ANTIBIOGRAM OF SOME GRAM NEGATIVE PATHOGENS TO EXTRACTS OF OKPETE (COSTUS AFAR) AND ISIKARA (SPONDIAS MOMBIN)

ABSTRACT

The antibiogram of some Gram negative pathogens (Escherichia coli, Salmonella and Shigella Species) to extract of Costus afar (Okpete) and Spondias mombin (Isikara) was evaluated. Bacteria Species were isolated from two plants (E. coli, Salmonella and Shigella). The Muller Hinton agar, Salmonella Shigella agar, Eosin Methylene Blue agar was used for the antimicrobial activity tests. The isolates were characterized culturally, phytochemically and biochemically. These bacteria Species were tested for their susceptibility in the different extracts (Okpete and Isikara). The phytochemical screening of the plant extracts, showed that the alkaloid was highly present and the cyanogenic glycoside was slightly present. The Antibiogram of the bacteria pathogen, Salmonella had the highest inhibition recorded with ethanol extract of Spondias mombin (17.67mm ± 0.58) while the least inhibition was with water extract of Costus afar (9.67mm ± 0.58). The minimum inhibition concentration showed values of 50mg/ml to 200mg/ml with variations among the extracts activity and ethanol extracts having the lowest concentration that affected inhibition. From the concentration of the relative potency of extracts, ethanol exhibited the highest potency of extracts while water exhibited the least potency of extracts. It is recommended that the potentials of these two plants in terms of health benefits should be harnessed.

TABLE OF CONTENTS

Cover page                                                                                                                              i

Certification                                                                                                                           ii

Dedication                                                                                                                              iii

Acknowledgements                                                                                                                iv

Table of Contents                                                                                                                   v

List of Tables                                                                                                                          vii

Abstract                                                                                                                                  x

CHAPTER ONE

Introduction                                                                                                                            1

1.1       Background of the Study                                                                                            1

1.2       Aim and Objectives                                                                                                    3

1.3       Objective of the Study                                                                                                4

CHAPTER TWO

Review of Related Literature                                                                                                 5

2.1       Costus afer Ker Gawl (Zingiberaceae)                                                                      5

2.2       Spondias mombin (Anacardiaceae)                                                                            6

2.3       Antibacterial Activity of Medicinal Plants                                                                 7

2.4       Antibiogram of Bacteria Species                                                                                14

2.5       Chemical Composition of the Stem Extract of Costus afer (Bush Cane                17

2.6       Chemical Composition of the Leaf Extract of Spondias mombin (Ichakara)                   21

2.7       Prevalence of Microorganism                                                                                    23

2.8       Isolation of Microorganism                                                                            24

2.9       Antibiotics Susceptibility of Microbial Contaminants                                   26

CHAPTER THREE

Materials and Methods                                                                                                           29

3.1       Source of Materials                                                                                                    29

3.2       Sample and Media Preparations                                                                                 29.

3.2.1    Preparation of Plant Samples                                                                                     29

3.2.2    Media Preparation                                                                                                      29

3.2.3    Preparation of Plant Extracts                                                                                      30

3.3       Confirmation of Gram Negative Pathogens                                                               30

3.4       Qualitative Phytochemical Screenings                                                                       31

3.4.1    Test for Saponin                                                                                                         31

3.4.2    Test for Steroids                                                                                                         31

3.4.3    Test for Flavonoids                                                                                                     31

3.4.4    Test for Tannin                                                                                                           32

3.4.5    Test for Cyanogenic Glycoside (HCN)                                                                      32

3.4.6    Test for Alkaloids                                                                                                       32

3.4.7    Test for Starch                                                                                                            32

3.5       Antimicrobial Sensitivity Test                                                                                   33

3.5.1    Determination of Relative Potency of Plant Extracts                                                33

3.6       Determinations of Minimum Inhibition Concentration (MIC)                                    34

3.7       Statistical Analysis                                                                                                     34

CHAPER FOUR

Results                                                                                                                                    35

CHAPTER FIVE

Discussion, Conclusion and Recommendation                                                                      43

5.1       Discussion                                                                                                                   43

5.2       Conclusion                                                                                                                  45

5.3       Recommendation                                                                                                       45

References                                                                                                                              46

Appendix

LIST OF TABLES

Table                       Title                                          Page

1:         Quantitative Phytochemical Screening of Okpete (Costus Afar) and Isikara (Spondias Mombin) Leaves                              36

2:         Antibiogram of Gram Negative Bacteria Pathogen towards Plant Extracts                       38

3:          Minimum Inhibition Concentrations of Plant Extracts                      40

4:         Concentration of Relative Potency of Extracts                            42

CHAPTER ONE

INTRODUCTION

1.1       Background of the Study

Plants provide a variety of resources that contribute to the fundamental needs of both human being and animals such as food, clothing and shelter. Among plants of economic importance are medicinal plants. Plants have been utilized as therapeutic agents since time immemorial in both organized and unorganized forms (Girach et al., 2003). The healing properties of many herbal medicines have been recognized in many ancient cultures. 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. 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 (Aibinu, 2004).

Numerous plants and herbs is the most ancient approach to healing known as the herbal medicines. Among these herbal medicines, Costus afar and Spondias mombin are not left out.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. 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 (Edeoga et al., 2005).

Spondias is a genus of flowering plants in the cashew family, Anacardiaceae. The genus consists of 17 described species, 7 of which are native to the Neotropics and about 10 are native to tropical Asia. They are commonly named hog plums, Spanish plums, libas in Bikol and in some cases golden apples for their brightly colored fruit which resemble an apple or small plum at a casual glance. They are only distantly related to apple and plum trees, however. A more unequivocal common name is mombins(Ayoka et al., 2008).Spondias mombin has several common names. Among the English speaking Caribbean islands it is known as yellow mombin or hog plum, while in In Ghana, it is hog plum or Ashanti plum. It is called "Akukor" in the Ewe language of Ghana. In Nigeria, the fruit is called Iyeye or Yeye in the Yoruba language, Ngulungwu in Igbo and Isada in Hausa (Adepoju and Oyewole, 2008).

Most of the plants and in ethnomedicines are said to possess bioactive constituents such as saponins, tannins, flavonoids, phenolics and other secondary metabolites which are capable of producing definite physiological action in the body(Sofowora, 1993). In Nigeria, the plant extracts is used as cough medicine. While it’s boiled root is applied to cuts and sores. 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. 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(Iwu, et al., 1993).

Investigations (studies) indicate that Costus aferandSpondias mombin 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.

1.2       Aim and Objectives

The aim of this project work was to study the antibiogram of some Gram -negative bacteria using extracts of Costus afar (Okpete) and Spondias mombin (isikara).

1.3       Objective of the Study

The objectives of this project work include the followings

i.      To produce extracts from Costus afarand Spondias mombin plant leaves using ethanol, water and methanol.

ii.     To isolate and identify some Gram -negative bacteria of established pathogenicity.

iii.   To test the sensitivity of the pathogens to the extracts of Costus afar and Spondias mombin.

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