1 UZOCHUKWU ANGELA N PG/MSc/11/60859 BIOREMEDIATION OF OIL SPILL POLLUTED SOIL USING OYESTA MUSHROOMS (FUNGI) IN ELEME, RIVERS STATE CENTRE FOR ENVIRONMENTAL MANAGEMENT AND CONTROL (CEMAC) Azuka Ijomah Digitally Signed by: Content manager’s Name DN : CN = Webmaster’s name O= University of Nigeria, Nsukka OU = Innovation Centre
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UZOCHUKWU ANGELA N PG/MSc/11/60859
BIOREMEDIATION OF OIL SPILL POLLUTED SOIL USING OYESTA MUSHROOMS (FUNGI) IN ELEME,
RIVERS STATE
CENTRE FOR ENVIRONMENTAL MANAGEMENT AND
CONTROL (CEMAC)
Azuka Ijomah
Digitally Signed by: Content manager’s Name
DN : CN = Webmaster’s name
O= University of Nigeria, Nsukka
OU = Innovation Centre
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UNIVERSITY OF NIGERIA, ENUGU CAMPUS
CENTRE FOR ENVIRONMENTAL MANAGEMENT AND CONTROL (CEMAC)
TOPIC
BIOREMEDIATION OF OIL SPILL POLLUTED SOIL USING OYESTA MUSHROOMS (FUNGI) IN ELEME, RIVERS STATE
A PROJECT
SUBMITTED IN PARTIAL FULFILMENT OF THE REQUIREMENT
FOR THE COURSE: EMC 651 (PROJECT)
BY
UZOCHUKWU ANGELA N PG/MSc/11/60859
SUPERVISOR: DR K.C. OGBOI
MAY,2014
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TABLE OF CONTENTS
Chapter One
1.1 Introduction 1.2 Background of Study 1.3 Statement of Problem 1.4 Aim and Objective 1.5 Research Question 1.6 Research hypothesis 1.7 Scope of Study 1.8 Limitations 1.9 Study Justification/Significance
Chapter Two
2.1 Conceptual Framework
2.2 Bioremediation (everything)
2.3 Definition of Parameters like oyesta mushroom,
biored, oil sp microbes, PH, conductivities,
Nitrogen etc K.PO4 Nitrogen, Particle Size,
Potassium, Total hydrocarbon, particle size
Chapter Three
3.1 Review of Related literature
3.2 Theoretical framework
3.3 Empirical Framework
Chapter Four
Study area
History people and culture
Location
Geographical Area Weather Climate
Ecological Problems Vegetation etc
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ABSTRACT
Bioremediation potential of oyster mushroom on crude oil impacted soil , in Eleme, Port-Hacourt Rivers State was studied following laboratory analysis of the soil sample for nutrient composition (N,K,PO4), total petroleum hydrocarbon (TPH), and soil textural composition, prior to , and after treatment with the oyster mushroom . Reference sample soil was analyzed for the same parameters. Soil samples were collected from the oil impacted soil every 4th day for 28 days of the bioremediation excercise . The crude oil impacted soil recorded a pH of 7.26 , electrical conductivity of 102.58us/cm , Nitrogen content of 0.28%, Phosphate content of 1.08ppm and Potassium content of 4.86ppm. The total petroleum hydrocarbon (TPH) was 284.65mg/kg. Post treatment analysis of the soil sample showed significant improvement in the nutrient compositions (N,0.98%, K,10.99ppm, PO4,3.08ppm) , reduction of the electrical conductivity and TPH(con,30.22, TPH, 20.42mg/kg). There were also improvements in the soil textural compositions. When the post treatment analytical values were compared with those of the reference soil , result showed significant closeness of the two values. The research proved that the oyster mushroom is a good bioremediating agent that should be explored and exploited in the management of oil polluted site.
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CHAPTER ONE
1.0 INTRODUCTION
1.1 BACKGROUND OF THE STUDY
Bioremediation refers to the use of microorganisms to degrade contaminants that pose
environmental and especially human risks. Due to its safety and convenience, it has become an
accepted remedy for cleaning of soil and water pollutants.
