Bioremediation Potentials of Hydrocarbonoclastic Bacteria Indigenous in the Oil Impacted Sites of Ogoniland, Rivers State, Nigeria

International Journal of Research and Innovation in Applied Science (IJRIAS) | Volume IV, Issue VIII, August 2019|ISSN 2454-6194
www.rsisinternational.org Page 107
Bioremediation Potentials of Hydrocarbonoclastic
Bacteria Indigenous in the Oil Impacted Sites of
Ogoniland, Rivers State, Nigeria
Wiri, .T.B, Desmond,.O., Mbato,.CI., Nabiradee, P.N.
Dept of Environmental Health, Rivers State College of Health Science and Management Technology Port Harcourt, Rivers State,
P.M.B.5039 Nigeria
Abstract:-Hydrocarbon pollution Remediation by Enhanced
Natural Attenuation method was adopted to remediate the
hydrocarbon impacted site in Ogoniland Rivers State, Nigeria .
The research lasted for 6 months. Samples were collected at
monthly intervals . samples were collected intermittently
between Feb 2019 to July 2019 . Mineral salt medium containing
crude oil was used as a sole source of carbon and energy for the
isolation of hydrocarbonoclastic bacteria. Samples were
collected from the four (4) local government that made up
Ogoniland and they includes Khana(k), Gokana (G),Tai (T),
Eleme (E) and transported immediately to the laboratory for
analysis. The microbial and physicochemical properties of the
soil samples varied with the different local government areas.
Seven bacteria genera were isolated from the samples from the
four locations, viz, Pseudomonas, Lactobacter, Micrococcus,
Arthrobacter, Bacillus, Brevibacterium and Mycobacterium
were isolated and identified. the seven isolate were indigenous in
the study area. Nutrient were added to identified plots of
hydrocarbon pollution polluted site within the four local
government and they were able degrade hydrocarbon within a
short of period of time. Reassessment of physicochemical
parameter impacted site was used to judge the bioremediation
potentials of microorganism.
Key words:- Bioremediation, Hydrocarbonoclastic, Bacteria,
potentials, Ogoniland, Rivers state
I. INTRODUCTION
ydrocarbon pollution in ogoniland is a global issue,the
land has been polluted with hydrocarbon which has
greatly affected health and agricultural activities of the people,
The extent of the pollution warranted the united nation
environmental programme (UNEP) recommended the clean
up of ogoniland.
Petroleum is at present Nigeria’s and indeed, the world’s most
important derived energy source ((Moffat and Linden, 2005;
Nigerian Environmental Study Action Team, 2006). However,
the growth and activities of petroleum and petroleum
associated industries in Nigeria and in other parts of the world
has led to increased oil pollution in our environment.
Petroleum in its natural state is called crude oil (Ukoli, 2003).
The greatest single environmental problem connected with
crude oil exploration in Nigeria is oil spillage both on-shore
and off-shore. The rate of oil spillage reported in the country
has been rising with a corresponding increase in petroleum
production.
The various genera that have been reported to contain
hydrocarbon-degrading species include Pseudomonas, Vibrio,
Corynebacterium, Arthrobacter, Brevibacterium,
Flavobacterium, Sporobolomyces, Achromobacte, Bacillus,
Aeromonas, Thiobacillus, Lactobacter, Staphylococcus,
Penicillium and Articulosporium. These organisms have been
isolated in large numbers from many oil polluted waters and
soils, but are found in less numbers in uncontaminated
environments.
Many studies have been conducted to isolate and characterize
hydrocarbon degraders from oil spill sites but little have been
done to determine the changes in soil mineral nutrients and
total petroleum hydrocarbon as bioremediation of the spill site
progresses. This study will provide information on the
effectiveness of microorganisms in eliminating oil pollutants
from the oil polluted areas of the Niger delta region in
Nigeria. Specifically, the experiment was designed to; isolate
and characterize hydrocarbon utilizing bacteria and fungi from
crude oil impacted soil samples and also determine the extent
to which the spilled crude oil has been degraded.
II. MATERIALS AND METHODS
Collection of Soil Samples
Soil contaminated samples used for the study were obtained
from oil impacted sites in Khana, Gokana, Tai and Eleme
Local Government Area of Rivers State, Nigeria. The sample
were obtained between Feb 2019 to July 2019. The samples
were collected aseptically using soil sampler to a depth of 20
cm, stored in sterile aluminum foils and transported to the
laboratory immediately for analysis. The sample were coded
using the first alphabet of the local government : K,T,G and E
respectively.
Enumeration of total heterotrophic bacteria and in the soil
samples 1 g of each sample was serially diluted (10-1
to 10-5
).
And were plated in duplicate on sterile nutrient agar plates,
using the pour plate method at 37°C for 24 h for the nutrient
agar plate, however colonies who shows slow growth rate
H

