GARRI AND AKAMU AS ALTERNATIVE CULTURE MEDIA FOR THE ISOLATION OF FUNGI

ABSTRACT

Newly obtained rhizome of Zingiber officinale (Ginger) and cloves of Allium sativum (Garlic) were obtained from the  market, they were put together and left at 25oC to permit air drying, milled to fine powder and then this powder were extracted (each alone) using distilled water and ethanol as solvents for the extraction into a sterilize air tight glass bottles. After that the extracts were examined for its antibacterial (inhibitory) effect toward clinical isolates of some gram negative organisms such as Escherichia coli and Salmonella typhi. Two kind of extract for garlic and two kind of extract for ginger (Aqueous and Ethanoic) were extracted/obtained and were then examined separately and compared with the antibiotics ceftriaxone. Agar well diffusion method was used to determine the antibacterial activity of the extract. The test isolates showed variable susceptibility to the garlic and ginger extract (Aqueous and Ethanoic) and to the antibiotics ceftriaxone used as control. The outcomes of the susceptibility experiment depicted that ethanoic extract of garlic and ginger (each alone) showed more inhibitory effect than aqueous which showed none at concentrations of 100%, 80% 60% and 40%. The inhibitory effect increased with an increase in the concentration of the extracts and decreased with a decrease in concentration. The minimum inhibitory effect of ethanolic ginger extract on E.coli was found to be 11 at 60% and 11 also at 60% in Salmonella typhi respectively while the minimum inhibitory concentration of ethanoic garlic extract was found to be 15.5 at 60% in E.coli and 12.6 at 60% concentration in Salmonella spp.

TABLE OF CONTENTS

Title page                                                                                                                                i

Certification                                                                                                                           ii

Dedications                                                                                                                             iii

Acknowledgements                                                                                                                iv

Table of contents                                                                                                                     v

List of tables                                                                                                                           vi

Lists of plates (figures)                                                                                                           vii

Abstract                                                                                                                                  viii

CHAPTER ONE

1.0     Introduction        .           .           .           .           .           .           .    .           .           1

1.1 Background of the study   .           .           .           .           .           .            .           .           1

1.2 Aim and objective of the study     .           .           .           .           .            .           .           2

 CHAPTER TWO

2.0     Literature Review           .           .           .           .           .           .    .           .           3

2.1 History of ginger in india             .           .           .           .           .            .           .           3

2.2 History of garlic   .           .           .           .           .           .           .            .           .           4

2.3 Medicinal property of garlic         .           .           .           .           .            .           .           5

2.4 Taxonomical review of ginger     .           .           .           .           .            .           .           6

2.5 Antioxidant Activity of Zingiber officinale (ginger) and other medicinal plants.         6

2.6 Antimicrobial Activity     .           .           .           .           .           .            .           .           9

CHAPTER THREE

MATERIALS AND METHOD

3.0     Materials             .           .           .           .           .           .           .    .           .           13

3.1   Collection of plant materials      .           .           .           .           .            .           .           13

3.2   Sterilization of materials            .           .           .           .           .            .           .           13

3.3   Source of test organism  .           .           .           .           .           .            .           .           13

3.4   Extracts preparation       .           .           .           .           .           .            .           .           13

3.4.1 Garlic Extract preparation         .           .           .           .           .            .           .           13

3.4.2 Ginger Extract preparation        .           .           .           .           .            .           .           14

3.5    Identification/confirmatory test for the Test Isolates    .           .            .           .           14

3.5.1 Gram staining    .           .           .           .           .           .           .            .           .           14

3.5.2 Catalase test       .           .           .           .           .           .           .            .           .           14

3.5.3 Coagulase test    .           .           .           .           .           .           .            .           .           15

3.5.4 Citrate test          .           .           .           .           .           .           .            .           .           15

3.5.5 Motility, Indole, Urease test (MIU)       .           .           .           .            .           .           15

3.5.6 Oxidase test       .           .           .           .           .           .           .            .           .           16

3.6   Media preparation           .           .           .           .           .           .            .           .           16

3.7   Antibacterial Sensitivity test      .           .           .           .           .            .           .           16

3.8   Determination of Minimum Inhibitory Concentration of plant extracts  .           .           16

CHAPTER 4  

RESULTS

4.1 Antibacterial Activity of Plant Extracts   .           .           .           .            .          .            17

