Nigerian Journal of Microbiology, Vol.

24(1): 1993 - 2000 2010

www.nsmjournal.org

EFFECTS OF FLOODING AND SEEPAGE FROM DRILLING BURROW PIT WASTES

ON ORASHI RIVER, EGBEMA, RIVERS STATE – NIGERIA.
1
Ogbonna, C.E., 2Njoku. H O 3Onyeagba, R.A. and. 3*Nwaugo, V.O.;
1
Dept of Environmental Resource Mgt, Abia State University, Uturu
2
Dept of Microbiology, University of Port Harcourt, Port Harcourt
3
Dept of Microbiology,Abia State University, Uturu
Abstract
The effects of flooding and seepage of drilling waste pit contents on Orashi River in Egbema was investigated using
physicochemical and microbiological parameters. Physicochemical parameters measured were temperature, pH,
salinity, total dissolved solids (TDS), total soluble solids (TSS), DO, BOD, SO4, PO4, CO3, and NO3. Most of the
parameters measured had their highest values at the discharge zone which reduced as distance increased along the
down steam course of the River. The up-stream samples (control) had the lowest values but concentration reduced as
one moved away from the discharge point downstream. The most prominent parameters were the BOD, TDS, TSS and
salinity which were above WHO and FEPA standards. Ca2+ and K+ ions did not show any significant change. Na+ and
Mg2+ which increased at the discharge zone, gradually decreased in the downstream samples. Bacteriological analysis
showed that the wastes caused increase in bioload of all the groups of bacterial species analyzed with the highest
values in downstream one (DS I) followed by downstream two (DS 11) before the discharge zone. Lowest bioloads
were observed in the up-stream samples. Groups of bacterial species analyzed include total heterotrophic, hydrocarbon
utilizers and coliforms. Results from screening tests for chrome-lignosulfonate utilization by the isolates showed that
Pseudomonas, Serratia and Saccharomyces species were the most prevalent and others utilized the substrate more
than Alcaligenes and Staphylococcus species. Bacillus, Escherichia and Micrococcus species showed very low
utilization while Enterobacter and Xanthomonas species were low in prevalence and did not utilize the substrate. The
drilling wastes therefore appeared to have influenced the physicochemical parameters and the microbial spectrum of
Orashi River.

Keys words: Over-flooding, drilling waste, bacteria, burrow pit

Corresponding Author: V.O. Nwaugo E-mail: vonwaugo@yahoo.com.

Introduction Disposal of the drilling wastes is

In petroleum production, various
types of wastes are generated. While some
are disposed off at the site of production,
others are disposed away from the site. Some
of these wastes include drilling wastes,
petroleum-produced water and natural gas
(Nwaugo, et al., 2006a; 2007a; Ekpo and
Nwaugo, 2008). Drilling wastes are a
complex mixture of drilling mud, drilling
fluid and drilling cuttings in varying
proportions. Drilling muds are formulated
from a mixture of complex organic and
inorganic chemicals, clay, barite, etc., in
liquid or oil base. Other chemical additives
may be added to modify its properties to
meet the specific purposes or site
requirements (Williams et al., 1981; Ifeadi et
al, 1985; Benker-Coker and Olumagan,
1996).
usually regulated by the type of ecological
zone involved e.g., offshore, swamp and land
(on shore). In the offshore area, the wastes
are discharged into the sea, In swamps, they
are deposited in swamp cuts, while on land,
pits and landfill systems are adopted
(Nwankwo and Irechukwu, 1981 Ifeadi et al,
1985, Oteri, 1985,). In most cases, the
disposal of drilling wastes does not meet the
regulatory standards (Ifeadi et al, 1985,
Okpokwasili and Odokuma, 1996) and this
results in environmental pollution. Seepage
in particular, through the wastes pollutes
underground and some surface water
(Enenchukwu and Okpokwasili, 2000).
With respect to land, surface water
serves several human uses including
recreational, domestic and occupational
purposes. Contamination of this fresh water
in any guise becomes a very serious problem

