This study, “ACTIVITIES OF CYTOCHROME OXIDASE IN THE RAT LIVER OF NORMAL AND PROTEIN UNDERNOURISHED RATS FED WITH DIET CONTAINING CASSAVA PEEL” contains concise information that will serve as a framework or guide for your project work. The project study is well-researched for academic purposes and are usually provided in complete chapters with adequate References. Keywords: ACTIVITIES OF CYTOCHROME OXIDASE IN THE RAT LIVER OF NORMAL AND PROTEIN UNDERNOURISHED RATS FED WITH DIET CONTAINING CASSAVA PEEL
TABLE OF CONTENTS Abstract CHAPTER ONE 1.0 Introduction 1.1 Aim of Study CHAPTER TWO 2.0 Literature review 2.1 Cassava peel 2.2 Cyanide 2.3 Respiratory Electron Transport Chain 2.3.1 The sequence of electron transport chain 2.3.2 Mitochondria 2.4 Cytochrome oxidase 2.5 Protein undernutrition 2.6 Liver Synthesis 24 2.6.2 Liver disease and blood supply 27 CHAPTER THREE 3.0 Materials and Methods 3.1 Materials 3.1.1 Chemicals and reagents 3.2 Methods 3.2.1 Animal managements 0 3.2.2 Feed composition 31 3.2.3 Body weight determination 32 3.2.4 Animal sacrifice 32 3.2.5 Isolation of mitochondria 33 3.3 Determination of complex IV (Cytochrome oxidase activity 34 3.4 Protein determination CHAPTER FOUR 4.0 Results CHAPTER FIVE 5.0 Discussion and Conclusion 5.1 Discussion 5.2 Conclusion References Appendix LIST OF FIGURES Figure 1 – The growth curve for normal and protein undernourished rat fed with diet containing cassava peel Figure 2 – The activities of cytochrome oxidase in normal and protein undernourished rat LIST OF TABLES Table 1 – Animal group Table 3- Showing organ weight to body weight ratio and feed Consumption ABBREVIATION ADP/ATP Adenosine 5-diphosphate/ Adenosine 5 triphosphate BSA Bovine serum albumin COX Cytochrome oxidase CuSO4 Copper sulphate EPA Environmental protection agency ETC Electron transport chain HCN Hydrogen cyanide LP Low protein Na2CO3 Sodium carbonate NaOH Sodium hydroxide NP Normal protein PU Protein undernutrition ABSTRACT The study was carried out to investigate the activities respiratory complex IV in the liver of normal and protein undernourished wistar rats fed with cassava peel. Twenty weaning male rats were grouped into four groups, five rats for each. Group A were fed with normal diets, Group B were fed with low protein diets containing 5% casein, Group C were fed with normal diets and coconut oil and Group D were fed with low protein diet and coconut oil. After feeding for a period of six weeks, the rats were sacrificed using cervical dislocation. The liver was harvested and homogenized in iced cold buffer and centrifuged at 4000rpm for 15 minutes. The pellets were discarded and the supernatant was stored. It was observed from this study that consumption of low protein diets significantly reduced the activities of cytochrome oxidase in the rat liver when compared to normal diet. However, the activities of cytochrome oxidase were significantly reduced in the rat liver when the rats were fed with normal and cassava peel diets compared to normal diet only. Also, low protein and cassava peel diets significantly reduced the activities of cytochrome oxidase in the rat liver when compared to normal diet. It was observed from this study that in the rat liver, the activities of cytochrome oxidase was significantly increased when the rats were fed with normal and cassava peel diets compared to low protein diet only. However, when the rats were fed with low protein diet and cassava peel, the diets significantly increased the activities of cytochrome oxidase compared to both normal and cassava peel and low protein diet only. This study shows that consumption of cassava peel significantly reduced the level of normal diets while it significantly increased the level of low protein diets. CHAPTER ONE 1.0 INTRODUCTION Cassava is the source of dietary food energy for the majority of the people living in the lowland tropics, and much of the sub-humid tropics of West and Central Africa (Tsegia et al, 2002). Therefore, its production and utilization must be given prime attention in food policy. Even though farmers have not yet attained the desired technical efficiency in cassava production as a result of weak access to external