Bioremediation processes typically involve many different microbe acting in parallel or sequence
to complete the degradation process. The ability of microbe to degrade a vast array of pollutants
makes bioremediation a technology that can be applied in different soil conditions.
A widely used approach to bioremediation involves stimulating a group of organisms in order to
shift the microbial ecology toward the desired process. This is termed. “Biostimulation.”
Biostimulation can be achieved through change in pH, moisture, aeration, or nutrient additions.
The other widely used approach is termed “Bioaugmentation” where organisms selected for high
degradation abilities are used to inoculate the contaminated site. These two approaches are not
mutually exclusive- they can be used simultaneously.
Eleme is a community in River State, one of the oil producing and agro-ecological areas in the
Niger-Delta region of Nigeria, a region with abundant natural resources including good weather
and fertile land for agriculture. Although the level of agriculture production in that region is very
low given the abundant resources endowment, it is the largest oil producing zone in the country.
It is the base of Nigerian oil and gas industry, generating over 90% of the nation’s economy
(Odjuvwlederhie et al, 2006). Oil exploration and activities have been concentrated in this Niger-
Delta region which has over 1000 production oil-wells and over 47,000km of oil and gas flow
lines (Ngobiri et al., 2007).
The negative impacts of these oil activities include destruction of wild life, loss of fertile soil,
pollution of air and water and damage to the ecosystem of the host communities (Aghlino, 2000).
The ecological problems observed as a result of oil spill include a brownish vegetation and soil
erosion, diminishing resources of the natural ecosystem, fertile land turned barren and adverse
effect on the life, health and economy of the people (Roberts, 1997).
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1.2 STATEMENT OF THE PROBLEMS
Oil spill is an unintentional release of liquid petroleum hydrocarbon into the environment as a
result of human activities. They are usually mostly caused by accidents involving oil tankers,
barges, refineries, pipelines and oil storage facilities. These accidents can be caused by human
mistakes or carelessness, deliberate acts by terrorist, militants or vandals and sometimes by
natural disasters such as earthquakes.
In Nigeria, the major cause of oil spill is lack of regular maintenance of the principles and
storage tanks. Most pipelines from the flow station are absolutely being more than 20 years old
making them subject to corrosion and leakage. Some of these pipes are laid above ground level
without adequate surveillance, exposing them to wear and tear and other dangers (Oyem, 2001).
Another major cause of oil spill here is sabotage which involves bunkering by some unpatriotic
Nigerians. They damage pipelines in the attempt to steal oil from them.
According to the annual report of the Department of Petroleum Resources, Abuja (1997), over
60000 spills have occurred in Nigeria during her 40 years of oil exploration. Between 1979 and
1996, the spill of 2.4x10 barrels of crude oil occurred from 647 incidents. Only 54706038 barrels
were of oil recovered while 182040666 barrels were lost to the ecosystem.
The growth of oil industry combined with population explosion and a lack of environmental
regulations have caused substantial damage to the environment of the Niger Delta. After several
years of ignoring or giving little or no attention to the adverse effect of oil spill, the Federal
Government of Nigeria along with the oil companies operating in the Niger Delta have began to
take steps to mitigate the damages. The role of the environmental agency in checking and
documenting oil-spills is getting stronger as the new wave of combating oil spill through
phytoremediation in dramatically unfolding in the remediation industry.
In Rivers State several incidents of oil spillage have occurred in the last few decades. One of the
major oil producing communities that has suffered the incidence of oil spill is Eleme. The spill
incidents that have occurred in Eleme include the following:
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1. Oil spill in Ogale due to pipeline vandalism that occurred in 2009. The resultant water
pollution has deprived the residents of the community their sources of livelihood as their
sources of potable water and farmlands were highly polluted.
2. Oil spill from the Okogbe tank truck explosion on 12 July 2012 which claims the life of 92
peoples who were scooping up spilled petrol, when the tanker caught fire.