1993
Ogbonna et al., 2010
(Nwaugo et al., 2006a). In Nigeria concrete
data concerning the effects of drilling wastes
in inland water bodies are very scanty (Ifeadi
et al., 1985; Benker-Coker and Olumagan,
1996).
Considering the increasing production
of oil in Nigeria and the accompanying
improper disposal of the drilling wastes so
generated, there is need for environmental
impact assessment of these practices.. This
work therefore examines the environmental
effects of over flooding and seepage of
drilling wastes pit contents into Orashi River,
Egbema, Rivers State, Nigeria.
Materials and Methods
Study Area
The study area is Egbema, Rivers
State. Egbema is located at the northern apex
of Rivers State and has all the features of a
typical tropical climate of the rainforest type.
The area has two distinct seasons – rainy and
dry seasons. While the rainy season spans
from March to September, the dry season
covers October to February.
Several oil companies operate in the
area, drilling several oil wells in the process
and the disposal of the wastes generated is
often improperly done. This is because no
cemented embankments are made for the
waste/burrow pits (Ifeadi et al., 1985).
The particular portion of study is a
recent well drilled about 85m from the banks
of Orashi River and the wastes were buried
within the same vicinity. The well-head
equipment (Christmas tree) was raised on a
metal platform to ensure safety during
flooding as the entire area is covered by
water during the floods . This work was
done between June, 2005 – September,
2006.
Sample Collection
The samples analyzed were obtained
from four (4) different sampling sites of the
Orashi River. One of the sites was upstream
(control), 120m up before the drilling waste
1994
Nig. J. Microbiol. 24(1): 1993 - 2000
pit zone. Second sample came from the
drilling waste pit zone i.e., the fallout point
(FP). Two downstream samples were
analyzed as downstream 1 (DS 1), 120m
away from fallout area and downstream 11
(DS 11), about 120m from downstream 1. At
each sampling area, water column was
collected in triplicate and pooled together to
make one sample. Samples were collected
thrice for physicochemical parameters
analysis at one month interval while ten
samples were collected for microbiological
analysis at two weeks interval within the
same period of study.
The water sampling bottles for the
analyses were washed clean and soaked in
acid-alcohol for 24 hours. They were rinsed
in sterile de-ionized water and kept for use.
With sterile disposable hand gloves, the
bottles were lowered into the water body
against the current 2-3 cm below the surface
to obtain water column samples. Using
similar bottles, the sediments were similarly
collected from the shores without diving into
the water. The collected samples were then
pooled as described earlier and taken to the
laboratories within 1-2 hours of collection.
Analysis of samples;
Physicochemical analysis;
The physicochemical parameters
measured include pH, temperature, salinity,
total dissolved solid (TDS), total soluble
solids (TSS), dissolved oxygen (DO) and
biological oxygen demand (BOD), sulphate
(SO4) nitrate (NO3) and phosphates (PO4).
The temperature and pH were measured at
the site (on the spot) using multipurpose
tester (Model Jenway HANNA 1910). The
DO and BOD were measured using the
Winklers titration method as described in
APHA (1992). The SO4, NO3, PO4, TDS and
TSS were determined using the
spectrophotometric method with HACH/D2/
2010 spectrophotometer at various
wavelengths, depending on the parameters.
Ogbonna et al., 2010
The metallic ions of Ca, K, Na and
Mg were determined using the flame atomic
absorption spectrophotometric (AAS) method
of APHA (1992). The oil content of the
samples was analyzed using the method of
Okpokwasili and Okorie, (1988).
Microbiological Analysis
The various samples were inoculated
on Tryptone soy agar using streaking
technique. The observed colonies were
subcultured to obtain pure isolates which
were subjected to macroscopy followed by
microscopy (after staining) and biochemical
tests as described by Chessbrough (2001) and
Cowan and Steel, (1976). They were cross-
checked according to Buchanan and Gibbons
(1976) for proper identification.
Further microbiological analyses were
done based on the effect of the drilling
wastes on some groups of bacteria. The total
heterotrophic bacteria (THB) were cultured
on Nutrient Agar, hydrocarbon utilizing
bacteria (HUB) on modified mineral salt
agar, and coliform bacteria (CB) on
McConkey agar as described by Okpokwasili
and Okorie (1988), and Nwokoro and
Okpokwasili (2003).
Screening of isolates for Chrome –
lignosulfonate utilization
Screening of isolates was done
according to Okpokwasili and Okorie (1988)
using modified Mineral Salt Medium of Mills
el al (1978). To 9.9 ml of the medium was
added 0.1 ml of the drilling waste from the
waste burrow pits. This was incubated at
room temperature for one week (7 day) and
then examined for growth using turbidity and
pH change as indicators.
Statistical analysis;
The various data generated in this
investigation were subjected to various
statistical analysis to find out if there were
significant differences among the sites. The
statistical methods applied include Chi-
1995
Nig. J. Microbiol. 24(1): 1993 - 2000
square, standard deviation and ANOVA
(Analysis of Variance).
Results
The values of the various
physiochemical parameters analyzed are
shown in Table 1. Most parameters measured
had their highest values in the fallout point
(FP). These include salinity, total dissolved
solids (TDS), total soluble Solids (TSS),
biological oxygen demand (BOD) and
carbonate (CO3). Others were sulphate (SO4),
phosphate (PO4), and nitrate (NO3). The
second highest values were seen in the
downstream 1 and downstream 11. Values of
these parameters from the upstream samples
were the lowest (Table 1). The values
showed statistical significance (P=0.05) in
the direction of fallout zone > downstream 1
> downstream 11 > upstream. In the case of
DO, more was seen in the up stream sample,
followed by the downstream II, and then
downstream I with the least in fallout point.
For metallic ions, only Na+ showed
statistical significance as the values changed
from 0.2 mg/l in the upstream (US) to 1.01 in
the fallout point (FP) but this decreased to
0.85 and 0.53 mg/l in DS 1 and DS 11
respectively. The oil contents also showed
the highest values in the FP (16.5 mg/l),
followed by the DS 1 11.3 and then 8.11 in
the DS 11. The upstream (US) samples had
the least values of 1.2 mg/l. Results obtained
showed that the pH changed from slightly
alkaline 7.05 in US, through 6.89 in DS 11,
6.62 in DS 1 and then to the least 6.31 in FP.
The temperature rose from 27.1oC in US to
29.3oC in FP decreasing gradually till 28.5oC
in DS 11.
The bioloads of the various groups of
organisms as shown on Table 2 indicated a
definite gradient. Highest values were
observed in the DS 1 followed by the DS 11
before the FP. The US had lowest values of
each bacterial group except coliforms with
