ANTIBACTERIAL ACTIVITY OF COSTUS AFER AND DIALIUM GUINEENSE ON SOME PATHOGENS

ABSTRACT

The antimicrobial activities of Costus afer and Dialium guineense extracts were tested against Staphylococcus aureus¸ Pseudomonas aeruginosa, Enterococcus faecalis, Escherichia coli and Salmonella typhi using disc agar diffusion technique. The ethanol extracts from both plants inhibited the growth of all the organisms used in this study except Salmonella typhi. It was further shown that the sensitivity of the organisms to the extracts did not only vary from one organisms to another but also in concentrations used. The most sensitive organism to Costus afer and Dialium guineense extract was Staphylococcus aureus, while the least was Escherichia coli. The inhibition zone measured 21.3 ± 1.13mm, 11.0 ± 1.73mm, 14.3 ± 2.08 and 19.3 ± 2.08mm against Staphylococcus aureus, Escherichia coli, Enterococcus faecalis and Pseudomonas aeruginosa, respectively at concentration of 100mg/ml. Thus while Staphylococcus aureus has the highest sensitivity, Escherichia coli has the least to Costus afer extract. The inhibition zones produced by Dialium guineense was 22.0 ± 3.16mm, 20.0 ± 4.36mm, 20.3 ± 2.08mm and 18.0 ± 2.00mm against Staphylococcus aureus, Escherichia  coli, Enterococcus faecalis and  Pseudomonas aeruginosa, respectively at 100mg/ml concentration. The Minimum Inhibitory Concentration (MIC) of extract of Costus afer was 12.5mg/ml, 25mg/ml, 12.5mg/ml and 12.5mg/ml against Staphylococcus aureus, Escherichia coli, Enterococcus faecalis and Pseudomonas aeruginosa, respectively Dialium guineense produced Minimuim Inhibitory Concentration (MIC) values of 12.5mg/ml, 25mg/ml, 25mg/ml and 12.5mg/ml against Staphylococcus aureus, Escherichia coli, Enterococcus faecalis and Pseudomonas aeruginosa, respectively. The result showed that the ethanol extracts of the plants produced appreciable inhibitory activity against the test organisms. These result support further research into the properties of the chemical substances producing these effects.

TABLE OF CONTENTS

Cover page i

Certification ii

Dedication iii

Acknowledgements iv

Table of Contents v

List of Tables vii

List of Figures viii

List of Plates ix

Abstract x

CHAPTER ONE

1.0 Introduction 1

1.1 Aims and Objectives 4

CHAPTER TWO

2.0 Literature Review 5

2.1 Phytomedicine 5

2.1.1 Characteristics of Phytomedicine 6

2.1.2 The Demand and Phytomedicine 6

2.2 Therapeutic Benefits of Phytomedicine 7

2.3 Plants with Medicinal Properties 9

2.4 Costus afer 10

2.5 Dialium guineense 14

2.6 Major group of Antimicrobial Compounds from Plants 19

CHAPTER THREE

3.0. Material and Method 23

3.1 Collection and Identification of Plant Material 23

3.2 Processing and Extraction of Plant Material 23

3.3 Antimicrobial Testing 24

3.4 Determination of Minimum Inhibitory Concentration (MIC) of Plant Extracts   25

3.5 Phytochemical Analysis 26

3.5.1 Qualitative Evaluation 26

3.5.2 Quantitative Determination 27

CHAPTER FOUR

4.0       Result 31

CHAPTER FIVE

5.1 Discussion 39

5.2  Recommendation 40

5.3 Conclusion 40

References 41

Appendix 47

LIST OF TABLES

Table Title Page

1.  Some medicinal plants, common names, location And their uses 9

2.  Diameter zone of inhibition (mm) produced by various

concentrations of Dailium guineense against test organisms 32

3.  Diameter zone of inhibition (mm) produced by various concentrations

of Costus afer against test organisms 33

4. Minimum inhibitory concentration and Minimum Bactericidal

Concentrations (MBC) of extract of Costus afer against test organisms 34

5.  Minimum inhibitory concentration and Minimum Bactericidal

Concentrations (MBC) of extract of Dailium guineense against

test organisms 35

6. Bioactive compound present in qualitative analysis of plant extract 37

7.  Quantitative analysis of bioactive compound present in plant extract

in (m/mg) 38

LIST OF FIGURES

Figure Title Page

1 Structure of alkaloids 21

2 Structure of phenols 21

3 Structure of saponin 21

4 Structure of tannin 22

5 Structure of flavonoid 22

6 Plant of Dialium guineense 47

7 Plant of Costus afer 47

LISTS OF PLATES

Plate Title Page

1  Zone of inhibition created by the two plants on Staphylococcus aureus (Sa),

salmonella typhi(st), Pseudomonas aeruginosa (Pa), Escherichia coli (Ec) and

Enterococcus faecalis (Ef). 36

CHAPTER ONE

3.0 INTRODUCTION

The use of higher plant and their extracts to treat infections is an age old practice in traditional African medicine. Traditional medicine practice using plants has been known for centuries in many part of the world (Sofowora, 1993). It is however observed that these practices vary from one country to another. The persistent increase in antibiotic-resistant strain of microorganisms has led to the development of more potent but also more expensive antibiotics, such as the third-generation fluoroquinolones and the cephalosporins (Aibinu, 2004; O’Brein et al., 1999 and Cohen, 2002). In most developing countries of the world, these antibiotics are not readily affordable, which makes compliance difficult. This calls for research into alternative sources of antimicrobials

Numerous plants and herbs is the most ancient approach to healing known as the herbal medicines.

