Analele Universitii din Craiova, seria Agricultur Montanologie Cadastru Vol.

XXXIX 2009
239
MBUNTTIREA BIOREMEDIERII SOLURILOR POLUATE CU IEI
PRIN UTILIZAREA UNUI PRODUS NATURAL BIODEGRADABIL
ENHANCED BIOREMEDIATION OF SOILS POLLUTED WITH CRUDE
OIL USING A NATURAL BIODEGRADABLE PRODUCT
MARIANA MARINESCU, M. DUMITRU, ANCA LCTUU, GEORGIANA PLOPEANU
Key words: bioremediation, crude oil, polluted soils, natural biodegradable product.
Cuvinte cheie: bioremediere, titei, soluri poluate, produs natural biodegradabil.
REZUMAT
Poluarea accidentala cu titei a devenit in zilele noastre un fenomen obisnuit si poate
cauza catastrofe ecologice si sociale. Bioremedierea solurilor poluate cu titei este limitata
de activitatea microorganismelor degradatoare de hidrocarburi.
Principalul scop al acestui studiu este acela de a creste bioremedierea solurilor
poluate cu titei prin adaugarea unui produs biodegradabil natural si a inoculului bacterian.
Biodegradarea a fost cuatificata prin analize de total hidrocarburi din petrol (THP).
Lucrarea prezinta date obtinute in urma procesului de biodegradare a unui sol
poluat artificial cu 5% si 10% petrol, tratat cu un produs natural biodegradabil si inocul
bacterian. Procesul de biodegradare are loc in timp pentru a reutiliza solul in scopuri
agricole.
ABSTRACT
Crude oil pollution accidents are nowadays become a common phenomenon and
have caused ecological and social catastrophes. Crude oil bioremediation of soils is
limited by the bacteria activity in degrading the spills hydrocarbons.
The aim of this study is to enhance the bioremediation of soils polluted with crude
oil by adding the natural biodegradable product and bacterial inoculum. Biodegradation
was quantified by total petroleum hydrocarbons (TPH) analyses.
The paper presents data obtained in biodegradation process of an artificial polluted
soil with 5% and 10% crude oil, treated with a natural biodegradable product and bacterial
inoculum. Biodegradation process takes time to reuse the soil in agricultural scopes.
INTRODUCTION
The development of petroleum industry, and also extractive and processing one,
including crude oil transport could lead to unpredictable secondary phenomena with
effects more or less harmful for environment and human life. One of this phenomenon is
soil pollution with crude oil, petroleum products with or without salty water, petroleum
residual products, resulted from crud oil exploitation and extraction (Ru, 1986).
In accordance with this phenomenon, starting with 1991, the Institute for Soil
Science, Agrochemistry and Environmental Protection ICPA, Bucharest initiated a study
in all country. This study had like propose to elucidate the pollution nature, its extension
and its impact on soil and microorganisms existing in soil (Toti, 1999).
In the study were identified and isolated the main heterotrophic bacteria and
microfungi biodegraders of crude oil and assigned some conditions to stimulate their
proliferation. Were selected some agro-pedo-improvements practices and biological
management to contribute and action on pollutant disappearance from soil (Voiculescu,
2002).
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All these research lead to the elaboration of a bioremediation method in-situ of soil
polluted with crude oil by using selected microorganisms, with a high efficiency in
biodegradation, low costs, without affecting soil characteristics, fertility (Toti, 2003).
Petroleum fuel and crude oil products represent the one of the most common
environmental contaminant. Common sources of these products are motor fuel station
underground storage tanks, home and commercial heating oil storage tanks, fuel
distribution centers, refineries, crude oil production sites and accidental spills (Bartha,
1986).
Based upon the available information on the chemistry and toxicology of petroleum
hydrocarbons, it is possible that petroleum hydrocarbon fuels and oils are mainly
composed of aliphatic and aromatic hydrocarbon compounds and petroleum aromatic
hydrocarbons generally appear to be more toxic than petroleum aliphatic compounds.
