The nutritional and chemical constituents of different extracts of Neem leaf (Azdirachta indica) and their effect on the haematology and histopathology of Wistar Albino Rats

                                                       ABSTRACT                                  

The nutritional and chemical constituents of different extracts of Neem leaf (Azdirachta indica) and their effect on the haematology and histopathology of Wistar Albino Rats were investigated. The samples analyzed includes processed and unprocessed leaves extract of Neem plant using cold water, hot water, 40% ethanol, 30%  methanol, 70% ethanol and methanol; And also,Wistar Albino Rats (25)  which were administered with extracts(processed and unprocessed) were divided into five(5) groups having five in each cage and one group as control. The phyotchemistry of the Neem leaves extract were determined in triplicates, qualitatively and quantitatively using standard methods. The proximate study carried out on the processed and unprocessed was observed  to contain carbohydrate ˃ moisture ˃ fat ˃ fibre ˃ protein ˃ ash.  Phytochemical studies on the leaves determined, detected  presence of alkaloids, flavonoids, saponnins, oxalate, phenol, tannin, steriod, glycosides, reducing sugar and carbohydrate. The results are presented as  mean ± STD of trplicate determination and means were compared using one-way ANOVA statitical analysis at 0.05 significance level. Rutin, ranging from 0.000 ± 0.01 to 77.543 ± 0.040,was observed to have high yield present in the processed and unprocessed leaves (14.290 ± 0.010 to 77.543 ± 0.040) while spartein was the lowest (0.002 ± 0.00 to 0.005 ± 0.00). The vitamins A,C,E and Riboflavin were determined, ranging from 0.89  ± 0.007 to 375.74 ± 1.22 amongst all samples,vitamin c was observed to be in high yield than any other of the vitamins. The total mineral concentration present in neem plant leaves extract were determined; and it was observed to contain Manganese , cobalt, zinc, copper, lead, nickel, sodium, potassium, magnesium and chromium  ranging from 0.003 to 23.529; But magnesium was observed to be in high yield ranging from 20.768 to 23.529.The haematological study of the animals treated using the processed leaves extract and after 24 hours of starvation were observed to have high count of platelets when compared with the controls(220 μl/kg to 350μl/kg);also the WBC count range was significantly decreased (4.0 to 14.0) /L when observed with the controls(13.00±0.00)/L,the RBC showed a significant increase, ranging from 4.5μl  to 7.2μl when compared with the control(4.5±0.00)/L.The HB count significantly increased from 10.7g/dl to 16.7g/dl when compared with the control(10.70±0.00)g/dL. The Histopathological study using liver and kidney,viewed under light microscope revealed spots and damages of the heart and liver,leading to distortion and haemorrhage. In conclusion,the consumption of alcoholic neem leaves extract during starvation and without dosage is toxic,harmful and can lead to death,especially when allowed to ferment due to bacteria activities than the hot water and tap water extract of neem leaves which is more safe to consumed with or without meal . 

 

 

TABLE OF CONTENTS

Title page                                                                                                                                                                              i

Certification                                                                                                                                                                         ii

Dedication                                                                                                                                                                            iii

Acknowledgement                                                                                                                                                              iv            

Abstract                                                                                                                                                                                 v

Table of contents                                                                                                                     vi-xi

List of Table                                                                                                                                                                         xii           

List of figure                                                                                                                                                                         xiii-xix   

List of abbreviation                                                                                                                                                       xx

CHAPTER ONE                                                                                                                

1.0. Introduction                                                                                                                                                                  1

1.1 Back Ground of Study                                                                                                                                                1             

1.2    Statement of the Problem                                                                                                                                          3

1.3. AIM AND OBJECTIVES                                                                                                                                           4

1.3.1.AIM                                                                                                                                                                             4

1.3.2. OBJECTIVES.                                                                                                                                                           4

1.4. SIGNIFICANCE OF STUDY                                                                                                                                     4

CHAPTER TWO

LITERATURE REVIEW                                                                                                                                                   5

2.1 Azadirachtaindica (Neem).                                                                                                                                       5          

2.2. Neem in Hindu Mythology                                                                                                                                        10

2.3. NEEM AS A WEED                                                                                                                                                    11

2.4    TAXONOMICAL CLASSIFICATION OF NEEM.                                                                                               11

2.5. Chemical Properties                                                                                                                                                    12

2.6. CHEMISTRY OF NEEM:                                                                                                                                          13                       2.7. USES AND BENEFITS OF NEEM TREE PARTS.                                                                                             16

 2.7.1  USES                                                                                                                                                                          16

 2.7.1.1.STORAGE                                                                                                                                                               16

2.7.1.2. INSECTICDES                                                                                                                                                      16

2.8.Cosmetic uses                                                                                                                                               16                           

2.9.VEGETABLE USE OF NEEM                                                                                                                                    17           

2.10.TRADITIONAL MEDICINAL USE OF NEEM                                                                                                       17

2.11. Name and uses of neem tree in the different African countries                                                                       18           

2.12.Panacea for All Diseases:                                                                                                                                        22

2.13.  Benefits of Neem Tree parts                                                                                                                                  22

2.13.1.Benefits of Neem leaves                                                                                                                                         22

2.13.1.1.Eye problems                                                                                                                                                        23

2.13.1.2.Viral Diseases                                                                                                                                                      23

2.14.Neem Effects against Bacteria                                                                                                                                                23

2.15.Neem Effects against Viruses                                                                                                                                  24

2.16.Neem Effects against Fungi                                                                                                                                     24

2.17.Neem Effects against Parasites                                                                                                                           25                               

2.17.1.De-worming                                                                                                                                                             26

2.17.2.Cardiac Care:                                                                                                                                                         26

2.17.3.Natural Pesticide:                                                                                                                                                  27

2.17.4.Fungal Diseases:                                                                                                                                                    27

2.17.5.Post-Parturition Disorder                                                                                                                                     27

2.17.6.Skin Care:                                                                                                                                                                28

2.18. neem and diabetes                                                                                                                                                    28

2.19. Uses of Neem Seeds and Fruits                                                                                                                               30

  2.19.1.Skin Problems                                                                                                                                                        30

2.19.2.Hair Care                                                                                                                                                                 30

 2.19.3.Eye and Ear Infections:                                                                                                                                        30

2.20.Other Ailments:                                                                                                                                                           31

2.20.1.Malaria:                                                                                                                                                                   31

2.20.2. neem cake organic manure                                                                                                                                  31

2.20.3.Components                                                                                                                                                            31

2.20.4.Use as a fertilizer                                                                                                                                                     32

2.20.5.Pest control                                                                                                                                                               32          

2.21.Neem Disadvantages:                                                                                                                                                               33

2.21.1.Strong Smell:                                                                                                                                                            33

2.21.2.Bitter Taste                                                                                                                                                               33

2.21.3.DANGER:                                                                                                                                                                 34

2.22.CHOLESTEROL                                                                                                                                                        34

