This project proposal aims to study the microbial and physicochemical composition of soil contaminated with petrol. Soil samples will be collected from a petrol station and analyzed to determine physicochemical parameters like pH and moisture content. Bacteria present will be enumerated and identified using techniques like gram staining. Isolates will be characterized based on tests such as coagulase. Their ability to biodegrade petrol will also be examined. Heavy metals in soil samples will be analyzed. The study expects to provide information on bacteria that can degrade oil and their physicochemical requirements to do so, contributing to oil spill cleanup efforts.

1. AN HND PROJECT PROPOSAL
ON
STUDIES OF MICROBIAL AND PHYSICOCHEMICAL COMPOSITION OF
SOIL CONTAMINATED WITH PETROL
BY
OLUSOLA ADEOLA TIMILEYIN
SO4/SLT/2015/2531
SUBMITTED TO THE DEPARTMENT OF SCIENCE LABORATORY
TECHNOLOGY (MICROBIOLOGY OPTION), FACULTY OF APPLIED
SCIENCES, RUFUS GIWA POLYTECHNIC, OWO, ONDO STATE.

2. INTRODUCTION
Soil is the most important component of the environment.
Pollution of the environment by petroleum products is an inevitable
consequence of oil production, transportation and distribution activities.
The presence of heavy metals in soil can affect the quality of food,
groundwater, micro-organisms activity, plant growth etc. (Antoaneta et al.,
2009).
Biodegradation of hydrocarbons by natural population of microorganisms
represents one of the primary mechanisms of eliminating petroleum
pollution from the environment (Facundo et al., 2001).

3. AIM AND OBJECTIVES
The aim of this project is to determine the effect of refined petrol on
bacteria population and physiochemical characteristics of the soil.
The objectives are to:
 To determine the physicochemical composition of petrol
contaminated soil samples.
 To determine the heavy metals of petrol contaminated soil samples.
 To isolate and identify bacteria in the soil sample collected.
 To identify bacteria that can degrade petrol in the soil samples.

4. MATERIALS AND METHODS
• Sample Collection
Five (5) soil samples will be collected from petrol station in Owo, Ondo State with the aid of
soil auger at depths 15-20cm into sterile black cellophane bags and well labelled. They will
be taken to Microbiology Research Laboratory for analysis according to Tanee et al. (2008).
• Determination of the physicochemical parameters
The physicochemical parameters to be determined are:
pH
soil texture and
moisture content.

5. • Enumeration of Bacteria Population
Nutrient agar (NA) will be used in the analysis by pour plating method.
• Identification of Bacteria
The following methods will be used:
Gram staining
• Bacteria Characterization
• The bacteria isolates will be characterized base on these tests:
• Morphological characteristics
• Coagulase test
• Catalase test
• Sugar fermentation (glucose, lactose and sucrose) (Holt et al., 1994)

6. • Biodegradation
The method proposed by Saratale et al. (2006) will be used.
A standard freshly grown pure culture will be inoculated in 250ml containing 150ml of mineral salt medium and
petrol oil of 200mg/100ml.
The flask will be incubated for 15days under shaking condition at 120rpm.
A control flask without inoculant will be prepared and incubated.
Total heterotrophic count and optical density will be monitored.
• Heavy Metal Analysis
The heavy metals to be determined are:
Lead (Pb),
Chromium (Cd),
Zinc (Zn),
Copper (Cu) and
Mercury (Hg) (Atlas, 2001).

7. EXPECTED CONTRIBUTIONS TO KNOWLEDGE
Oil-degrading bacteria are abundant in soils contaminated with used
petroleum products. This can be exploited for large oil-spill clean-up
campaigns.
This study will also provide information on the physicochemical
requirements for optimum degradation by these bacteria.

8. REFERENCES
• Antoaneta E, Alina B, Georgescu L (2009). Determination of heavy metals in soils using
XRF Technique. Rom. J. Phys. 55(7-8):815-820.
• Facundo, J., M.R.., Vanessa, H.R and Teresa, M.L. (2001). Biodegradation of diesel oil
in soil by a microbial consortium. Water Air Soil Pollut. 128: 313-320.
• Gowd, S.S., Reddy, M.R.M. and Govil, P.K. (2010), Assessment of heavy metal
contamination in soil at Jajmau (Kanpur) and Unnao Industrial areas of the Ganga
plain, Uttar Pradesh, India, Journal of Hazardous materials, 174:113-121.
• Holt, J.G., Krieg, N.R., Sneath, P.H.A., Stanley, J.T., William, S.T. (1994). Bergey’s Manual
of Determinative Bacteriology. Williams and Wilikins, Baltimore, USA. p786-792.
• Nelson, R.J. and Sommers, T.G. (1996). Environment. Saunders College Publishing,
Philadelphia. USA. p. 569.
• Saratale, R., Saghi, M.H., Ghafari, H.R. and Hadi, M. (2006). Biosorption of iron from
Aqueous solution by dried biomass of activated sludge. Iran Journal of Environmental
Health Science and Engineering, 6(2):107-114.
• Tanee, F.B.G and Kinako, P.D.S (2008). Comparative studies of biostimulation and
phytoremediation in the mitigation of crude oil toxicity on tropical soil. Journal of
Applied Science and Environmental Management, p. 143 – 147.