3. The several incidents of oil spill in Ogali and Agbonchia that drastically contaminated the
soil in the communities.
In response the remedial actions that have been carried out to mitigate or clean up oil spill in soil
and rivers of Eleme community are:-
1. The use of skimmers to adsorb spilled oil accumulated on the river surface. There were
polyethylene mop like pads which were placed on water surface of the affected rivers.
2. In-situ burns of spilled oil (slicks) on water surface – this was done in controlled delineated
areas by fire resistant booms.
3. Spray of chemical dispersants from planes into the oil slicks on top of the river water – in
order to break down the oil into small droplets which are more susceptible to natural
degradation.
4. Scrapping of the top soil to remove contaminated areas.
Meanwhile the gaps and inadequacies that that arose from the application of the measures stated
above are as follows:
1. The clean up method above affects air quality through the smoke and residuals
2. Some heavy crude oil compounds are left behind (they do not burn well) such as sticky
asphaltenes from which road tars are made.
3. It has indicated that applying dispersants has toxic effects on coral reefs and other marine
life.
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4. Also scrapping of the stop of soil degrades the soil by removing the nutrients for plant
growth and removes the fertility of the soil and renders it unsuitable for agriculture.
5. Introduction of contaminant to the soil
This therefore, requires that more suitable and environment friendly methods should be
developed and applied in the Niger Delta. This study examines the application of bio-
remediation as an alternative for oil spill clean up in the region.
1.3 AIM AND OBJECTIVES OF THE STUDY
The aim of this research is to examine the effectiveness of bioremediation in the oil spill
contaminated soil using Oyesta Mushrooms (fungi) in Eleme Community in the Niger Delta. sss
In order to achieve the aim , the specific objectives of the study are as follows:
1. To determine the nature and extent of damage caused by oil spill on the quality of the soil in
Eleme community
2. To demonstrate the treatment of contaminated soil using Oyesta Mushrooms (fungi)
3. To recommend further ways to improve the application of the bioremediation in the
treatment of oil spill contaminated soil in the study area.
1.4 RESEARCH QUESTIONS
The following questions are raised in order to carry out the study:
1. What is the nature and extent of oil spilled contamination on the soil in Eleme community?
2. What is the extent of the damages caused by oil spill on the quality of the soil in Eleme
community?
3. What are the parameters that measure the pollutants in the oil spilled soil in Eleme?
4. How can the treatment of contaminated soil using bio-remediation mechanism be carried
out?
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5. In what ways can the application of the bioremediation in the treatment of oil spill
contaminated soil in the study area be improved?
1.5 HYPOTHESIS
The hypothesis formulated to guide the investigation in this study includes:
1. There is no significant change after the treatment of oil spilled polluted soil.
2. There is no significant change in bioremediation of oil spill soil in Eleme Community.
3. There is no significant improvement in the bioremediation of oil polluted soil in Eleme.
1.6 SCOPE OF STUDY
This study is restricted to bioremediation and its role as strategy for oil spill clean up. This was
conducted in Eleme community in River state of the Niger-Delta. The research entails an
analysis of oil spill in Eleme and the application of remediation of oil spill. It also considered the
conventional techniques used for bio-remediation, factors that affect duration of bioremediation
and the microorganisms used as bio-remediator. It also assessed the parameters for measuring
soil quality. Based on the results recommendations was made.
1.7 LIMITATIONS
Some of the limitations that encountered in the course of the study include:
The unavailability of proper records of up-to-date oil spills in the study area. However, the data
in the Department of Petroleum Resources Annual Report (1997) on oil spill was used.
The resident made it difficult for researcher to elicit information due to certain secrets they
would not want to disclose. This is informed by the long standing hostility between the oil host
communities and the oil companies. Psychological problems, poor retrieval of existing
information and obsolescence of materials are other constraints. The researcher was forced to
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contend with the non-chalant attitude of staff in certain oil companies and related agency in
providing the researcher with the necessary materials.