Petroleum hydrocarbon products are mixtures of over 250 hydrocarbon
compounds. The various product mixtures produced by the manufacturers are based upon
physical and performance-based criteria and not specific formulas. As a result, the product
compositions can vary depending upon, in part, the crude oil refined to produce the
product, the type of product, the season of the year, and any performance additives
(Alexander, 1994).
Because high concentrations (>1%) of petroleum hydrocarbons in soil have
demonstrated phytotoxic properties, as well as the uncertainty about the fate and transport
of high concentrations of petroleum in soils and upon human health, the specific fuel
nondefault closure levels have maximum limits applied. It is generally recognized that the
toxicity, (human and environmental), of petroleum products increases as the molecular
weights of the compounds decrease (Pepper, 1996).
MATERIALS AND METHODS
The main objective of the present study was to enhance the bioremediation method
of the soils polluted with crude oil by using the natural biodegradable product ECOSOL. It
has been achived an experiment in Green House. The soil was artificial polluted with 5%
and 10% crude oil, then was applied the treatment consisting in ECOSOL and bacterial
inoculum.
The experiment has 11 experimental variants with soil polluted 5% and 10% crude
oil, treated with 50g, 100g and 200g ECOSOL/20 kg polluted soil, inoculated and
uninoculated with bacteria selected according to the following experimental scheme:
V
1
, control (unpolluted soil);
V
2
, polluted soil with 5% crude oil;
V
3
, polluted soil with 10% crude oil;
V
4
, polluted soil with 5% crude oil + 50 g ECOSOL;
V
5
, polluted soil with 5% crude oil + 50 g ECOSOL + bacterial inoculum;
V
6
, polluted soil with 5% crude oil + 100 g ECOSOL;
V
7
, polluted soil with 5% crude oil + 100 g ECOSOL + bacterial inoculum;
V
8
, polluted soil with 10% crude oil + 100 g ECOSOL;
V
9
, polluted soil with 10% crude oil + 100 g ECOSOL + bacterial inoculum;
V
10
, polluted soil with 10% crude oil + 200 g ECOSOL;
V
11
, polluted soil with 10% crude oil + 200 g ECOSOL + bacterial inoculum.
The soil used in experiment is a cambic chernozem sampled from Teleorman area.
Chemical characteristics of the soil used in the experiment are presented in table 1.
Analele Universitii din Craiova, seria Agricultur Montanologie Cadastru Vol. XXXIX 2009
241
Table 1
Chemical characteristics of the soil
Soil type pH Organic Carbon (%) Total Nitrogen (%) C/N ratio
Calcic
chernozem
8,09 2,99 0,279 12,5
The chemical characteristics of the natural biodegradable product ECOSOL used in
the experiment are presented in table 2.
Table 2
Chemical characteristics of the natural biodegradable product ECOSOL
Natural
biodegradable
product
Total
Nitrogen
(%)
Organic
Carbon
(%)
Phosphorous
(%)
Potassium
(%)
Sodium
(%)
ECOSOL 0,935 23,72 0,39 3,32 4,97
The inoculum was developed from microorganisms that occur naturally in the soil
with the addition of crude oil. The microorganisms use the petroleum hydrocarbons as a
source of carbon. It was stimulated the growth of the microbial population by adding the
fibers provided from celluloid wastes.
The density of crude oil used in artificial pollution of soil is been calculated with the
following formula:
The value obtained for the crude oil used in the experiment is:
= 836,9 kg/m
3
This value shows that is a light crude oil.
In the first experimental year was observed an increase of biodegradability rate in
accordance with the applied treatment, so the experiment continued. In the second
experimental year was used a plant to observe the influence of crude oil on plant growth
and also to enhance the bioremediation by phytoremediation. The plant used was maize.
RESULTS AND DISCUSSIONS
Petroleum fuel or crude oil contaminated sites have been characterized by the total
of all the petroleum hydrocarbons, called total petroleum hydrocarbons (TPH). In this
paper are presented the influence of crude oil on plant growth.