lowest bioload in the FP (3.9 X 102 cfu/ml).
Ogbonna et al., 2010
The values obtained are as shown in Table 2;
for THB count, it was DS 1 (5.2 x 104) > 4.9
x 104 (DSII) > 3.3 x 104 (US). The same
trend continued with the other bacterial
groups but with lower biolaods. In all the
sampling points, the order was THB > HUB
> CB except in the upstream (US) where CB
was higher than the HUB. The other
observations were similar.
In the prevalence of each observed
organism (Table 3), a similar trend as
observed in bioloads was also recorded. In
the US samples, most prevalent organism
was Bacillus species (80%) followed by
Saccharomyces species (60%) while the least
were Serratia, Pseudomonas and Alcaligenes
species with 30% each. On the other hand,
the highest prevalence of each organism was
observed in the DS 1 samples except
Streptococcus species which did not change
much (Table 3). The highest prevalence was
in DS 1, followed by DS 11, the FP and then
the US. This was statistically significant (P <
0.05).
Results obtained from the screening
of chrome lignosulfonate utilization (Table 4)
showed that Enterobacter and Xanthomonas
species did not metabolize chronic
lignosulphate as there was no change in
turbidity and pH. On the other hand
Saccharomyces, Pseudomonas and Serratia
species utilized the test substrate. These
organisms showed high turbidity (growth)
and change in pH (towards acidity).
Discussion
The analysis of the physicochemical
parameters at the various sampling points
showed some variations in some parameters
while a few did not really change. Salinity,
TDS, TSS, BOD and DO showed extensive
increase from the FP till the DS II indicating
adverse effects of the wastes. Among the
metallic elements only Na+ showed statistical
change from 0.4 (US) to 1.01 (FP) mg/ml.
Similar situation had earlier been reported by
1996
Nig. J. Microbiol. 24(1): 1993 - 2000
Nwaugo et al, (2007a, 2009). Again, Carlton
and Darby (1985) reported that Drilling –
operations affected the physicochemical
properties of Alabama Mobile Bar Waters, a
fact also expressed by Wills (2000) and
Richards and Love (2002). This is similar to
the findings of this study. However, the little
oil observed in the upstream showed that
other activities upstream had added the oil.
All the observed parameters which changed
in values showed a common gradient. Most
of them had lower values in the upstream but
increased in the fallout point and decreased
again from DS 1 to DS 11. This showed the
diluting effects of the water as one moved
away from the Fallout Zone.
Several researchers working on oil
related matters have at various places and
time isolated most of the organisms observed
in this study. Pseudomonas, Alcaligenes,
Serratia, Saccharomyces and Micrococcus
species have been reported in oil
contaminated sites (Enemchukwu and
Okpokwasili, 2000, Nnubia and Okpokwasili,
1993, 1997; Benker-Coker and Olumagan
1996, Spain and Vanveld, 1983; Nwaugo et
al., 2007b). However, not much report have
been made of Xanthomanas, Enterobacter,
Eschericha and Streptococcus species in oil
polluted area but their presence in natural
water bodies had been a major issue
(Nwaugo et al., 2006b, Nwachukwu and
Otukanefor, 2003 and Okpokwasili and
Akujuobi, 1996). Ogbulie (1996) had also
reported the presence of these organisms and
others in a fish-farm in the study area. Some
of these organisms have also been associated
with human activities (Prescott et al, 2001).
This further explains the presence of such
organisms as Saccharomyces, Eschericha,
Micrococcus and Staphylococcus species as
Orashi Rivers serves several purposes for the
Egbema people including recreational
domestic, occupational and food processing
(Cassava Fermentation).
Ogbonna et al., 2010
The prevalence of each of the
observed organisms varied according to the
sampling point. The down stream (DS 1) had
the highest prevalence, followed by the
downstream (DS 11) and the upstream (US)
before the fallout zone (FP). This showed
that the organisms were present in the water
but in low level before the pollution. The
high prevalence observed in the DS 1 showed
that the drilling wastes from the FP was
diluted which enhanced microbial growth in
the DS I. Nwaugo et al., (2006b) had earlier
observed this dilution effects in abandoned
quarry pit waters. With the dilution, the
levels of the concentrations of waste water
were moderated, resulting in the utilization of
the drilling wastes as nutrients by the
microorganisms. Organisms which could not
metabolize the wastes utilized the less toxic
intermediates produced by the drilling wastes
utilizers.
Observations in the bioloads of the various
groups of organisms investigated agreed with
the prevalence explained above. Highest
bioloads were observed in the DS 1 but
decreased in DS 11. This showed that the
wastes, though foreign, were metabolized as
nutrients hence the increases in numbers, but
the nutrient decreased in DS 11 resulting in
lower bioload. The most adversely affected
were the coliforms while the most
beneficiaries were the total heterotrophic
bacteria (THB). The hydrocarbon utilizing
bacteria (HUB) also increased extensively in
line with the increase in oil content. Richard
and Love (2002) and Wills (2000) reported
this increase in oil contents. The lignosulfate
in the drilling wastes served as nutrients for
both the HUB and THB. The general increase
1997
Nig. J. Microbiol. 24(1): 1993 - 2000
in THB was because THB encompass all
groups of organisms (Chessbrough, 2001).
Screening of isolates for chorme-
lignosulfate further explained the observation
in the prevalence and bioload. While
Pseudomonas, Serratia, Saccharomyces
Staphylococcus and Alcaligenes species have
at various times and places been implicated
in oil spillage and its remediation, other
organisms are not common in oil polluted
habitats. These were the major organisms
which utilized the drilling wastes. Bacillus,
Micrococcus and Staphylococcus species
only metabolized very little of the substrate
while Xanthomonas and Enterobacter species
could not. Atlas (1981), Benker-Coker and
Olumagen (1996),Okpokwasili and Okorie
(1988) and Nwaugo et al, (2007b)had
implicated these organisms too. Analysis of
utilization of Chrome-lignosulfate showed a
change of pH towards acidity. Metabolism of
the substrate turned the medium acidic. The
level of acidity was commensurate with the
level of chrome-lignosulfate utilization as
shown by Pseudomonas, Serratia and
Sacharonmyces species along with
Staphylococcus and Alcaligenes species
showed much change in pH (towards acidity)
Xanthomonas and Enterobacter species
which could not utilize the substrate, could
not change the pH of the medium. This
observation agrees with Nweke and
Okpokwasili (2004) Okpokwasili and Okorie
(1988) Enenchukwu and Okpokwasili (2000)
and Nwaugo et al,(2009).
In conclusion, the drilling waste
seepage and over flooding of the drilling
waste pit caused extensive modification of
the physicochemical and bacterial spectrum
of the Orashi River.
 Ogbonna et al., 2010 Nig. J. Microbiol. 24(1): 1993 - 2000

