ANTIBACTERIAL AND PHYTOCHEMICAL SCREENING OF EXTRACT OF RICINUS COMMUNIS AND MANNIOPHYTON FULVUM

ABSTRACT

This study was carried out to determine the phytochemical properties and investigate the antibacterial activities of the ethanolic extracts of Manniophyton fulvum and Ricinis communis against Escherichia coli, Staphylococcus aureus, Enterococcus faecalis and Salmonella typhi using disc diffusion method and agar dilution methods. in vitro inhibition was observed in ethanol extract of Manniophyton fulvum which produced inhibition zone of 16 and 14.5 mm against Enterococcus faecalis  and Salmonella typhi respectively, while at a similar concentration for Ricinus communis, zone sizes of 13mm, 14mm and 12mm were produced against S. aureus, E. faecalis and Salmonella typhi respectively. Among the two plant extracts tested, Ricinis communis inhibited Staphylococcus aureus at concentrations of 5000μg/ml and 10,000μg/ml while Manniophyton fulvum did not inhibit S. aureus at a similar concentration. No inhibitory effect by the extracts was observed for E. coli at the concentration of 10000μg/ml, 5000μg/ml and 2000μg/ml. The minimum inhibitory concentration (MIC) of the plant materials ranged between 12.5mg/ml and 50mg/ml; and 25mg/ml to 100mg/ml for M. fulvum and R. communis respectively. M. fulvum prevented growth of S. typhi at the lowest concentration of 12.5mg/ml. The phytochemical compounds observed in the plant extracts include flavonoids, tannins, terpenoids, saponins, and alkaloids. These findings provide scientific evidence of traditional use of medicinal plants and also indicate the potential of these plants for the development of antimicrobial agents.

TABLE OF CONTENTS

Title Page                                                                                                                                 i

Certification                                                                                                                             ii

Dedication                                                                                                                               iii

Acknowledgements                                                                                                                 iv

Table of Contents                                                                                                                     v

List of Tables                                                                                                                            vi

Abstract                                                                                                                                    vii

CHAPTER ONE

1.1 Introduction                                                                                                                       1

1.2 Objectives of the Study                                                                                                     4

CHAPTER TWO

LITERATURE REVIEW     

2.1 Medicinal Plants                                                                                                                5

2.3 Morphological Description of Ricinus communis                                                               6

2.4 Habitat and Growth of Ricinus communis                                                                          7

2.5 Phytochemical Constituents of Ricinis communis                                                              8

2.6 Medicinal Uses of Ricinus communis                                                                                8

2.7 Morphological Description of M. fulvum

2.8 Habitat and Growth of Manniophyton fulvum

2.9 Phytochemical Constituents of Manniophyton fulvum                                                       10

2.10 Medicinal Uses of Manniophyton fulvum                                                                                    11

2.11 Brief Description of the Test Organisms                                                                          11

2.11.1 Escherichia coli                                                                                                            11

2.11.2 Staphylococcus aureus                                                                                                             12

2.11.3 Enterococcus faecalis                                                                                                   13

2.11.14 Salmonella typhi                                                                                                         13

CHAPTER THREE

MATERIALS AND METHODS

3.1 Source of Plant Materials                                                                                                   14

3.2 Sample Preparation                                                                                                            14

3.3 Extraction of Samples                                                                                                        14

3.4 Evaporation of Samples                                                                                                     14

3.5 Phytochemical Screening                                                                                                   15

3.5.1 Tannins Test                                                                                                                   15  

3.5.2 Terpenoids (Salkowski Test)                                                                                           15

3.5.3 Saponins Test                                                                                                                 15

3.5.4 Flavonoids (Alkaline Reagent Test)                                                                                15

3.6 Antibacterial Activity                                                                                                         16

3.6.1 Preparation of Stock Solution of Extract                                                                                     16

3.6.2 Reactivation of Stock Culture of Test Organisms                                                           16

3.6.3 Agar Dilution Susceptibility Testing                                                                                16

3.6.4 Determination of MIC and MBC Values

CHAPTER FOUR

4.1 RESULTS                                                                                                                         18

CHAPTER FIVE

5.1 Discussion                                                                                                                                      25

5.2 Conclusion                                                                                                                        27

5.3 Recommendation                                                                                                                27

References  

LIST OF TABLES

TABLE                                   TITLE                                                                                     PAGE

4.1          Phytochemical Screening of M. fulvum and R. communis Leaf Extract                 20

4.2          Diameter Zones of Inhibition (mm) produced by Ethanol extracts of

               M. fulvum against selected test organisms                                                               21

4.3          Diameter Zones of Inhibition (mm) produced by Ethanol extracts of R.

              communis against selected test organisms                                                                22

4.4          Agar Dilution Susceptibility Testing of Ricinus communis and M. fulvum                         23

4.5          MIC and MBC values (mg/ml) of ethanol extract of M. fulvum and R. communis

                 against susceptible Organisms                                                                                     24

