ABSTRACT
The leaf extracts of Mitracarpus villosus were screened for their phytochemical properties and anti-bacterial effects. Aqueous, methanol, ethyl acetate and n-hexane were used as solvents for extraction of the leaf sample. The leaf sample were also screened qualitatively and quantitatively for their bioactive constituents. The aqueous, methanol, ethyl acetate and n- hexane extracts of the leaf of M. villosus were concentrated at 100mg/ml, 50mg/ml, 25mg/ml and 12.5mg/ml respectively. The antibacterial activities of the leaf sample were tested against Candida albicans, Trichophyton mentagrophytes, Microsporum auduounii and Aspergillus flavus. The MIC and MFC of the extracts was also determined for all the bacterial species. The result revealed that there were no significant differences in bioactive constituents of both plants qualitatively and quantitatively. Ethyl acetate extract of Mitracarpus villosus had highest zone of inhibition of 24.50±0.71mm and 26.00±0.00mm on Candida albicans, 24.00±0.00mm and 24.00±0.00mm on Trichophyton mentagrophytes while aqueous extract inhibition was 20.00±.00mm and 22.00±0.00mm on Candida albicans, 18.00±0.00mm and 18.00±0.00mm on Trichophyton mentagrophytes respectively. No zone of inhibition was produced in Microsporum auduouinii and A. flavus in all the solvent extracts used. The combined effects of Mitracarpus villosus plant extracts using the same solvents (aqueous, methanol, ethyl acetate and n–hexane) showed significant differences in all the solvent extracts at 100mg/ml of aqueous (23.00±0.00mm and 20.00±0.00mm), methanol (19.00±0.00mm and 18.00±0.00mm), ethyl acetate (28.00±0.00mm and 26.00±0.00mm) and n-hexane (20.00±0.00mm and 17.00±0.00mm) on Candida albicans and Trichophyton mentagrophytes respectively. No zone of inhibition was shown in Aspergillus flavus and Microsporum auduouinii. The MIC and MFC ranged from 50 – 6.25mg (MIC) and 100 – 6.25mg (MFC). Thus the traditional claims of the uses of the plants as anti-bacterial agents were therefore justified.
CHAPTER ONE
1.0 INTRODUCTION
1.1 Background of the Study
Medicinal plants have been used for centuries as remedies for human diseases because they contain components of therapeutic value (Nostro et al., 2000; Tanaka et al., 2002). According to the World Health Organization (WHO) in 2008, more than 80% of the World’s population relies on traditional medicine for their primary health care needs. Traditional medicine is an important part of African cultures and local medicinal systems vary among cultural groups and regions (Makhubu, 2006). Herbs are now very popular in developing countries on account of improved knowledge about the safety, efficiency and quality assurance of ethno- medicine (Makhubu, 2006). In recent years, secondary plant metabolites (phytochemicals) have been extensively investigated as a source of medicinal agents. Thus it is anticipated that phytochemicals with good anti-bacterial activity will be used for the treatment of bacterial infections. This is because according to Arora and Keur (1999), the success story of chemotherapy lies in the continuous search for new drugs to counter the challenges posed by resistant strains of micro-organisms. Studies indicate that in some plants, there are many substances such as peptides, tannins, alkaloids, essential oils, phenols and flavonoids among others which could serve as sources of antimicrobial production. These substances or compounds have potentially significant therapeutic applications against human pathogens including bacteria, bacterial and viruses (Arora and Keur, 1999, Okigbo and Omodamiro, 2006). The development of microbial resistance to the available antibiotics has led researchers to investigate the antimicrobial activity of medicinal plants (Bisignano et al., 1996, Hammer et al.,1999). Antibiotic resistance has become a global concern (Westh et al., 2004) as the clinical efficacy of many existing antibiotics is being threatened by the emergence of multi-drug resistant pathogens (Bandow et al., 2003). Natural products either as pure compounds or as standardized plant extracts provide unlimited opportunities for the development of novel drugs because of the great diversity in their chemical structure. There is a continuous and urgent need to discover new antimicrobial compounds with diverse chemical structure and novel mechanisms of action for new and re-emerging infectious diseases (Rogas et al., 2004).
