ANTIBACTERIAL EFFECTS OF GONGRONEMA LATIFOLIUM (UTAZI) LEAF AND JATROPHA CURCAS (BARBADOS) LEAF EXTRACTS ON SOME CLINICAL BACTERIAL ISOLATES

ABSTRACT

Methanol and water extracts of J. curcas and G. latifolium were prepared in different concentration of 0.5mg/l to 5mg/l. Methanol extract showed more inhibitory properties over water extract. Gongrenema latifolium exhibited high inhibitory effect than J.curcas. The diameter of zone of inhibition of J.curcas range from 6mm to 10mm and 6mm to 14mm for water and methanol extracts respectively while the diameter zone of inhibition for G. latifolium range from 6mm to 10mm and 6mm to 26mm for water and methanol extract respectively. The minimum inhibitory concentration of J.curcas on E. coli was 0.5mg/l and 2.5mg/l on S. aureus and Salmonella typhi. The minimum inhibitory concentration (MIC) of G. latifolium on E. coli was 1mg/l while S. aureus and Salmonella typhi was 2.5mg/l. Therefore, the result obtained indicated that the extracts of Gongronema latifolium and Jatropha curcas possess antibacterial activities against Staphylococcus aureus, Salmonella typhi and Escherichia coli. Jatropha curcas are  efficacious for the remedy of boils, carbouncle and other entero-infection cause by Staphylococcus aureus, E. coli and Salmonella typhi.   

TABLE OF CONTENTS

Title page                                                                                                                                i

Certification                                                                                                                            ii

Dedication                                                                                                                               iii

Acknowledgement                                                                                                                  iv

Table of contents                                                                                                                    v

List of Tables                                                                                                                          viii

List of plants                                                                                                                           ix

List of plates                                                                                                                           x

Abstract                                                                                                                                   xi

CHAPTER ONE

1.0           Introduction                                                                                                              1

1.1           Aim and Objectives                                                                                                    4

CHAPTER TWO     

2.1        Literature Review                                                                                                 4

2.1.1      Phytomedicine                                                                                                                        4

2.1.2    Characteristics of Phytomedicine                                                                               5

2.2        The Demand for Phytomedicine                                                                          5

2.3       Benefits of Phytomedicines                                                                                        5

2.3.1    Therapeutic benefits                                                                                                   5

2.3.2    Plants with Medicinal Values                                                                                     7

2.4       Main Leaves of Study                                                                                                 9

2.4.1    Jatropha curcas (origin and distribution)                                                                  9

2.4.2      Cultivation                                                                                                                9

2.4.3      Medicinal properties                                                                                                 10

2.5       Gongronema latifolium (Utazi) (Origin and distribution)                                         10

2.5.1    Traditional and medicinal uses                                                                                   11

2.6       Biology and Pathogeneicty of Test Organism                                                            12

2.6.1    Escherichia coli                                                                                                          12

2.6.2      Staphylococcus aureus                                                                                               13

2.6.3    Salmonella typhi                                                                                                         14

2.7       Major groups of antimicrobial compounds from plants                                             15

2.8      Mechanisms of bacterial resistance to antibiotics                                                  19

2.8.1      Actions of some antibiotic during cell wall synthesis                                                20

CHAPTER THREE

3.1        Materials and Methods                                                                                      21

3.2        Collection of plant sample                                                                                    21

3.2.1      Materials used                                                                                                             21

3.2.2      Media used                                                                                                                  21

3.2.3      Reagents used                                                                                                             21

3.3           Preparation of the pant extract                                                                                   22

3.3.1      Methanol extract preparation                                                                                     22

3.3.2      Cold water  extract preparation                                                                                  22

3.4           Isolation of microorganisms                                                                                       24

3.5           Gram staining                                                                                                             24

3.6           Biochemical tests                                                                                                        24

3.7           Sterility test of the Jatropha crucas and Gongronema latifolium leaves extracts 29

3.8           Preparation Of Disc                                                                                                    29

3.8.1      Disc diffusion method                                                                                                29

3.8.2      Determination of antibacterial properties of the extracts                                           30

3.8.3      Determination of minimum inhibitory concentration                                                30

3.9           Phytochemical Analysis                                                                                             30

CHAPTER FOUR

4.0       Results                                                                                                                        32

4.1       Interpretation of Zone of the Inhibition                                                                     32

CHAPTER FIVE

5.0       Discussion, Conclusion and Recommendation                                                      41

5.1       Discussion                                                                                                                   41

5.2       Conclusion                                                                                                                  43

5.3       Recommendation                                                                                                       44

References

Appendix 1: Culture Media

Appendix 2: Sensitivity Plates

LIST OF TABLES

Table                                                                                Page

1          Triple sugar iron agar test                                                                                           28

2          Morphological and Biochemical Characterization                                                    33

3          Diameter of Zone of Inhibition of Different concentration of the extract             34

4          Diameter of zone of inhibition of different concentration of the extract             35

5          Antibiotic of Plant extract on tested bacterial isolates                                               38

6          Phytochemical screening of Jatropha curcas and Gongronema latifolium                    40

LIST OF PLANTS

Plant                                                                                            Page

1               Gongronema latifolium                                                          23

2               Jatropha curcas                                                                   23

LIST OF PLATES

Plate                                                                                     Page

1               Salmonella typhi                                                  36

2               E. coli                                                                                  37

3               Staphylococcus aureus                                                       37

CHAPTER ONE

1.0        INTRODUCTION

The treatment and control of disease by the use of available medicinal plants in a locality will continue to play significant role in medical health care implementation in the developing countries. The intractable problems of antimicrobial resistance has led to the resurgence of interest in herbal products as sources of noble compound to suppress or possible eradicate the ever-increasing problems of emergence of newer diseases though to be brought under control (Wurochekker et al., 2007). There is a growing interest in plants with antimicrobial activity. Scientist are increasingly becoming involved in the screening of such plants with the aim of establishing their potential antimicrobial effects and identifying the compounds responsible for the antimicrobial properties (Aibinu et al., 2007; Ndukwe et al., 2007). Traditional medicine using plants extract continues to provide health coverage for over 80% of the world’s population, especially in developing countries (WHO, 2002).

