post graduate work

1. BYBY
EZE CHINWEEZE CHINWE
POSTGRADUATE DIPLOMA IN
ENVIRONMENTAL
MANAGEMENT TECHNOLOGY
Supervised by
DR J.D NJOKU
EffEct of palm oil mill EffluEnt onEffEct of palm oil mill EffluEnt on
mill EffluEnt on soil samplEs inmill EffluEnt on soil samplEs in
isiala mbano lgaisiala mbano lga

2. INTRODUCTION
Palm oil mill effluent (POME) is the voluminous liquid
waste that comes from the sterilization and clarification
sections of the oil palm milling process. The raw effluent
contains 90-95% water and includes residual oil, soil
particles and suspended solids
Raw POME has an extremely high content of degradable
organic matter, which is due in part to the presence of
unrecovered palm oil
Oil palm cultivation and processing like other
agricultural and industrial activities raise environmental
issues
Palm oil mill effluent is a highly polluting material and
much research has been dedicated to means of alleviating
its threat to the environment.

3. INTRODUCTION CONTD.
The POME discharged is objectionable and could pollute streams, rivers, or
surrounding land. Raw POME has Biological Oxygen Demand (BOD) values
averaging around 25,000 mg/litre, making it about 100 times more polluting
than domestic sewage (Fang et al., 1999).
In Isiala Mbano LGA, palm oil production is one of the major socioeconomic
activities of inhabitants in the area .As a result of the abundance of palm trees
(Elaies guineensis), large quantities of palm fruit are harvested and processed
and during procession, large quantities of palm oil effluents are discharged onto
the soil in its raw form by small-scale operators
The palm oil industry contributes 83% of the largest polluter in some palm oil
producing countries; the situation is probably similar in other palm oil
producing countries
It has been observed that most of the POME produced by the small-scale
traditional operators undergoes little or no treatment and is usually discharged
into the surrounding environment and so raises the need to look at the effect of
raw POME on the soil in the study area.

4. OBJECTIVES OF THE STUDY
The aim of the study is to determine the microbial
characteristics of palm oil effluent disposal on soil samples.
The objectives of this study therefore are:
To carry out sampling via visual inspection of POME and
non-POME sites
To collect soil samples from POME and non-POME sites
with sterile polythene bags and soil augers.
To collect samples by air drying and sieving. The air-dried
and sieved samples will be used to analyze for various
parameters
To carry out Physico-chemical analysis of soil samples from
POME and non-POME sites.
To make recommendations on how best to handle POME in
the study area.

5. SCOPE OF THE STUDY
The study is limited to a selected and
representative area, Ogbor Ugiri Isiala
Mbano LGA in Imo State, focusing
on effect of Palm Oil Mill Effluent
on soil.

6. MATERIALS AND
METHODS•Field survey and soil sampling were carried out.
•visual inspection of the sampling sites was conducted and the
differences between the sites in terms of vegetation, presence of
constitution, soil colour, odour, e.t.c. were observed and noted.
•Sampling was done five times from each location using the
quadrant approach (A plot of 25 m by 25m was delineated in the
two soil communities) after which five soil samples were
randomly collected using the soil auger.
• Samples used for the study were collected at a depth of 0-20cm
from a discharge point near the Palm Oil Processing Mill in
Umuokohia village, Ogbor-Ugiri, Isiala Mbano, Nigeria. Soil
samples, collected at the same depth from a normal garden soil
from the same village, 1km away from the discharge point, served
as the control

7. MATERIALS AND METHODS CONTD.
•Both sites have similar soil parent materials, topography, and climate.
•The soil samples were put in autoclaved glass jars with labels, which
were immediately sealed and kept on ice packs before being transported
to the lab.
•The collected soil samples were air-dried for seven (7) days in the
laboratory, grinded separately into fine size using a mortar and pestle
and , pretreated and analyzed for various parameters.
•Glassware and media used were sterilized by autoclaving
•Physico-Chemical Analyses were carried out for the following
Parameters :
•pH, Conductivity, total suspended soil, total dissolved solid, acidity,
alkalinity, Chloride, Hardness, sulphate, phosphate, nitrate, NH4,
Calcium, magnesium, sodium, potassium, dissolved oxygen demand,
biological oxygen demand etc

8. MATERIALS AND METHODS CONTD.
Microbiological Analysis of the Soil Samples.
Isolation from soil samples using the method as described by Holt et al.,
(1994). analysis of the soil samples were carried out according to the
methods of Oyeleke & Manga (2008) and Rabah et al, (2008). Bacterial
isolates were identified and characterized using standard biochemical tests
(Cheesebrough, 2006). The fungal isolates were identified according to
Oyeleke & Okusanmi (2008) based on the colour of aerial hyphae and
substrate mycelium, arrangement of hyphae, conidial arrangement as well as
morphology.
The tests employed include colonial, morphological characteristics, gram
stain, motility, Catalase, methyl red, Voges- Proskaeur, Indole production,
urease activity, H2S and gas production, citrate utilization, glucose, sucrose,
and lactose utilization tests.
The media used in this study were Nutrient agar (Fluka Biochemica,
Germany), MacConkey agar (Antec), and Sabourand dextrose agar (Fluka
Biochemica, Germany). All the media were prepared and sterilized
according to manufacturer’s specifications.Statistical analysis was carried
out

