IDENTIFICATION AND EVALUATION OF IMMUNO-MODULATORY PROPERTIES OF PREBIOTICS EXTRACTED FROM VERNONIA AMYGDALINA
ABSTRACT
Prebiotics are non-digestible food substances that when consumed provide a beneficial physiological effect on the host, by selectively stimulating the favourable growth or activity of a limited number of indigenous bacteria. Most known prebiotics are complex carbohydrates commonly found in fruits, vegetables and plant products. Vernonia amygdalina, commonly called bitter leaf is widely consumed in Africa especially in Nigeria. The leaf extract has been reported to contain some prebiotic potential.
This study was aimed at identifying and evaluating immuno-modulatory properties of prebiotics extracted from the leaf. The aqueous extract was separated into fractions by Liquid-liquid fractionation and the prebiotic compounds were identified through Thin Layer Chromatography (TLC).The evaluation of immuno-modulatory effects was conducted by monitoring the effects of oral administration of the aqueous extract on the haematological and lipid profiles of Rattus norvegicus. The animals were distributed into six groups with six animals in each group. One group was designated control while the other five groups were administered different concentrations of the extract. The control group was administered distilled water and the other groups 50, 100, 200, 400 and 800 mg/kg of the extract respectively, twice daily for three weeks. Blood was collected through the orbital-plexus of the rats to evaluate the effect of the extract on CD4 cells, packed cell volume (PCV), white blood cell count (WBC), haemoglobin (Hb) levels and lipid profile. Five fractions designated E1 to E5, were obtained from the extract.
Thin Layer Chromatography analysis of these fractions revealed the presence of fructo-oligosaccharides namely D-fructose, D-arabinose, D-glucose, D-xylose, D-mannose and D-galactose suggesting that these are the prebiotic compounds of the plant extract. The aqueous extract of the plant showed a significant (p<0.05) increased effect on White blood cells (WBC), CD4 cells, High Density Lipid- Cholesterol (HDL-C) and Total Cholesterol (TC). There was significant (p<0.05) reduction on Low Density Lipid-Cholesterol (LDL-C) and no significant (p>0.05) increased effect on haemoglobin (Hb) and packed cell volume (PCV) levels. The results of this study show that Vernonia amygdalina leaf extracts contain fructo-oligosaccharides and other prebiotic compounds and can play important roles in the modulation of the immune system.
CHAPTER ONE
1.0 Introduction And Literature Review
1.1 Introduction
Vernonia amygdalina, a member of the Asteraceae family, is a widely used local vegetable in Nigeria, Uganda and other African countries. It grows in a range of ecological zones in Africa and the Arabian Peninsula (Bonsi et al., 1995). The leaf is commonly called “bitter leaf” in English, “Olubu or Onugbo” in Igbo, “Ityuna” in Tiv, “Oriwo” in Edo, “Ewuro” in Yoruba, “Chusar-doki” in Hausa, “Etidot” in Efik, “Omubirizi” or “Omululuza” in West and Central Uganda; “Olusia” in Luo, Kenya. It is used in various food preparations and in ethnomedicine for the treatment of malaria and gastrointestinal infections. It is a shrub of 2-5 m tall with petiolate leaves of about 6.0mm wide (Ojiako and Nwanjo, 2006). It is up to 20 cm long and its bark is rough. The bitter taste of the leaf has been attributed to the presence of anti-nutritive principles like saponins, alkaloids, tannins and glycosides (Buttler and Bailey, 1973). There have been several reports on its antimicrobial, antiplasmodial, antitumor, antioxidant and antihelminthic properties (Jisaka et al., 1993; Izevbigie, 2003; Farombi, 2003; Ehiagbonare, 2007). Aqueous leaf extracts of V. amygdalina have been previously reported to have prebiotic properties (Ukwah and Ezeonu, 2008; Ezeonu and Ukwah, 2009; Ezeonu et al., 2012).
For many years, medicine depended exclusively on leaves, flowers and barks of plants, until the 1970s when synthetic drugs came into use (Conway, 1973). In orthodox medicine, a plant may be subjected to several chemical processes before its active ingredients are extracted, while in traditional medicine, a plant is simply eaten raw, cooked or infused in water or native wine or even prepared as food (Sofowora, 1992; Jones, 1996; Reynolds, 1996). Plant extracts are potential sources of novel antimicrobial compounds (Lis-Balchin and Deans, 1997) especially against bacterial pathogens, but it is necessary to investigate those plants scientifically which have been used in traditional medicine to improve the quality of healthcare. Plants extracts are usually composed of many phytochemical constituents.
Prebiotics are non-digestible substances that when consumed provide a beneficial physiological effect on the host, by selectively stimulating the favourable growth or activity of a limited number of indigenous bacteria (Roberfroid, 2007; Mitchell, 2010). These prebiotic substances survive digestion in the stomach and reach the colon where they are metabolized by the bacteria, thereby directly providing the host with energy and metabolic substrates (Wang and Gibson, 1993; Cummings et al., 2001). Efficient prebiotics usually have a specific fermentation in the colon and have the ability to alter the faecal microflora composition towards a more beneficial community structure (Kolida et al., 2002; Chakraborti, 2011). Thus, prebiotics exert their beneficial effects on the host indirectly; by stimulating functions of the intestinal microflora. These include regulation of bowel function, modulation of gut immunity, inhibition of pathogens and anticarcinogenic properties.
