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Department of Microbiology, Federal University of Rio Grande do Sul, Sarmento Leite, 500, CEP 90050-170, Porto Alegre, RS, Brazil
Department of Inorganic Chemistry, Federal University of Rio Grande do Sul, Bento Gonalves, 9500, CEP 91500-970, Porto Alegre, RS, Brazil
Department of Soils, Federal University of Rio Grande do Sul, Bento Gonalves, 7712, CEP 91540-000, Porto Alegre, RS, Brazil
a r t i c l e i n f o
a b s t r a c t
Article history:
Received 13 December 2013
Received in revised form
7 June 2014
Accepted 24 August 2014
Available online 3 October 2014
Keywords:
Bioaugmentation
Biostimulation
Bioremediation
Oily sludge
Soil landfarming
1. Introduction
Petroleum processing generates considerable amounts of a
residue named oily sludge. One of the main issues faced by reneries and petrochemical industries is related to the safe disposal
of this residue, since its destination and/or inappropriate treatment
can cause serious impact to the environment and potential risk to
human health (Cameotra and Singh, 2008; Xu and Lu, 2010). Oil
sludge has been classied by the United States Environmental
Protection Agency (US EPA) as a hazardous organic complex (Ubani
et al., 2013). Usually, the oily sludge contains water, sand, oils,
grease, organic compounds, chemical elements, and metals. Among
the organic compounds present, the most common are alkenes,
cyclic alkenes, benzene, toluene, ethyl-benzene, xylenes and polycyclic aromatic hydrocarbons (PAHs) and phenols (Kriipsalu et al.,
2008). Polycyclic aromatic hydrocarbons (PAHs) consist in a
Landfarming
soil
Oily
sludge
Methodology
Humidity
pH
SMP index
Oil and
greases
Organic
carbon
Nitrogen
Total
Phosphorus
Total
Potassium
15%
6.4
6.8
1.75%
ND
ND
ND
48%
Gravimetry/105 C
pH in water 1:1
e
Soxhlet extraction
9.5%
88%
0.14%
0.08%
0.06%
0.01%
0.08%
<0.01%
Kjeldahl
Humid digestion nitric-perchloric/ICP;
OES
Humid digestion nitric-perchloric/ICP;
OES
339
340
341
Fig. 1. Estimate of the number of total heterotrophic microorganisms (log MPN/g of soil) in soils contaminated with (a) 1.5% and (b) 6% and of total hydrocarbon degrading microorganisms (log MPN/g of soil) in soils contaminated with (c) 1.5% and (d) 6% during 90 days in open system.
Fig. 2. Estimate of the number of total heterotrophic microorganisms (log MPN/g of soil) in soils contaminated with (a) 1.5% and (b) 6% and of total hydrocarbon degrading
microorganisms (log MPN/g of soil) in soils contaminated with (c) 1.5% and (d) 6% during 90 days in closed system.
342
pez et al.,
rings present higher resistance to microbial attack (Lo
2008). Marn et al. (2005) showed that the CO2 emissions gradually decrease while the most labile hydrocarbon fractions disappear, remaining only the more recalcitrant fraction.
It was veried that after the thirtieth day the accumulated
CeCO2 production had started to stabilize for all evaluated treatments. This indicates that the mineralization of the oily sludge
occurred markedly in a period of 30 days. Beyond 70 day processing
a reduction in the production of CeCO2 was observed in all tested
treatments, indicating a possible inhibition of the catabolic activity
of the local microorganisms.
Biostimulation and bioaugmentation treatments in contaminated soils at 1.5% showed higher mineralization values than the
natural attenuation treatment at the end of the period. Higher
accumulation productions of CeCO2 after 90 day treatment
occurred in the experiments BA(NP) (3010.1 63.9 mg/kg) and
BS(NP) (2825.9 55.7 mg/kg), which values differ statistically
from the natural attenuation treatment (2014.5 36.7 mg/kg).
The period of high CeCO2 production in experiments with
contaminated soils at 6% oily sludge was higher than that in
contaminated soils at 1.5%, probably due to the higher amount of
hydrocarbons to be metabolized and microbial competence in
metabolizing hydrocarbons.