2.23. USE AS CONTRACEPTIVE   IN BOTH MALE AND FEMALE:                                                   35

2.23.1.NEEM OIL - DIRECT USE:                                                                                                                                   36          

2.24.NEEM - CONTRACEPTIVE FOR MEN:                                                                                                              36

2.25.Insect Repellent:                                                                                                                                                         37

2.26.Flea Repellent :                                                                                                                                                          37

2.27.Uses of Neem Bark                                                                                                                                                    38

2.27.1.Oral Health                                                                                                                                                              38

2.27.2.Fever                                                                                                                                                                         38

2.27.3.Pile                                                                                                                                                                            38

2.28.Uses of Neem Flower                                                                                                                                                 38-39

2.29. Other Medical Benefits of Neem Tree                                                                                                                   39

2.29.1.Arthritis:                                                                                                                                                                   39

2.29.2. rheumatism                                                                                                                                                             40

2.29.3.Chronic Fatigue:                                                                                                                                                      41

2.29.4.Cancer:                                                                                                                                                                     41

2.30.Neem and anti-microbial activity:                                                                                                                          42-43    

2.31.Detoxifying Effects                                                                                                                                                    43

2.32.Gastric Health                                                                                                                                                             43

2.33.Pore Size                                                                                                                                                                       44

2.34.Phytochemicals                                                                                                                                                          44

2.34.1. antioxidant                                                                                                                                                              45

2.35.flavonoids                                                                                                                                                                    46

2.35.1.Definition of Flavonoids:                                                                                                                                      46

2.35.2. health benefits to humans                                                                                                                                    48

2.35.3.Food sources:                                                                                                                                                           49

2.35.4.Interactions:                                                                                                                                                             50

2.36.Classification of Flavonoids                                                                                                                                    50-53

2.36.1.Flavanols and Flavonols                                                                                                                                       53

2.36.2.Flavanones and Flavones                                                                                                                                     54

2.37.Anthocyanins                                                                                                                                                              58

2.38.Tannin:                                                                                                                                                                         63

2.39.Terpenene                                                                                                                                                                    63

2.40. Antioxidant Vitamins in Neem                                                                                                                               64

2.41. Alkaloids:                                                                                                                                                                    65

2.42. Neem and Environmental Protection                                                                                                                    68

2.42.1. Afforestation                                                                                                                                                          68

2.43.Vitamins and Minerals in Neem Plant                                                                                                                    69

2.43.1.Vitamins:                                                                                                                                                                  69

 

2.44.2.Minerals:                                                                                                                                                                   69

2.45.Anti-Vitamins.                                                                                                                                                             71

2.46.Herbal Preparation                                                                                                                                                     72

2.47. Traditional Herbal Extraction Methods                                                                                                                                73

2.48.Method of Administration                                                                                                                                        73

2.49. Haematological Parameters                                                                                                                                    74

2.49.1.Haematocrit (Packed Cell Volume):                                                                                                                   74

2.49.2. Elevated Haematocrit Levels                                                                                                                              74

2.49.3.Lowered Haematocrit Levels                                                                                                                               75

2.50.Platelet Count:                                                                                                                                                            75

2.51.Differential Leucocyte Count                                                                                                                                  76

2.52. Red Blood Cell Count                                                                                                                                              78

2.53.Histology                                                                                                                                                                      79

                                                                      

CHAPTER THREE

 3.0  Materials And Methods                                                                                                                                             81

3.1.  Materials                                                                                                                                                                      81

3.1.1 Reagents Used For The Study                                                                                                                                 81

3.1.2. APPARATUS                                                                                                                                                            81

3.2. Methods                                                                                                                                                                        82

3.2.1. Sample Collection and Preparation                                                                                                                      82

3.2.1.1. Sample Collection,Drying ,Cutting and Grinding                                                                                            82

3.2.1.2 .Sample Preparation                                                                                                                                              82

3.3.Proximate                                                                                                                                                                       83

3.3.1.Crude Fibre Determination                                                                                                                                      83

3.3.2.Crude Protein Determination                                                                                                                                  83:

3.3.3.Crude Fat Determination:                                                                                              85

3.3.4.Moisture Content Determination                                                                                   85-86

3.3.5.Determination of Ash:                                                                                                    86

3.6.Determination of Carbohydrate                                                                                        87

3.4.Phytochemical Study                                                                                                                                                   87

3.4.1. Extraction and Phytochemical Determination                                                                                                   87

3.5.Mineral Study                                                                                                                                                                88

3.5.1. Digestion                                                                                                                                                                    88

3.6. Vitamins                                                                                                                                                                        89 - 90

3.7.Haematological Study                                                                                                                                                                 91

3.7.1.Animals used for The Study                                                                                                                                    91

3.7.2.Haematological Parameters Analyzed                                                                                                                 93

3.7.2.1  Haematocrit or Packed Cell Volume (Pcv) Determination.                                                                          93

3.7.2.2.White Cell Count (Wyngarden Method,1985).                                                                                                                94

3.7.3.Differential White Blood Cell Count( by Beckman Coulter method)                                                             95

3.7.4.Platelet count by Haemacytometer counting method:                                                   96

3.7.4.1.RBC Count (Haemacytometer counting method):                                                       97

3.8. Histological Study:                                                                                                           98

3.8.1 Fixation:                                                                                                                         98

3.8.2.Embedding:                                                                                                                    99

3.8.3.Sectioning:                                                                                                                     99

3.8.4.Staining:                                                                                                                        99

3.8.5.Mounting:                                                                                                                       100

3.9.Statistical Analysis oF Data                                                                                                                                           100

CHAPTER FOUR: RESULTS AND DISCUSSION

4.0.Results:                                                                                                                                                                           101

4.1. Proximate Analysis                                                                                                                                                     101-104

4.2.    Phytochemistry                                                                                                                                                         105

4.5.  Qualitative                                                                                                                                                                   105

4.3.Quantitative Analysis                                                                                                                                                  107-110

4.3    Vitamins:                                                                                                                                                                     111

4.4.     Minerals                                                                                                                                                                     112

4.5. Haematological Study                                                                                                                                                                113

4.6.   Histopathological  Study                                                                                                                                  114-198

 4.1. Discussion:                                                                                                                                                                  199-204

CHAPTER FIVE:

5.0. Conclusion and Recommendation                                                                                                                          205                                                                                                                                                                                    

5.1. Conclusion                                                                                                                                                                    205

5.3.Recommendation:                                                                                                                                                       206

5.4.Contribution to Knowledge                                                                                                                                         206

Reference

APPENDIX

 

 