In figure 1 are presented the maize plants (was choosed one repetition) in each
experimental variant after one month from emergence. The figure shows that in all the
plots were the soil was polluted with 10% crude oil, the plants havent rise up even the
treatment was applied.
In the plots were the soil was polluted with 5% the plants rise up with delay, the
plant are at least two times lower than control, and differences between treatments can be
observed after one month.
Analele Universitii din Craiova, seria Agricultur Montanologie Cadastru Vol. XXXIX 2009
242
Figure 1 Maize plant in each experimental variant after one month from appearence
In table 3 are presented the characteristcs of plants after 5 months from seeding, in
the second experimental year of bioremediation and the first one when a plant was used in
experiment.
The number of plant/plot, the height of plants, the number of leaves and the
biomass are proportional with the treatment. As it can be observed the control presents
values very high for all the parameters analysed, the plots with soil polluted with 5% crude
oil have values lower then control and the plots with soil polluted with 10% crude oil have
no value because the plants did not rise up.
Analele Universitii din Craiova, seria Agricultur Montanologie Cadastru Vol. XXXIX 2009
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Table 3
Evolution of plant biomass in all experimental variants and repetitions according with the applied treatment
No. Experimental variant Repetition
Plants/plot
(number)
Height
(cm)
Leaves
(number)
Biomass
(g)
1 R1 4 138 12 99
2 R2 5 140 13 113
3
V
1
, control (unpolluted soil)
R3 5 139 12 107
4 R1 4 46 7 6
5 R2 4 53 9 4
6
V
2
, polluted soil with 5% crude oil
R3 4 57 8 5
7 R1 - - - -
8 R2 - - - -
9
V
3
, polluted soil with 10% crude oil
R3 - - - -
10 R1 5 57 7 7
11 R2 5 50 7 5
12
V
4
, polluted soil with 5% crude oil + 50 g
ECOSOL
R3 5 51 8 3,5
13 R1 3 57 8 5
14 R2 4 56 9 5
15
V
5
, polluted soil with 5% crude oil + 50 g
ECOSOL + bacterial inoculum
R3 5 52 7 6,5
16 R1 5 68 9 7
17 R2 4 47 8 6,5
18
V
6
, polluted soil with 5% crude oil + 100
g ECOSOL
R3 5 56 8 5
19 R1 5 83 8 8
20 R2 5 48 9 5
21
V
7
, polluted soil with 5% crude oil + 100
g ECOSOL + bacterial inoculum
R3 5 56 8 6,5
22 R1 - - - -
23 R2 - - - -
24
V
8
, polluted soil with 10% crude oil +
100 g ECOSOL
R3 - - - -
25 R1 - - - -
26 R2 - - - -
27
V
9
, polluted soil with 10% crude oil +
100 g ECOSOL + bacterial inoculum
R3 - - - -
28 R1 - - - -
29 R2 - - - -
30
V
10
, polluted soil with 10% crude oil +
200 g ECOSOL
R3 - - - -
31 R1 - - - -
32 R2 - - - -
33
V
11
, polluted soil with 10% crude oil +
200 g ECOSOL + bacterial inoculum
R3 - - - -
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From treatment point of view, can be analysed all the parameters for the soil
polluted with 5% crude oil. The number of plant/plot, the height of plants, the number of
leaves and the biomass are increasing with the treatments applied.
CONCLUSIONS
The experimental study leads to the following preliminary conclusions:
In the plots were the soil was polluted with 5% the plants rise up with delay, the
plant are at least two times lower than control, and differences between
treatments can be observed after one month.
In all the plots were the soil was polluted with 10% crude oil, the plants havent
rise up even the treatment was applied.
The number of plant/plot, the height of plants, the number of leaves and the
biomas are proportional with the treatment. For all the parameters analysed the
control presents values very high, the plots with soil polluted with 5% crude oil
have values lower then control, proportionaly with the treatment apllied and the
plots with soil polluted with 10% crude oil have no value because the plants did
not rise up.
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