2011 Sriwijaya University FMIPA 14113-57

Journal of Science Research Volume 14, Number 1 (D)

1411



The utilization of grass Fimbrisylis sp. In the Soil Bioremediation Process at
Different Concentrations of Petroleum Waste

Sri Pertiwi E, Hary Widjajanti, Bambang Yudono, dan Hary Wahyudi
faculty of Biological Science, University of Sriwijaya, South Sumatra, Indonesia

Abstract: The research aims to investigate the effect of Fimbristylis sp. to the population of bacteria and
the decreasing of TPH concentration in bioremediation process. The experimental design is factorial randomized
complete design consists of 2 factors. Factor 1 is petroleum sludge concentration, it consists of 5 levels; C0:0%,
C1:5%, C2:10%, C3:15% dan C4: 20%. Factor 2 is Fimbrisylis sp plant, it consists of 2 levels; R0 without
Fimbrisylis sp plant and R1 using Fimbrisylis sp plant. There are 10 combinations; each treatment is repeated 3
times. The results showed that Fimbrisylis sp plant affected the bacteria population and the decreasing of TPH
concentration; however the increasing of TPH concentration effected Fimbrisylis sp plant growth.

KEYWORDS: bioremediation, Fimbristylis sp, petroleum sludge
E-mail: yudonob@hotmail.com
January 2011


1 INTRODUCTION

he petroleum industry is an industry that is likely to
cause negative effects to the environment around.
Petroleum hydrocarbons (petroleum) is one of the
pollutants with a very wide spread and can contaminate
the soil, underground water, beaches, sediment, and water
surface
[1]
. The content of the total petroleum
hydrocarbon (TPH)15% in petroleum waste shall be
prepared. It is based on the decision by the Ministry of
Environmen number 128 of 2003.
Most of the technology or conventional recovery
methods (both chemical and physical) needs a lot of cost
and decreased soil fertility and a negative impact on the
ecosystem. Bioremediation is a technology that is effective
and friendly to the environment.
At the end of the bioremediation process,
activity and number of hydrocarbon degrading
bacteria population has decreased so it takes
additional energy. This is supported by research
reports Estuningsih et al [3] which states that the
TPH conditions towards the end of the
bioremediation process decreased only 25%.
Therefore, it needs an additional energy source for
hydrocarbon degrading bacteria.
According to frick et al (4) states that in the process
bioremediasi several herbs associated with microbes to
lower or cleanse contaminant. Plant of root exudate
provides such as nutrients, enzymes, and oxygen for
microbes in rhizosfer. This means that a plant induces an
increase in the population of microbes and the
degradation of organic contaminants may increase in
rhizosfe.
Fimbristylis sp. utilization in the process of
bioremediation can be considered this plant allegedly
capable of being fitoremediasi or agent able to reduce
hydrocarbon pollution. It is based on the research of
Estuningsih et al
[3]
Who claimed that at the end of the
process of bioremediasi seen the presence of cooperation
with phyto alg and higher plants it is characterized by
the proliferation of algae and phyto Fimbristylis sp.
Substances from the roots of the plants represent one of
the main energy source for microbes and its effects on soil
dirt mikroflora. Microbes in rhizosfer stimulated by
increased concentration of chemicals that act as a source
of energy, rhizosfer is a phenomenon confirmed by the
quantitative increase in microbial populations
[5].
Based on research Isnaini [6] which states the ratio
of sludge and top soil with a ratio (1:1) or use 50% of
the total composition sludge decreased slow TPH is
32.6%, while the ratio of (2:1) reduction in TPH only
27.45%. Munawar [7] states that wastes too much on
the ground or terrestrial environment can inhibit the
absorption of nutrients by plant roots.
This study aims to examine the use of grass
Fimbristylis sp in bioremediation technology
using indigen bacteria, at various concentrations
of petroleum waste.
This study is expected to be used to enhance
the bioremediation technology with utilizing
grass Fimbristylis sp.