                                                       ABSTRACT                                  

The nutritional and chemical constituents of different extracts of Neem leaf (Azdirachta indica) and their effect on the haematology and histopathology of Wistar Albino Rats were investigated. The samples analyzed includes processed and unprocessed leaves extract of Neem plant using cold water, hot water, 40% ethanol, 30%  methanol, 70% ethanol and methanol; And also,Wistar Albino Rats (25)  which were administered with extracts(processed and unprocessed) were divided into five(5) groups having five in each cage and one group as control. The phyotchemistry of the Neem leaves extract were determined in triplicates, qualitatively and quantitatively using standard methods. The proximate study carried out on the processed and unprocessed was observed  to contain carbohydrate ˃ moisture ˃ fat ˃ fibre ˃ protein ˃ ash.  Phytochemical studies on the leaves determined, detected  presence of alkaloids, flavonoids, saponnins, oxalate, phenol, tannin, steriod, glycosides, reducing sugar and carbohydrate. The results are presented as  mean ± STD of trplicate determination and means were compared using one-way ANOVA statitical analysis at 0.05 significance level. Rutin, ranging from 0.000 ± 0.01 to 77.543 ± 0.040,was observed to have high yield present in the processed and unprocessed leaves (14.290 ± 0.010 to 77.543 ± 0.040) while spartein was the lowest (0.002 ± 0.00 to 0.005 ± 0.00). The vitamins A,C,E and Riboflavin were determined, ranging from 0.89  ± 0.007 to 375.74 ± 1.22 amongst all samples,vitamin c was observed to be in high yield than any other of the vitamins. The total mineral concentration present in neem plant leaves extract were determined; and it was observed to contain Manganese , cobalt, zinc, copper, lead, nickel, sodium, potassium, magnesium and chromium  ranging from 0.003 to 23.529; But magnesium was observed to be in high yield ranging from 20.768 to 23.529.The haematological study of the animals treated using the processed leaves extract and after 24 hours of starvation were observed to have high count of platelets when compared with the controls(220 μl/kg to 350μl/kg);also the WBC count range was significantly decreased (4.0 to 14.0) /L when observed with the controls(13.00±0.00)/L,the RBC showed a significant increase, ranging from 4.5μl  to 7.2μl when compared with the control(4.5±0.00)/L.The HB count significantly increased from 10.7g/dl to 16.7g/dl when compared with the control(10.70±0.00)g/dL. The Histopathological study using liver and kidney,viewed under light microscope revealed spots and damages of the heart and liver,leading to distortion and haemorrhage. In conclusion,the consumption of alcoholic neem leaves extract during starvation and without dosage is toxic,harmful and can lead to death,especially when allowed to ferment due to bacteria activities than the hot water and tap water extract of neem leaves which is more safe to consumed with or without meal . 

 

 

TABLE OF CONTENTS

Title page                                                                                                                                                                              i

Certification                                                                                                                                                                         ii

Dedication                                                                                                                                                                            iii

Acknowledgement                                                                                                                                                              iv            

Abstract                                                                                                                                                                                 v

Table of contents                                                                                                                     vi-xi

List of Table                                                                                                                                                                         xii           

List of figure                                                                                                                                                                         xiii-xix   

List of abbreviation                                                                                                                                                       xx

CHAPTER ONE                                                                                                                

1.0. Introduction                                                                                                                                                                  1

1.1 Back Ground of Study                                                                                                                                                1             

1.2    Statement of the Problem                                                                                                                                          3

1.3. AIM AND OBJECTIVES                                                                                                                                           4

1.3.1.AIM                                                                                                                                                                             4

1.3.2. OBJECTIVES.                                                                                                                                                           4

1.4. SIGNIFICANCE OF STUDY                                                                                                                                     4

CHAPTER TWO

LITERATURE REVIEW                                                                                                                                                   5

2.1 Azadirachtaindica (Neem).                                                                                                                                       5          

2.2. Neem in Hindu Mythology                                                                                                                                        10

2.3. NEEM AS A WEED                                                                                                                                                    11

2.4    TAXONOMICAL CLASSIFICATION OF NEEM.                                                                                               11

2.5. Chemical Properties                                                                                                                                                    12

2.6. CHEMISTRY OF NEEM:                                                                                                                                          13                       2.7. USES AND BENEFITS OF NEEM TREE PARTS.                                                                                             16

 2.7.1  USES                                                                                                                                                                          16

 2.7.1.1.STORAGE                                                                                                                                                               16

2.7.1.2. INSECTICDES                                                                                                                                                      16

2.8.Cosmetic uses                                                                                                                                               16                           

2.9.VEGETABLE USE OF NEEM                                                                                                                                    17           

2.10.TRADITIONAL MEDICINAL USE OF NEEM                                                                                                       17

2.11. Name and uses of neem tree in the different African countries                                                                       18           

2.12.Panacea for All Diseases:                                                                                                                                        22

2.13.  Benefits of Neem Tree parts                                                                                                                                  22

2.13.1.Benefits of Neem leaves                                                                                                                                         22

2.13.1.1.Eye problems                                                                                                                                                        23

2.13.1.2.Viral Diseases                                                                                                                                                      23

2.14.Neem Effects against Bacteria                                                                                                                                                23

2.15.Neem Effects against Viruses                                                                                                                                  24

2.16.Neem Effects against Fungi                                                                                                                                     24

2.17.Neem Effects against Parasites                                                                                                                           25                               

2.17.1.De-worming                                                                                                                                                             26

2.17.2.Cardiac Care:                                                                                                                                                         26

2.17.3.Natural Pesticide:                                                                                                                                                  27

2.17.4.Fungal Diseases:                                                                                                                                                    27

2.17.5.Post-Parturition Disorder                                                                                                                                     27

2.17.6.Skin Care:                                                                                                                                                                28

2.18. neem and diabetes                                                                                                                                                    28

2.19. Uses of Neem Seeds and Fruits                                                                                                                               30

  2.19.1.Skin Problems                                                                                                                                                        30

2.19.2.Hair Care                                                                                                                                                                 30

 2.19.3.Eye and Ear Infections:                                                                                                                                        30

2.20.Other Ailments:                                                                                                                                                           31

2.20.1.Malaria:                                                                                                                                                                   31

2.20.2. neem cake organic manure                                                                                                                                  31

2.20.3.Components                                                                                                                                                            31

2.20.4.Use as a fertilizer                                                                                                                                                     32

2.20.5.Pest control                                                                                                                                                               32          

2.21.Neem Disadvantages:                                                                                                                                                               33

2.21.1.Strong Smell:                                                                                                                                                            33

2.21.2.Bitter Taste                                                                                                                                                               33

2.21.3.DANGER:                                                                                                                                                                 34

2.22.CHOLESTEROL                                                                                                                                                        34

2.23. USE AS CONTRACEPTIVE   IN BOTH MALE AND FEMALE:                                                   35

2.23.1.NEEM OIL - DIRECT USE:                                                                                                                                   36          

2.24.NEEM - CONTRACEPTIVE FOR MEN:                                                                                                              36

2.25.Insect Repellent:                                                                                                                                                         37

2.26.Flea Repellent :                                                                                                                                                          37