Table 1: Values of physicochemical parameters of the various water samples examined.

Parameter Upstream Fallout Point Downstream I Downstream II
pH 7.02 + 0.03a 6.31 + 0.05ab 6.62 + 0.04ab 6.80 + 0.03ab
o a b b
Temp C 27.1 + 0.03 29.3 + 0.4 29.1 + 0.03 28.5 + 0.03ab
Salinity 0.18 + 0.02a 0.81 + 0.04a 0.01 + 02ab 0.39 + 0.03ab
TDS mg/l 260 + 10.3a 2200 + 2.10b 2100 + 1.03b 1800 + 1.04b
a b b
TSS mg/l 67 + 1.1 95 + 1.03 90 + 1.02 85 + 1.01ab
DO 7.1 + 0.5a 4.9 + 0.4b 5.4 + 0.4ab 6.1 + 0.5ab
a b ab
BOD 60 + 0.9 80 + 1.1 75 + 1.03 67 + 1.02ab
CO3 3.7 + 0.03a 5.1 + 0.05b 41 + 0.03ab 3.9 + 0.02a
a b b
PO4 0.08 + 0.01 0.30 + 0.2 0.29 + 0.02 0.20 + 0.03b
a b b
NO3 1.04 + 0.01 0.7 + 0,03 0.63 + 0.03 0.50 + 0.02b
Cl 1.2 + 0.03a 128 + 0.04a 1.96 + 0.04ab 1.22 + 0.03a
a b ab
Na mg/l 0.4 + 0.03 1.01 + 0.06 0.85 + 0.04 0.63 + 0.04ab
K mg/l 0.4 + 0.02a 0.5 + 0.03 a 0.48 + 0.04a 0.43 + 0.03a
a b b
Mg mg/l 0.4 + 0.02 1.10 + 0.03 1.03 + 0.03 0.93 + 0.02ab
Oil Content 1.2 + 10.3a 16.6 + 0.09a 11.3 + 0.07b 8.1 + 0.04a
Values followed by the same alphabets are not significantly different but those followed by different alphabets are significantly
different.

