Towards Sustainable Indigenous Mahogany Production in Ghana
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PD 528/08 Rev.1 PD 528/08 Rev.1 (F): Towards sustainable indigenous Mahogany production in Ghana: Phase II, refining the silviculture “tool kit” and practical training for industrial-foresters and community farmers Antimicrobial potential and phytochemical properties of four species of African mahogany 1, 2 1 2 Ofori E. Opuni-Frimpong E. and Amissah-Eshun, R. 1) Forestry Research Institute of Ghana 2) Accra Polytechnic ABSTRACT Traditional healers’ use different species of plant parts for healing purposes, however, very little scientific research to support claims of effectiveness have been undertaken. The objective of this study was to determine the phytochemical properties in four species of African mahogany (Khaya grandifoliola, khaya senegalensis, Khaya anthotheca and khaya ivorensis) and to determine its antimicrobial potential. Standard procedures were followed in the extraction and analysis of both aqueous and ethanolic extracts of stem bark of the African mahogany. The study identified the presences of Glycoside, Tannins, Saponins, Anthraquinones in the stem bark of all four species of the African mahogany, Terpenes are absent in Khaya anthotheca but was present in the other species. Aqueous and Ethanolic extracts were evaluated for potential antimicrobial activities against medically important bacterial and fungal strain. An antimicrobial activity was determined in the extracts using agar disc diffusion method. At a concentration of 20mg/ml, the zone of inhibition (mm) ranges from 13.5±0.50 – 24.0±0.00, for both extracts of African mahogany whiles ciprofloxacin (10µg/ml) was also in the range of 14.0±0.00 – 36.0±1.00 against all organism tested. The results showed a broad spectrum of activity against Staphylococcus aureus, Streptococcus pyogenes, Bacilius subtilise, Escherichia coli, Pseudomonas aeruginosa, Salmonella typhi and Candida albicans. The results also identified ethanolic extract to have higher zone of inhibition than that of the aqueous extract. The analysis was carried out in the Pharmacognosy department of Kwame Nkrumah University of Science and Technology. 1.0 INTRODUCTION 1.1 BACKGROUND Herbal medicines have been known to man for centuries. Therapeutic efficacy of many indigenous plants for several disorders has been described by practitioners of traditional medicine. Antimicrobial properties of medicinal plants are being increasingly reported from different parts of the world (Ramasamy et al., 2009). The healing of diseases in Ghana and other part of the world make use of numerous plants species as a source of medicine. Some of these plants and their parts used in the healing processes are fairly known. Information on the use and the properties of these plants are kept secret by these traditional healers (Dokosi, 1969). About 61% of new drugs developed between 1981 and 2002 were based on natural products and they have been very successful, especially in the areas of infectious disease and cancer (Cragg, 2005). The World Health Organization estimates that plant extracts or their active constituents are used as folk medicine in traditional therapies of 80% of the world's population (Shaik et al., 1994). The harmful microorganisms can be controlled with drugs and these results in the emergence of multiple drug-resistant bacteria and it has created alarming clinical situations in the treatment of infections. The pharmacological industries have produced a number of new antibiotics; resistance to these drugs by microorganisms has increased. In general, bacteria have the genetic ability to transmit and acquire resistance to synthetic drugs which are utilized as therapeutic agents (Towers et al., 2001). Natural products of higher plants may give a new source of antimicrobial agents with possibly novel mechanisms of action (Shahidi, 2004). The effects of plant extracts on bacteria have been studied by a very large number of researchers in different parts of the world (Reddy et al., 2001). In an effort to expand the spectrum of antibacterial agents from natural resources, four species of African Mahogany were studied. These species include Khaya grandifoliola, Khaya ivorensis, Khaya anthotheca and Khaya senegalensis of the family Meliaceae. They are found in the humid forest of Africa, ranging from Cote d’Ivoire, through Ghana and Nigeria to Gabon (Irvine, 1961). K. ivorensis naturally occurs mainly within the moist semi-deciduous forest zone of Ghana, the distribution of K. anthotheca spans across three ecological zones (moist semi- deciduous forest zone, dry semi-deciduous forest zone and the transitional zone between dry semi-deciduous forest zone and Savannah zone) (Ofori et al., 2007). Khaya grandifoliola is naturally found in the dry