T
14113-2

2. Research methods
1.1 Time and place
The research was carried out for 4 months which is
housed in a glass house and the Laboratory of
Microbiology and Genetics and Biotechnology
Laboratory, Department of Biology, Faculty of
Mathematics and Natural Sciences, University of
Sriwijaya, Inderalaya.

1.2 materials and Tools
The tools used in this study is aluminum foil,
autoclave, stir bar, Bunsen, Petri dishes, colony
counters, large bucket 30 pieces (diameter 30 cm),
erlenmeyer, glass beaker, measuring cup, hot
plate, incubators, needle ose, camera digital,
cotton, rubber, paper label, paper filter, magnetic
stirrer, oven, water bath, pH meter, serological
pipette, Pasteur pipette, centrifuges, shakers, 20 D
spectrophotometer, analytical balance, and test
tubes.
The material used is 70% alcohol, distilled water,
which consists of starter bacteria: Pseudomonas
pseudoalcaligenes, Bacillus sphaericus, Bacillus
sphaericus var. Rotans, Bacillus megaterium,
autotraphicus Xanthobacter, Bacillus mycoides,
Bacillus cereus var. albolactis were isolated from
petroleum waste PT Pertamina EP Field Jambi Sungai
lilin, n-hexane, sawdust, grass Fimbristylis sp, soil, and
fertilizer NPK, KCL, and TSP.

2.3 Research Design
The experimental design used in this study was a
completely randomized design (CRD) with factorial
pattern consists of 2 factors: Factor I. The
concentration of waste oil (C), which consists of 5
levels ie: C0: no waste oil (control), C1: C2 5%: 10%
C3: C4 15%: 20%. Moderate use of Factor II
Fimbristylis grass sp (R) which consists of two levels
ie: R0: without using grass Fimbristylis sp. R1: using
grass Fimbristylis sp weighing 100 g. Total there are
10 treatment combinations and each repeated 3 times
so that it will produce 30 units of the experiment.
2.4 Works
Sampling
Sampling was conducted in petroleum waste PT
Pertamina EP Jambi field Sungai Lilin South Sumatra.
Taken as a "purposive random sampling" of an oil
production waste containment.

Bioreactor Preparation
Each bioreactor incorporated materials that have been
prepared with the following composition: soil, fertilizer
KCL, TSP, and NPK as well as a bulking agent such as
sawdust, add the oil waste in accordance with the
treatment, and then inserted into the bioreactor. Each
bioreactor was inoculated 5% starter containing the
bacteria were cultured indigen mix (minimum bacterial
population 106cfu/ml culture), stirring until all ingredients
rata.Bioreaktor randomly placed in the greenhouse.
Moisture and pH of the media in the bioreactor set every
day.

Planting plants

Do Bioremediation using bacteria indigen for 4 weeks.
Furthermore, the bioreactor Fimbristylis sp planted grass in
the vegetative phase. Grass comes from the location of waste
treatment at PT Pertamina EP Jambi field Sungai Lilin
South Sumatra, so that the process of adaptation of grass
could run well. overall bioremediation process was carried
out for 8 weeks.

Measurement of Total Petroleum Hydrocarbon
(TPH)
Observed conditions initial TPH and TPH end of the study.
Measurement of Total Petroleum Hydrocarbons using
spectrophotometric method.

2.5 Variable of Observations

All variables were measured at the end of the study
observation (8 weeks), the observation variables include: (1)
The number of bacterial populations; (2) The percentage
degradation of Total Petroleum Hydrocarbon (TPH); (3) the
growth of grass Fimbristylis sp.

Calculation of the total population of bacteria

At the end of the study calculated through the number of
bacteria colonies using colony counter. Of each treatment
was calculated in units of the number of bacterial cfu / gram,
the calculation of the number of bacterial cells to use cup
count method (pour plate method).

