Quantitative Assessment, Mycochemical Constituents And Chitosan Production Of Pleurotus Ostreatus (Jacq.ex Fr) P. Kumm Produced On Different Substrates

ABSTRACT

The study involved the Quantitative Assessment, Mycochemical Constituent and Chitosan Production of P. ostreatus produced from different substrates. The substrates include Andropogon gayanus (A.G), Pennisetum purpureum (P.P), Dactylis chlorometa (D.C) and the combinations of the substrates which included A.G + D.C, A.G + P.P and P.P + D.C. On the morphological yield, the highest fruit-body number was obtained from A.G (40.25±32.07^a) in 1st flush and D.C (21.67±14.15^a) in 2nd flush while the least was obtained from A.G+P.P (10.25±8.14^a and 9.67±4.73^a ) from both flushes . The largest cap diameter was obtained from A.G + P.P (5.45±0.68^abcm and 4.96±0.30^a) respectively from both flushes while the least was from A.G + D.C (3.65±0.16^a cm and 3.89±1.58^a) respectively from both flushes. The longest average stipe length in 1st flush was produced in D.C (2.85±0.37^bcm) and 2nd flush was in A.G + P.P (2.69±0.52^a cm) while the lowest in both flushes was produced in A.G + D.C (2.03±0.25^a cm and 2.09±1.07^a cm) respectively. A.G had the highest biological efficiency (50.2%) while P.P+D.C had the lowest biological efficiency (18.3%). The proximate compositions, vitamins, minerals, bioactive compositions of the fruit-bodies of P. ostreatus were analyzed. Moisture contents of the fruit-bodies ranged from (9.29±0.23% to 8.26±0.12%), ash ranged from (5.52±1.04% to 3.36±0.41%), fibre ranged from (11.31±0.52% to 8.37±1.47%), fat ranged from (2.87±0.88% to 0.88±0.28%), protein ranged from (17.76±0.06% to 13.07±1.27%) and carbohydrates ranged from (63.21±0.95% to 56.98±1.42%). For the vitamin composition, vitamins B₁ ranged from (0.033±0.14 to 0.026±0.12 mg/100g), B₂ ranged from (0.024±0.48 to 0.016±0.47 mg/100g), B₃ ranged from (0.2150.19 to 0.160±0.31 mg/100g), C ranged from (15.39±0.18 to 10.77±1.18 mg/100g) and A ranged from (4.70±0.29 to 3.93±0.24mg/100g). For the Mineral compositions, calcium ranged from (305.96±0.29 to 203.46±0.24mg/100g), magnesium ranged from (15.63±0.08 to 12.74±0.12 mg/100g), potassium ranged from (352.37±0.74 to 220.49±0.22 mg/100g), sodium ranged from (12.63±0.08 to 9.28±0.39 mg/100g) and phosphorus ranged from (233.49±0.17 to 155.23±0.86 mg/100g). For bioactive contents, tannin ranged from (0.26±0.11 to 0.13±0.07%), saponin ranged from (0.35±0.10 to 0.14±0.08%), flavonoid ranged from (0.18±0.11 to 0.09±0.07%), HCN ranged from (3.93±0.08- 2.32±0.11%) and Phenols ranged from (0.15±0.08 to 0.05±0.15%). The result of physicochemical properties of chitosan was investigated, the highest chitosan yield was obtained from A.G (14.77±1.25^ag) while the lowest was from P.P (12.48±2.14^fg), the highest degree of deacetylation was obtained from A.G + D.C (85.78±1.89^a%) while the lowest was from A.G + P.P (84.20±1.43^d%), the highest percentage of acetylation was obtained from substrate with (13.39±2.13^a%) while the lowest was from A.G (10.31±2.33^a%), the highest molecular weight was obtained from A.G + P.P (6482.49±4.71^ag/mol) while the least was from A.G+ D.C (6255.19±3.27^d), and the highest viscosity was obtained from P.P (5.24±2.47^aml/g) while the least was from P.P + D.C (4.13±2.57).  These results indicated that grass substrates are suitable for quantitative growth and yield of oyster mushroom. The results obtained indicated that the appreciable levels of mycochemical constituents of mushroom as affected by different substrates and its combinations are of good quality, it is edible and recommended for maintenance of good health. The physicochemical properties of chitosan result showed that chitosan from mushroom is cheaper, simple to obtain, less cost, no limitations of site and season and posses significant potentials for industrial and agricultural use. 

TABLE OF CONTENTS

Title page                                                                                                                                i

Declaration                                                                                                                              ii

Certification                                                                                                                            iii

Dedication                                                                                                                              iv

Acknowledgement                                                                                                                  v

Table of contents                                                                                                                    vi

List of tables                                                                                                                           viii

List of Plates                                                                                                                           ix

List of figures                                                                                                                         x

Abstract                                                                                                                                  xi

CHAPTER 1:            INTRODUCTION

1.1       Background of the study                                                                                            1

1.2       Statement of problems                                                                                                3

1.3       Justification of the study                                                                                            4

1.4.      Aim and objective                                                                                                       6

CHAPTER TWO: LITERATURE REVIEW

2.1       Growing of Mushroom on Substrates                                                             7

2.2       Related Works:                                                                                                           8

2.3       Chitosan Structure                                                                                                      10

2.4       Properties of Chitosan                                                                                              11

2.5     Features of Fungal Chitin and Chitosan Production                              13

2.6       Industrial Applications of Chitosan                                                                            14

2.6.1    Applications of chitosan in various industries include                                               14

2.6.2    Waste water treatment and biosorption of heavy and toxic metals

from the effluents                                                                                                       14