2.27.Uses of Neem Bark                                                                                                                                                    38

2.27.1.Oral Health                                                                                                                                                              38

2.27.2.Fever                                                                                                                                                                         38

2.27.3.Pile                                                                                                                                                                            38

2.28.Uses of Neem Flower                                                                                                                                                 38-39

2.29. Other Medical Benefits of Neem Tree                                                                                                                   39

2.29.1.Arthritis:                                                                                                                                                                   39

2.29.2. rheumatism                                                                                                                                                             40

2.29.3.Chronic Fatigue:                                                                                                                                                      41

2.29.4.Cancer:                                                                                                                                                                     41

2.30.Neem and anti-microbial activity:                                                                                                                          42-43    

2.31.Detoxifying Effects                                                                                                                                                    43

2.32.Gastric Health                                                                                                                                                             43

2.33.Pore Size                                                                                                                                                                       44

2.34.Phytochemicals                                                                                                                                                          44

2.34.1. antioxidant                                                                                                                                                              45

2.35.flavonoids                                                                                                                                                                    46

2.35.1.Definition of Flavonoids:                                                                                                                                      46

2.35.2. health benefits to humans                                                                                                                                    48

2.35.3.Food sources:                                                                                                                                                           49

2.35.4.Interactions:                                                                                                                                                             50

2.36.Classification of Flavonoids                                                                                                                                    50-53

2.36.1.Flavanols and Flavonols                                                                                                                                       53

2.36.2.Flavanones and Flavones                                                                                                                                     54

2.37.Anthocyanins                                                                                                                                                              58

2.38.Tannin:                                                                                                                                                                         63

2.39.Terpenene                                                                                                                                                                    63

2.40. Antioxidant Vitamins in Neem                                                                                                                               64

2.41. Alkaloids:                                                                                                                                                                    65

2.42. Neem and Environmental Protection                                                                                                                    68

2.42.1. Afforestation                                                                                                                                                          68

2.43.Vitamins and Minerals in Neem Plant                                                                                                                    69

2.43.1.Vitamins:                                                                                                                                                                  69

 

2.44.2.Minerals:                                                                                                                                                                   69

2.45.Anti-Vitamins.                                                                                                                                                             71

2.46.Herbal Preparation                                                                                                                                                     72

2.47. Traditional Herbal Extraction Methods                                                                                                                                73

2.48.Method of Administration                                                                                                                                        73

2.49. Haematological Parameters                                                                                                                                    74

2.49.1.Haematocrit (Packed Cell Volume):                                                                                                                   74

2.49.2. Elevated Haematocrit Levels                                                                                                                              74

2.49.3.Lowered Haematocrit Levels                                                                                                                               75

2.50.Platelet Count:                                                                                                                                                            75

2.51.Differential Leucocyte Count                                                                                                                                  76

2.52. Red Blood Cell Count                                                                                                                                              78

2.53.Histology                                                                                                                                                                      79

                                                                      

CHAPTER THREE

 3.0  Materials And Methods                                                                                                                                             81

3.1.  Materials                                                                                                                                                                      81

3.1.1 Reagents Used For The Study                                                                                                                                 81

3.1.2. APPARATUS                                                                                                                                                            81

3.2. Methods                                                                                                                                                                        82

3.2.1. Sample Collection and Preparation                                                                                                                      82

3.2.1.1. Sample Collection,Drying ,Cutting and Grinding                                                                                            82

3.2.1.2 .Sample Preparation                                                                                                                                              82

3.3.Proximate                                                                                                                                                                       83

3.3.1.Crude Fibre Determination                                                                                                                                      83

3.3.2.Crude Protein Determination                                                                                                                                  83:

3.3.3.Crude Fat Determination:                                                                                              85

3.3.4.Moisture Content Determination                                                                                   85-86

3.3.5.Determination of Ash:                                                                                                    86

3.6.Determination of Carbohydrate                                                                                        87

3.4.Phytochemical Study                                                                                                                                                   87

3.4.1. Extraction and Phytochemical Determination                                                                                                   87

3.5.Mineral Study                                                                                                                                                                88

3.5.1. Digestion                                                                                                                                                                    88

3.6. Vitamins                                                                                                                                                                        89 - 90

3.7.Haematological Study                                                                                                                                                                 91

3.7.1.Animals used for The Study                                                                                                                                    91

3.7.2.Haematological Parameters Analyzed                                                                                                                 93

3.7.2.1  Haematocrit or Packed Cell Volume (Pcv) Determination.                                                                          93

3.7.2.2.White Cell Count (Wyngarden Method,1985).                                                                                                                94

3.7.3.Differential White Blood Cell Count( by Beckman Coulter method)                                                             95

3.7.4.Platelet count by Haemacytometer counting method:                                                   96

3.7.4.1.RBC Count (Haemacytometer counting method):                                                       97

3.8. Histological Study:                                                                                                           98

3.8.1 Fixation:                                                                                                                         98

3.8.2.Embedding:                                                                                                                    99

3.8.3.Sectioning:                                                                                                                     99

3.8.4.Staining:                                                                                                                        99

3.8.5.Mounting:                                                                                                                       100

3.9.Statistical Analysis oF Data                                                                                                                                           100

CHAPTER FOUR: RESULTS AND DISCUSSION

4.0.Results:                                                                                                                                                                           101

4.1. Proximate Analysis                                                                                                                                                     101-104

4.2.    Phytochemistry                                                                                                                                                         105

4.5.  Qualitative                                                                                                                                                                   105

4.3.Quantitative Analysis                                                                                                                                                  107-110

4.3    Vitamins:                                                                                                                                                                     111

4.4.     Minerals                                                                                                                                                                     112

4.5. Haematological Study                                                                                                                                                                113

4.6.   Histopathological  Study                                                                                                                                  114-198

 4.1. Discussion:                                                                                                                                                                  199-204

CHAPTER FIVE:

5.0. Conclusion and Recommendation                                                                                                                          205                                                                                                                                                                                    

5.1. Conclusion                                                                                                                                                                    205

5.3.Recommendation:                                                                                                                                                       206

5.4.Contribution to Knowledge                                                                                                                                         206

Reference

APPENDIX

 

 

 

 

 