The natural attenuation, biostimulation and bioaugmentation
treatments with lower amounts of nutrients showed high
Fig. 4. Accumulated production of CeCO2 (mg/kg of soil) during 90 days in closed system in soils contaminated with (a) 1.5% and (b) 6%.
isolated from the landfarming soil and from the studied oily sludge
and selected in liquid medium according to the hydrocarbon
degradation capacity and biosurfactant production. The isolates
showed capacity of growing and mineralization in contaminated
soils at 1.5% and 6%. According to Nasseri et al. (2010) the use of
native microbiota for bioaugmentation is preferred since these
microorganisms have higher adaptation capacity to the pollutant
than exogenous microorganisms and genetically modied microorganisms which can lead to gene transference, which is undesirable (Nasseri et al., 2010). The use of microbial consortiums can be
interesting when an increase of biodegradation rate and extension
of the hydrocarbons that constitute the oily sludge is required, in
view of their complex composition of alkanes, aromatic compounds, resin fractions and asphaltenes. Mixed populations with a
large range of enzymatic capacities and co-metabolism are required
for mineralization of oily sludge as opposed to individual microorganisms, which are able to mineralize only a limited range of
pez et al., 2008).
hydrocarbon substrates (Gallego et al., 2007; Lo
The contamination level and the treatments, as well as the
interaction of these factors, showed inuence on the CeCO2 production rate. In average, higher mineralization rates were obtained
in contaminated soils at 6%. It can be observed that the best condition to obtain the highest oily sludge mineralization rates in
closed microscosms corresponds to bioaugmentation and biostimulation treatments using lower nutrient concentration in
contaminated soils with 6% of oily sludge.
3.3. Total petroleum hydrocarbons degradation
Table 2 shows the biodegradation rates achieved in different
treatments in open and closed microcosms in soils contaminated
with 1.5% and 6% of oily sludge.
Analyzing the systems, it can be observed that higher biodegradation rates were obtained using the closed microcosms
(p < 0.05). The closed microcosms presents the better control of
abiotic losses as well as a better soil humidity control as some
advantages. During the experimental period it was veried that the
open microcosms presented a higher rate of water evaporation,
requiring more frequent periods of humidity correction in order to
Table 2
TPH degradation (%) obtained in different treatments applied to soils contaminated
with 1.5 and 6.0% in open (O) and closed (C) system.
Experiment
System
Contamination
level
Treatment
1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
17
18
19
20
O
O
O
O
O
O
O
O
O
O
C
C
C
C
C
C
C
C
C
C
1.5%
1.5%
1.5%
1.5%
1.5%
6.0%
6.0%
6.0%
6.0%
6.0%
1.5%
1.5%
1.5%
1.5%
1.5%
6.0%
6.0%
6.0%
6.0%
6.0%
NA
BS(NP)
BA(NP)
BS(NP)
BA(NP)
NA
BS(NP)
BA(NP)
BS(NP)
BA(NP)
NA
BS(NP)
BA(NP)
BS(NP)
BA(NP)
NA
BS(NP)
BA(NP)
BS(NP)
BA(NP)
52.7
60.4
57.0
67.2
58.7
70.3
78.7
74.6
76.7
72.8
59.7
74.9
72.4
72.2
74.3
74.9
78.8
79.6
84.1
81.3
0.5a
3.5ab
0.2a
4.2bdi
0.3ab
2.4cd
1.3cjk
1.4 cdk
3.2cjk
0.0cdk
1.5ai
2.5eghcd
3.9 egcd
4.4egcd
0.0egcd
2.3 fgcd
0.4fgc
2.8fgc
0.9 fhj
0.5fgk
*
Equal letters indicate that the experiments do not presented signicant differences (p > 0.05) for the studied response.
343
344
Scholarship. This work was nanced with resources from the Laboratory of Biodeterioration and Biofuels and from the Post Graduation Programs in Agricultural and Environmental Microbiology
and Soil Sciences of the Universidade Federal of Rio Grande do Sul e
Brazil.
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