CHAPTER ONE

            1.1              INTRODUCTION

An antimicrobial is a compound that kills or inhibits the growth of microbes such as bacteria. Such a compound is said to have antibacterial activity (Jagessar et al., 2008).  Medicinal herbs are a rich source of antimicrobial agents (Karinge, 2006; Bandaranayake et al., 2006; Mosihuzzanman and Chowder, 2008).  A wide range of extracts from medicinal plant parts are used as raw drugs and possess varied medicinal properties (Gisesa, 2004; Egwaikhide and Gimba, 2007). Primitive people learned by trial and error to distinguish useful plants with beneficial effects from those that were toxic or non-active and also which combinations or processing methods had to be used to gain consistent and optimal results (Jagessar et al., 2008). This reliance on herbal medicine has proven to be effective in the treatment of long term illness namely diabetes, malaria and pneumonia where it is seen to have lesser side effects and a cheaper form of medicine and preventive measure against diseases (Jagessar et al., 2007). In spite of the great advances observed in modern medicine in recent decades, plants still make an important contribution to health care (Ravikumar et al., 2010). In comparison with modern medicine, herbal medicines cost less, are more often used to treat chronic diseases and the occurrence of undesirable side effects seems to be less frequent (Jagessar et al., 2007). Several factors have contributed to the growth of the use of traditional herbs worldwide, among which are: preference of consumers for natural therapies, concern regarding undesirable side effects of modern medicines and the belief that herbal drugs are free from side effects, great interest in alternative medicines; preference of populations for preventive medicine due to increasing population age, the belief that herbal medicines might be of effective benefit in the treatment of certain diseases where conventional therapies and medicines have proven to be inadequate; tendency towards self-medication, improvement in quality, proof of efficacy and safety of herbal medicines and high cost of synthetic medicines (Ravikumar et al., 2010; Ceylan and Fung, 2004; De, 2004; Rattanachaikunsopon and Phumkhachorn, 2010).

The problem of antibacterial resistance to commonly used antibiotics has led to a search for newer and alternative compounds for the treatment of drug-resistant infections. Several findings on chemotherapeutic potentials of plants have shown that they can be sources of antimicrobial compounds of value and typical examples of such plants are Ricinus communis and Manniophyton fulvum. Presently there are global problems of antibiotic resistance to infections coupled with the emergence of new and re-emerging diseases. There is also a belief that the use of plants for medicinal purposes has been associated with fewer side effects. There is therefore a need to search for more efficacious and cost-effective antimicrobial agents of natural origin to complement the existing synthetic antimicrobial drugs that are becoming less potent against pathogenic microorganisms.

From another standpoint, antibiotic resistance has become a global concern (Westh et al., 2004). There has been increasing incidence of multiple resistances in human pathogenic microorganisms in recent years largely due to indiscriminate use of commercial antibiotic drugs commonly employed in the treatment of infectious diseases. This has forced scientists to search for new antimicrobial substances from various sources like the medicinal and herbal plants. Search for new antibacterial agents should be continued by screening many plant families. Recent work revealed the potential of several herbs as sources of new drugs (Iwu, 2002).

Medicinal plants were found to contain various phytochemical compounds which are used as natural medicines to treat common bacterial infections. The potential for developing antimicrobials from medicinal plants appears rewarding as it may lead to development of phytomedicine against microbes.

In the past few decades, the antimicrobial properties of various plant species of family Euphorbiaceae have been well studied by several workers such as Acalphya indica (Hiremath et al., 1993); Croton urucurana (Marize et al., 1997), Alchornea cordiflora (Ebi, 2009) and Tetracarpidium cocnophorum (Ajaiyeoba and Fadare, 2006). The screening of plant extracts and plant products for antimicrobial activity has shown that higher plants represent a potential source of novel antibiotic prototypes (Afolayan, 2003).

According to literature, little studies have been carried out to investigate the antibacterial activity and phytochemical properties of Ricinus communis and Manniophyton fulvum. Ricinus communis L (Euphorbiaceae) popularly called castor plant is a soft wooden small tree, wide spread throughout tropicsand warm temperatures regions of the world (Parekh and Chanda, 2007). The seeds are the source of castor oil, used as carthartic and also for lubrication and illumination. The oil is used in the manufacture of seabic acid, surface coatings, disinfectants, cosmetics and pharmaceuticals preparation (Bringi et al., 1985). The seeds of Ricinus communis are biochemically composed of various macromolecules: the fat which is about 15 to 25% consists of about 40 to 53% of fixed oil comprising glycosides of ricinoleic, isoricinoleic, stearic and dihydroxy stearic acids (Lin and Areinas, 2007). Ricin is a well-known poisonous compound that elicits violent purgative action in man (Trease and Evans, 2002). Also, the seeds contain about 25% protein with 10 to 20% carbohydrates, 2.2% ash and 5.1 to 6.5% moisture (Verscht et al., 2006).

In the traditional system of medicines, euphorbiaceae plants are used to treat various microbial diseases such as diarrhea, dysentery, skin infections and gonorrhea (Ajibesin et al., 2008). The effects of plant extract on bacteria have been studied by a very largenumber of researches in different parts of the world (Ates and Erdogrul, 2003). There are several reports in the literature regarding the antibacterial activity of crude extracts prepared from plants (El-seedi et al., 2002; Rojas et al., 2003; Duraipandiyan et al., 2006; Parekh and Chanda, 2006).

            1.2              OBJECTIVES OF THE STUDY

The main objective of this study was to investigate the antibacterial activity and phytochemical properties of Ricinus communis and Manniophyton fulvum.

The specific objectives of the study were:

a.       To evaluate and determine the antibacterial activity ofRicinus communis and Manniophyton fulvum against selected test organisms

b.      To determine the MIC and MBC values of the extracts against susceptible organisms

c.       To screen the plant materials for phytochemical components.

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