The calculation of impairment TPH

The concentration of hydrocarbons beginning and end of
treatment was calculated by spectrophotometric method.
Further reduction in TPH value is defined as the TPH value
was eliminated / removed after a given treatment condition.
TPH degradation results calculated with the following
formula:

The decrease in the value of TPH = TPHstart - TPHend


Fimbristylis sp Growth


Sp Fimbristylis growth observed by measuring the wet
weight of the plant, on the morning before the plants wilt
(loss of water). Measurements were taken at the end of the
study [8].

2.6 Analysis of Data

Data obtained from the observation that the amount of
degradation of the bacterial population and the percentage of
variance analysis is not performed while the grass growth
analysis of variance, if there is a significant area of Duncan's
14113-3

multiple test at level of 0.05.


3. RESULTS AND DISCUSSION

Total population of bacteria (cfu / g)
Interaction between the concentration of waste oil and the
use of real grass does not affect the amount of bacteria
population. The number of bacterial populations on the
interaction of these two factors showed no difference in
treatment. This is presumably because the grass is not an
exudate main energy source for the bacteria. Bacteria use
carbon from waste oil as a primary energy source, exudates
released by the roots of grass are used as additional
nutrients. The extra nutrients will be used if available, but
if not available the growth will continue.
Medium concentration of petroleum waste treatment
significantly affect the amount of Bacterial population.
Further test results DNMRT 5%, the effect of various
concentrations of waste oil in any treatment of bacterial
population numbers presented in Table 1

Table 1: Effect of various concentrations of petroleum
waste to the average number of bacterial populations

Treatment Total bacterial
populations (Concentration
crude oil) (cfu/gram)

C0 (0%) 3,8 10
11a

C1 (5%) 6,0 10
11b

C2 (10%) 5,6 10
11b

C3 (15%) 4,6 10
11a

C4 (20%) 4,4 10
11a

Based on Table 1 it can be seen that the number of
bacterial Populations between treatments C0 (0%) or a
control Showed different results are not real with C3
treatment (15%) and C4 (20%) and C1 Significantly
different from treatment (5%) and C2 (10%). Because this
is at no additional controls so that no waste petroleum
petroleum wastes as the main energy source so that the low
number of bacteria. Being the addition of petroleum waste
15% and 20%, waste oil is added to this treatment was
thought to be high enough for the bacteria to be less
optimum growth. Being the addition of waste oil at
Concentrations of 5% and 10%, Able to support bacterial
growth until the end of the observation. The concentration
of waste oil were added still be tolerated and utilized by
microbes.
Utilization of grass significantly affect the amount of
bacteria in each treatment population. Further test results
DNMRT 5% effect on the amount of grass utilization
bacterial population are presented in Table 2.

Of Table 2. Showed a significant difference between the
number of bacterial populations using the treatment and do
not use lawn grass. This is because the grass giving
substances needed by the bacteria, according to the opinion
of Lay (1992) that the part where there is a meeting
between the roots and the soil is called the rhizosphere, the
microbial population
Table 2: Effect of utilization of grass to total bacterial population

The use of grass Factor Total population
bacteria at week 7
(cfu / g)
R0 (Without using grass) 4,4 x 10
11a
R1 (Using grass) 5,4 10
11b

in this section is much higher than other parts. Bacteria
dominate the rhizosphere, growth is supported by nutrients
released root tissues of plants. For examples of amino acids,
vitamins, and other nutrients that the bacteria are able to
grow better and the amount of bacterial populations in the
rhizosphere more of these.

The decrease in the value of total petroleum
hydrocarbons (TPH).

