ASSESSMENT OF THE INVITRO ANTI BACTERIAL ACTIVITY OF PTEROCAPUS SANTTALINOIDES AND AGERATUM CONYZOIDES

Abstract

The antibacterial and phytochemical screening of A. conyzoides and P. santalinoides were carried out to find the medicinal importance of these plants among the people of Ikwuano L.G.A in Abia State. Agar well diffusion method was used to measure the zone of inhibition and broth dilution assay to assess the Minimum Inhibitory Concentration (MIC) of the anti-bacterial activity testing of the ethanol extracts of these plant extracts against selected susceptible organisms. The four test bacteria used were Staphylococcus aureus (ATCC 25923), Escherichia coli (ATCC 25922), Pseudomonas aeruginosa (ATCC) and Salmonella typhi. The methanol extract of A. conyzoides produced a diameter of zone of inhibition of 12.00mm against Staphylococcus aureus, 00.00mm against Pseudomonas aeruginosa, 00.00mm against Salmonella typhi while Methanol extract of P. santalinoides produced a diameter of zone of inhibition of 14.00 mm against Staphylococcus aureus, 10.00mm against Pseudomonas aeruginosa, 11.00mm against Salmonella typhi. Both plants extract had no effect on Escherichia coli. The phytochemical screening of these plants leaves showed the presence of alkaloids, tannins, Saponins, phenol and flavonoids. The research work suggests the use of A. conyzoides and P. santalinoides in herbal medicine.

TABLE OF CONTENTS

Title page                                                                                               i

Certification                                                                                                                           ii

Dedication                                                                                                                              iii

Acknowledgement                                                                                                                  iv

Table of content                                                                                                                      v

List of Tables                                                                                                                          iv

Abstract                                                                                                                                  viii

CHAPTER ONE

1.0 Introduction                                                                                                                      1

1.2 Aim and Objective                                                                                                           3

CHAPTER TWO

2.0 Literature review                                                                                                              4

2.1 Medicinal Plants                                                                                                               4

2.2 Ageratum conyzoides                                                                                                        6

2.3 Phytochemicals Properties of A. Conyzoides                                                                   7

2.3.1 Insecticidal activity                                                                                                       7

2.3.2 Anthelmintic activity                                                                                                     7

2.3.3 Antidiabetic activity                                                                                                      7

2.4 Pterocarpus santalinoids.                                                                                                 8

2.5 Phytochemicals Properties of Pterocarpus santalinoides                                                            9

2.5.0 Test organisms                                                                                                               9

2.5.1 Escherichia coli                                                                                                                                                                         9

2.5.2 Pseudomonas aeruginosa                                                                                              10

2.5.3 Staphylococcus aureus                                                                                                  10

2.5.4 Salmonella typhi                                                                                                            11

CHAPTER THREE

3.0 Materials and methods                                                                                                      12

3.1 Collections and identification of research plant                                                               12

3.1.1 Plant Leaves Preparation                                                                                               12

3.1.2 Extraction procedure of plant materials                                                                        12

3.2 MEDIA                                                                                                                             13

3.2.1 Preparation of Media                                                                                                     13

3.3 Preparation Stock Solution of Extract                                                                              13

3.4 Test Organism                                                                                                                  13

3.4.1 Reactivation of Test Organism                                                                                      14

3.5 Antibacterial bioassay of crude extracts                                                                          14

3.6 Determination of Minimum Inhibitory Concentration (MIC)                                          14

3.7 Determination of Minimum Bactericidal Concentration (MBC)                                     15

3.8 Phytochemical Screening                                                                                                 16

3.8.1 Alkaloids                                                                                                                       16

3.8.2 Flavonoids                                                                                                                     16

3.8.3 Tannins                                                                                                                          16

3.8.4 Terpenoid                                                                                                                      16

3.8.5 Saponin                                                                                                                          16

CHAPTER FOUR

4.0 Results                                                                                                                              18

CHAPTER FIVE

5.1 Discussion                                                                                                                                                                                                 23

5.2 Conclusion                                                                                                                        25

5.3 Recommendation                                                                                                             25

REFERENCES                                                                                                                     26

LIST OF TABLES

Table

4.1      Diameter zone of inhibition (mm) produced by Methanol extracts of A. conyzoides and P. santalinoides against some test organisms.                  19

