NUTRIENT AND BIOACTIVE POTENTIALS OF Sphenostylis stenocarpa and Glycine max AND THEIR EFFECTS ON PERSISTENT PHYSICALLY INDUCED STRESS OF ADULT MALE WISTAR RATS

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TABLE OF CONTENT

TITLE PAGE…………………………………………………………………… i                 APPROVAL PAGE……………………………………………………………….. ii CERTIFICATION……………………………………………………………………. iii DEDICATION……………………………………………………………….….iv ACKNOWLEDGEMENT…………………………………………………………..…v LIST OF TABLES……………………………………………………………….……..xi         LIST OF FIGURES.………….……………xii                              LIST OF PLATES……………………………………………………….xii ABSTRACT……………………………………………………………..……xiii

CHAPTER ONE: INTRODUCTION

1.1: Background to the Study……………………..……………………… 1

1.2 Statement of the Problem………………………………………………… 4

1.3 Broad Objective of the study…………….……………………………………. 6

1.4   Significance of the Study ……………….……………………… 7

CHAPTER TWO:  LITERATURE REVIEW

2.0: Concept of African Yam Bean and Soy Bean………………………………… 9

2.1:   Classification African Yam Bean and Soy Bean….…………………… 11

2.1.2: African Yam Bean……………………….………………. 11

2.1.3: Soy Bean…………………………………………..…………… 11

2.2:  Potentials of African Yam Bean and Soy Bean……………….………… 12 

2.2.1: African Yam Bean…………………………………………..…….. 12

2.2.2: Soy Bean…………………………………………………………….. ….15

2.3:   Anti-Nutritional Factors………………………………………………..… 16

2.3: Antioxidants in Fruits and Vegetables…………………………….. …17

2.4: Antioxidants in Legumes……………………………………………….. 18

2.5: Physical Stress……………………………………………………….. 20

2.6: Chemistry of Stress………………………….…………………………… 22

2.7: Antioxidants……………………………………………………………………. 23

2.7.1: Effects of Antioxidant………………………………………………… ….24

2.8: Enzymatic Antioxidant Parameters…………………………………… 26

2.8.1: Lipid Peroxidation…………………………………………………. 26

2.8.2: Catalase Activity……………………………………………………….27

2.8.3: Superoxide dismuthase…………………………………………   27

2.8.4: Glutathione proxidase………………………………………………….. ..27

2.9: Non-Enzymatic Antioxidant Parameters…….……………………………. 29

2.9.1: Vitamin A……….…………………………………………………………. 29

2.9.2: Vitamin C…….………………………………………………………………. 29

2.9.3: Vitamin E…….…………………………………………………………… 30

2.9.4: Glutathione….……………………………………………………. 30

2.10:   Vitamins………………….……………………………………………….. 31

2.10.1:   Ascorbic Acid……………………….……………………………….. 32

2.10.2:    Vitamin E………………………….………………………………… 32

2.10.3:   Vitamin A……………………………………………………………… 33

2.11:     Minerals…………………………………………………………….. 33

2.11.1:   Iron………………………………………………………………….. 34

2.11.2:   Magnesium………………………………………………………….. 34

2.11.3:  Calcium………………………………………………………………… 34

2.11.4: Selenium……………………………………………………………… 35

2.11.5: Zinc……………………………………………………………………… 36

2.11.6: Copper………………………………………………………………… 37

2.12:   Phytochemistry…………………………………………………………….. 38

2.13:    Phytochemical Constituents in Plants………………………….. 39 

2.13.1: Alkaloids……………………………………………………….. 39

2.13.2: Flavonoids……………………………………………………………… 40

2.13.3: Tannins……………………………………………………………………… 40

2.13.4: Steroids………………………………………………………………… 41

2.13.5: Saponins……………………………………………………………… 41

2.13.6: Terpenoids……………………………………………………………… 41

Abstract

This study was carried out to examine the efficacy of roasted Sphenostylis stenocarpa and Glycine max flours on persistent physically induced stress in wistar albino rats. Thirty two (32) male wistar rats weighing between 130-192g were distributed into eight (8) groups of four (4) rats each to acclimatize to feed and environment. Six hundred (600g) of S. stenocarpa and G. max used in this study were purchase from local markets in Enugu State. Group 1 to 6 were induced physical stress persistently for 2 hours in week 1, and 3 hours in week 2 daily. Group 1 to 3 and groups 4 to 6 were treated with roasted 500mg/kg bw, 1000mg/kg bw and 2000mg/kg of S. stenocarpa and G. max flours respectively. Group 7 (stress control) and group 8 (normal control) were not treated but were fed with feed and water ad libitum. Chemical analysis was done to determine the proximate, phytochemical, minerals, vitamins and antinutrient compositions of the samples. Blood samples were collected from the animals on the 7th and 14th day through ocular puncture. They were used to determine enzymatic and non enzymatic antioxidants parameters. Data obtained from the study were subjected to statistical analysis and the results were presented as mean and standard deviation. Differences between mean were determined by ANOVA and pos hoc multiple comparisons. Results showed that Sphenostylis stenocarpa and Glycine max had protein (22.29% and 