In addition to these, prebiotics also have the advantage of relative ease of manufacture because they can be either directly extracted from natural sources or be produced by partial acid or enzymatic hydrolysis of polysaccharides or by transglycosylation reactions (Macfarlane et al., 2006; Falcao-e- Cunha et al., 2007).
The role of prebiotics in modulation of bowel function has been widely studied and reported (Gibson and Roberfroid, 1995; Hamilton-Miller, 2004; Lomax and Calder, 2009), but more recent studies have focused on their protective role against infections and diseases (Lomax and Calder, 2009; Chakraborti, 2011; Licht et al., 2011). The most widely studied prebiotics are inulin and non-digestible oligosaccharides such as oligofructose (Watzl et al., 2005; Leenen and Dieleman, 2007; Guarner, 2007; Lomax and Calder, 2009). These complex carbohydrates are non- digestible by the gastric juice, pancreatic and brush border enzymes and selectively stimulate the growth of intestinal microflora. These carbohydrates are commonly found in fruits, vegetables and plant products (Menne et al., 2000; Rastall et al., 2005).
In the large intestine, prebiotics, in addition to their selective effects on intestinal bacteria, influence many aspects of bowel function through fermentation (Cummings et al., 2001). These substances are fermented by bacteria such as Bifidobacteria and Lactobacilli in the colon, to produce short chain fatty acids (SCFA) such as propionic, lactic, acetic, folic, and butyric acids. Hydrogen gas (H2) and Carbon dioxide (CO2) are also major products of prebiotic metabolism (Wang and Gibson, 1993; Cummings et al., 2001; De Vuyst et al., 2005; Saulnier et al., 2007). Besides the stimulation of intestinal bacteria, fermentability is an important property for evaluating or screening potential prebiotics, according to Cummings et al (Cummings et al., 2001). However, recent studies show that there may be other candidate-prebiotics such as xylitol, sorbitol, mannitol and lactulose (Chakraborti, 2011).
Pharmaceutical and nutritional industries are exploring more natural treatments for health conscious consumers as natural treatments have been effective since immemorial and are staging a comeback and natural ‘renaissance’ is happening all over the globe. The emerging area of prebiotics points towards the holistic role of nature in the health and nutrition of human beings.
CHAPTER TWO
2.0 Literature Review
2.1 Introduction
The review of relevant literature is the main topic of this chapter. A literature review covers theoretical and methodological contributions to a certain issue in addition to the state of knowledge at the time of writing. It provides information about the state of the art in relation to the subject you are writing about. It examines the body of work on the chosen subject. The literature evaluation in this study comprises the …
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TABLE OF CONTENTSPRELIMINARY PAGES
- Title page
- Approval page
- Dedication
- Acknowledgement
- Table of Contents
- Abstract
INTRODUCTION
- 1.1 Introduction
- ⋮
LITERATURE REVIEW
- 2.1 Introduction
- ⋮
CHAPTER THREE
MATERIALS AND METHODS
- 3.1 Introduction
- ⋮
CHAPTER FOUR
RESULTS AND DISCUSSION
- 4.1 Introduction
- 4.2 Results
- ⋮
- 4.3 Discussion of Findings
CHAPTER FIVE
SUMMARY, CONCLUSION AND RECOMMENDATION
- 5.1 Introduction
- 5.2 Summary
- 5.3 Conclusion
- 5.4 Recommendation
REFERENCES
Disclaimer for Complete Material Utilization
The displayed research work titled "Identification and Evaluation of Immuno-Modulatory Properties of Prebiotics Extracted From Vernonia Amygdalina" is stated as a research guideline towards accomplishing your assigned project / seminar research work. All the research materials on this website are ONLY for research purposes and should be used as a guideline in developing your research work. For no reason should you copy word for word as projectlist.com.ng will not be liable for any who copied the material. By ordering the complete research guideline, it signifies that you've accepted our terms of service.
PRELIMINARY PAGES
- Title page
- Approval page
- Dedication
- Acknowledgement
- Table of Contents
- Abstract
INTRODUCTION
- 1.1 Introduction
- ⋮
LITERATURE REVIEW
- 2.1 Introduction
- ⋮
CHAPTER THREE
MATERIALS AND METHODS
- 3.1 Introduction
- ⋮
CHAPTER FOUR
RESULTS AND DISCUSSION
- 4.1 Introduction
- 4.2 Results
- ⋮
- 4.3 Discussion of Findings
CHAPTER FIVE
SUMMARY, CONCLUSION AND RECOMMENDATION
- 5.1 Introduction
- 5.2 Summary
- 5.3 Conclusion
- 5.4 Recommendation
REFERENCES
Disclaimer for Complete Material Utilization
The displayed research work titled "Identification and Evaluation of Immuno-Modulatory Properties of Prebiotics Extracted From Vernonia Amygdalina" is stated as a research guideline towards accomplishing your assigned project / seminar research work. All the research materials on this website are ONLY for research purposes and should be used as a guideline in developing your research work. For no reason should you copy word for word as projectlist.com.ng will not be liable for any who copied the material. By ordering the complete research guideline, it signifies that you've accepted our terms of service.
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