1.8 STUDY JUSTIFICATION / SIGNIFICANCE
This research without doubt has a lot of significances, which will be of great benefit to oil
producing communities, professionals, academics, industrialist, the government, businessmen
and the general public who are interested in the bioremediation of oil polluted soil using oyster
mushroom fungi and the economic value of the mitigated soil.
The study unveils practices of bioremediation and the famers who cultivate and sell mushroom.
The researcher exposed various methods used to mitigate farmland that was polluted by oil spill
and recover the land that has marked off due to pollution into fertile agricultural farmland. The
study there for exposed the various ways one can make himself economical stable through
bioremediation . The environmental protection that bioremediation render to the environ will
arouse the interest of anybody who is environmentally friendly and this study reveals such
responsibilities of those people or companies engaged on bioremediation taken upon themselves.
This study will of course serves as an eye opener to the governments and oil companies who
may never have considered the activities of bioremediation important and will there for guide
policy making in the area of environmental cleanup. Moreover the use of oyster mushroom as
remediator as oil spill cleanup has a lower or no environmental impact to compare to other
means of oil spill cleanup like use of chemicals.
Finally, the study will be beneficial to undergraduates and postgraduate students who are
intending to carry out further research on bioremediation of oil spill pollution. international
organizations interested in alleviation poverty and improving environmental condition of the
developing countries will find this study very useful.
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CHAPTER TWO
CONCEPTUAL FRAMEWORK
For proper understanding of this study, different definition and meaning of bioremediation and
other related or relevant topics as they relate to bioremediation was explored. Meanwhile the
researcher used oyster mushroom (fungi) to decontaminate or remediate oil spill polluted soil in
Eleme community because the mycelium secretes extracellular enzymes and acids that break
down lignin and cellulose, the two main building blocks of plant fiber. These are organic
compounds composed of long chains of carbon and hydrogen, structurally similar to many
Graph 1: show the concentration of total petroleum hydrocarbon (TPHmg/kg)
TPHmg/kg
Graph 1 shows the concentration of total petroleum hydrocarbon (TPHmg/kg) in the polluted
site. It also shows that as the day of treatment increases the total concentration of petroleum
hydrocarbon decreases.
0
284.65 282.9
256.15
220.18
202.26
180.01
72.18
20.42
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Graph 2: shows the concentration of Nitrogen (N%)
N% Graph 2 shows the concentration of nitrogen (N%) in the site, it indicate that as the day fo
treatment increases, the concentration of nitrogen increases.
Graph 3: shows the concentration of Potassium (Kppm)
Kppm
1.09
0.28 0.34
0.66
0.84
0.92
0.28
0.96 0.98
11.33
4.86 4.88 5.10
6.22
8.14
9.98 10.12 10.99
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Graph 3 shows the concentration of the potassium in the polluted site, it also indicates that as the
day of treatment increases, the potassium contents in the soil increases too.
Graph 4: shows the total conductivity of the soil cond.us/cm
Graph 4 shows the total conductivity (cond.us/cm) of the soil. This entails that as the day of
treatment increases the conductivity of the soil decreases.
6.2 THE HYPOTHESIS TESTING
The hypothesis formulated at the beginning of this research was tested using simple T test and
ANOVA.
A statistical hypothesis test is a method of making decision using data, whether from a control
experiment or an observational study (not controlled) in statistics. The critical region of
Cond.µs/cm
25.6
102.58 102.44 98.22
70.15
48.54
40.1 36.18
30.22
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hypothesis testing is the set of all out comes which causes the null hypothesis to be rejected in
favour of the alternative hypothesis.
The critical region is usually denoted by later c or z.
HO: state there is no significant different, that is the oil spill in Eleme community in Rivers State
have no effect on their soil.
HI: States that there is statistical. Significant different between the oil spill on soil of Eleme in
Rivers State, that is to say that oil spill in Rivers State has statistical effect in the soil of Rivers
State Nigeria.
