TABLE OF CONTENTS
Title page
Certification
Dedication
Acknowledgments
Table of Contents
List of Figures
Abstract
CHAPTER ONE: INTRODUCTION AND LITERATURE REVIEW
1.1 Introduction
1.1.1 Statement of Problem
1.1.2 Objectives
1.2 Literature Review
1.2.1 Environmental factors influencing yeasts
1.2.2 Temperature
1.2.3 Light and solar radiation
1.2.4 Pressure
1.2.5 Chemical factors
1.2.6 Nutrients
1.2.7 Water
1.2.8 Oxygen relations
1.2.9 Acidity and pH
1.3.0 Antimicrobial compounds
1.3.1 Yeast responses to stress
1.3.2 Response to heat stress
1.3.3 Response to osmostress
1.3.4 Response to salt stress
1.3.5 Response to oxidative stress
1.3.6 Stress-protection mechanisms
1.3.7 Trehalose accumulation
1.3.8 Synthesis of molecular chaperones
1.3.9 Antioxidant proteins and other molecules
CHAPTER TWO: MATERIALS AND METHODS
2.1 Sample Collection
2.2 Isolation of Yeasts from the Honey Sample
2.3 Preparation of Inocula
2.4 Tolerance to Sodium Chloride (NaCl)
2.5 Selection of Working Isolates
2.6 Identification of Osmophilic Yeast
2.6.1 Colony morphology
2.6.2 Cell morphology
2.6.3 Biochemical characteristics of NaCl tolerant yeast
2.6.4 Preparation of inocula for the study of growth under stress on the isolates
2.6.5 Effect of heavy metals on the growth of the isolates
2.6.6 Effect of food preservatives on the growth of the isolates
2.6.7 Effect of temperature on the growth of the isolates
2.6.8 Effect of pH on the growth of the isolate
2.6.9 Preparation of phosphate buffer
2.7 Preparation of phosphate-buffered Sabouraud dextrose broth
2.8 Effect of ultraviolet radiation on the growth of isolates
2.9 Data Analysis
CHAPTER THREE: RESULTS
3.1 Isolation of Organisms
3.2 Tolerance of the Isolates to NaCl
3.3 Growth Responses of the Isolates to NaCl
3.4 Growth Responses of the Isolates to Different Concentrations of AgNO3
3.5 Growth Responses of the Isolates to Different Concentrations of HgCl2
3.6 Growth Responses of the Isolates to Different Concentrations of Sorbic Acid
3.7 Growth Responses of the Isolates to Different Concentrations of Benzoic Acid
3.8 Effect of Temperature on Growth of the Isolates
3.9 Effect of pH on the Growth of the Isolates
3.10 Effect of Ultra Violet (UV) Radiation on Growth of the Isolates.
CHAPTER FOUR: DISCUSSION AND CONCLUSION
4.1 Discussion
4.2 Conclusion
REFERENCES
APPENDICES
ABSTRACT
Yeasts are continuously exposed to series of changes within their natural habitat in the environment. Their growth and proliferation are readily affected by certain factors in their environment. This study was designed to isolate osmophilic yeast from honey and to evaluate the effect of some stress factors such as temperature, pH, NaCl, heavy metals and preservatives on the growth of yeast. The honey samples for this study were collected from Ogige market in Nsukka. The yeasts were isolated, identified and characterized using standard microbiological techniques while the growth of the isolates at different concentration of stress factors were determined using spectrophotometer. The osmophilic yeasts identified as Candida sp., Debaryomyces sp., Rhodotorula sp. andZygosaccharomyces sp. were isolated from honey samples. These isolates displayed varying degrees of responses to different stress factors. There was no significant difference (P≥0.05) in the growth of the isolates at the different concentrations of NaCl. The isolates had the highest growth at 0% NaCl and least growth at 20% NaCl. The growth of the isolates was also affected by increase in concentration of heavy metals. In the absence of HgCl2 and AgN03, the isolates showed very high growth while there was no apparent growth at 2.5% HgCl2 and AgNO3. The isolates also showed the highest growth at 0%
benzoic and sorbic acids and least growth at 1.0% and 1.5% benzoic acid and sorbic acid, respectively. All the isolates showed the highest growth at temperature of 30oC after 96 h of incubation. The optimum pH for the growth of Debaryomyces sp. was 4.0 while that of Rhodotorula sp.,Zygosaccharomyces sp. and Candida sp. was 5.0. There was significant difference (p≤0.05) in the growth of isolates at different exposure times to uv with higher growth recorded at 0 mins and the least observable colonies were obtained at 60 mins. The results showed that yeasts can tolerate different stress conditions and also indicated that sorbic and benzoic acids can be useful in the preservation of food products whereas increase in exposure time to uv radiation can be a good method of inhibiting growth of yeast.
CHAPTER ONE
1.0 INTRODUCTION AND LITERATURE REVIEW
1.1 Introduction
Yeasts are continuously exposed to a series of changes in the environment within their natural habitat. These changes may occur suddenly or may take place over an extended period of time and they can contribute a single specific change or combination of changes. These changes may be in form or physical, chemical or biological and can always represent stresses to a cell. These stresses will require specific response mechanism in order to protect and adapt the cell to the new conditions. Such conditions are generally referred to as stress conditions and the adaptation mechanisms they evolve as stress responses (Hohman and Mager, 2003).
Environmental parameters change ceaselessly and thus force most of the microorganisms to adapt to mildly stressful conditions. The organisms adapt to these changes in the following ways physiological, genetic and epigenetic adaptations. These adaptive measures lead to phenotypic modification which makes the organism to survive. The stress parameters include changes in temperature, pH, osmotic pressure, concentration of waste, ions, and sugars as well as exposure to extremes of radiation, toxic chemicals, oxidative conditions and nutrient starvation (Hohman and Mager, 2003)
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