ANTIMICROBIAL EFFECTS OF THE LEAF EXTRACTS OF CHROMOLAENA ORDORATA (SIAM WEEDS) ON SOME HUMAN PATHOGENS

ABSTRACT

Antimicrobial effects of Chromolaena odorata (commonly called Siam weed), were tested on Salmonella typhi, Pseudomonas aeruginosa, Staphylococcus aureus and Candida albicans. Different solvents such as methanol and chloroform were used for the leaf extraction. Test organism Pseudomonas aeruginosa and  Salmonella typhi were susceptible to methanol extracts of Chromolaena odorata at concentration of 25, 50 and 100mg/ml with zone of inhibition of 15mm, 19mm and 22mm respectively for Salmonella typhi and 29mm, 32mm and 36mm for Pseudomonas aeruginosa, while the chloroform extract produced similar concentration of 18mm, 20mm and 22mm for Salmonella typhi  and 19mm, 20mm and 21mm for Pseudomonas aeruginosa.  The Minimum Inhibitory Concentration (MIC) of methanol extract against Salmonella typhi and Pseudomonas aeruginosa were 12.5mg/ml and 3.12mg/ml respectively. The MIC of chloroform extract against Salmonella typhi and Pseudomonas aeruginosa were 6.25mg/ml and 6.25mg/ml. For the pathogen Staphylococcus aureus and Candida albicans, no MIC was carried out because there was no zone of inhibition seen on the plate with 100% concentration. This justifies the therapeutic use of Chromolaena odorata. This research work establishes a good support to the use of these plants in herbal remedies and for the development of new drugs.

TABLE OF CONTENTS

 Title page i

Certification ii

Dedication iii

Acknowledgements iv

Table of Contents v

List of Tables vii

List of Plates viii

Abstract ix

CHAPTER ONE

1.1 Introduction 1

1.2 Aims and Objectives 3

CHAPTER TWO

2.0 Literature review 4

2.1 Traditional uses of Chromolaena ordorata 6   

2.2 Phytochemicals and Chemotherapy 6

2.3 Phytochemical Screening 7

2.4.1 Nutritional value of Chromolaena ordorata 7

2.4.2 Antimicrobial Susceptibility testing 8

2.4.3 Antibacterial activity of the plant extracts 8

2.4.4 Antifungal activity of the plant extracts 9

CHAPTER THREE

3.0 Materials and methods 10

3.1 Materials 10

3.2 Methods 10

3.2.1 Sample collection 10

3.2.2 Preparation of extracts 10

3.2.3 Methanol extract preparation 11

3.2.4 Chloroform extract preparation 11

3.2.5 Preparation of stock solution of extracts 11

3.2.6 Media used 11

3.2.7 Media preparation 12

3.2.8 Collection of test bacterial isolates 12

3.2.9 Preparation of paper disc 12

3.2.10 Paper disc diffusion methods 12

3.2.11 Determination of Minimum Inhibitory Concentration 13

CHAPTER FOUR

4.1 Results 14

CHAPTER FIVE

5.0 Discussion and conclusion

5.1 Discussion 22

5.2 Conclusion

23

References  24  

LIST OF TABLES

TABLES TITLE        PAGES

  1 Zone Of Inhibition Of Methanol Extract On The

Test Organisms.                                 15

  2 Zone Of Inhibition Of Chloroform Extract On

The Test Organisms.                             16

  3 Zone Of Inhibition Produced By The Antibiotics

Used As Controls                             17

  4 The Minimum Inhibitory Concentration Of Methanol

 Plant Extracts Against Susceptible Organisms.       18

  5 The Minimum inhibitory Concentration Of Chloroform

Plant Extract Against Susceptible Organisms.     