From this research work, the researcher is trying to know if there is relationship between oil spill
and bioremediation that is if bioremediation can remediate the effect of oil spill in the soil of
Eleme of Rivers State.
Hypothesis 1
Ho: There is no significant change after the treatment of oil spilled polluted soil.
Hi: There is a significant relationship/change after the treatment of total petroleum hydrocarbon
oil spill in the soil.
The researcher used the total petroleum hydrocarbon concentration to test the first hypothesis as
follows.
T = x - µ S/√ X = 176.3
µ = 284.65
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n = 7
S = √Σ(Xi – x-2
n – 1 = S √ (282.90 – 176.3)2 +…….+ (20.42 – 176.3)2
7 – 1 = 96.2
t = 176.3 – 284.65 - 108.35 96.2/√7 = 36.36 = 2.98 Decision Rule: If /t/cal < ttab we therefore accept the null hypothesis and conclude that there is a
significant relationship. While ttab = 1.90 using α 0.05 or 5% to check since /t/cal = 2.98 > ttab =
1.90, we therefore reject Ho and conclude that there is a significant change after the treatment of
total petroleum hydrocarbon in the soil.
Hypothesis 2 testing:
Ho: There is no significant change in bioremediation of soil oil spill soil in Eleme community.
Hi: There is a significant change in bioremediation of oil spill soil in Eleme community. The
researcher tested this hypothesis by testing the Nitrogen percent/content in the
bioremediated soil.
X = 0.71 µ = 0.28 N = 7 S = 0.29 t = 0.71 – 0.28 0.29/√7 = 0.43 0.11
= 3.91
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/t/ = 3.91 > ttab = 1.9, thus we reject Ho and conclude that there is a significant change –
treatment of oil spilled soil brought about change and increase in N% content of the soil which
aid the growth of living things.
Hypothesis 3
Ho: There is no significant improvement in the bioremediation of oil polluted soil in Eleme.
Hi: There is a significant improvement in the bioremediation of oil polluted soil in Eleme.
The researcher tested hypothesis 3 by testing the concentration of the following Nitrogen (N%),
PH-value, phosphorous as phosphate (PO43-), potassium (k).
ANOVA TABLE Source of variation DF SS MS F – ratio Treatment 4-1=3 101.52 101.52
3 = 33.84 33.84 2.58 = 13.12
Error 12-4 = 8 20.66 20.66 8 = 2.58
Total 12-1 = 11 122.18
Decision Rule: Reject Ho if Fcal > Fα, k-1, n-k otherwise accept Ho checking with α 0.05.
F0.05, 3, 8 = 4.07
Decision
Since Fcal = 13.12 > F0.05, 3, 8 = 4.07, we therefore reject the Ho and conclude that there is a
significant improvement after the soil has been treated.
6.3 DISCOVERING
Since 0.8 is approximately one (+1) we could conclude that Bioremediation has perfect
relationship with oil spill in general.
Since Bioremediation has perfect relationship with oil spill,
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1. We can conclude that bioremediation can be use to control the adverse reaction of oil
spill occurred in the soil
2. Bioremediation is used to boost the soil fertility
3. It can also be used to control the ecosystem if well cultured.
4. It can also be used to boost the economy of oil producing states like Eleme in Rivers
state.
5. It can also help to create job opportunity for the teaming youths.
The result from the hypothesis testing revealed that there is a statistical significant relationship
between oil spill and bioremediation in oil producing area in River State (Eleme community).
Oil spill if not proper managed can also result to the death of wild life and aquatic animals
(biodiversity) as well as loss of future cultivable land and low fertility in the soil leading to low
production.
HO: No statistical significant different
HI: Significant different
Since there is relationship between bioremediation and oil spill.
HO: That is null hypothesis is rejected in favour of alternative hypothesis.