2.6.3    Chitosan and its derivatives for food applications                                                     15

2.6.4    Chitosan-Based emulsions for food quality preservation                                           15

 2.6.5   Chitosan in agriculture and agro allied industries                                                       15

2.6.6    Chitosan seed treatments                                                                                            16

2.6.7    Chitosan as soil amendment for control of plant diseases                                          17

2.6.8    Chitosan from the cell wall of Pleurotus spp                                                              17

CHAPTER 3:            MATERIALS AND METHODS

3.1       Description of Study Area                                                                                          21

3.2       Source of Materials                                                                                                     21

3.3       Preparation of the Substrates for Cultivation                                                             21

3.4       Inoculation and Incubation                                                                                         22

3.5       Fruiting and Harvesting                                                                                              24

3.6       Measurements of Parameters                                                                                      25

3.6.1    Stipe Lenght (Height)                                                                                                 25

3.6.2    Cap (pileus) Diameter                                                                                                 25

3.6.3    Fresh Weights                                                                                                             25

3.6.4    Dry Weight.                                                                                                                25

3.6.5    Number of Fruit Bodies                                                                                              25

3.6.6    Biological Efficiency                                                                                                  25

3.7       Mycochemical Components                                                                                        26

3.8       Determination of Vitamins                                                                                         26

3.8.1    Determination of Vitamin A (Retinol)                                                                       26

3.8.2    Determination of Vitamin B₁ (Thiamin)                                                                     26

3.8.3    Determination of Vitamin B₂ (Riboflavin                                                                  26

3.8.4    Determination of Vitamin C (Ascorbic Acid)                                                            27

3.8.5    Determination of Vitamin B₃ (Niacin)                                                                        27

3.9       Determination of Mineral                                                                                           28

3.10     Proximate Analysis                                                                                                     28

3.10.1  Determination of crude protein                                                                                  28

3.10.2  Moisture content                                                                                                         28

3.10.3  Determination of Ash contents                                                                                   29

3.10.4  Determination of carbohydrate                                                                                   29

3.10.5  Determination of fats                                                                                                  29

3.10.6  Determination of crude fibre                                                                                      30

3.11     Determination of Bioactive Compounds                                                                    30

3.11.1  Determination of saponins                                                                                          30

3.11.2  Determination of phenols                                                                                           31

3.11.3  Determination of flavonoids                                                                                       31

3.11.4  Determination of tannins                                                                                            31

3.11.5  Hydrogen cyanide (HCN) determination                                                                   32

3.12     Extraction of Chitin and Production of Chitosan                                                      32

3.13     Determination of Physiochemical Properties of Chitosan Extracted                         33

3.13.1  Determination of intrinsic viscosity and molecular weight (MW) of chitosan           33

3.13.2  Deacetylation                                                                                                              33

3.13.3  Acid hydrolysis, distillation and titration                                                                   34

            Statistics                                                                                                                      34

CHAPTER 4:                RESULT AND DISCUSSION

4.1       Results                                                                                                                        35

4.1.1   Morphological characteristics of the fruit-bodies and the yield of P. ostreatus

on different substrates and substrate combinations                                                    35

4.1.2    Myco-chemical components of Pleutorus ostreatus                                                   43

4.1.2.1 Proximate composition of Pleutorus ostreatus                                                           43

4.1.2.2 Vitamin composition                                                                                                   46

4.1.2.3 Mineral composition                                                                                                   49

4.1.2.3 Bioactive composition                                                                                    52

4.1.2.4 Chitosan yield and its physicochemical properties                                                     55

4.2       Discussion                                                                                                                   57

CHAPTER 5: CONCLUSION AND RECOMMENDATION

5.1       CONCLUSION                                                                                                         65

5.2       RECOMMENDATION                                                                                             66

REFERENCES                                                                                                                      68

APPENDICES                                                                                                                       78

LIST OF TABLES

                                                                                                                                                  Pages

2.1       Comparison between Fungal Chitosan Production from Mycelia and

Crustaceans Sources                                                                                                   13

4.1       The Morphological Characteristics and the Effects of the Substrates and

Substrates Combination on the Pleutorus ostreatus                                                   35

4.2       Effect of Different Substrates and Substrate Combination on the Proximate

          Composition of Pleutorus ostreatus                                                                              43

4.3       Effect of Different Substrates and Substrate Combination on the Vitamin

          Composition (mg/100g) of Pleutorus ostreatus                                                            46

4.4       Effect of Different Substrates and Substrate Combination (mg/100g) on the

           Mineral Composition of Pleutorus ostreatus                                                               49

4.5       Effect of Different Substrates and Substrate Combination (%) on the Bioactive

Composition of Pleutorus ostreatus                                                                            52

4.6       Chitosan Yield and its Physico-chemical Properties from P. ostreatus

Cultivated from Different Substrates                                                                         55

LIST OF PLATES

                                                                                                                                                Pages

3.1       Substrate Inoculation with the Mushroom Spawns                                                    23

3.2       Perforated Buckets Laid Out In Mushroom House before Covering with               

             Black Polythene                                                                                                         24

4.2       Fruiting of the Mushrooms from Perforated Buckets                                                41

4.3       Fruit Bodies (side view)                                                                                            42

4.4       Fruit Bodies (bunches harvested)                                                                               42

LIST OF FIGURES

                                                                                                                                                Pages

2.1       Chemical Structures of Cellulose, Chitin and Chitosan                                             11

4.1       Effects of Substrates and Substrates Combinations on the Biological

Efficiency of the P. ostreatus.                                                                                                39