EVALUATION OF ANTIBACTERIAL ACTIVITY OF GARCINIA KOLA AND DENNETTIA TRIPETALA AGAINST SOME BACTERIA

ABSTRACT

Garcinia kola and Dennettia tripetala were examined for their antibacterial activity against clinical isolates (S. aureus and E.coli). The samples (Garcinia kola and (Dennettia tripetala) were examined using different solvents (water and ethanol) and Agar Well diffusion technique. The minimum inhibitory concentration and minimum bactericidal concentration of the samples was also determined. The invitro antibacterial activity results showed that both the water and ethanol extract of the samples exhibited inhibitory activity against the test isolates. The ethanol extract of both Garcinia kola sample and Dennettia tripetala sample exhibited greater antibacterial activity that the water extracts of the samples. From the result of this study, Dennettia tripetala sample was more effective than the Garcinia kola sample ranging from 30mm at 200mg/ml to 18mm at 12.5mg/ml using ethanol , and 25mm at 200mg/ml to 12mm at 12.5mg/ml using water extract against S. aureus and 35mm at 200mg/ml to 20mm at 25mg/ml using ethanol extract, 30mm at 200mg/ml to 25mm at 100mg/ml using water extract against E.coli. The research findings showed that the traditional plant leaves used in this study exhibit varying degree of antibacterial activity.

TABLE OF CONTENTS

Title page                                                                                                                                i

Certification                                                                                                                           ii

Dedication                                                                                                                              iii

Acknowledgement                                                                                                                  iv

Table of contents                                                                                                                    v

List of tables                                                                                                                           vii

Abstract                                                                                                                                  viii

CHAPTER ONE

 1.0 Introduction                                                                                                                     1

1.1 Antimicrobials from phytomedicines                                                                               6

1.2 Present use of plants as antimicrobials                                                                            7

1.3 Aims and objectives                                                                                                         6

CHAPTER TWO

 2.0 Literature Review                                                                                                                        8       

2.1 Garcinia kola (bitter kola)                                                                                                                         8

 2.2 Dennetia tripetala (pepper fruit)                                                                                      17 

CHAPTER THREE     

3.0 Materials and Methods                                                                                                     21    

3.1 Collection of plant materials                                                                                            21

3.2 Sample collection                                                                                                             21

3.3 Media used                                                                                                                        21

3.4 Sterilization                                                                                                                      21

3.5 Identification of bacterial isolates                                                                                    21

3.6 Agar well diffusion technique                                                                                          22

3.6.1 Broth dilution technique                                                                                                22

3.6.2 Determination of minimum inhibitory concentration (mic)                                          23

and minimum bactericidal concentration.     

 3.7 Confirmatory test                                                                                                             23

3.7.1 gram staining                                                                                                                 23

3.7.2 Catalase test                                                                                                                   24

3.7.3 Coagulase test                                                                                                                24

3.7.4 Citrate test                                                                                                                      24

3.7.5 Motility, indole, urease test (MIU)                                                                                25

3.7.6 Triple sugar iron test                                                                                                      25

3.7.7  Oxidase test                                                                                                                  26

3.7.8 Determination of antibacterial activity                                                                         26

CHAPTER FOUR

4.0 Result                                                                                                                                27                                                                                                                   

CHAPTER FIVE

5.0 Discussion, Conclusion and Recommendation                                                                32

5.1 Discussion                                                                                                                         32

5.2 Conclusion                                                                                                                        33

5.3 Recommendation                                                                                                             33

References                                                                                                                              34

LIST OF TABLES

Table 1: Diameter zones of inhibition (mm)/concentration (mglml) of                               28

Pepper fruit and bitter kola against S.aureus                                                             

Table 2: Diameter zones of inhibition (mm)/concentration (mglml) of                               29

Pepper fruit and bitter kola against E. coli                                                                

Table 3: MIC, MBC of S.aureus pepper fruit and bitter kola                                                30

Table 4: MIC, MBC of E. coli pepper fruit and bitter kola                                                    31

