ABSTRACT

The aim of the study was assessing the antibacterial effect of Methanol and Aqueous extracts of ginger and non-antibiotic drugs against Escherichia coli, Staphylococcus aureus and Pseudomonas aeruginosa, Enterococcus faecalis. The antibacterial activities of extracts were evaluated using the disk diffusion method as well as agar well diffusion method; the inhibitory zones were recorded in millimeters. The minimal inhibitory concentration (MIC) of the plant extracts against E. coli, S. aureus and P. aeruginosa, E. fecaelis and S. typhi were assessed using microdilution method. The synergistic effect between plants and extraction of antibiotics and / or Non-antibiotic drugs was assessed using disk diffusion method. The results of this study showed that methanolic extracts used against E. coli, S. aureus and P. aeruginosa E. fecaelis and S. typhi were showed antimicrobial and synergistic effect with most antibiotics better than methanolic and aquatic extracts. The results of this study showed that there is a decrease in MIC in case of methanolic extract of E. faecalis against S. typhi and the methanol and aquatic extract of ginger against S. aureus varying from 6.25 to 3.125 mg/ml, and the methanol extract of E. faecalis against P. areuginosa (6.25 mg/ml).the phytochemical results shows alkaloid 1.92, Saponin 1.08, Terpenoid 2.28 and Flavonoid 0.56.  Thereby, our results indicate the possibility of using these extracts in the treatment of bacterial infections, and the results of this study was encouraging, despite the need for clinical studies to determine of the real effectiveness and potential toxic effects in vivo. These results was revealed the importance of plant extracts when associated with antibiotic and Non-antibiotic drugs in control of bacteria.

TABLE OF CONTENT

Title page                                                                                                                               i

Certification                                                                                                                          iii

Dedication                                                                                                                             iv

Acknowledgement                                                                                                                v

Table of Content                                                                                                                   vii

Abstract                                                                                                                                 ix

CHAPTER ONE

INTRODUCTION                                                                                                              1

1.1 Background of the Study                                                                                                            1

1.2 Aims and Objectives of the Study                                                                                  5

CHAPTER TWO

REVIEW OF LITERATURE                                                                                          

2.1 Literature Review                                                                                                            5

2.2. Origin and Geographical Location                                                                                 5

2.3 Medicinal Properties of Ginger                                                                                       6

2.4 Antimicrobial Susceptibility Testing Method                                                                  7

2.5 Medicinal Properties of the Plants                                                                                   8

2.6 Phytochemical Constituents of Ginger                                                                            10

2.7 Chemical Composition of Ginger                                                                                    11

2.8 Overview of Test Organism                                                                                            13

CHAPTER THREE

RESEARCH METHODOLOGY                                                                 

3.1 Materials                                                                                                                          18

3.2 Collection, Preparation and Extraction                                                                           18

3.3 Preparation of Stock Extract Culture of Test Organism                                                 19

3.4 Preparation of Stock Solution of Extract                                                                                    19

3.5 Reactivation of Stock Culture of Test Organism                                                            19

3.6 Antimicrobial Testing in the Agar Well Diffusion Method                                            19

3.6 Determination of Minimum Inhibitory Concentration                                                    20                                          

3.7 Quantitative Analysis                                                                                                      20

3.7.1 Phytochemical analysis                                                                                     20

CHAPTER FOUR

RESULTS AND DISCUSSION

4.1      Result                                                                                                                          22

CHAPTER FIVE

CONCLUSION, SUMMARY AND RECOMMENDATION

5.1 Conclusion and Summary                                                                                                25

5.2 Recommendation                                                                                                             25

5.3 Suggestion for future study                                                                                             25

CHAPTER ONE

INTRODUCTION

1.1       Background of the Study

There has been a great swing from the use of antibiotics to the use of remedial plant (Shahid and Hussain, 2014). A relatively small percentage of about 500.000 kind of plant are used as flavors, drugs and food(Ekwenye and Elegalam, 2005). Plant species have been used for centuries as remedies for human diseases. Ginger has been used as remedy for a range of illnesses including diarrhoea, stomach aches, nausea asthma, and respiratory disorders because ginger contain components of therapeutic values. The approval of traditional medicine as a reserve form of health care and the progress of microbial resistance to the existing antibiotics has directed researchers to explore antimicrobial activity of medicinal plants (Kaushik and Goyal, 2011) Ginger extract derived from the rhizomes of Zingiber officinale Roscoe (Adel and Prakash, 2010), is a famous spice used in the regular diet in many Asian countries(Habib et al., 2008; Shihet al., 2014), Chemical analysis of ginger illustrations that it consist from  more than  400 different compounds. The major constituents in ginger rhizomes are lipids (3–8%), terpenes, phenolic compounds, and carbohydrates (50–70%) (Shahid and Hussain, 2014), that have anti-inflammatory, antioxidant, anti-angiogenesis anti-artherosclerotic and anti-cancer properties. Ginger has direct anti-microbial activity and thus can be used in treatment of bacterial infections(Islam et al., 2014) The potential active chemical constituents of ginger include: Sesquiterpenes compounds like bisapolene, zingiberene, zingiberol, sesquiphellandrene, curcurmene, Phenolic compounds like shogaols and gingerols., and Other compounds  like 6-dehydrogingerdione, galanolactone, gingesulfonic acid, zingerone, geraniol, neral, monoacyl digalactosyl glycerols, gingerglycolipids (Islam et al., 2014).

