Comparative Study of the Protective Effect of Granulated Sugar and Garri Meal on Petroleum-induced Changes of Lipid Profiles in Albino Rats ( Rattusnorvegicus )

: Crude oil contaminated diet has been reported to induced changes in lipid profile of rats. In this study, 60 albino rats in 6 groups were fed with crude oil contaminated sugar-garri diets at varied concentrations to monitor petroleum-induce lipid profile changes using standard methods. The lipid profiles estimated were total cholesterols, triglycerides, high density lipoprotein cholesterol and Low density lipoproteincholesterol.The data showed that, the levels of total cholesterol (Mmol/l) increased from 2.89+0.08 in sugar fed rats compared with 3.05+0.12 in Gari fed rats. The concentration of triglycerides(Mmol/l) reduced from 1.10+0.51 in sugar fed rats to 1.06 + 0.03 in Gari fed rats while the high density lipoprotein cholesterol(Mmol/l) concentration of 2. 01+0.08 was significantly higher than 1.89 + 0.03 in Gari fed rats (P<0.05).There was no significant difference in Low density lipoprotein cholesterol (Mmol/l) concentrations of 0.68+0.11 in sugar fed rats compared with 0.83 + 0.10 in Gari fed rats. The effect of pre-treatment of the diet with sugar reduced petroleum-induce changes in lipid profile than gari during the exposure of the albino rats to the petroleum diet suggesting that sugarcould possibly ameliorate petroleum-induce changes in lipid profile as shown by the significantlowering of the cholesterol and Low density lipoprotein cholesterol than gari in albino rats.

Crude oil has been described as a complex mixture of over 6000 potentially different hydrocarbons and metal (Edwards, 1989) and the chemical composition varies between geologic formations (Coppock et al.,1995).Exposure of humans and animal to crude oil, which is increasing in terms of the environmental levels, and application to body, may be toxic.Crude oil is used in folkloric medicine in the Niger-delta area of Nigeria for the treatment of various ailments including stomach up-set, wound, and burns (Orisakwe, et al., 2000).The route of administration is mostly oral and external application for burns and wounds.The ingestion of petroleum hydrocarbon has been reported to induce oxidative stress (Val and Almeida-Val, 1999) through the generation of free radical (Achuba and Osakwe, 2003).It has been established that free radical generation with subsequent oxidative modification leads to lipid peroxidation (Halliwell, 1994) that damages critical cellular macromolecules such as DNA, lipids and proteins (Breimer 1990;Romero et al 1998;Souza et al, 1999); that results in inactivation of antioxidant enzymes (Pigeolet et al., 1990).Hydrocarbons and other constituents of petroleum and petrochemical products like other xenobiotics are metabolized in the liver to a greater extent (Sims, 1980).Achuba (2005) had reported that total cholesterol and LDLcholesterol were insignificantly increased while HDL-cholesterol and triglyceride significantly decreased in rabbits fed petroleum contaminated diet and the ingestion of crude petroleum contaminated diet imposed a reciprocal relationship between HDLcholesterol and LDL-cholesterol in the plasma of Rabbit.Nearly all of the energy needed by the human body is provided by the oxidation of carbohydrates and lipids.The sugar cane is one of the most important sucrose sources, containing until 20% weight sucrose (Glazer and Nikaido, 1995).Sucrose hydrolysis produces a fructose and glucose equimolar mixture named inverted sugar, which has higher edulcorant power.The inverted sugar is incorporated more easily in industrial preparations and has more added value than sucrose (Chou and Jasovsky 1993).Sugar has been reported to reduce haematoxicity caused by petroleum (Braide et al., 2011a).
Cassava was transplanted from Brazil to Africa by the Portuguese nearly three centuries ago (Bourdoux et al., 1980).Gari processing consisted of peeling, shredding, fermenting and dehydrating (light roasting) in sacks for 3-4 d, sifting, gelatinizing and roasting.Gari is made by grating cassava (Mannihot esculenta Crantz) and then dehydrating the fibre in sacks for 3 days after which the residue is roasted.
Cassava is a staple food in human diets in over 80 countries (Gomez et al., 1988).Gari a starchy food prepared from cassava (Mannihot esculenta Crantz)tubers is one of the most popular staple foods of the people of the rain forest belt of West Africa and contains mainly starch-20% amylase and 70% amylopectin having lost the soluble carbohydrates (i.e.glucose and sugar) during processing.Gari has been reported to reduce enzymes induction caused by petroleum through the phenomenon of glucose effect (Braide et al., 2011b).
The aim of this paper was to compare possible protective roles of sugar and gari diets against petroleum induced changes in lipids using cholesterol, triglycerides, high density lipoprotein cholesterol and Low density lipoproteincholesterol as indicators.Animal studies: Preliminary study was carried out by authors to ascertain the sugar and gari concentrations that will cause glucose effect by feeding rats with various concentrations of sugar and gari, then observing the concentration of sugar and gari with the lowest cholesterols concentration.The albino rats were fed contaminated diet mixed with 20% sugar and 20% gari adlibitum for three weeks, slaughtered and blood samples taken for analysis.The concentrations of crude oil used were 3.88, 7.75, 15.51, 31.01 and 62.02g/kg while the last group was fed rat diet with distilled water adlibitum to serve as control.Preliminary investigation had established that this concentration of crude oil was tolerable to the albino rats on a prolonged basis without any drastic effect.