were further allow for 48h at 37° and were afterwards
enumerated
Enumeration of Hydrocarbonoclastic Bacteria
1 ml from dilutions of 10-5
were plated in duplicate on pre
dried mineral salt agar using the spread plate technique, the
10-3 and 10-4 dilutions were plated . A filter paper saturated
with sterile crude oil was aseptically placed on the inverted
Petri dishes and the culture plates were incubated for 72h at
37°C . Plates yielding significant colonies were enumerated .
Isolation and Characterization of Hydrocarbon-Utilizing
Bacteria
Colonies of different hydrocarbon eating bacteria were
selected at random using a sterile wire loop and subcultured,
by streaking on nutrient agar plates to obtain pure colonies.
All the isolates were characterized using the techniques of
Chesborogh (2000)
Determination of Physicochemical Properties of
Contaminated Soil
Analyses of physicochemical properties of the soil samples
were performed according to the American Society ASTM
(1998).
Determination of Total Petroleum Hydrocarbon
FTI Spectrophotometer was used for the test. Prior to analysis
the soil samples were extracted with carbon tetrachloride and
treated with 2% deactivated silica gel. The equipment was
calibrated with isooctane/octane in carbon tetrachloride. TPH
concentrations in the samples were determined by using the
stored calibration graphmoisture Content determination
A weighed amount of the soil sample was placed in a weighed
crucible and dried at 105°C in the oven until a constant weight
was reached. From the difference in weight, the percentage
moisture content was calculated.
Phosphate Content Determination
Samples were extracted with 25% acetic acid and the extract
run on the Unicam UV/visible spectrophotometer at a
wavelength of 700 nm. A spike sample was analyzed in every
batch of analysis. A standard was analyzed after every batch
of samples and the first value of the standard was used to plot
the means control chart.
Nitrate and pH Determination
Soil samples were extracted with sodium acetate in the
presence of sulphuric acid and measured at a wavelength of
470 nm using Unicam UV/visible spectrophotometer. To
assess the pH, electrodes of a multimeter were dipped into a
mixture of soil sample and deionized water. The pH values of
the samples were subsequently read on the multimeter.
Statistical Analysis of Data
All the data obtained were subjected to statistical analysis of
variance (ANOVA) using SPSS program.
III. RESULTS
Microbial Counts and Identification
Hydrocarbonoclastic bacteria isolated were, Brevibacterium
Lactobacter, Arthrobacter, Bacillus, Pseudomonas,
Mycobacterium and Micrococcus were isolated and identified.
TABLE 1: TOTAL HETEROTROPHIC BACTEIA THB
SAMPLE DILLUTIONS 10-3
10-4
KHANA (K) 3.0X105
2.7X105
GOKANA (G) 2.41X105 1.7X105
TAI (T) 1.90X105 1.5X105
ELEME (E) 2.30X1O5 2.50X105
TABLE 2 HYDROCARBONOCLASTIC BACTERIA ISOLATED FROM
THE FOUR LOCATIONS
SAMPLE
ISOLATES K G T E
Mycobacterium + + + +
Pseudomonas + + + +
Lactobacter, + + + +
Micrococcus + + + +
Arthrobacter + + + +
Bacillus + + + +
Brevibacterium + + + +
TABLE 3. PHYSICOCHEMICAL PARAMETER OF OIL POLLUTED
SOILBEFORE BIOREMEDIATION
Parameter Sample Location
PH
K G T E
7.00 7.01 6.50 6.90
Nitrogen 0.28mg/kg
0.26mg/k
g
0.31mg/k
g
0.33mg/
kg
Phosphorus 0.33mg/kg
0.35mg/k
g
0.30mg/k
g
0.34mg/
kg
Total Organic
Carbon (TOC)
0.8% 0.7% 0.8% 0.7%
Total hydrocarbon
content (THC)
2,845mg/g
2,885mg/
g
2,950mg/
g
2,740mg
/g
Total Petroleum
Hydrocarbon
(TPH)
3,475mg/g
3,375mg/
g
3,500mg/
g
3,380mg
/g
Polycyclic
Aromatic
Hydrocarbon(PAT
H)
15.65mg/g
15.40mg/
g
15.35mg/
g
15.29mg
/g
TABLE 3. PHYSICOCHEMICAL PARAMETEROF OIL POLLUTED
SOIL AFTER BIOREMEDIATION
PARAMETER SAMPLE LOCATION
PH
K G T E
4.00 4.01 4.50 3.90
Nitrogen
5.01g/k
g
4.45mg/kg 3.22mg/kg
2.9.5mg/k
g
Phosphorus
0.55mg/
kg
0.50mg/kg 0.70mg/kg 0.61mg/kg