inputs such as fertilizers and herbicides (Ezedinma et al, 2006), the widescale adoption of high yielding varieties and the resulting increase in yield have shifted the problem of the cassava sector from supply (production) to demand issues, such as finding new uses and markets for cassava. Estimates of industrial cassava use in Nigeria suggest that approximately 16 percent of cassava root production was utilized as chips in animal feed, 5 percent was processed into a syrup concentrate for soft drinks and less than 1 percent was processed into high quality cassava flour used in biscuits and confectionery, dextrin, adhesives, starch, and hydrolysates for pharmaceuticals and seasonings (Ene, 1992). Cassava roots provide an ideal raw material for many of these types of food products, since they are easy to process and have a bland flavour. However, fresh cassava roots are bulky and costly to carry and, in addition, are likely to rot within a few days of harvesting. Cassava roots contain cyanide, which is poisonous. Different varieties of cassava contain different amounts of cyanide. Roots that contain high amounts of cyanide normally taste bitter and should not be eaten raw, while the roots of ‘sweet’ varieties contain low levels of cyanide and can be eaten raw. Cassava peels derived from garri processing are normally discarded as wastes and allowed to rot in the open, thus resulting in health hazards. As a rough estimate, about 10 million tonnes of cassava are processed into garri annually in Nigeria alone. Since these peels could make up to 10% of the wet weight of the roots, they constitute an important potential resource for animal feeds if properly processed by a bio-system (Antai and Mbongo, 1994). The peels contain toxic levels of cyanogenic glycosides, while the liquid contains a heavy load of microorganisms capable of hydrolyzing the glycosides. The resulting products of fermentation of cassava peels with squeezed out water can be dried and used as animal feeds (Tweyongyere and Katongole, 2002). Cyanide originates primarily from anthropogenic sources in the environment, but cyanide is also released from biomass burning, volcanoes, and natural biogenic processes from higher plants, bacteria, and fungi (ATSDR, 2006). Cyanide is also a component of tobacco smoke and can be present at high concentrations in structural fires (Steinmaus et al., 2007; Brauer et al., 2006; Tsuge et al., 2000; Brandt-Rauf et al., 1988). Cyanide compounds are used in a number of industrial processes, including mining, metallurgy, manufacturing, and photography, due to their ability to form stable complexes with a range of metals. Cyanide has been employed extensively in electroplating, in which a solid metal object is immersed in a plating bath containing a solution of another metal with which it is to be coated, in order to improve the durability, electrical resistance, and/or conductivity of the solid (Brandt-Rauf et al., 1988). Cytochrome c oxidase (COX), transfers electrons from cytochrome c to molecular oxygen and pumps protons across the inner mitochondrial membrane. It is comprised of thirteen subunits: the 3 largest are encoded by mtDNA and the other 10 by nDNA (Manfredi et al., 1995). The mtDNA-encoded subunits have two heme containing cytochrome prosthetic groups (cytochromes a and a3), as well as three copper atoms (located in the CuA and CuB sites). Although isolated COX deficiency due to mutations in mtDNA-encoded genes has been associated with myopathies and multisystemic disease, no pathogenic mutations in the nuclear encoded subunits of COX have been found (Michel et al., 1998). 1.1 Aim of study The aim of this study is to investigate the activities of respiratory complex IV in the rat liver of normal and protein undernourished rat fed with diet containing cassava peel Objectives · To isolate mitochondrial and prepare crude homogenate from the liver tissue · To compare the effect of normal and protein undernourished rats fed with diet containing cassava peel
Keywords: ACTIVITIES OF CYTOCHROME OXIDASE IN THE RAT LIVER OF NORMAL AND PROTEIN UNDERNOURISHED RATS FED WITH DIET CONTAINING CASSAVA PEEL
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