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CHAPTER SEVEN
7.1 DISCUSSION, CONCLUSION AND RECOMMADATION
1. Bioremediation of the oil polluted soil using oyster mushroom was studied. Preliminary
analysis of the crude impacted soil revealed the following; a pH of 7.26, Conductivity of
102.58 us/cm, percentage Nitrogen of 0.285%, Phosphate content of 1.08ppm Potassium
content of 4.86ppm, Total Petroleum Hydrocarbon of 284.65mg/kg. The fertility of soil is a
function of its physico - chemical and biological parameters. The values of Nitrogen,
Potassium and Phosphate of the polluted soil were significantly low if compared with the
reference standard from the same geographical location. The Total Petroleum Hydrocarbon
[TPH] content of 284.65 in the polluted site was significantly high when compared with the
control site Total Petroleum Hydrocarbon value of 18.10mg/kg.
The Bioremediation exercise lasted for 28 days with soil samples collected every
4days. The pH of the post treated sample ranged between 7.10 to 5.0. The aim of any
Bioremediation exercise is to redeem a negatively impacted soil to near originality. Since
the pH of the reference soil sample was 4.84, the pH of the remediated soil at the 28th day
was 5.00, a very close value to that of the reference soil, supporting the efficacy of the
oyster mushroom as a good bioremediation agent.
2. The pH showed an inverse relationship with the period of bioremediation. As the days
increased the pH decreased from the alkaline range towards the acidic range. The normal
flora and fauna began the normal symbiotic activities, bringing the pH to near the reference
pH . The total petroleum hydrocarbon [TPH] of the treated soil ranged from 282.92mg/kg in
day 1 to 20.42mg/kg in day 7. At the day 7, representing 28th day of the treatment, the total
petroleum hydrocarbon reduced significantly when compared with value prior to the
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commencement of the bioremediation, a difference of 264.23, representing 92.83%
reduction. The total petroleum hydrocarbon showed inverse relation with the period of
treatment. As the period of treatment increased from day 1, the concentration of total
petroleum hydrocarbon decreased, indicative of progress of bioremediation.
The soil nutrients, Nitrogen, Phosphate, and Potassium showed direct relationship
with the period of treatment, as they increased alongside with the period of treatment, and
approached the value of the reference soil. The Nitrogen content increased from 0.28% in
pretreatment analysis to 0.98% in post treatment analysis, a difference of 0.7, representing
71.42%. The value of Phosphate increased from 1.08ppm in pre treatment analysis to
3.08ppm at the 28th day of the treatment, a difference of 2.0, representing 185.2% increase.
The Potassium content increased from 4.86 in the pretreated sample to 10.99ppm at the
28day of bioremediation excercise, a difference of 6.13, representing 126.13% improvement
The soil textural composition improved as the treatment period increased from
day 1 to day 7, with each value approaching that of the reference site. The improved post
treatment values of the nutrients and total petroleum hydrocarbon were correlated with the
observed life activities in the site. The swam of flies gnats, and birds increased as the
treatment days increased, and at the 28th day, the little of the flies and gnats were seen as the
green grass covered the soil surface in a matted form, indicating the redemptive efficacy of
oyster mushroom.
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CONCLUSION
Bioremediation of oil impacted soil using oyster mushroom was successful . The polluted soil
was redeemed to near that of the control site as the physcio -chemical and textural compositions
of the two showed very close values. some of the parameters analyzed after treatment recovered
over 100% improvement, supportive of the successfulness of the remediation excercises . A plot
of total petroleum hydrocarbon concentration showed an inverse relationship, explaining the fact
that the oyster mushroom were using the crude petroleum as their source of energy for growth
and development, so that as the period of treatment increases, the hydrocarbon content of the soil
decreased.
RECOMMENDATION
The use of oyster mushroom for bioremediation of oil impacted agricultural farmland is an
environmental friendly remediation excercise , cheap and does not require much skilled
manpower. it is a positive action that should be explored and exploited in the remediation of
crude oil impacted soil, especially in the mashy soil environment of the coastal areas of the Niger
Delter States.
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Source: field study May, 2014
Oil spill on Ogale Community soil
Oil spill on Ogale Community soil
Oil spill on Ekporo Community
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