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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