9. LOCATION DO, mg/l PH COD,
mg/l
BOD5,
mg/l
PO4
-3
mg/l TOC (%) K, mg/l N, mg/l EC(dSm-
1
)
Oil and
grease,m
g/l
Organic
matter
(%)
Bulk
density(g
cm-3
)
vegetatio
n
AI 1.582 3.940 5543.40 5463.10 8.324 3.40 30.0 34.0 0.19 97.234 2.25 1.998 bare
A2 1.787 4.001 3345.20 4830.40 8.234 3.34 36.0 29.0 0.18 77.108 2.23 1.900 bare
A3 1.002 5.450 2340.50 3630.20 7.678 3.43 32.0 20.0 0.19 50 .002 2.30 1.872 bare
A4 1.987 5.780 2134.60 3200.00 6.345 3.40 29.0 16.0 0.20 44.801 2.58 1.805 bare
A5 2.512 7.435 1231.00 2254.33 5.184 3.50 9.50 17.0 0.24 39.878 1.88 1.605 little
B1 5.845 6.930 209.500 61.700 5.123 2.34 7.40 9,70 0.27 7.78 1.28 0.826 little
B2 5.897 6.890 225.200 22.600 5.112 2.25 5.70 6.00 0.25 6.56 1.56 0.923 grown
B3 5.765 5.654 234.200 23.900 5.012 2.22 5.00 5.40 0.27 5.23 1.65 0.922 grown
B4 6.234 6.347 221.900 34.400 5.109 2.13 4.98 7.60 0.26 9.34 1.87 0.820 grown
B5 5.098 5.876 289.000 49.670 4.345 2.10 4.92 5.70 0.23 7.45 1.35 0.825 grown
All parameters in mg/l except pH, organic matter (%) and bulk density (gcmAll parameters in mg/l except pH, organic matter (%) and bulk density (gcm22
) DO= dissolved oxygen, PO) DO= dissolved oxygen, PO44
-3-3
= phosphates, TOC= Organic Carbon, BOD== phosphates, TOC= Organic Carbon, BOD=
biochemical oxygen demand, COD= chemical oxygen demand, EC= electrical conductivity. All values at mean temp of 42biochemical oxygen demand, COD= chemical oxygen demand, EC= electrical conductivity. All values at mean temp of 4200
cc
TABLE 4.1 PHYSICO-CHEMICAL PARAMETER OF POME (A) AND NONE POME (B) SOIL
SAMPLES

10. Figure 4.1 Chart Representation of Fungal % Occurrence in Both Locations
A=Aspergillus niger B= Aspergillus flavus C= Fusarium species D= Penicillin species, E=Mucor.

11. Counts represent means of triplicate samples, cfu/g: coliform forming units per gramme THBC= TOTAL HETEROTROPHIC
BACTERIAL COUNT, THFC=TOTAL HETEOTROPHIC FUNGAL COUNT, HDBC=HYDROCARBON DEGRADING BACTERIAL
COUNT, HDFC=HYDROCARBON DEGRADING FUNGAL COUNT,
LOCATION A1 A2 A3 A4 A5 B1 B2 B3 B4 B5
THBC, x106
cfu/ml
1.36 2.01 2.15 2.40 2.42 1.79 1.80 1.60 1.50 1.76
THFC, x104
cfu/ml
3.05 3.00 2.89 2.49 1.34 1.40 1.46 1.04 1.42 1.22
HDBC,
x106
cfu/ml
1.00 1.10 1.28 1.29 1.40 0.90 0.70 0.80 1.00 0.90
HDFC,x104
cfu/ml
2.52 2.34 2.00 1.92 0.90 0.55 0.68 0.75 0.60 1.00
MEAN
COUNTX105
3.80 5.43 5.82 9.33 9.61 6.77 6.30 6.05 6.30 6.73
TABLE 4.2 VIABLE COUNTS OF BACTERIA AND FUNGI ISOLATED
FROM LOCATIONS

12. Table 4.3 ANOVA TABLE
Data Summary
Sample
Locations
A B
N 5 5
- X 33.99 32.15
Mean 6.798 6.43
- X2
257.1983 207.1083
Variance 6.5336 0.0959
Std.Dev. 2.5561 0.3098
1.1431 0.1385

13. Source of
variation
Df SS MS F P
Y-Y among
groups
1 0.3386 0.3386 0.1 0.759923
Y-Y within
groups
8 26.5181 3.3148
Y-Y total 9 26.8566
ANOVA TABLE