                                                       ABSTRACT                                  

The nutritional and chemical constituents of different extracts of Neem leaf (Azdirachta indica) and their effect on the haematology and histopathology of Wistar Albino Rats were investigated. The samples analyzed includes processed and unprocessed leaves extract of Neem plant using cold water, hot water, 40% ethanol, 30%  methanol, 70% ethanol and methanol; And also,Wistar Albino Rats (25)  which were administered with extracts(processed and unprocessed) were divided into five(5) groups having five in each cage and one group as control. The phyotchemistry of the Neem leaves extract were determined in triplicates, qualitatively and quantitatively using standard methods. The proximate study carried out on the processed and unprocessed was observed  to contain carbohydrate ˃ moisture ˃ fat ˃ fibre ˃ protein ˃ ash.  Phytochemical studies on the leaves determined, detected  presence of alkaloids, flavonoids, saponnins, oxalate, phenol, tannin, steriod, glycosides, reducing sugar and carbohydrate. The results are presented as  mean ± STD of trplicate determination and means were compared using one-way ANOVA statitical analysis at 0.05 significance level. Rutin, ranging from 0.000 ± 0.01 to 77.543 ± 0.040,was observed to have high yield present in the processed and unprocessed leaves (14.290 ± 0.010 to 77.543 ± 0.040) while spartein was the lowest (0.002 ± 0.00 to 0.005 ± 0.00). The vitamins A,C,E and Riboflavin were determined, ranging from 0.89  ± 0.007 to 375.74 ± 1.22 amongst all samples,vitamin c was observed to be in high yield than any other of the vitamins. The total mineral concentration present in neem plant leaves extract were determined; and it was observed to contain Manganese , cobalt, zinc, copper, lead, nickel, sodium, potassium, magnesium and chromium  ranging from 0.003 to 23.529; But magnesium was observed to be in high yield ranging from 20.768 to 23.529.The haematological study of the animals treated using the processed leaves extract and after 24 hours of starvation were observed to have high count of platelets when compared with the controls(220 μl/kg to 350μl/kg);also the WBC count range was significantly decreased (4.0 to 14.0) /L when observed with the controls(13.00±0.00)/L,the RBC showed a significant increase, ranging from 4.5μl  to 7.2μl when compared with the control(4.5±0.00)/L.The HB count significantly increased from 10.7g/dl to 16.7g/dl when compared with the control(10.70±0.00)g/dL. The Histopathological study using liver and kidney,viewed under light microscope revealed spots and damages of the heart and liver,leading to distortion and haemorrhage. In conclusion,the consumption of alcoholic neem leaves extract during starvation and without dosage is toxic,harmful and can lead to death,especially when allowed to ferment due to bacteria activities than the hot water and tap water extract of neem leaves which is more safe to consumed with or without meal . 

 

 

TABLE OF CONTENTS

Title page                                                                                                                                                                              i

Certification                                                                                                                                                                         ii

Dedication                                                                                                                                                                            iii

Acknowledgement                                                                                                                                                              iv            

Abstract                                                                                                                                                                                 v

Table of contents                                                                                                                     vi-xi

List of Table                                                                                                                                                                         xii           

List of figure                                                                                                                                                                         xiii-xix   

List of abbreviation                                                                                                                                                       xx

CHAPTER ONE                                                                                                                

1.0. Introduction                                                                                                                                                                  1

1.1 Back Ground of Study                                                                                                                                                1             

1.2    Statement of the Problem                                                                                                                                          3

1.3. AIM AND OBJECTIVES                                                                                                                                           4

1.3.1.AIM                                                                                                                                                                             4

1.3.2. OBJECTIVES.                                                                                                                                                           4

1.4. SIGNIFICANCE OF STUDY                                                                                                                                     4

CHAPTER TWO

LITERATURE REVIEW                                                                                                                                                   5

2.1 Azadirachtaindica (Neem).                                                                                                                                       5          

2.2. Neem in Hindu Mythology                                                                                                                                        10

2.3. NEEM AS A WEED                                                                                                                                                    11

2.4    TAXONOMICAL CLASSIFICATION OF NEEM.                                                                                               11

2.5. Chemical Properties                                                                                                                                                    12

2.6. CHEMISTRY OF NEEM:                                                                                                                                          13                       2.7. USES AND BENEFITS OF NEEM TREE PARTS.                                                                                             16

 2.7.1  USES                                                                                                                                                                          16

 2.7.1.1.STORAGE                                                                                                                                                               16

2.7.1.2. INSECTICDES                                                                                                                                                      16

2.8.Cosmetic uses                                                                                                                                               16                           

2.9.VEGETABLE USE OF NEEM                                                                                                                                    17           

2.10.TRADITIONAL MEDICINAL USE OF NEEM                                                                                                       17

2.11. Name and uses of neem tree in the different African countries                                                                       18           

2.12.Panacea for All Diseases:                                                                                                                                        22

2.13.  Benefits of Neem Tree parts                                                                                                                                  22

2.13.1.Benefits of Neem leaves                                                                                                                                         22

2.13.1.1.Eye problems                                                                                                                                                        23

2.13.1.2.Viral Diseases                                                                                                                                                      23

2.14.Neem Effects against Bacteria                                                                                                                                                23

2.15.Neem Effects against Viruses                                                                                                                                  24

2.16.Neem Effects against Fungi                                                                                                                                     24

2.17.Neem Effects against Parasites                                                                                                                           25                               

2.17.1.De-worming                                                                                                                                                             26

2.17.2.Cardiac Care:                                                                                                                                                         26

2.17.3.Natural Pesticide:                                                                                                                                                  27

2.17.4.Fungal Diseases:                                                                                                                                                    27

2.17.5.Post-Parturition Disorder                                                                                                                                     27

2.17.6.Skin Care:                                                                                                                                                                28

2.18. neem and diabetes                                                                                                                                                    28

2.19. Uses of Neem Seeds and Fruits                                                                                                                               30

  2.19.1.Skin Problems                                                                                                                                                        30

2.19.2.Hair Care                                                                                                                                                                 30

 2.19.3.Eye and Ear Infections:                                                                                                                                        30

2.20.Other Ailments:                                                                                                                                                           31

2.20.1.Malaria:                                                                                                                                                                   31

2.20.2. neem cake organic manure                                                                                                                                  31

2.20.3.Components                                                                                                                                                            31

2.20.4.Use as a fertilizer                                                                                                                                                     32

2.20.5.Pest control                                                                                                                                                               32          

2.21.Neem Disadvantages:                                                                                                                                                               33

2.21.1.Strong Smell:                                                                                                                                                            33

2.21.2.Bitter Taste                                                                                                                                                               33

2.21.3.DANGER:                                                                                                                                                                 34

2.22.CHOLESTEROL                                                                                                                                                        34

2.23. USE AS CONTRACEPTIVE   IN BOTH MALE AND FEMALE:                                                   35

2.23.1.NEEM OIL - DIRECT USE:                                                                                                                                   36          

2.24.NEEM - CONTRACEPTIVE FOR MEN:                                                                                                              36

2.25.Insect Repellent:                                                                                                                                                         37

2.26.Flea Repellent :                                                                                                                                                          37

2.27.Uses of Neem Bark                                                                                                                                                    38

2.27.1.Oral Health                                                                                                                                                              38

2.27.2.Fever                                                                                                                                                                         38

2.27.3.Pile                                                                                                                                                                            38

2.28.Uses of Neem Flower                                                                                                                                                 38-39

2.29. Other Medical Benefits of Neem Tree                                                                                                                   39

2.29.1.Arthritis:                                                                                                                                                                   39

2.29.2. rheumatism                                                                                                                                                             40