CHAPTER ONE

1.0      INTRODUCTION

The study of higher plant for the purpose of detecting antimicrobial agents in their tissues is of comparatively recent origin and the early investigation in this area focused on those plants that have found application in the age-old practice or their blind usage as therapeutics for human and animal diseases (Benjamine et al., 1983; Okigbo and Nmeka, 2005). The use of complementary medicines increased the interest of pharmacologist and herbalists over the past decade. Historically, medicinal plants have provided a source of inspiration for novel drug compounds, as plant derived medicines have made large contributions to human health and wellbeing. On the other hand, there is an increment of herbal products all over the world, in USA, it reached 38% between 1990 and 1997 (Eisenberg et al., 1990). According to Arora and Keur (1999), the success story of chemotherapy lies in the continuous search of new drugs to counter the challenges posed by resistance strains of microorganisms. The investigation of certain indigenous plants for their antimicrobial properties mayyield useful results. Many studies indicate that in some plants there are many substances such as peptides, alkaloids, essential oils, phenols, coumarines and flavonols which confer antimicrobial properties to them. These compounds have potentially significant therapeutic application against human pathogens, including bacteria, fungi or virus (Arora and Keur, 1999; Okigbo and Igwe, 2007). The search for natural products to cure diseases represents an area of great interest in which plants have been the most important sources because of the prevalence of microbial resistance to existing synthetic drugs. In addition, in order to halt the trend of increased emerging and microbial resistance infectious diseases, it will require a multi prolonged approach that includes the development of new drugs (Iwu et al., 1999, Okigbo and Ajalie, 2005). Thus, evaluating plants from the traditional African system of medicine provide us with the clues as to how they can be used in the treatment of diseases (Okigbo et al., 2005). The world Health organization (WHO) has estimated that up to 80% of the world’s populations rely on plants for their primary Health care. In Nigeria, a 1985 WHO survey estimated that up to 75% of the population patronizes traditional medicine (Omoseyindemi, 2003). Many of the drugs used in modern medicine were initially used in in crude form in traditional or folk healing practices or other purposes that suggest potentially useful biological activity (Iwu et al., 1999). However, it has become necessary to investigate the anti microbial effect of these plants and drugs derived from them. Investigation of medicinal potentials of plants therefore may lead to the development of plant-based drugs. The use of plants in the maintenance of good health is well reported (Burkill, 1995; Edeoga and Eriata, 2001; Moerman, 1996). It has also been reported that the bases of many modern pharmaceuticals used today for the treatment of various ailments are plants and plant based products (Kambba and Hassan, 2010). Plants have been generally utilized for the treatment of diseases worldwide. About 80% of the world populations depend on plants based medicine for their health care (WHO, 2001). WHO (1996), also observed that the majority of the population in the developing countries still rely on herbal medicines to meet their health need. The use of plants and plant based products to meet societal health need stems from the fact that indiscriminate use of commercial antimicrobial drugs commonly utilized in the treatment of infectious diseases has led to the development of multiple drug resistance (Gupta et al.,2008), the adverse effect on host, associated with the use of conventional antibiotics (Gupta et al., 2008), the safety and cost effectiveness of the use of plants in traditional as well as in modern medicine (Koche et al.,2011), and high cost, adulteration and increasing toxic side effects of these synthetic drugs (Shariff, 2001). Thus there has been the need to develop alternative antimicrobial drugs from medicinal plants for the treatment of infectious diseases; because antimicrobials of plant origin have been found to have enormous therapeutic potential (Werner et al., 1999). Furthermore, the studies by Perumalsamy and Ignacimuthu (2000) showed that antimicrobials from plant origin are effective in the treatment infectious diseases and on the other hand simultaneously mitigates many of the side effects that are linked with synthetic antimicrobials. Many of these indigenous plants contain bioactive compounds that exhibit physiological activities against bacteria and other microorganisms and are also used as precursors for the synthesis of useful drugs. Thus the use fullness of these plant products in medicine is due to the presence of bioactive substances such as alkaloids, tannins, flavonoids, phenolic compounds, steroids, resins and other secondary metabolites which they contain and are capable of producing definite physiological action in the body (Bishnu et al, 2009; Edeoga et al., 2005). Phytochemicals are known to carry out important medicinal roles in the body. Alkaloids are known to have a powerful effect on animal physiology. They play some metabolic role and control development in living system (Edeoga and Eriata, 2001). They are also used as starting materials in the manufacture of steroidal drugs and carry out protective function in animals, thus are used as medicine especially steroidal alkaloids (Maxwell et al., 1995; Stevens et al., 1992). Isolated pure plant alkaloids and their synthetic derivatives are used as basic medicinal agent for their analgesic, antispasmodic and bacterial effect (Ogukwe, et al., 2004). Flavonoids are known to carry out antioxidant, protective effects and inhibit the initiation, promotion and progression of tumors (Kim et al., 1994; Okwu, 2004). Isoflavones, some kind of flavonoids are phytoestrogen which effectively modulate estrogen levels in human (Okwu and Omodamiro, 2005). A type of flavonoid anthocyanin helps in reducing the incidence of cardiovascular diseases, cancer, hyperlipidemias and other chronic diseases (de Pascual-Teresa and Sanchez-Ballesta, 2008). Phenolic compounds in plants are potentially toxic to the growth and development of pathogens (Singh and Sawhney, 1988). Research reports also show that phenolic compounds carry out potent antioxidant activity and wide range of pharmacologic activities which include anti- cancer, antioxidant and platelet aggregation inhibition activity (Rein et al., 2000; Rice – Evans et al., 1996;). Saponins play essential roles in medicine. These include serving as expectorant and emulsifying agent (Edeoga et al., 2009) and having antifungal properties (Osuagwu et al., 2007). Tannins are reported to inhibit pathogenic fungi (Burkill, 1995). They are also associated with many human physiological activities such as stimulation of phargocytic cells and host mediated tumor activity and a wide range of infective actions (Haslam, 1996). Steroid containing compounds are of importance in pharmacy due to their role in sex hormones (Okwu, 2001). Steroids such as equine estrogen are implicated in the reduction of risks of coronary heart andneurodegenerative diseases in healthy and young postmenopausal women (Perrella et al., 2003). At low concentration tannins show antimicrobial, cytotoxic and astringent properties (Zhu et al., 1997; Ijeh et al., 2004). The phytochemical screening of some plants has been carried out and they are found to be rich in alkaloids, phenols, flavonoid, saponin and tannins (Osuagwu et al., 2007; Iniaghe et al., 2009; Ganjewala et al., 2009; Omoyeni and Aluko, 2010). The antimicrobial activities of plants have been reported (Arshad et al., 2010; Kamba and Hassan, 2010, Koche et al., 2011). They are therefore used in the treatment of many diseases such as rheumatism, diarrhea, malaria, elephantiasis, cold, obesity, dysentery, high blood pressure, malnutrition, gonorrhea and others (Burkill, 1995; Edet et al., 2009; Akuodor et al, 2010).