Ginger (Zingiber officinale), belongs to the family Zingiberacae, and is a perennial herb with thick tuberous rhizomes. The natural products are found to be more effective with least side effects as compared to commercial antibiotics so that reason they are used an alternated remedy for treatment of various infections. (Tepe et al., 2004). Spices are defined as plant substances used to enhance flavor, they include leaves(mint and coriander), flower (clover), bulbs (Ginger, turmeric), fruits(black pepper), stem (cinnamon), rhizomes (ginger and turmeric). Medicinal plants produce certain bioactive molecules which show both antibacterial and antifungal activities. (Chopra et al., 2002). Many medicinal plants produce antioxidant and antimicrobial properties which protect the host from cellular oxidation reactions and other pathogens highlighting the importance of search for natural antimicrobial drugs. (Mothana and Lindequist, 2004, Bajpai et al., 2005; Wojdylo et al., 2007). Most of the foods borne bacterial pathogens are sensitive to extracts from plants such as Ginger, mustard, onion and oregano. Gram positive bacteria are more sensitive to antimicrobial compounds in spices than Gram negative bacteria (Lawson, 2006).

Ginger is therapeutically effective because of its oil and water soluble   organosulfur compounds, thiosulfinates is mainly responsible for its antibiotic activity as Hughes and Lawson, (2001) reported that if extract is free from thiosulfinates the antimicrobial capacity will be lost. Ginger has antibacterial and antifungal activity and contains powerful sulfur and numerous phenolic compounds. (Benkeblia, 2004).Previously conducted researches confirmed that ginger is not only effective against many Gram positive and Gram negative bacteria but also possess antiviral and antifungal activity. (Whitemore and Naidu; 2000, Rosss et al., 2001; Tsao and Yin., 2001, Martin and Ernst., 2003). Antibacterial activity may vary between different strains of same species and moreover depends on the form that is used such as dried, fresh or extracted and also on the harvesting seasons and geographical area as well. (Nanasombat et al., 2005, Cervenka et al., 2006).The ginerols (an essential oil) have antibacterial properties. It has been showed that ginger extracts have medicinal properties, antibacterial activity. Also, Ginger inhibited the growth of Staphylococcus aureus and Streptococcus pyogenes (Malu et al., 2009).

Scientific investigations for medicinal plants have been initiated in many countries because of their contributions to health care. It was cleared that the primary benefits of using plant derived medicines are relatively safer than synthetic alternatives, offering profound therapeutic benefits and more affordable treatment. Current social trends in health care showed a definite movement towards the use of natural remedies like medicinal plants away from chemotherapeutic regimens (Selvamoha et al., 2012). The use of crude extracts of plants parts and phytochemicals, of known antimicrobial properties, can be of great significance in the therapeutic treatments. Also, use of plant essential oils in both food and pharmaceutical industries has been developed interestingly; systematic examination of plant extracts for these properties has become increasingly important. The use of natural plant antimicrobial compounds is important not only in the preservation of food but also in the control of microbial growth in disease condition (Rasool, 2013). In recent studies, extracts of various parts of medicinal plants were found to have broad spectrum antimicrobial activities against pathogenic organisms (Oyetaro, 2008).

Antibacterial agents are effective in curing diseases because of their selective toxicity against pathogenic microbes without causing any harm to the cells of the host (Thawal et al., 2013). Some studies concluded that the spices may be very valuable because bacteria develop resistance to conventional antibiotics (Gull et al., 2012). Antibiotic resistance has become a global concern, therefore there has been an increasing incidence of multiple resistances in human pathogenic microorganisms largely due to indiscriminate use of commercial antimicrobial drugs commonly employed in the treatment of infectious diseases (Menghani, 2011).

Thus, this study is aimed at investigating the antibacterial activity of aqueous and methanol extract of ginger.

1.2  Aims and Objectives of the Study

The aims and objectives of the study are as follows:

        i.            To determine the minimum Inhibitory concentrate (MIC) and Minimum Bacterial Concentration (MBC) of the extracts against the test organisms

      ii.            To determine / evaluate the phytochemical composition of Z. officinale.

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