2.29.3.Chronic Fatigue:                                                                                                                                                      41

2.29.4.Cancer:                                                                                                                                                                     41

2.30.Neem and anti-microbial activity:                                                                                                                          42-43    

2.31.Detoxifying Effects                                                                                                                                                    43

2.32.Gastric Health                                                                                                                                                             43

2.33.Pore Size                                                                                                                                                                       44

2.34.Phytochemicals                                                                                                                                                          44

2.34.1. antioxidant                                                                                                                                                              45

2.35.flavonoids                                                                                                                                                                    46

2.35.1.Definition of Flavonoids:                                                                                                                                      46

2.35.2. health benefits to humans                                                                                                                                    48

2.35.3.Food sources:                                                                                                                                                           49

2.35.4.Interactions:                                                                                                                                                             50

2.36.Classification of Flavonoids                                                                                                                                    50-53

2.36.1.Flavanols and Flavonols                                                                                                                                       53

2.36.2.Flavanones and Flavones                                                                                                                                     54

2.37.Anthocyanins                                                                                                                                                              58

2.38.Tannin:                                                                                                                                                                         63

2.39.Terpenene                                                                                                                                                                    63

2.40. Antioxidant Vitamins in Neem                                                                                                                               64

2.41. Alkaloids:                                                                                                                                                                    65

2.42. Neem and Environmental Protection                                                                                                                    68

2.42.1. Afforestation                                                                                                                                                          68

2.43.Vitamins and Minerals in Neem Plant                                                                                                                    69

2.43.1.Vitamins:                                                                                                                                                                  69

 

2.44.2.Minerals:                                                                                                                                                                   69

2.45.Anti-Vitamins.                                                                                                                                                             71

2.46.Herbal Preparation                                                                                                                                                     72

2.47. Traditional Herbal Extraction Methods                                                                                                                                73

2.48.Method of Administration                                                                                                                                        73

2.49. Haematological Parameters                                                                                                                                    74

2.49.1.Haematocrit (Packed Cell Volume):                                                                                                                   74

2.49.2. Elevated Haematocrit Levels                                                                                                                              74

2.49.3.Lowered Haematocrit Levels                                                                                                                               75

2.50.Platelet Count:                                                                                                                                                            75

2.51.Differential Leucocyte Count                                                                                                                                  76

2.52. Red Blood Cell Count                                                                                                                                              78

2.53.Histology                                                                                                                                                                      79

                                                                      

CHAPTER THREE

 3.0  Materials And Methods                                                                                                                                             81

3.1.  Materials                                                                                                                                                                      81

3.1.1 Reagents Used For The Study                                                                                                                                 81

3.1.2. APPARATUS                                                                                                                                                            81

3.2. Methods                                                                                                                                                                        82

3.2.1. Sample Collection and Preparation                                                                                                                      82

3.2.1.1. Sample Collection,Drying ,Cutting and Grinding                                                                                            82

3.2.1.2 .Sample Preparation                                                                                                                                              82

3.3.Proximate                                                                                                                                                                       83

3.3.1.Crude Fibre Determination                                                                                                                                      83

3.3.2.Crude Protein Determination                                                                                                                                  83:

3.3.3.Crude Fat Determination:                                                                                              85

3.3.4.Moisture Content Determination                                                                                   85-86

3.3.5.Determination of Ash:                                                                                                    86

3.6.Determination of Carbohydrate                                                                                        87

3.4.Phytochemical Study                                                                                                                                                   87

3.4.1. Extraction and Phytochemical Determination                                                                                                   87

3.5.Mineral Study                                                                                                                                                                88

3.5.1. Digestion                                                                                                                                                                    88

3.6. Vitamins                                                                                                                                                                        89 - 90

3.7.Haematological Study                                                                                                                                                                 91

3.7.1.Animals used for The Study                                                                                                                                    91

3.7.2.Haematological Parameters Analyzed                                                                                                                 93

3.7.2.1  Haematocrit or Packed Cell Volume (Pcv) Determination.                                                                          93

3.7.2.2.White Cell Count (Wyngarden Method,1985).                                                                                                                94

3.7.3.Differential White Blood Cell Count( by Beckman Coulter method)                                                             95

3.7.4.Platelet count by Haemacytometer counting method:                                                   96

3.7.4.1.RBC Count (Haemacytometer counting method):                                                       97

3.8. Histological Study:                                                                                                           98

3.8.1 Fixation:                                                                                                                         98

3.8.2.Embedding:                                                                                                                    99

3.8.3.Sectioning:                                                                                                                     99

3.8.4.Staining:                                                                                                                        99

3.8.5.Mounting:                                                                                                                       100

3.9.Statistical Analysis oF Data                                                                                                                                           100

CHAPTER FOUR: RESULTS AND DISCUSSION

4.0.Results:                                                                                                                                                                           101

4.1. Proximate Analysis                                                                                                                                                     101-104

4.2.    Phytochemistry                                                                                                                                                         105

4.5.  Qualitative                                                                                                                                                                   105

4.3.Quantitative Analysis                                                                                                                                                  107-110

4.3    Vitamins:                                                                                                                                                                     111

4.4.     Minerals                                                                                                                                                                     112

4.5. Haematological Study                                                                                                                                                                113

4.6.   Histopathological  Study                                                                                                                                  114-198

 4.1. Discussion:                                                                                                                                                                  199-204

CHAPTER FIVE:

5.0. Conclusion and Recommendation                                                                                                                          205                                                                                                                                                                                    

5.1. Conclusion                                                                                                                                                                    205

5.3.Recommendation:                                                                                                                                                       206

5.4.Contribution to Knowledge                                                                                                                                         206

Reference

APPENDIX

 

 

 

 

 