Jatropha species belong to the family Euphorbiaceeae and are used in traditional for folk medicine to cure ailments in Africa, Asia and Latin America. Jatropha curcas is commonly called physic nut Barbados nut and purging. Previous studies have reported that the plants exhibits bioactive activities for  fever, mouth infections, jaundice, guinea worm sores and joint rheumatic (Oliver-Bever, 2000). Aiyelaagbe (2001) reported that anti-parasitic activity of the sap and crushed leaves of J. curcas. The water extract of the branches also strongly inhibited HIV induced cytopathic effects with low cytotoxicity. Previous works have shown that many Jatropha species possess antimicrobial activities (Aiyelaagbe, 2001). The caparison of phytochemicals of old and new growth leaves from Nine trees used medically in Nigeria shows that Jatrophacurcas contains saponin, tannin, steroid, alkaloid, phenols and flavonoid (Fasola and Egunyomi, 2005). J. curcas leaves have significant variability in their morphology. In general, the leaves are green to pale green alternate to sub-opposite and three to five labeled with a spiral phyllotaxis (Nahar and Ozore-hampton, 2011). Herbs that have tannin as their main components are astringent in nature and are used for treating intestinal disorders such as diarrhoea and dysentery (Kan, 2002).

Gongronemalatifolium, known as Utazi in the south-eastern and Adkekein the South Western part of Nigeria, is a tropical rainforest plants which belongs to the family Asclepiadaceae (Ugochukwu and Baddy, 2002; Ugochukwu et al., 2003). It is a climber with a tuberous base found in deciduous forests from Guinea-Bissau to western Cameroons. Various parts of these pants, particularly the stems and leaves are used as chew-sticks or liquor in places such as sierra-0leone. The liquor, usually obtained after the plants is sliced and boiled with lime juice or infused in water for over three days, is frequently taken as a purge for colic and stomach pains as well as to treat symptoms connected with worn infection (Okafor, 1975). The plants have also been widely used in folk medicine for maintaining healthy blood glucose levels (Okafor, 1987, 1989). The plants leaves have been found very efficacious as an anti-diarrhoea and anti-tussive (Sofowora, 1982; Iwu, 1993).

            Staphylococcus is a group of bacteria that can cause a number of diseases as a result of infection of various tissues of the body. Staphylococcus-related illness can range from mild and requiring no treatment to severe and potentially fatal. Staphylococcus bacteria look like a bunch of grapes or little round berries. They are gram positive, facultative anaerobic, usually un-encapsulated cocci (Todar, 2008). Over 30 different types of Staphylococcus can infect humans, but must infections are caused by Staphylococcus aureus. Staphylococci can be found normally in the nose and on the skin (and less commonly in other locations) of around 35 to 30% of healthy adults and in 25% of hospital workers (Enright et al., 2002). In the majority of cases, the bacteria do not cause disease. However, damage to the skin or other injury may which may allow the bacteria to overcome the natural protective mechanisms of the body, leading to infection. Leaching to infection, Staphylococcus aureus constitute a major public health threat, being one of the most common cause of hospital and community acquired infections (Aires-de-Sousa et al., 2006). These organisms are carried by people on skin, in boils, pimples and throat infections. Spread when carriers handle food. S. aureus bacteria produce toxins (poisons) at warm temperatures. Meat, poultry, salads, cheese-eggs, custards and cream filled desserts are susceptible food. Food poisoning symptoms include vomiting, diarrhoea, nuasea and abdominal cramps lasting up to 1 to 2 days. Staphylococcus infections are contagious until the infection has resolved. It is on record that of all Staphylococcus aureus infections in the U.S are resistant to penicillin, tetracycline and erythromycin and Vancomycin. The same can be said to the organisms in most parts of the world. According to Prasad et al., (2008), the emergence of multi-drug resistant bacterial strains throughout the globe limits the effectiveness of current drugs and significantly limits treatment, leading to prolonged infections. The increasing resistance of bacteria to antibiotics is kindly due to the misuse and over prescription of drugs. As resistance to antibiotic spreads, the development of new antimicrobial agents has to be expedited if the problem is to be contained. There is therefore a need to develop new antibiotics to delay or prevent the arrival of post-antibiotics to delay or prevent the arrival of a post-antibiotic era.

1.1        AIM AND OBJECTIVES

1.     To determine the Minimum Inhibitory Concentrations (MIC) of the plants extracts (Jatropha curcas and Gongrenema latofolium) against the Escherichia coli, Salmonella typhi and Staphylococcus aureus.

2.     To compare the degree of effectiveness of the cold water and methanolic extracts against Escherichia coli, Salmonella typhi and Staphylococcus aureus.

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