14. TABLE 4.4: MICROSCOPIC MORPHOLOGY, CULTURAL
CHARACTERISTICS AND % OCCURRENCE OF FUNGAL
ISOLATES FROM SOIL SAMPLES
Organism Microscopic
morphology
Cultural characteristics Occurrence (%)
POME soil NON pome soil
Aspergillus niger Has septate hyphae with
long and smooth
conidiophores, large ,
round unbranched
sporangiophores
Golden reverse side,
creamy and brownish
mycelium, powdery
27 30
Aspergillus flavus Colourless ,long, erect
swollen conidiophores
and septate hyphae
Colonies are Greenish
yellow colour with
creamy edge
18 20
Fusarium species Dark pigmented
conidiophores, spherical
Powderish and creamy
colonies
22 10
Penicillin species Fruity mycelium,
branched conidiophores
with white margin
Greenish and filamentous
colonies
11 27

15. TABLE 4:5 BIOCHEMICAL TESTS OF BACTERIA ISOLATES FROM SOIL
SAMPLES
Organism Gram
reaction
Cultural
characteri
stics
motility oxidase Catalase citrate coagulase urease indole Occurrence (%)
POME soil NON POME
Pseudomona
s sp
Negative
rods
Blue green
colonies
+ + + + - - - 21 10
Lactobacillus
spp
15
Serratia sp Negative
rod
Large gray
colonies
+ + + + - - - 7
Bacillus sp Positive
rod
Milky
white
colonies
+ + + + - -/+ - 34 27
Staphylococc
us sp
Positive
cocci
- - + - + - - 17
Corynebacter
ium sp
Positive
rod
- - + + - - + 9
E.coli -rods Small
white
colonies
+ + - - + 5
Proteus Spp Positive
rods
Large
milky
white
+ - + + + - 4 7
Kleb
pneumoniae
-rods Large gray
colonies
+ - - + + - 8 5
Streptococcu
s
+ cocci - - - - - - 8 10
micrococcus 8 5
(+ =positive; - negative reactions)

16. FIG 4.2 Chart Representations of Microorganisms isolated from Both Locations
A=Pseudomonas, B= Lactobacillus, C=Serratia, D=Bacillus E=, Staphylococcus, F= Escherichia, G= Proteus,
H=Klebsiella, I=Streptococcus, J=Micrococcus, K= Corynebacterium

17. Conclusions
•Due to the oil-palm effluent discharge noticeable in locations A, the color of the soil was dark
brown, damp and odiferous while that of the non – POME site, locations (B) was observed to be
brown, dry and free of odour. An impenetrable layer of the soil makes it very difficult for
vegetative cover to exist.
• As shown in the ANOVA table, there was significance in the mean square among groups due to
POME effect on the samples from locations A. Higher concentration of the effluent significantly
reduced the soil bacterial population in the soil. The soil pH however remained in acidic
conditions at all levels of palm oil mill effluent pollution probably due to acidic nature of applied
effluent. Palm Oil processing gives rise to high values of COD, which indicate the recalcitrance
of chemicals that have escaped biodegradation.
• It was also noticed that the soil acidity is increased as raw POME is discharged but the pH
seems to increase as biodegradation takes place. In addition, the increase of electrical
conductivity in the present study was likely due to the loss of weight and release of other mineral
salts such as phosphate and ammonium ions through the decomposition of organic substances as
reported by Wong et al., (2001). The relatively high DO reported in this study may be due to the
high temperature and duration of bright sunlight, which influenced the percentage of soluble
gases (O2 and CO2) in the effluent (Chow 1991). It is apparent from the analyses that POME
significantly and substantially increases the soil nutrient levels in the soil. When soil is polluted,
the physiochemical properties are affected which may decrease its productive potentials.

18. Palm oil mill effluent application to soil can result to some beneficial soil chemical and physical
characteristics, such as increases in organic matter, organic carbon, major nutrients (e.g. N, P),
water-holding capacity and porosity (Rupani et al, 2010) .However, it brings about undesirable
changes such as decreases in pH, and increases in salinity. Additionally, the decomposition of
POME by soil microbes could have induced oxygen depletion in the surface soil, thereby
inhibiting aerobic microbial activityIt therefore follows that palm oil mill wastes should be well
cured before they are disposed of on soils, as research has confirmed their efficiency in use as
fertiliser (composting)(Chan, 1980).
There is therefore the need to monitor the effects of these wastes on the soil as the level of
influence will vary .The organic substance of POME is generally biodegradable; therefore,
treatment by biodegradable process could be suitable, which are based on anaerobic, aerobic, and
facultative processes (Radziah, 2001). Although POME is a land and aquatic pollutant when
discharged directly into the environment; it is amenable to biodegradation .
Conclusions

19. References
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Research, 2 (12): 656-662 Orji, M.U., Nwokolo, S.O., and Okolo, I. (2006). Effect of
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Zakaria Z.Z, A. Khalid and A.B Hamdan(1994). “Guidelines on land application of
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20. Thanks for
listening!