Total Organic
Carbon (TOC)
115.5% 100.5% 200.1% 185.7%
Total
hydrocarbon
content (THC)
2,845m
g/g
2,885mg/g 2,950mg/g 2,740mg/g
Total
Petroleum
Hydrocarbon
(TPH)
70.5mg/
g
75.5mg/g 80.7mg/g 68.0mg/g
Polycyclic
Aromatic
Hydrocarbon
(PATH)
2.65mg/
g
3.40mg/g 3.35mg/g 4.29mg/g
IV. DISCUSSION
The results of the study indicated that the total heterotrophic
bacteria (THB) counts varied over a period of time. Result
shows significant differences in physicochemical parameter
before and after bioremediation. The differences in the
physicochemical parameter was use to ascertain the degree or
potentials of hydrocarbon utilizing effects of the isolated
microorganism. The counts of hydrocarbonoclastic bacteria
were higher in hydrocarbon polluted soil than non impacted
soil, this may be as a result of the fact the hydrocarbon
provides a source of carbon and nitrogen needed for the
proliferation of the hydrocarbonoclast. The pH values of
crude oil polluted soil were lower as compared to those of
crude oil free soil. The decrease in pH value may be due to
increased degradation of crude oil by microorganisms in the
soil, res. Both nitrogen and phosphorus levels were high in
hydrocarbon impacted soil than oil free soil. This agrees with
the finding of (A.K. Onifade and F.A. Abubakar (2007)), who
reported increase in nitrogen and phosphorus contents of a
crude oil polluted soil.
Conclusively, the study shows that there was residual crude
oil in the soil after 6 months of research. The study also shows
that the physiochemical properties of the soil were
significantly affected. Finally, the study has proven that
Ogoniland has indeginous microbial population that has the
potentials to remediate hydrocarbon pollution.
REFERENCES
[1]. ASTM, 1998.Standard Guide for Remediation of Soil by Natural
Attenuation at Petroleum Release Sites E-1943-98.ASTM, West
Conshohocken, Pennsylvania.
[2]. Atlas, R.M., 1995. Bioremediation of petroleum pollutants. Int.
Biodeteriorat. Biodegradat., 35: 317-327.
[3]. Ijah, J.J. and O.P. Abioye, 2003.Assessment of physicochemical
and microbiological properties of soil 30 month after kerosene
spill. J. Res. Sci. Manage., 1: 24-30.
[4]. A.K. Oniade and F.A.Abubakar (2007) characterization of
hydrocarbon degrading microorganism isolated from crude oil
contaminated soil and remediation by natural attenuation .journal
of microbiology, 2:149-155

Bioremediation Potentials of Hydrocarbonoclastic Bacteria Indigenous in the Oil Impacted Sites of Ogoniland, Rivers State, Nigeria

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Bioremediation Potentials of Hydrocarbonoclastic Bacteria Indigenous in the Oil Impacted Sites of Ogoniland, Rivers State, Nigeria