Table 2; Bioload of the various groups of organisms observed in the water examples (cfu/ml).
Group of organism Upstream Fallout Point Downstream I Downstream II
THB 3.1 X 104 3.3 X 104 5.2 X 104 4.9 X 104
HUB 2.1 X 102 3.5 X 103 4.3 X 104 3.6 X 104
3 2
CB 2.3 X 10 3.9 X 10 4.3 X 104 3.1 X 103
*Values reported are averages of three samplings.
THB = total heteroltrophic bacterial;CB = coliform bacteria ;HUB = hydrocarbon utilizing bacteria

Table 3: Prevalence of Microbial species in the various water samples types analyzed
Upstream Fallout point Down stream I Down stream II
NE NO % NE NO % NE NO % NE NO %
Saccharamyces sp 10 6 60 10 5 50 10 6 60 10 7 70
Bacillus sp. 10 8 80 10 6 60 10 7 70 10 8 80
Pseudomonas sp 10 3 30 10 6 60 10 8 80 10 6 60
Alcaligenes sp. 10 3 30 10 5 50 10 7 70 10 7 70
Escharichia coli 10 5 50 10 4 40 10 7 70 10 6 60
Enterobacter sp. 10 4 40 10 4 40 10 5 50 10 5 50
Seiratia sp. 10 3 30 10 4 40 10 6 60 10 4 40
Staphylococcus sp 10 4 40 10 5 50 10 6 60 10 5 50
Micrococus sp 10 4 40 10 5 50 10 7 70 10 6 60
Streptococcus sp 10 4 40 10 3 30 10 3 30 10 3 30
NE=No. of samples examined; NO= No. of isolates observed; %=Percentage

Table 4: Utilization of Chrome-lignosultate by isolated microbial species

Organisms Turbidity Initial pH Final pH
Saccharamyces sp +++ 7.50 6.21 + 0.04a
Bacillus sp. + 7.50 7.30 + 0.03b
Pseudomonas sp +++ 7.50 6.26 + 0.04a
Alcaligenes sp. ++ 7.50 6.61 + 0.04a
Escherichia coli + 7.50 7.33 + 0.02b
Enterobacter sp. - 7.50 7.50 + 0.01c
Seiratia sp. +++ 7.50 6.24 + 0.03a
Staphylococcus sp ++ 7.50 6.57 + 0.03ab
Micrococus sp + 7.50 7.20 + 0.04b
Streptococcus sp + 7.50 7.22 + 0.04b
Xanthomonas sp - 7.50 7.50 + 0.01c
KEY + = Scanty turbidity ++ = moderate growth
- = No observed turbidity +++ = High turbidity

1998
Ogbonna et al., 2010
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