36.79%), fibre (12.70% and 3.50%), and carbohydrate (60.5% and 22.62%) respectively. Phytochemical and antinutrient compositions of the legumes contained bioactive chemical substances with strong presence of alkaloids (29.76mg.100g-27.98mg/100g), flavonoids (32.68mg/100g-22.03mg/100g), glycosides (0.88mg/100g-0.87mg/100g), tannins (0.53mg/100g-296.63mg/100g) and phytates (0.53mg/100g-38.30mg/100g) respectively. Pro-vitamins A (87.75mg/100g-207.67mg/100g), vitamin C (272.87mg/100g-38.30mg/100g) and vitamin E (81.60mg/100g-276.01mg/100g) ranges were detected in both samples. The mineral contents indicated the presence of Mg (83.43mg/100g-82.87mg/100g), Fe (16.53mg/100g-7.64/mg/100g) Ca (72.03mg/100g-263.20/mg/100g) in both samples while Se (12.60), Zn (0.80) and Cu (42.30) were found to be present in S. stenocarpa. A significant (p < 0.05) decrease of 0.55mg/dl-0.82mg/dl was observed in the serum malondaidehyde (MDA) concentration of rats in the treated groups when compared to the control groups (1.21mg/dl-1.35mg/dl) while serum catalase (1.74mg/dl – 4.21mg/dl) and superoxide dismutase (1.07 IU/L – 1.133 IU/L) of the treatment groups differed significantly (p > 0.05) compared to control groups (1.06 IU/L -1.08 IU/L). The gluthatione peroxidase (GPx) activity in all the treated groups had a significant (p < 0.05) change. There was increase of 0.47mg/dl – 0.52mg/dl and decrease of 0.36mg/dl – 0.43mg/dl of Vitamin C activity in the groups treated with S. stenocarpa and in the group treated with G. max,respectively. A significant (p < 0.05) decrease was observed in vitamin E (0.06mg/dl – 0.08mg/dl) activity in all the treated groups compared to control groups (0.08mg/dl – 0.09mg/dl). There was an increase of 0.54mg/dl – 1.77mg/dl in vitamin A activity in all the treated groups when compared to stress group (0.27mg/dl). The glutathione (GSH) activity in all the experimental groups differed. Apart from stress control group that had decrease percentage change (0.04%) in body weight, all the other groups of the experimental rats had increase percentage change (1.11- 6.90%) in body weights. The antioxidant contents of S. stenocarpa and G. max were able to reduce behavioral changes in rats that were physically stressed after treatment. The results of antioxidant enzymes showed that these legumes could be used to scavenge free radicals in the system which often leads to risks of various diseases associated with persistent physical stress.

Key words: Bioactive potentials, Sphenostylis stenocarpa, Glycine max, Antioxidants, Induced stress.

CHAPTER ONE

INTRODUCTION

1.1: Background to the Study

African yam bean (Sphenostylis stenocarpa)and soy bean(Glycine max) belong to the legume family. Leguminous plants play important role in human nutrition. They provide a significant amount of food in developing countries (Okaka, Akobundu & Okaka, 2002). Leguminous seeds are important source of protein energy and other nutrients in the diet of large population groups and the world. They form an excellent source of thiamine and contributing appreciable quantities of the other water soluble vitamin. African yam bean is mainly cultivated in West, East, Central Africa, and in Malawi and Zimbabwe (Utter, 2007). In Nigeria it is locally known as “odudu” in Abia State, “ekpakpani” in Urhobo, Delta state; “illoloegwa” in Edo state (Nwosu, Onwuamanam, Onuegbu, Ogueke & Ojukwu, 2012) and ijiriji and Azamu in Nsukka and Ezeagu local government areas of Enugu state respectively.

Soy bean protein has increased attention in recent years among consumers, and researchers (Asif & Acharya, 2013). Botanically, soy bean belongs to the order Rosaceae, family Leguminosae, the genus Glycine and the cultivar Glycine max. African yam bean is one of the leguminous crops that contribute about 18% of the world protein requirements (Olawumi et al., 2013). The deficiency of protein in the diet cause a disease known as “kwashiorkor” a condition characterized by depigmentation of the hair in children and oedema. In order to bridge the protein supply gap that exists today, scientists have focused their attention on the lesser known protein crops among which is the African yam bean (Okaka & Okaka, 2001). Legumes are cheaper than animal products such as meat, fish, poultry and egg. They are consumed world wide as a major source of cheap protein and, especially in the developing countries where consumption of animal protein may be limited as a result of economic, social, cultural or religious factors.