                                                       ABSTRACT                                  

The nutritional and chemical constituents of different extracts of Neem leaf (Azdirachta indica) and their effect on the haematology and histopathology of Wistar Albino Rats were investigated. The samples analyzed includes processed and unprocessed leaves extract of Neem plant using cold water, hot water, 40% ethanol, 30%  methanol, 70% ethanol and methanol; And also,Wistar Albino Rats (25)  which were administered with extracts(processed and unprocessed) were divided into five(5) groups having five in each cage and one group as control. The phyotchemistry of the Neem leaves extract were determined in triplicates, qualitatively and quantitatively using standard methods. The proximate study carried out on the processed and unprocessed was observed  to contain carbohydrate ˃ moisture ˃ fat ˃ fibre ˃ protein ˃ ash.  Phytochemical studies on the leaves determined, detected  presence of alkaloids, flavonoids, saponnins, oxalate, phenol, tannin, steriod, glycosides, reducing sugar and carbohydrate. The results are presented as  mean ± STD of trplicate determination and means were compared using one-way ANOVA statitical analysis at 0.05 significance level. Rutin, ranging from 0.000 ± 0.01 to 77.543 ± 0.040,was observed to have high yield present in the processed and unprocessed leaves (14.290 ± 0.010 to 77.543 ± 0.040) while spartein was the lowest (0.002 ± 0.00 to 0.005 ± 0.00). The vitamins A,C,E and Riboflavin were determined, ranging from 0.89  ± 0.007 to 375.74 ± 1.22 amongst all samples,vitamin c was observed to be in high yield than any other of the vitamins. The total mineral concentration present in neem plant leaves extract were determined; and it was observed to contain Manganese , cobalt, zinc, copper, lead, nickel, sodium, potassium, magnesium and chromium  ranging from 0.003 to 23.529; But magnesium was observed to be in high yield ranging from 20.768 to 23.529.The haematological study of the animals treated using the processed leaves extract and after 24 hours of starvation were observed to have high count of platelets when compared with the controls(220 μl/kg to 350μl/kg);also the WBC count range was significantly decreased (4.0 to 14.0) /L when observed with the controls(13.00±0.00)/L,the RBC showed a significant increase, ranging from 4.5μl  to 7.2μl when compared with the control(4.5±0.00)/L.The HB count significantly increased from 10.7g/dl to 16.7g/dl when compared with the control(10.70±0.00)g/dL. The Histopathological study using liver and kidney,viewed under light microscope revealed spots and damages of the heart and liver,leading to distortion and haemorrhage. In conclusion,the consumption of alcoholic neem leaves extract during starvation and without dosage is toxic,harmful and can lead to death,especially when allowed to ferment due to bacteria activities than the hot water and tap water extract of neem leaves which is more safe to consumed with or without meal . 

 

 

TABLE OF CONTENTS

Title page                                                                                                                                                                              i

Certification                                                                                                                                                                         ii

Dedication                                                                                                                                                                            iii

Acknowledgement                                                                                                                                                              iv            

Abstract                                                                                                                                                                                 v

Table of contents                                                                                                                     vi-xi

List of Table                                                                                                                                                                         xii           

List of figure                                                                                                                                                                         xiii-xix   

List of abbreviation                                                                                                                                                       xx

CHAPTER ONE                                                                                                                

1.0. Introduction                                                                                                                                                                  1

1.1 Back Ground of Study                                                                                                                                                1             

1.2    Statement of the Problem                                                                                                                                          3

1.3. AIM AND OBJECTIVES                                                                                                                                           4

1.3.1.AIM                                                                                                                                                                             4

1.3.2. OBJECTIVES.                                                                                                                                                           4

1.4. SIGNIFICANCE OF STUDY                                                                                                                                     4

CHAPTER TWO

LITERATURE REVIEW                                                                                                                                                   5

2.1 Azadirachtaindica (Neem).                                                                                                                                       5          

2.2. Neem in Hindu Mythology                                                                                                                                        10

2.3. NEEM AS A WEED                                                                                                                                                    11

2.4    TAXONOMICAL CLASSIFICATION OF NEEM.                                                                                               11

2.5. Chemical Properties                                                                                                                                                    12

2.6. CHEMISTRY OF NEEM:                                                                                                                                          13                       2.7. USES AND BENEFITS OF NEEM TREE PARTS.                                                                                             16

 2.7.1  USES                                                                                                                                                                          16

 2.7.1.1.STORAGE                                                                                                                                                               16

2.7.1.2. INSECTICDES                                                                                                                                                      16

2.8.Cosmetic uses                                                                                                                                               16                           

2.9.VEGETABLE USE OF NEEM                                                                                                                                    17           

2.10.TRADITIONAL MEDICINAL USE OF NEEM                                                                                                       17

2.11. Name and uses of neem tree in the different African countries                                                                       18           

2.12.Panacea for All Diseases:                                                                                                                                        22

2.13.  Benefits of Neem Tree parts                                                                                                                                  22

2.13.1.Benefits of Neem leaves                                                                                                                                         22

2.13.1.1.Eye problems                                                                                                                                                        23

2.13.1.2.Viral Diseases                                                                                                                                                      23

2.14.Neem Effects against Bacteria                                                                                                                                                23

2.15.Neem Effects against Viruses                                                                                                                                  24

2.16.Neem Effects against Fungi                                                                                                                                     24

2.17.Neem Effects against Parasites                                                                                                                           25                               

2.17.1.De-worming                                                                                                                                                             26

2.17.2.Cardiac Care:                                                                                                                                                         26

2.17.3.Natural Pesticide:                                                                                                                                                  27

2.17.4.Fungal Diseases:                                                                                                                                                    27

2.17.5.Post-Parturition Disorder                                                                                                                                     27

2.17.6.Skin Care:                                                                                                                                                                28

2.18. neem and diabetes                                                                                                                                                    28

2.19. Uses of Neem Seeds and Fruits                                                                                                                               30

  2.19.1.Skin Problems                                                                                                                                                        30

2.19.2.Hair Care                                                                                                                                                                 30

 2.19.3.Eye and Ear Infections:                                                                                                                                        30

2.20.Other Ailments:                                                                                                                                                           31

2.20.1.Malaria:                                                                                                                                                                   31

2.20.2. neem cake organic manure                                                                                                                                  31

2.20.3.Components                                                                                                                                                            31

2.20.4.Use as a fertilizer                                                                                                                                                     32

2.20.5.Pest control                                                                                                                                                               32          

2.21.Neem Disadvantages:                                                                                                                                                               33

2.21.1.Strong Smell:                                                                                                                                                            33

2.21.2.Bitter Taste                                                                                                                                                               33

2.21.3.DANGER:                                                                                                                                                                 34

2.22.CHOLESTEROL                                                                                                                                                        34

2.23. USE AS CONTRACEPTIVE   IN BOTH MALE AND FEMALE:                                                   35

2.23.1.NEEM OIL - DIRECT USE:                                                                                                                                   36          

2.24.NEEM - CONTRACEPTIVE FOR MEN:                                                                                                              36

2.25.Insect Repellent:                                                                                                                                                         37

2.26.Flea Repellent :                                                                                                                                                          37

2.27.Uses of Neem Bark                                                                                                                                                    38

2.27.1.Oral Health                                                                                                                                                              38

2.27.2.Fever                                                                                                                                                                         38

2.27.3.Pile                                                                                                                                                                            38

2.28.Uses of Neem Flower                                                                                                                                                 38-39

2.29. Other Medical Benefits of Neem Tree                                                                                                                   39

2.29.1.Arthritis:                                                                                                                                                                   39

2.29.2. rheumatism                                                                                                                                                             40

2.29.3.Chronic Fatigue:                                                                                                                                                      41

2.29.4.Cancer:                                                                                                                                                                     41

2.30.Neem and anti-microbial activity:                                                                                                                          42-43    

2.31.Detoxifying Effects                                                                                                                                                    43

2.32.Gastric Health                                                                                                                                                             43

2.33.Pore Size                                                                                                                                                                       44

2.34.Phytochemicals                                                                                                                                                          44

