ABSTRACT
The high cost of microbial culture media necessitates the screening and production of alternative media from cheap local materials. Mycological culture media has a great role to play in the growth and sporulation of fungi. In this study, the feasibility of using nutrient sources from Garri (a tuber product) and Akamu (a cereal product) to cultivate fungal species was investigated together with a conventional media-Sabouraud Dextrose agar (SDA) which served as control. Soil samples were serially diluted up to the 6th dilution and were inoculated into the SDA, Akamu and Garri medium using pour plate and spread plate techniques from the 4th dilution tube. Growth of the fungal species was observed to be slightly lower in the alternative media but was about the same to the control (SDA). The isolates were identified based on preliminary morphology and microscopic examination. They were species of Aspergillus, Penicillium and Rhizopus. From the percentage mean of each isolate, 89% of Penicillium grew on SDA. This was closely followed by Garri with 76% and Akamu with 32%. The frequency of isolation of Aspergillus was 38% on SDA, 37% on Garri and 65% on Akamu. Rhizopus recorded 64% in SDA, 57% in Garri and 16% in Akamu. SDA has been reported to be a better media for the cultivation of fungi but however, results from this study shows that hot-water made Garri and Akamu can compete favourably with SDA. Colony radical growth and sporulation of fungi were optimal for Aspergillus on Akamu (21mm) after 72hrs followed by Penicillium on SDA (18.2mm) after 72hrs The results from this research work shows that alternative media produced from maize and cassava (cheap raw materials) can be used for the cultivation of fungi in scenarios where acquiring conventional media is difficult.
Table of Contents
Title Page i
Certification ii
Dedication iii
Acknowledgement iv
Table of Contents v
List of Tables vii
List of Plates viii
Abstract ix
Chapter One
1.0 Introduction 1
1.1 Aims and Objectives 3
Chapter Two
2.0 Literature Review 4
2.1 Media as a Research tool 4
2.1.1 Media Sterilization & Aseptic Techniques 6
2.2 Garri 6
2.2.1 Cassava: Raw Material for Garri 8
2.2.2 Cyanide Detoxification 10
2.2.3 Potentials and Future of Cassava as Animal and Microbial Feed 11
2.3 Ogi/Akamu 11
2.4 Fungi 12
2.4.1 Penicillium 14
2.4.2 Rhizopus 15
Chapter Three: Materials and Methods
3.0 Materials and Methods 16
3.1 Materials 16
3.2 Methodology 16
3.2.1 Sterilization of Materials 16
3.2.2 Preparation of Media 16
3.2.3 Preparation of Inoculums Size 17
3.2.4 Inoculation of plates 17
3.2.5 Incubation 18
3.2.6 Isolation and Identification 18
3.2.7 Purification of Isolates 18
3.2.8 Lactophenol Cotton Blue test 19
Chapter Four: Results
4.0 Results 20
Chapter Five: Discussion and Conclusion
5.1 Discussion 26
5.2 Conclusion 28
References 30
LIST OF TABLES
1 Frequency/Occurrence of Various Fungal Isolates on Garri, Akamu and SDA. 21
2 Colony Morphology and Microscopic description 22
3 Number of Each Isolate on Garri, Akamu and SDA 23
4 Mean Colony Diameter (mm) of Each Isolate on Garri, SDA and Akamu after 48 and 72 Hrs of Incubation. 24
5 % Mean of Each Isolate on Akamu, Garri and SDA 25
LIST OF PLATES
I Plate showing growth of fungi on Akamu 35
II A plate showing growth of Aspergillus on Akamu 34
III Plate showing growth of fungi on SDA 36
CHAPTER ONE
1.0 Introduction
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.
Pelezar et al, 1993, defined a culture medium simply 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.
The growth of microorganisms in an artificial 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 sulphur, suitable pH, temperature, relative humidity, inorganic salt and water.
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 concotion 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 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, 1999).
1.1 Aims and Objectives
The aim of this research project is to investigate the feasibility of using garri and akamu as an inexpensive but effective alternative culture medium to conventional mycological media for isolating and identifying moulds.
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