Semi-deciduous and the moist Semi-deciduous zone (Irvine, 1961). These two ecological zones are characterized by high rainfall, short dry season and rich soil nutrient. These zones also have a wider range of environmental conditions which Khaya grandifoliola is adapted to in the natural forest (Hall and Swaine, 1981). Khaya senegalensis is also a transitional and Savannah zone tree species 1.2 PROBLEM STATEMENT In Ghana and in most developing countries, extracts from indigenous plants species are employ for the treatment of several diseases. The herbal medicines produced from these extracts are very important for the economies of these countries. The traditional knowledge of employing these local plants for medicine are not normally supported by scientific information and also not usually written down (Dokosi, 1966). This work seeks to determine and document the effect of extracts from the African mahogany on some clinically important microbes. 1.3 JUSTIFICATION Antibiotics are one of the most important drugs in fighting bacterial infections and have greatly benefited the health of human since their introduction. However, over the past few decades, the health benefits of antibiotics are under threat as many commonly used antibiotics have become less and less effective against certain bacterial infections. This is due to emergence of drug- resistant bacteria. It is therefore essential to search for newer drugs to which bacteria will have lesser resistance. In many developing countries, traditional medicine is one of the primary healthcare systems (Farnsworth, 1993). Drugs derived from natural sources play significant role in the prevention and treatment of human diseases. Scientific knowledge on the use of indigenous plant such as African Mahogany is limited, there is therefore the need to investigate the antimicrobial properties of African Mahogany extracts for proper use in the treatment of illness. 1.4 RESEARCH OBJECTIVES The research objectives were: 1. Determination of the phytochemical properties of the African Mahogany (Khaya grandifoliola, Khaya ivorensis, Khaya anthotheca and Khaya senegalensis). 2. Determination of the antimicrobial potential on some clinical important microbes. 1.5 RESEARCH QUESTION Can African Mahogany be a potential source of antimicrobial substance against disease causing organisms? 3.0 METHODOLOGY 3.1 COLLECTION OF PLANT MATERIALS The bark of stem of the four (4) African Mahogany species (Khaya grandifoliola, Khaya ivorensis, Khaya anthotheca and Khaya senegalensis) were harvested from moist semi- deciduous forest zone, dry semi-deciduous forest zone and the transitional zone between dry semi-deciduous forest zone and Savannah zone of Ghana. The samples were air-dried and then send to Kwame Nkrumah University of Science and Technology in department of Pharmacognosy for extraction and analysis. 3.2 PREPARATION OF PLANT EXTRACT The extraction of the African Mahogany was carried out using known standard procedures (Harbome, 1973). The plant material was dried in shade and powdered in a mechanical grinder. About 25.0g of the powder from the plant material was first extracted with 900 ml petroleum ether at temperature 60-80°C for 72 hours, followed by 900 ml of ethanol by using a Soxhlet extractor. The extracts were filtered using Whatman filter paper (No.1) while hot and concentrated under vacuum and reduced pressure using rotary evaporator, and dried in a desiccator to obtained dried powdered extracts. The aqueous extract of each sample was prepared by soaking 15g of the dry powdered sample in 100ml of double distilled water (DD) for 24 hours. The extract centrifuged for 10 min. at 2500 rpm, then filtered using Wattman filter paper No. 42. 3.3 PRELIMINARY PHYTOCHEMICAL SCREENING The extracts were subjected to preliminary phytochemical testing to detect for the presence of different chemical groups of compounds. The petroleum ether and the ethanol plant extracts were screened for the presence of saponins, tannins, alkaloids, flavonoids, triterpenoids, steroids, glycosides, anthraquinones, coumarin, as described in literatures, (Khandelwal, 2009 and Kumar et al, 2009). One gram of dry powdered extract was dissolved in 5ml DD. A standard procedure for identification of the various classes of active chemical constituent was used (Abdul-Aziz, 2005). Alcoholic extraction are similar to aqueous extraction expect using 70% ethanol instead of DD H2O (Okogum, 2000). The preliminary detection of some chemical composition were performed such as flavonids that l0g of dry powdered extract (A) was dissolved in 5ml of ethanol 95% filtered; then Aliquot of 10ml of KOH 50% was added to 10ml ethanol 50% (B), equal volumes from (A) and (B) solution were mixed, a yellow color was developed indicating the presence of flavonoid (Jaffer et al, 1983). Glycosides