Among these herbal medicines, Dialium guineense and Costus afer are not left out.

Dialium guineense belong to the kingdom plantae, family of facaceae and of genus Dialium. It   is commonly called black velvet or velvet tarimand  (English),  Icheku (Ibo,  Eastern  Nigeria),  Awin  (Yoruba,  Western  Nigeria), Tamarinier  noir  (French)  (Ezeja et  al.,  2011).  It is a woody plant that occurs in the rain forest region of West Africa and can grow up to 10 to 20m. The tree possess densely, hairy leafy crown, smooth greyish bark and whitish  flowers  which  bears  densely  velvet  black  fruits that are more or less circular and flattened enclosing dry, brownish, sweet acidic edible pulp.  In Nigeria, the tree flowers from September to October, and fruits from October to January (Keay, 1989).

The  ripe  fruits  of  the  plant  are  chewed  among  some women  in  southeast  Nigeria  to  improve  lactation  and check  genital  infection  (Nwosu,  2000).  Among  the  Esan tribe  of  Edo  State in  Nigeria,  the  twig  or  bark  is  chewed for oral hygiene and stomach ache (Idu et al., 2009). The genus Dialium comprises  five  species  in  West Tropical Africa but Dialium guineense,  Dialium    dinklagel and Dialium packyphyllum are  represented in Nigeria (Omotayo, 1999). Traditionally, Dialium guineense leaves and stem bark are used as remedies for   diarrhea,   severe   cough, bronchitis, wound, stomach aches, malaria, fever, jaundice, antiulcer and haemorrhoids (Ogu    and Amiebenomo, 2012).

Okwu and Okeke reported in their studies that the plant contains saponin which is presumed to add to the cleaning effect of teeth and at the same time prevent cavies and plaques on the teeth of the users (Okwu et al., 2003).

Costus afer belong to the group of perennial herbaceous plants in the family Costaccae described by Linnaeous as a genus in 1753. It is one among the 150 species of stout, perennial and rhizomatus herbs of the genus Costus (Edeoga et al., 2000). The leave, stem, seed and rhizomes are harvested from wild plant and they contain metabolites. The plant is used as a remedy for cough, inflammation, arthritis as laxatives, apparent, purgative, diuretic, in rheumatism and treatment of several other diseases (Awonters et al., 1978).

In Nigeria, the plant extracts is used as cough medicine. While it’s boiled root is applied to cuts and sores. (Awouters, et al., 1978). It has been shown to reduced carageenan – induced edema in the rats paw, and check diarrhoea caused by arachidonic acid and castor oil. (Iwu et al., 1993).It  ameliorate all signs associated with adjuvant-induced polyarthritis in rats and dose of 100-300mg/kg daily is well tolerated in human (Iwu, et al., 1993). In Ohafia Abia State, Nigeria and in some part of Cameroon, the decoction of the plant is administered to diabetic patient to alleviate the clinical signs (Dioka, et al., 2002). An infusion of the inflorescence is taken to treat stomach complaints. A stem decoction, mashed stem or the pounded fruit, sometimes, mixed with sugar cane juice, are taken to treat cough, respiratory problem and sore throat (Iwu et al., 1993). In Nigeria, a cold water extract of the stem is taken to treat small epileptic attacks. Rhizome pulp is applied to abscess and ulcers to heal them applied to teeth to cure toothache and mixed with water to treat diarrhoea and amoebic dysentery. A rhizome decoction or the rawrhizome is taken to treat leprosy and venereal diseases. (Iwu et al., 1993). Leaf sap is used as eye drops and as nose drops to treat headache with vertigo. It is also used to treat edema and fever. Stem sap is applied to treat urethral discharges, venereal diseases, jaundice and to prevent miscarriage. Stem sap is acid and rubefacient and burns on open wound is applied to different skin ailments. The stem decoction is widely taken to treat rheumatoid arthritis. An infusion of the dried aerial parts is taken to treat hypertension. The powdered stem is used as enema to treat worms and haemorrhoids. The pulped stem taken in water is strongly diuretic (Iwu, et al., 1993). Other species of the genus also have medicinal effects, e.g.  Costus lucanustanus is used in the Ivory Coast for the treatment of impending abortion. It also exhibits uterine relaxant activity. Costus schlechteri and Costus afer  are used in the treatment of diabetes mellitus.

Investigations (studies) indicate that Dialium guineense andCostus afer leaf extract has significant antimicrobial properties, and thus substantiates its popular and wide traditional applications in diverse ailments. These plants may therefore be exploited as a potential preservative in the pharmaceutical and food industries.

1.1 AIMS AND OBJECTIVES

1. The aim of this study is to assess the antimicrobial activities of the ethanol extracts of Dailium guineense and Costus afer against some bacterial pathogens.

2. To determine the minimum inhibitory concentration and minimum bactericidal concentrations of susceptible test bacteria.

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