INFLUENCE and histological analysis were carried out after


Achuba ,F. I .
Department of Biochemistry, Delta State University, PMB 1, Abraka Nigeria. [email protected]

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The toxicity of petroleum hydrocarbon across the
living systems is now a common knowledge among the scientific community. What
is lacking is a mini-scale antidote that can be adopted by the inhabitants of
crude oil producing areas of the world. This was the reason for this study. The
study is comprised forty eight female rats divided into six groups of eight
rats each. Rats in control group were fed with diet without any treatment while
rats in groups 2 and 3 were fed with diets treated with known amount of Elaesis guineensis leaf respectively.
Rats in group 4 were fed with crude oil contaminated diet. Rats in groups 5 and
6 were fed with contaminated diet mixed with known amount of ground Elaesis guineensis leaf. Biochemical and
histological analysis were carried out after three and six months respectively.
The results show that pretreatment of crude oil contaminated diet with Elaesis guineensis leaf tend to restore values of lipid peroxidation, xanthine oxidase
activity, superoxide dismutase activity and catalase activity close to control
values. Histological examination indicates
protective effect of Elaesis guineensis leaf against deleterious effect of
crude oil on the kidney.   Thus, it is pertinent to state that
there exist potentials in the use Elaesis
guineensis leaf in the treatment of crude oil toxicity. And indeed setting
a fresh agenda for further serious scientific investigations




Keywords: Catalase, Crude oil, Kidney, Lipid
peroxidation, Elaesis guineensis,
Superoxide dismutase .Xanthine oxidase,




 1.0 Introduction

 Humans and animals get exposed to crude oil or
its byproducts when these chemicals are released into the surroundings during
oil exploration activities, equipment failures, corrosion, illegal bunkering,
usage, oil theft and illicit refining 1-3. Crude oil stimulates oxidative
stress in animals 4, 5. Lipid peroxidation, xanthine oxidase superoxide
dismutase (SOD) and catalase activities are part of oxidative stress indices
6. Lipid peroxidation elicits oxidative damage in plants and animals and its
value in conjunction with alterations in the level of antioxidants represent a
measure of oxidative stress. Similarly, the activity of xanthine oxidase is a
defense mechanism as well as measure of oxidative stress in animals 6. Report
has it that the deleterious action of crude oil on the kidney is based on
oxidative stress 7.

 Byproducts of the Elaesis guineensis tree are
important medicinally. This is because the leaf juice have wound healing
property while the sap is used as laxative 8.This is due to   compounds rich in medicinal and antioxidant
properties inherent in Elaesis guineensis
leaf 9, 10.  The antioxidant action is
attributed to the presence of phytochemicals (flavonoid, tannin and phenols) in
the leaves of Elaesis guineensis tree
11. In fact, Elaesis guineensis leaf extract contains more antioxidative
phenolic compounds than various green tea extracts 12. Therefore, Elaesis
guineensis leaf extract is a potential source of functional food ingredient,
based on reports of its health benefit 13 .This study is aimed at evaluating
the protective potentials of Elaesis
guineensis leaf against crude oil contaminated diet induced nephrotoxicity
in rats.

2.0 Materials
and methods

The crude oil used for this study was obtained from
Nigeria National Petroleum Corporation (NNPC) Warri, Delta State, Nigeria. The
palm leaf used was obtained from Elaeis guineensis tree in Obiaruku,
Delta state, Nigeria Forty eight (48) female albino wistar rats with weights
ranging from 0.088kg to 0.182 kg obtained from the animal house of Department
of Anatomy, Delta State University, Abraka, Nigeria were used for this study.
The rats were housed in a standard wooden cage made up of wire gauze, net and
solid woods and left to acclimatize for one week on grower’s marsh and tap
water at laboratory temperature of 28o C and
12 hour day/ night regime. After the acclimatization period, the rats
were weighed and grouped.

2.1 Preparation
of leaf powder.

leaves of Elaeis guineensis were isolated from the stock and
sun- dried. The dried leaf was then ground with domestic kitchen blender into a
fine powder and stored in a clean and sealed plastic container

2.2 Treatment
of animals

The forty eight (48) female albino wistar rats were
assigned to six (6) groups according to their weights, with eight rats in each
group. Rats in the control, Group 1 were fed with grower’s marsh only. Rats in
Group 2 were fed with grower’s marsh treated with 5g of powdered Elaeis
guineensis leaf. Group 3 rats were fed with grower’s marsh treated 10g of powdered
Elaeis guineensis leaf. Group 4 rats were fed with grower’s marsh
contaminated with crude oil (4ml per 100g of feed).This
concentration of crude oil in diet was established to be tolerated by the rats
over a long period in a preliminary study. Rats in Group 5 were fed
grower’s marsh contaminated with crude oil (4ml per 100g of feed) plus 5g of
powdered palm fronds. While rats in Group 6 were fed with crude oil
contaminated marsh (4ml per 100g of feed) plus 10g of powdered palm leaves. The
rats in each group were allowed access to clean drinking water while the
experiment lasted. The feeds were prepared fresh daily and stale feed remnants
were discarded regularly. This was done every morning
between the hours of 8 am – 9 am and each group provided with 400 g of the
respective diet. The animals in each group were exposed to their
respective diets for three and six months respectively. The National Institute of health guide for the care and use
of laboratory animals (NIH, 1985) was adhered to in the course of the



2.3 Collection
of samples

After three months, four rats were sacrificed in each
group and the kidneys collected. Five grams (5.0 g) of the kidneys were weighed
in chilled conditions and homogenized with 5ml of normal saline in a mortar. The mixture was diluted with 45 ml of buffered saline (pH
7.4) before it was subjected to centrifugation at 2, 500 rpm and the
supernatant was transferred into plastic tubes and stored at – 4o C in the refrigerator before
used for analysis within forty eight hours. This same procedure was adopted
after six months exposure period.

2.4 Determination of lipid peroxidation and xanthine oxidase activity

The activity of
xanthine oxidase in the kidney of rats was measured using the method of
Bergmeyer et. al. 14, a reaction based on the oxidation of xanthine to uric
acid, a molecule that absorbs light maximally at 290 nm. A unit of activity is
that forming one micromole of uric acid per minute at 25oC. Lipid
peroxidation in the kidney of rats was measured by the thiobarbituric acid
reacting substances TBARS, method of Gutteridge and Wilkins 15.Total
superoxide dismutase activity was assayed using the method of Misra and
Fredorich 16. Catalase was assayed as reported by Rani et al. 17  

2.5Statistical Analysis

of variance (ANOVA) and post Hoc Fisher’s test for multiple comparison
were carried out using version 20 of   statistical package for social science (SPSS) to
determine statistical significant differences between means. P values