1.1    ANTIMICROBIALS FROM PHYTOMEDICINES

A vast number of plant species have been screened for antimicrobial actions (Ogunlana and Ramstad, 1975; Leven et al., 1979; Nair and Burke, 1990; Okigbo and Mmeka, 2006). In recent times, interest in this area has increased tremendously due partly, to certain disadvantages that are associated with the use of many synthetic antimicrobial agents and to the rising incidences of multidrug resistance against these agents (Odama et al., 1997). These shortcomings include their toxicity, the ability of organisms to develop resistance to the drugs previously known to be effective, and loss of potency of the drug with time. On the other hand, the merits of herbal medicine over orthodox drugs include; minimal or no side effects on the organic functioning of the body, consistent potency, and the fact that they are well absorbed and distributed in the area of infection (Cheij, 1988; Nkere, 2003; Okigbo and Omodamiro, 2006). The role of plants in herbal medicine as the major remedy in traditional medicinal system has been in medical practice for thousands of year. The use of plants have made great contribution to maintaining human health and thus, a majority of the world’s population in developing countries still relies on herbal medicine to meet their needs (WHO, 1991).  Nigeria is one of the countries rich in rare and useful herbs, thus providing a vast area of medicinal plant research for novel drug development (Sofowara, 1993; Okigbo and Mmeka, 2006). Many of these plants are designated weeds and are used as potherbs. Their healing powers as claimed by local medicinal practitioners range from headache through skin diseases to gonorrhea and syphilis (Akobundu, 1987; Burkill, 1997). Other ailments treated with these medicinal plants include asthma, cough, diarrhoea, malaria, diabetes, bleeding, childcare, healing of wounds and sores and tooth extraction. Some of these medicinal plants are used as styptic and as simple laxative as cure for dysentery. They can be given to pregnant women or nursing mothers in food for various medicinal reasons. They are also good sources of pesticide (Burkill, 1997; Gill, 1992; Okwute, 1992; Okigbo and Nmeka, 2005; Okigbo and Ogbonnaya, 2006). Many of these plants have leaves that are consumed as leafy vegetables. These vegetables have nutritional potentials that can be harnessed for dietary purposes of man (Edeoga and Gomina, 2000; Okigbo and Mmeka, 2008). The plant parts used include the leaves, roots, barks and stems. Some of these plants are also used as animal feeds on by local farmers who do not have money to buy expensive compounded commercial feed (Edeoga and Erita, 2001). Though a lot of the screened plants that have antimicrobial properties have not been used in modern medicine, their usage in traditional medical practice is fairly very high (Iwu et al., 1999).

1.2 PRESENT USE OF PLANTS AS ANTIMICROBIALS

It is estimated that today, plant materials are present in, or have provided the model for 50% of Western drugs (Robbers et al., 1996). Many commercially proven drugs used in modern medicine were initially used in crude form in traditional or folk healing practices, or for other purposes that suggested potentially useful biological activity. The primary benefits of using plant derived medicines are that they are relatively safer than synthetic alternatives, offering profound therapeutic benefits and are more affordable (Iwu, 1999; Okigbo and Mmeka, 2006).

1.3 AIMS AND OBJECIVES

The aim of this research is to evaluate the antibacterial activity of Garcinia kola and Dennetia Tripetala on some clinical pathogens.

OBJECTIVES

1.     To determine the minimum inhibitory minimum bactericidal concentration Garcinia kola and Dennetia Tripetala against some clinical isolation.

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