Adebowale and Sanni (2009) regarded African yam bean as underutilized crop due to its low esteem and lack of detailed information on its compositional analysis. The seed is highly prized food legume in south eastern Nigeria (Asoiro & Ani, 2011) owing to high crude protein content. It ranks well among neglected crops and can contribute to food security (Adewale and Odo, 2013). The dry seeds when cooked are very delicious and it can be roasted and eaten with palm kernel as snacks or boiled and eaten with local seasoning, even converted to paste for the production a local type of moi moi. In West Africa, they are often preferred over other grain legumes (Obizoba & Nnam, 1992 and Okpara & Omaliko, 1995). There is a wide spread use of Africa yam bean in Nigeria diet, especially in southern states (Obizoba & Nnam, 1992). Nwokolo (1996a) reported that the Igbo people extensively explored the crop as a good source of dietary protein in feeding the displaced and severely malnourished refugees during the Nigerian civil war. The seed is one of the organs of economic importance providing food for human and livestock (Potter, 1992 & Nwokolo, 1996b). Nutritionally, African yam bean and soy bean are used extensively in various dietary preparations and has potentials for supplementing the protein requirement of many families throughout the year (Ajayi, 2011).

African yam bean and soy bean are important ingredients of a balanced human diet in many parts of the world due to their high nutritive contents. The rapidly growing of food industry, which constantly demands new ingredients, has drawn researchers to legume components (bioactive compounds) (Adebowale and Sanni, 2009). The increasing demand for plant-based diet in lieu of expensive animal based- protein have been expounded by several researchers (Adebowale and Sanni, 2009). Therefore, there is the need to intensify research efforts aimed at identifying new legume utilization. A reappraisal of beneficial effects of legume seed dietary intake is the basis for various health claims. The food components involved in health claims are numerous: lipids, vitamins, oligosaccharides, minerals, fibers, flavonoids, small organic compounds and less frequently, proteins and peptides (Esan, & Fasasi, 2013). Many of these biologically active compounds originate from the plant kingdom, to which a safe and healthy attribute has traditionally been associated to legumes. In most cases the ability to prevent disease is based on rather generic effects such as antioxidant, anti-inflammatory, anti-ageing, detoxification and antigenicity. Bioactive compound in African yam bean and soy bean could be used in the formulation of functional foods and nutraceuticals to prevent damage related to oxidative stress in human diseases. Studies (Klu, Amoatey, Bansa & Kumaga, 2001) have shown that the lesser known legumes such African yam bean together with other convectional legumes can be used in combating malnutrition. Therefore, research on such crops merit significant consideration.

Stress is defined as the force applied to a given area of biological tissue. Stress has been shown to decrease impairments, functional limitations, disability, and pain in a variety of patient populations (Brown & Hollosz, 1991). When one feel threatened, nervous system responds by releasing a flood of stress hormones, including adrenaline and cortisol, which rouse the body for emergency action. During stress response, heart rate increases, blood pressure rises, breathing quickens and muscles tighten. These physical changes increase strength and stamina, speed reaction time, and enhance focus. Study has shown a negative effect stress has on the immune system (Kalat, 2013). Stress affects the immune system in many ways. The immune system protects the body from viruses, bacteria, and anything that the body does not recognize. The immune system sees these as intruders and it sends messages to attack. The white blood cells, leukocytes, are very important to the immune system (Kalat, 2013). 

1.2 Statement of the Problem

Antioxidants prevent free radicals induced tissue damage by preventing the formation of radicals, scavenging them or by promoting their decomposition. Free radicals damage contributes to the etiology of many chronic health problems such as cardiovascular and inflammatory disease, cataract, anemia and cancer. The search for effective, non-toxic natural compounds with antioxidant activity has been intensified in recent years. In addition to endogenous antioxidant defense system, consumption of dietary and plant derived antioxidants appears to be a suitable alternative. Dietary and other components of leguminous plants form major sources of antioxidants (Lobo, Patil, Phatak & Chandra, 2010).

            Diet and lifestyle choices are the major factors contributing to the risk of stress and diseases which is responsible for more death. A diet based approach is required to abolish stress and health problems which will secure a solution against the existing problem. Recent studies suggest that plant based diet may be more effective in the treatment and management of the chronic health problems (Lobo et al., 2010). In order to bridge the gap of research in roasted African yam bean and soy bean flours and arrest the limited information on the stress and chronic health problems; the present study was carried out to investigate the nutrient and bioactive potentials of Sphenostylis stenocarpa and Gycine max and their effects on persistent physically stress induced adult male wistar rats.  During each day of individual’s life, an individual is exposed to oxidative stress. A dietary ingredient derived from plant-based diet has been confirmed to relieve stress (Milesi, Lacan, Brosse, Desor and Notin, 2009). Their study show that plant based diet rich in antioxidant.

NUTRIENT AND BIOACTIVE POTENTIALS OF Sphenostylis stenocarpa and Glycine max AND THEIR EFFECTS ON PERSISTENT PHYSICALLY INDUCED STRESS OF ADULT MALE WISTAR RATS