MATERIAL AND METHODS
Biochemicalstudies: The cholesterol is determined after enzymatic hydrolysis and oxidation.The indicator quinoneimine is formed from hydrogen peroxide and 4-aminoantipyrine in the presence of phenol and peroxides (Allainet al 1974).
Ten microlitre (10 l µ ) of sample, control, standard and distilled water was pipette into respective test tube then 1000 l µ of cholesterol working reagent was added.It was mixed and incubated for 5 minutes at 37 0 C. The absorbance of the sample was measured against the reagent blank at 520nm.The concentration of sample was calculated using the absorbance of sample against absorbance of standard multiplied by concentration of standard.
The triglycerides are determined after enzymatic hydrolysis with lipases.The indicator is a quinoneimine formed from hydrogen peroxide, 4aminophenazone and 4-chlorophenol under the catalytic influence of peroxidase (Buccolo and David 1973).
Ten microlitre (10) l µ of sample, control, standard and distilled water was pipetted into respective test tube then 1000 l µ of triglyceride reagent was added.
It was mixed and incubated for 5 minutes at 37 0 C. The absorbance of the sample was measured against the reagent blank at 520nm.The concentration of sample was calculated using the absorbance of sample against absorbance of standard multiplied by concentration of standard.
Low density lipoproteins (LDL and VLDL) and chylomicron fractions are precipitated quantitatively by the addition of phosphotungstic in the presence of magnesium ions.After centrifugation, the cholesterol

RESULTS AND DISCUSSION
The cholesterol concentration for control in sugar treated albino rats was 1.64 +0.(Omoregie, 1998).The result showed a reduction in cholesterol concentration of sugar fed compared with gari fed.This suggests that feeding on 20%sugar tend to reduce cholesterol in petroleum treated rats than gari treated rats.Achuba (2005) had reported that feeding rabbits with diets containing various concentrations of crude petroleum caused significant dose related increases in cholesterol suggesting that ingestion of petroleum contaminated diet could predispose humans to cardiovascular diseases while the result of this study has shown that sugar diet reduced cholesterol concentration more than gari diet.Adegoke et al. (2013) reported that feeding sugar and gari can cause synthesis of total and HDL cholesterols but the increase is more pronounced in gari than sugar.Braide et al (2011c) in their study reported that sugar diet reduced enzymes induction in petroleum treated rats more than gari.This was attributed to cyanogenic component present in gari.Ezeji et al (2009) had suggested that cyanide affects some important enzymes of the mitochondtrial electron transport system which are used as markers of the organelle.(2005) reported significantly decreased triglyceride in rabbits fed petroleum contaminated diet but the result of this study showed that sugar fed rats increased triglycerides more than the gari fed rats.In humans, hypertriglyceridemia can be induced endogenously by a high fat diet (Austin et al., 1998) or it can be carbohydrate induced (Knittle andAhrens 1964, Reavenet al 1965).A high triacylglycerol (TAG) flux rate was observed in subjects fed HCLF diet (Abbott, et al., 1990).The TAG increase observed in humans on a HCLF diet may have been from de novo lipogenesis (Parks, and Hellerstein 2000).Ben- David et al (2001) reported that the ingestion of petroleum caused reduction in blood glucose causing shift in the demand for metabolic substrate to lipid, the supplementation of the diet with sugar and gari reversed the reduction in blood glucose hence the shift in demand for lipid will be reversed causing increase triglycerides.The increase in triglycerides concentrations of sugar more than gari might be as a result of some other components like cyanogens present in gari.(2005) had reported that the ingestion of crude petroleum contaminated diet imposed a reciprocal relationship between HDL-cholesterol and LDLcholesterol in the plasma of Rabbit.The decrease in HDL-cholesterol with a corresponding increase in LDL-cholesterol is the primary risk factor for coronary heart disease (Mckee andMckee, 1999, Glew, 1997).Achuba (2005) Comparative Study of the Protective Effect of Granulated Sugar ADEGOKE, O. ADEBAYO; BAMIGBOWU, E. OLUGBENGA; GEORGE-OPUDA M. IBITOROKO; BRAIDE, A. SOLOMON; OKEKE, U. CHUKWUEBUIKE; EKWUSA V. OBIAGERI Effect of Gari and Sugar on cholesterol and triglycerides in albino rats treated with petroleum ADEGOKE, O. ADEBAYO; BAMIGBOWU, E. OLUGBENGA; GEORGE-OPUDA M. IBITOROKO; BRAIDE, A. SOLOMON; OKEKE, U. CHUKWUEBUIKE; EKWUSA V. OBIAGERI Table 1

Table 2
Effects of gari and sugar on HDL cholesterol and LDL cholesterol in albino rats treated with petroleum Packed cell volume (PCV) and white blood cell count (an indicator of possible blood damage) but supplementation of the diet with 20% gari further decreased the haemoglobin (Hb), Packed cell volume (PCV) and white blood cell count observed in petroleum fed albino rats.Therefore, this study has shown that sugar diet appeared to ameliorate petroleum-induced changes in lipid profile compared with gari.This may as a result of OBIAGERI cyanogens present in gari which is absent in granulated sugar used in this study.

Table 3
effect of Sugar and Gari on lipid concentration in albino rats treated with petroleum Conclusion:This study has shown that sugar diet appeared to ameliorate petroleum-induced changes in lipid profile compared with gari.The effect of pretreatment of the diet with sugar reduced petroleuminduce changes in lipid profile than gari during the exposure of the albino rats to the petroleum diet suggesting that sugar could possibly ameliorate petroleum-induce changes in lipid profile as shown by the significant lowering of the cholesterol and Low density lipoprotein cholesterol than gari in albino rats.