Interaction treatment of various concentrations of petroleum
waste and the use of grasses significantly affect TPH
impairment. This is because even though the plants are
autotrophs and can not directly utilize the carbon from
petroleum waste, but can work with bacteria or microbes in
utilizing the hydrocarbon waste. According to Frick et al [4],
the general degradation of the petroleum hydrocarbons may
be done indirectly by plants. Plants will cooperate with
microbes and depending on the type of pollutants, microbes
and their environment.
Further test results DNMRT 5% interaction effect per-
imposition of various concentrations of petroleum waste and
grass utilization Fimbristilys sp on decreasing the TPH
values are presented in Table 3.
In Table 3 it can be observed also that the higher the
addition of hydrocarbon waste, then the greater the decline
in value of TPH. This is probably due to the higher
petroleum waste, the more available carbon sources.
Allegedly there is cooperation between the living bacteria
are applied on a per grass (rhizosphere) and grass
Fimbristilis sp in utilizing existing carbon in the waste oil.
That waste is degraded by bacteria with some of its enzymes
and will produce CO2 and H20 as well as energy. Instead
plant will utilize CO2, H2O and energy to make the process
of metabolism. Plants in this regard will be issued
Fimbristilys sp root exudates which can be used by bacteria
to metabolize the results will be used to improve activities
including waste degrade petroleum hydrocarbons. This is in
accordance with that raised by Wenzell that the rhizosphere
is an excellent habitat for microbial growth because the roots
of the plants provide a variety of organic materials which
generally stimulate microbial growth. Organic materials that
can be issued by root: root exudates such as sugars, amino
acids, organic acids, fatty acids and sterols, growth factors,
Nuk-leotida, flavonon, enzymes and miscellaneous. The
main enzymes produced by the roots is oksidoreduk-tage,
hydrolase, lyase, and while the transferase enzyme produced
by microbes in the rhizosphere is Selu-lase, dehydrogenase,
urease, phosphatase and sulfatase.

Fimbristylis sp Growth.

From the research that has been carried out and obtained the
following results are presented in Table 4.
Table 4 shows the presence of waste oil can inhibit the
growth of grass Fimbristilys sp, so the more the addition of
14113-4

petroleum Llimbah heavy influence on the plants.
According Fitter and Hay [5] can mempen-fects of
pollutants such plants inhibit the enzyme, inhibiting cell
division and loss of substrate repirasi and O2 deficiency.
Besides pollution causes biochemical changes in the level
of the cell is then followed by physiological changes,
contaminants individually and in combination cut down on
the growth of cambium, roots and part of the re-productive.
Weight gain that can be said Seba-gai indicator of
growth, this research has not been good grass growth. It
can be seen from the weight gain is still low. The existence
of petroleum waste too much will inhibit plant growth
Fimbristylis sp so that growth becomes stunted. Munawar
[7] states that the more levels of petroleum waste contained
in an environment will lead to endangerment of biota in the
environment, such as waste containing aliphatic and
aromatic hydrocarbons having high weight molecular.

4. CONCLUSION
Based on the research that has been carried out, it can be
concluded as follows:

Table 3: The decrease in the average value of the interaction TPH concentrations petroleum waste treatment and utilization
of grass Fimbristylis sp


Using 0% effluent 5% effluent 10% effluent 15% effluent 20% effluent
Grass concentration concentration concertration concentration concertration
(C0) (C1) (C2) (C3) (C4)
Without 0
a
2,5
b
3,6
c
4,3
d
5,1
e

using grass
Using grass 0
a
2,7
b
3,7
c
4,6
de
6,1
f



Table 4: Effect of utilization of grass to total bacterial
population

Treatment of late wet weight (grams)
C0 (0%) 226,3 39,1
C1 (5%) 163,7 7,5
C2 (10%) 151 14
C3 (15%) 134,3 7,6
C4 (20%) 108,3 16,9

1. Utilization Fimbristylis sp. increase the number of the
bacterial population.
2. Utilization of grass Fimbristylis sp increase Total
Petroleum Hydrocarbons impairment (TPH) higher
compared to the treatment that does not use grass
Fimbristylis sp.
3. Growing number of waste oil were added then the
growth Fimbristylis sp. the lower

5. Counsel
1. Further research should be conducted pulled nai
Cyperacae use other types as they may be able to
perform a more optimal bioremediation process.
2. More attention to the time of application and
Fimbristylis sp bacterial associations in a
bioremediation process.

ACKNOWLEDGEMENTS
To PT Pertamina EP Jambi over the cost of this study.

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