4.2       MIC and MBC values (mg/ml) of the extracts against susceptible

organisms                 20

4.3       Quantitative Phytochemical constituents of plant extracts                    21

4.4       Qualitative  phytochemical screening of the plants             22

 CHAPTER ONE

1.0       INTRODUCTION

The importance of plants in traditional medicine and as raw materials in pharmaceutical industries cannot be overemphasized and its use to treat diseases is almost universal among under developing countries. A number of traditions came to dominate the practice of herbal medicine at the end of the twentieth century. Many of the pharmaceuticals currently available to physicians have a long history of use as herbal remedies, including opium, asprin, digitals and quinine ((Fukuyama et al., 2012). The use of medicinal plants is increasing worldwide in view of the tremendous expansion of traditional medicine and a growing interest in herbal treatments. Plants are used in medicine to maintain and augment health-physically, mentally and spiritually as well as to treat specific conditions and ailments. It has been found that countries in Africa, Asia and Latin America use traditional medicine to help meet some of their primary health care needs. In Africa, for example, up to 80 percent of the population uses traditional medicine for primary health care (Akinyemi et al., 2018).

The use of medicinal plants cuts across gender, social and racial classes including the developing and developed countries of the world. Due to the increasing popularity herbal plants, stakes in the local and international world markets are also rapidly increasing and the annual sale has approached US $62 billion. An important driver in this wide upsurge is as a result of patronage and use which boils down to low cost, the  wide acceptance due to its status of being a natural product with the acclaim of low toxicity,  efficacy in certain challenging diseases, flexibility in its accessibility, easy preparation and use (Fasinu et al., 2012).

These herbal medicines may be produced from any part of the plant. They are commonly made from the leaves, roots, bark, seeds, and flowers. The herbs are eaten, swallowed, drunk, inhaled, or applied to the skin (WHO, 2016). Herbal medicine includes preparations of any biologically active natural ingredient or products that consist largely of medicinal plants or herbal materials. Most recipes often contain materials such as bee products and funga as well as minerals (bentonite), ash, shells, insects and animal parts are all used for the maintenance of health and management of various diseases. Medicinal plants come with numerous health benefits while some can cause adverse health effects. The toxic and pharmacologic effects elicited by some medicinal plant have been linked to the activities of the secondary metabolites. In most cases, medicinal plants have been appropriately used, misused and sometimes misunderstood. The use of these plants should be investigated for better understanding of their properties, safety and efficiency. Due to the increasing interest in plants as source of agents to fight microbial pathogens and the wide spread incidence of antibiotic resistance of pathogenic microbes in particular, there is still necessity for discovering new and effective therapeutic agent (Chintamunnee and Mohomoodally, 2012).

A number of plant species are being used in various human health around the world. Plant species contain active ingredients such as alkaloids, phenols, tannins, cryogenics, glycocides, terpeniods. These ingredients have been used and found effective as sweeteners, anti-infections and anti-bacterials ((Fukuyama et al., 2012). It has been estimated that more than 400 traditional plants or plant derived products have been used for the management of type 2 diabetes across the world. Plant derived agents are also being used for the treatment of cancer. Several anticancer agents including vincristine, taxol, vinblastine, derivatives, irinotecan and topotecan and etoposide derived from epipodophyllotoxin are in clinical use worldwide. More so, it was used as condiments or seasoning in food, which in turn provides some health value to humans (Adepoju, 2017).

It also constitutes an integral part of the culture of many societies of the world. Many medicinal plants and herbal recipes have a long standing traditional history of community uses and claims of health benefits. Scientific research has shown that medicinal plants contain complex chemical compounds that are responsible for the pharmacological activities, which corresponds to health benefits and/or toxicity they elicit (Amoo et al., 2012). Medicinal plants have been used as prophylaxes for the passive maintenance of health as well as for radical treatment of varieties of mild to serious diseases (Parasuraman, 2014).

Plants promise a source of natural antimicrobial agents. It has been reported that the antimicrobial activity of plants is related with the defence mechanism against microorganism (Fukuyama et al., 2012). Other applications for natural antioxidants may include bioactive nutraceuticals, bio-pharmaceuticals, and food additives. In relation to that, the extraction, characterization and utilization of natural antioxidants are intensively performedto find potent candidatesin combating the aging process (Amoo et al., 2012).

1.2       AIM AND OBJECTIVE

1.         To screen two medicinal plants (Pterocarpus samtalinoides and Ageratum coryzoides) for antimicrobial activity against some bacterial pathogens.

2.         To carry out phytochemical screening of the ethanolic extracts of the medicinal plants.

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