LIST OF PLATES

PLATES TITLE PAGES

 1  Diameter of Zone of Inhibition of the Methanol and

Chloroform extract of Chromolaena ordorata On

Salmonella typhi.     20

 2 Diameter of Zone of Inhibition of the Methanol and

Chloroform extract of Chromolaena ordorata On

 Pseudomonas aeruginosa.                          21

 CHAPTER ONE

1.1 Introduction

Medicinal plants have been used for centuries as remedies for human diseases because they contain chemical components of therapeutic value (Nostro et al., 2000). Plants for decades have been a valuable source of natural products for maintaining human health, especially within -depth investigation for their natural therapeutic potentials. According to Santos, et al., (1995) several varieties of drugs can be derived from medicinal plants. There is a continuous and urgent need to discover new antimicrobial compounds with diverse mechanisms of action and chemical structures that can be used against novel and re-emerging infectious diseases (Rojas, et al., 2003). The abundance of plants on the earth’s surfaces has led to an increasing interest in the investigation of different extracts obtained from traditional plant as potential sources of new antimicrobial agents. Therefore, researchers are increasingly turning their attention to complementary medicine looking for new ways to develop better drugs against microbial infections (Benkeblia, 2004). Many plants have been used because of their antimicrobial traits, which are due to compounds synthesized as secondary metabolites of plant. These products are known by their active substances, for example, the phenolic compounds which are a part of the essential oils (Jansen, et al.,1987) as well as tannin (Saxena, et al., 1994). Medicinal values of plants lie in their component phytochemicals such as alkaloids, tannins, flavonoids and other phenolic compounds, which produce a definite physiological action on the human body (Daniel, 1999).Secondary metabolites (also called specialized metabolites) is a term for pathways and small molecule products of metabolism that are not absolutely required for the survival of the organism, many of which are antibiotics and pigments. Plants synthesize varieties of phytochemicals such as alkaloids, phenolics, terpenoids, glycosides etc.

Chromolaena odorata (Siam weeds) is a known toxic weed that is widespread over many parts of the world including Nigeria. Chromolaena odorata is a species from the family of Asteraceae. The weed goes by many common names including Siam weed, devil weed, French weed, communist weed, hagonoy, cohoy etc. The native range of Chromolaenais in the Americas, extending from Florida (USA) to Northern Argentina. Away from its native range, Chromolaenais an important weed in tropical and subtropical areas extending from west, central and southern Africa to India, Sri Lanka, Bangladesh, Laos, Cambodia, Thailand, southern China, Taiwan, Indonesia etc (Umukoro and Ashorobi, (2006); Hung, et al., 2011). Nevertheless, the plant has also been incriminated in the illness and death of cattle and goats in Karnataka, India. This weed was probably introduced into Nigeria about 50 years ago and found along road-sides, waste and fallow lands. Chromolaena odorata was first identified in Central America and Vietnam and formerly called Eupatorium odorata. It is a diffused scrambling shrub that is mainly a weed of plantation crops and pasture of southern Asia and West Africa. It forms a bush 3-7 metre in height when growing in the open according to (Nyananyo, 2006). The plant is locally called “bienqua” among the Ijaws in the Niger Delta region of Nigeria where it is believed to posses healing potentials for wounds and treatment of pile ailment (Hung, et al., 2011). A Concoction of the leaf is used as a cough remedy and as an ingredient with lemon grass and guava leaves for the treatment of malaria. Other traditional medicinal uses include anti-diarrheal, astringent, antispasmodic, antihypertensive, anti-inflammatory, diuretic tonic, antipyretic and heart tonic (Vital and Windell, 2009). The fresh leaves and extract of Chromolaena. odorata are a traditional herbal treatment in some developing countries for burns, soft tissue wounds and skin infections. A formulation prepared from the aqueous extract of the leaves has been licensed for clinical use in Vietnam (Ayyanar and Ignacimuthu, 2009).

In Nigeria the local use of the leaf extracts of Chromolaena odorata for sore throat and treatment of pile, burns and wounds have been documented (Anwannil and Atta, 2006)

1.2 Aims And Objectives

i. To determine the inhibitory activities of the extracts against S.aureus, P. aeruginosa C. albicans and S. typhi

ii. To determine the Minimum Inhibitory Concentration (MIC) of the extracts against S. aureus, P. aeruginosa, C. albicans and S. typhi.

iii. To extract active ingredients of C. ordorata using chloroform and methanol as solvents.

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