The research into biologically active compounds from natural sources has always been of great interest for scientists looking for new sources of useful drugs against infectious diseases. Mitracarpus villosus is widely employed in traditional medicine in West Africa for headaches, toothache, amenorrhoea, dyspepsia, hepatic diseases, venereal diseases and leprosy (Bisignano et al., 2000). Among the folkloric uses, the juice of the plants is applied topically for the treatment of skin diseases (infectious dermatitis, eczema and scabies). Daiziel, (1937); Kerharo and Adam, (1974) observed that a lotion and a skin ointment made with the aerial part of M. villosus are used for skin infections or skin diseases and other infectious.
Previous studies by Moulis et al. (1992) reported the isolation of pentalogin from fresh, aerial parts of Mitracapus villosus which demonstrates a potent anti-bacterial activity against Candiba albicans and Trichophyton soudanense. Other investigations (Sanogo et al., 1996) showed that different extracts of M. villosus exhibited broad antibacterial and anti-bacterial activity against standard strains and clinical isolates of Staphylococcus aureus and C. albicans responsible for common skin infections. More recently, Germano et al. (1999) reported the hepato protective effects of Mitracarpus villosus decoction on tetrachloromethane (CCl4) induced hepatotoxicity in vivo as well as in vitro using isolated hepatocytes.
1.2 Statement of Research Problem
The use of Mitracarpus villosus have been on the basis of trial and error in different communities in Africa without any scientific basis. Plant parts have been used in different locations for the treatment of different ailments which sometimes bring about conflicting results. Often traditional healers use plants according to their analogy and morphological similarities to the ailment being treated. For example, plants containing red juice are used to treat ailments connected to menstruation problems and bleedings (Neuwinger, 2000). There is therefore the need to ascertain the basis for the claims of the efficacy of the plants used locally in ethnomedicine.
1.3 significance of the study
The increasing resistance to antibiotics has resulted in the research to form new organic molecules from plants with antimicrobial properties for treating diseases since some microorganisms have developed resistance to many orthodox drugs (Sofowora, 2006). There is the need to find an alternative approach in the treatment of infectious diseases. Using local plants will be a welcome development as the cost will be minimal.
The leaves extracts of Mitracarpus villosus have been reported in the treatment of various ailments such as ulcer, cancer, skin diseases e.t.c. It is therefore important to scientifically investigate these plant parts to ascertain their therapeutic potentials.
Determination of their chemical composition as well as antimicrobial efficacy against specific pathogens is important in the recognition of this plant as a potent commercial medicinal plant. Tests can determine its efficacy against a pathogen and thus, establish the minimal dosage required for the treatment of ailments.
1.4 Aim of Study
To evaluate the phytochemical constituents and anti-bacterial of leave extracts of Mitracarpus villosus.
1.5 Objectives
1. To obtain the methanol, ethyl acetate, n- hexane and aqueous extracts of leaf of Mitracarpus villosus.
2. To determine qualitatively and quantitatively secondary metabolites present in the methanol, ethyl acetate, n- hexane and aqueous extracts of leaf of Mitracarpus villosuss.
3. To assess the anti-bacterial activities of the aqueous, methanol, ethyl acetate and nhexane extracts of single and combined leaf of E. heterophylla and M. villosus on some selected microorganisms.
4. To determine the Minimum Inhibitory Concentration (MIC) and Minimum Antibacterial Concentration (MFC) of aqueous, methanol, ethyl acetate and n-hexane extracts of single and combined plants of E. heterophylla and M. villosus against different species of isolated bacterial.
1.6 Hypotheses
i. There are no significant differences qualitatively and quantitatively between secondary metabolites present in the aqueous, methanol, ethyl acetate and n-hexane extracts plant of E .heterophylla and M. villosus.
ii. There are no significant differences in the anti-bacterial activities of the aqueous, methanol, ethyl acetate and n- hexane extracts of single and combined leaf E. heterophylla and M. villosus on some selected microorganisms.
iii. There are no significant differences in the MIC and MFC of aqueous, methanol, ethyl acetate and n-hexane extracts of single and combined leaf E .heterophylla and M. villosus against different species of isolated bacterial microorganisms
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