3% In order to investigate the optimum conditions for the
w/w AgNO3 was used to monitor the abiotic loss of hydro- biodegradation of oil sludge, soil was supplemented with
carbons. The treatment unit with inorganic nutrients nutrients, and inorganic amendments, bulking agent and a
received 24 g of ammonium nitrate and 4.37 g of dipotas- bacterial consortium as inoculum. Results showed a corre-
sium hydrogen phosphate as an additional supplement. The sponding influence due to the different amendments in the
experimental units containing bacterial consortium received remediation of oil sludge-contaminated soil (Table 1).
approximately 106 colony forming units (cfu)/g of soil as Addition of inorganic nutrients produced little effect on
inoculum. Homogenizations of soil, oily sludge, compost, oil removal compared to the soil amendment without
bacteria and wheat bran in different reactors were carried out inorganic nutrients. The soil microbial population played
in a stainless steel blending machine. The moisture level of a major role in the treatment of hydrocarbons. Addition of
the different soil mixtures was maintained at 20%. organic compost instead of the inorganic nutrients did not
The bacterial population in the unsterilized soil was 103 enhance the removal of petroleum hydrocarbon compared
cfu/g. The oil sludge contained 8% moisture, 26% ash and to the treatment unit containing inorganic nutrients, indi-

Table 1
Relative percent biodegradation of oily sludge and bacterial count in soil
Time after sludge application (days)
15 30 45 60 75 90
Sample no. Treatment (Percent degradation and cell number)a
1 Soil + oil sludge (abiotic control) ± ± ± ± ± ±
2 Soil + oil sludge 1 (6  103) 4 (2  104) 12 (6  104) 18 (2  105) 24 (3  105) 25 (5  105)
3 Soil + oil sludge + compost 2 (8  104) 6 (3  105) 16 (1  106) 3 (3  106) 27 (6  106) 28 (9  106)
4 Soil + oil sludge + activated sludge 2 (3  105) 7 (3  106) 16 (8  106) 24 (1  107) 27 (2  107) 29 (3  107)
5 Soil + oil sludge + bacterial consortium 2 (1  107) 8 (1  108) 19 (2  108) 28 (1  109) 34 (4  109) 40 (2  1010)
6 Soil + oil sludge + inorganic nutrients 4 (6  107) 13 (2  109) 27 (6  1010) 44 (5  1011) 54 (2  1012) 65 (3  1012)
+ bacterial consortium
7 Soil + oil sludge + wheat bran 5 (1  109) 15 (3  1010) 28 (3  1011) 45 (3  1012) 56 (2  1013) 72 (6  1013)
+ bacterial consortium
a
Cell number (cfu/g) in parentheses.
N. Vasudevan, P. Rajaram / Environment International 26 (2001) 409±411 411

cating the lack of suitable hydrocarbon-degrading strains in strates (Rahman et al., 1999). The results of the present
the compost. study indicated that the use of bulking agent played an
In the case of the treatment unit containing inorganic important role in the bioremediation of oil-contaminated
nutrients, nearly 66% oil degradation was recorded. The soil. In general, tillage of soil might enhance the contact
initial bacterial count of 5  106 cfu/g had increased to between oil and bacterial populations thereby enhancing
3  1012 cfu/g in 90 days (Table 1). During the period of 90 the bioremediation process.
days, up to 76% of petroleum hydrocarbons were degraded
in bulked soil compared to other amendments. An increase
of 32% over the control unit without wheat bran addition References
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