2.34.1. antioxidant                                                                                                                                                              45

2.35.flavonoids                                                                                                                                                                    46

2.35.1.Definition of Flavonoids:                                                                                                                                      46

2.35.2. health benefits to humans                                                                                                                                    48

2.35.3.Food sources:                                                                                                                                                           49

2.35.4.Interactions:                                                                                                                                                             50

2.36.Classification of Flavonoids                                                                                                                                    50-53

2.36.1.Flavanols and Flavonols                                                                                                                                       53

2.36.2.Flavanones and Flavones                                                                                                                                     54

2.37.Anthocyanins                                                                                                                                                              58

2.38.Tannin:                                                                                                                                                                         63

2.39.Terpenene                                                                                                                                                                    63

2.40. Antioxidant Vitamins in Neem                                                                                                                               64

2.41. Alkaloids:                                                                                                                                                                    65

2.42. Neem and Environmental Protection                                                                                                                    68

2.42.1. Afforestation                                                                                                                                                          68

2.43.Vitamins and Minerals in Neem Plant                                                                                                                    69

2.43.1.Vitamins:                                                                                                                                                                  69

 

2.44.2.Minerals:                                                                                                                                                                   69

2.45.Anti-Vitamins.                                                                                                                                                             71

2.46.Herbal Preparation                                                                                                                                                     72

2.47. Traditional Herbal Extraction Methods                                                                                                                                73

2.48.Method of Administration                                                                                                                                        73

2.49. Haematological Parameters                                                                                                                                    74

2.49.1.Haematocrit (Packed Cell Volume):                                                                                                                   74

2.49.2. Elevated Haematocrit Levels                                                                                                                              74

2.49.3.Lowered Haematocrit Levels                                                                                                                               75

2.50.Platelet Count:                                                                                                                                                            75

2.51.Differential Leucocyte Count                                                                                                                                  76

2.52. Red Blood Cell Count                                                                                                                                              78

2.53.Histology                                                                                                                                                                      79

                                                                      

CHAPTER THREE

 3.0  Materials And Methods                                                                                                                                             81

3.1.  Materials                                                                                                                                                                      81

3.1.1 Reagents Used For The Study                                                                                                                                 81

3.1.2. APPARATUS                                                                                                                                                            81

3.2. Methods                                                                                                                                                                        82

3.2.1. Sample Collection and Preparation                                                                                                                      82

3.2.1.1. Sample Collection,Drying ,Cutting and Grinding                                                                                            82

3.2.1.2 .Sample Preparation                                                                                                                                              82

3.3.Proximate                                                                                                                                                                       83

3.3.1.Crude Fibre Determination                                                                                                                                      83

3.3.2.Crude Protein Determination                                                                                                                                  83:

3.3.3.Crude Fat Determination:                                                                                              85

3.3.4.Moisture Content Determination                                                                                   85-86

3.3.5.Determination of Ash:                                                                                                    86

3.6.Determination of Carbohydrate                                                                                        87

3.4.Phytochemical Study                                                                                                                                                   87

3.4.1. Extraction and Phytochemical Determination                                                                                                   87

3.5.Mineral Study                                                                                                                                                                88

3.5.1. Digestion                                                                                                                                                                    88

3.6. Vitamins                                                                                                                                                                        89 - 90

3.7.Haematological Study                                                                                                                                                                 91

3.7.1.Animals used for The Study                                                                                                                                    91

3.7.2.Haematological Parameters Analyzed                                                                                                                 93

3.7.2.1  Haematocrit or Packed Cell Volume (Pcv) Determination.                                                                          93

3.7.2.2.White Cell Count (Wyngarden Method,1985).                                                                                                                94

3.7.3.Differential White Blood Cell Count( by Beckman Coulter method)                                                             95

3.7.4.Platelet count by Haemacytometer counting method:                                                   96

3.7.4.1.RBC Count (Haemacytometer counting method):                                                       97

3.8. Histological Study:                                                                                                           98

3.8.1 Fixation:                                                                                                                         98

3.8.2.Embedding:                                                                                                                    99

3.8.3.Sectioning:                                                                                                                     99

3.8.4.Staining:                                                                                                                        99

3.8.5.Mounting:                                                                                                                       100

3.9.Statistical Analysis oF Data                                                                                                                                           100

CHAPTER FOUR: RESULTS AND DISCUSSION

4.0.Results:                                                                                                                                                                           101

4.1. Proximate Analysis                                                                                                                                                     101-104

4.2.    Phytochemistry                                                                                                                                                         105

4.5.  Qualitative                                                                                                                                                                   105

4.3.Quantitative Analysis                                                                                                                                                  107-110

4.3    Vitamins:                                                                                                                                                                     111

4.4.     Minerals                                                                                                                                                                     112

4.5. Haematological Study                                                                                                                                                                113

4.6.   Histopathological  Study                                                                                                                                  114-198

 4.1. Discussion:                                                                                                                                                                  199-204

CHAPTER FIVE:

5.0. Conclusion and Recommendation                                                                                                                          205                                                                                                                                                                                    

5.1. Conclusion                                                                                                                                                                    205

5.3.Recommendation:                                                                                                                                                       206

5.4.Contribution to Knowledge                                                                                                                                         206

Reference

APPENDIX

 

 

 

 

 

 

 

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APA

EMELIEZE, N., BENJAMIN, A , MARY, E , CHARITY, O , EMMANUEL, A & PETER, A (2019). The nutritional and chemical constituents of different extracts of Neem leaf (Azdirachta indica) and their effect on the haematology and histopathology of Wistar Albino Rats. Afribary. Retrieved from https://afribary.com/works/full-work-pub-article

MLA 8th

EMELIEZE, NNEKA, et. al. "The nutritional and chemical constituents of different extracts of Neem leaf (Azdirachta indica) and their effect on the haematology and histopathology of Wistar Albino Rats" Afribary. Afribary, 25 Feb. 2019, https://afribary.com/works/full-work-pub-article. Accessed 24 Nov. 2024.

MLA7

EMELIEZE, NNEKA, AMADI BENJAMIN , EMELIEZE MARY , OGUNKA CHARITY , AGOMUO EMMANUEL and AND PETER . "The nutritional and chemical constituents of different extracts of Neem leaf (Azdirachta indica) and their effect on the haematology and histopathology of Wistar Albino Rats". Afribary, Afribary, 25 Feb. 2019. Web. 24 Nov. 2024. < https://afribary.com/works/full-work-pub-article >.

Chicago

EMELIEZE, NNEKA, AMADI BENJAMIN , EMELIEZE MARY , OGUNKA CHARITY , AGOMUO EMMANUEL and AND PETER . "The nutritional and chemical constituents of different extracts of Neem leaf (Azdirachta indica) and their effect on the haematology and histopathology of Wistar Albino Rats" Afribary (2019). Accessed November 24, 2024. https://afribary.com/works/full-work-pub-article