4.2 Minimum Inhibitory Concentration Result          .           .           .            .           .           17

CHAPTER 5

5.0  Discussion and Recommendation                        .           .           .      .           .           .           26

5.1 Conclusion            .           .           .           .           .           .           .            .           .           27

REFERENCES      .         .         .         .         .         .         .          .         .         28

LIST OF TABLES

Table                            Title                                                                Page

 1.  Morphological Identification of E.coli and Salmonella typhi      .      .      .      .      18

 2.  Biochemical Characterization of the Test Isolates    .      .      .      .      .     .     .      19

 3.  Antibacterial Activity of the Extracts with Diameter zones of inhibition       .        20

 4.  Minimum Inhibitory concentration on E.coli by the two plant extracts     .      .      21

5.   Minimum Inhibitory Concentration on Salmonella typhi    .        .      .      .      .      22

LIST OF FIGURES

Plate                                  Title                                              page    

1.  Aqueous/Ethanolic Ginger Extract   .    .   .   .   .    .    .   .   .   .   .   .   .   .   .   23

2.  Aqueous Ginger/Garlic Extract with E.coli       .    .    .   .   .   .   .   .   .   .   .   24 

3.  Plates showing zones of inhibition    .    .   .   .   .    .    .   .   .   .   .   .   .   .   .   25

CHAPTER ONE

1.0       INTRODUCTION

A culture medium is simply defined as any material in which microorganisms find nourishment and can reproduce themselves. Microorganisms need nutrients, a source of energy and certain environmental conditions in order   to grow and reproduce. In the environment, microorganisms adapt to the habitats most suitable for their needs while in the laboratory, these requirements must be met by a culture medium (Simin, 2011). Microorganisms can obtain energy directly from sunlight while carbon can be made available in organic form such as carbohydrate.

Media are used for selective and differential cultivation of microorganisms. When a medium is being prepared for microbial growth, consideration must be given to the provision of carbon and energy sources and other growth factors that are essentials for the organisms. Microorganisms can obtain energy directly from sunlight (autotrophs) while carbon can be made available in organic forms such as carbohydrayes or inorganic forms such as carbon dioxide (Madagan et al., 2010)

The growth of microorganisms in an artificial culture medium is influenced by several physical and chemical factors. Microbiological studies depend on the ability to grow and maintain microorganisms under laboratory conditions by providing suitable culture media that offer favorable environmental condition including good carbon source, nitrogen source such as protein, enzymes vitamins, mineral elements such as phosphorus and sulfur, suitable pH, temperature, relative humidity, inorganic salt and water (Okorondu et al., 2013).

 For a microbiological media to fulfill its specific purpose, it must contain all the substances and compounds necessary for the growth and reproduction of the organism. Various substances have been combined into nutritive concoction and have successfully been used to isolate important microorganisms from materials such as water, soil, food, clinical specimens and dairy products (Okorondu et al., 2011). An optimal nutrient culture medium should provide not simply adequate growth, but the best possible growth (Meletiadis et al., 2001). The knowledge of the source of nutrients that encourage the growth of microorganisms is a useful determinant factor in considering the availability of the enzyme present in the microorganism which can be industrially useful.

Fungi are a group of eukaryotic spore-bearing, achlorophyllous organism that generally reproduce asexually and sexually. They are important in nutrient recycling department of nature (Khalid et al; 2006). Fungi due to their competitive saprophytic ability expressed by fast mycelial growth, spore production, presence of efficient and extensive system of powerful enzymes are able to utilize complex polysaccharides and protein as their carbon and nitrogen sources (Wubah, 2009).

The increasing cost of microbial culture media has necessitated the continuous search for more readily available alternative culture media using local raw materials (potatoes, groundnut, cereals, cassava, etc) at an affordable price. Different media for the growth and isolation of microorganisms have been reported from different substrates (Famurewa and David, 2008). Plant materials have been used to recover both fungi and bacteria from different sample sources such as maize, sorghum, groundnut, cassava, local food stuff waste etc.

1.1 AIMS AND OBJECTIVES

This research is therefore aimed at using the different types of garri and akamu as culture media for  the isolation of fungi.

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