Toxicity of Abuleila (Detarium senegalense) Fruit Flour to Rats By Hanan Mohamed Neimtalla Gibreil B.V.Sc. (2003), M.V.Sc. (2009) University of Khartoum A Thesis Submitted to the University of Khartoum in Fulfillment of the RequirementsFor the Ph.D. Degree in Veterinary Science (Toxicology). Supervisor: Professor Ahmed El Amin Mohammed Department of Pharmacology and Toxicology Faculty of Veterinary Medicine University of Khartoum May, 2015 Gibreil, Hanan M. N.,Ph.D. Thesis,2014 لاي ذؼاٌٝ: )فأنبتنا فيها حبا * وعنبا وقضبا * وزيتىنا ونخﻻ * وحذائق غلبا * وفاكهت وأبا * هتاعاً لكن وﻷنعاهكن( سىرة عبس اﻻياث )27 - 32( ii Gibreil, Hanan M. N.,Ph.D. Thesis,2014 DEDICATION I Would like to dedicate this research lovingly to: My kind Parents, who support me in everything. My Husband, who helped me to finished this research. My Sisters and brother, who encouraged me to reach this goal. My Son and Daughter, who were a great source of support, and Every person, who gave me a hint to exact my trail. Hanan August , 2014 iii Gibreil, Hanan M. N.,Ph.D. Thesis,2014 CONTENTS Item Page List of Tables vii List of Figures ix List of Plates x List of Abbreviations xi Acknowledgements xii Abstract xiii Arabic abstract xvi INTRODUCTION 1 CHAPTER ONE LITERATURE REVIEW 3 1.1 Ethnotoxicology 3 1.2 Food trees 4 1.2.1 Adansonia digitata 4 Phytochemistry 4 Food uses 5 Medicinal uses 5 1.2.2 Ziziphus spina-christa 6 Phytochemistry 6 Food uses 6 Medicinal uses 7 1.2.3 Hyphaene thebaica 7 Food uses 8 Medicinal uses 8 1.2.4 Balanites aegyptiaca 8 Phytochemistry 9 Food uses 9 Medicinal uses 9 1.3 Fabales food trees 10 1.3.1 Family Fabaceae 10 Taxonomy and description 11 Uses 12 1.3.2 Caesalpinioideae 13 1.3.3 Detarieae 13 1.3.3.1 Tamarindus indica 13 Food uses 14 Medicinal uses 14 1.3.3.2 Detarium 14 1.3.3.2.1 Detarium macrocarpum 15 1.3.3.2.2 Detarium microcarpum 15 Phytochemistry 15 iv Gibreil, Hanan M. N.,Ph.D. Thesis,2014 Food uses 16 Medicinal uses 16 1.3.3.2.3 Detarium senegalense 17 Taxonomy 17 Distribution 17 Description 18 Nutrients composition 19 Proximate analysis 20 Phytochemical analysis 20 Nutritional uses 21 Medicinal uses 22 Toxicity 24 CHAPTER TWO MATERIALS AND METHODS 26 2.1 Materials and experimental designs 26 2.1.1 Toxicity of dietary Abuleila (D. senegalense) fruit flour to rats 26 2.1.1.1 Animals, housing and management 26 2.1.1.2 Test plant 26 2.1.1.2.1 Fruit flour preparation 27 2.1.1.2.2 Fruit flour proximate analysis 27 2.1.1.2.3 Microminerals 27 2.1.1.2.4 Preliminary phytochemical screening 28 2.1.1.3 Administration and dose rates 29 2.1.1.4 Data collected 29 2.1.2 Toxicity of Abuleila (D. senegalense) fruit flour water extract to rats 29 via s/c route 2.1.2.1 Animals, housing and management 29 2.1.2.2 Fruit flour extraction 29 2.1.2.3 Administration and dose rates 30 2.1.1.4 Data collected 30 2.1.3 Toxicity of Abuleila (D. senegalense) fruit flour water extract to rats 30 via i/p route 2.1.3.1 Animals, housing and management 30 2.1.3.2 Administration and dose rates 31 2.1.3.3 Data collected 31 2.2 Methods 31 2.2.1 Haematological methods 31 2.2.1.1 Red blood cells count (RBCs) 31 2.2.1.2 Packed cell volume (PCV) 32 2.2.1.3 Haemoglobin concentration (Hb) 32 2.2.1.4 Red blood cell haematostatistics 32 2.2.1.4.1 Mean corpuscular volume (MCV) 32 2.2.1.4.2 Mean corpuscular haemoglobin concentration (MCHC) 32 2.2.1.5 White blood cell count (WBCs) 33 2.2.1.6 Differential leukocyte count 33 2.2.2 Chemical methods 33 v Gibreil, Hanan M. N.,Ph.D. Thesis,2014 2.2.2.1 Serum metabolites 33 2.2.2.1.1 Glucose 33 2.2.2.1.2 Total protein 34 2.2.2.1.3 Albumin 35 2.2.2.1.4 Urea 35 2.2.2.2 Serum enzyme activities 36 2.2.2.2.1 Aspartate aminotransferase (Glutamicoxaloacetic transaminase, L, 36 Aspartate = 2 – OXO-glutarate aminotransferase E.C.2, 6,1.1,AST). 2.2.2.2.2 Alanine amino transferase, (Glutamic pyruvic transaminase, 36 L- aspartate,2-oxoglutamate ALT) 2.2.2.3 Test plant macrominerals 37 2.2.2.4 Test plant microminerals 37 2.2.3 Histopathological methods 39 2.3 Statistical analyses 39 CHAPTER THREE 40 RESULTS 3.1 Toxicity of dietary Abuleila (D. senegalense) fruit flour to rats 40 3.1.1 Clinical signs 40 3.1.2 Body weight changes 40 3.1.3 Haematological findings 41 3.1.4 Changes in serum constituents 42 3.1.5 Post mortem findings 43 3.1.6 Histopathological findings 43 3.2 Toxicity of Abuleila (D. senegalense) fruit flour water extract to 46 rats via subcutaneous route 3.2.1 Clinical sings 46 3.2.2 Body weight changes 46 3.2.3 Haematological findings 47 3.2.4 Changes in serum constituents 48 3.2.5 Post mortem findings 50 3.2.6 Histopathological findings 50 3.3 Toxicity of Abuleila (D. senegalense) fruit flour water extract to rats 51 via i/p route 3.3.1 Clinical sings 51 3.3.2 Body weight changes 51 3.3.3 Haematological findings 52 3.3.4 Changes in serum constituents 54 3.3.5 Post mortem findings 54 3.2.6 Histopathological findings 55 CHAPTER FOUR DISCUSSION 58 4.1 Conclusion 62 4.2 Suggestions for future work 62 REFERNCES 63 APPENDIX 80 vi Gibreil, Hanan M. N.,Ph.D. Thesis,2014 List of Tables Table Page No. Title 1 Proximate percent crude chemical components (dry-matter basis) 27 of Detarium senegalense fruit flour. Micromineral components (dry-matter basis) of Detarium 2 28 senegalense fruit flour. 3 Preliminary phytochemical screening of Detarium senegalense 28 fruit flour extract. 4 Average (mean ± s.d.) body weight changes of experimental rats 40 fed Detarium senegalense fruit flour for 6 weeks. 5 Average (mean ± s.d.) blood values of experimental rats fed 41 Detarium senegalense fruit flour for 6 weeks. 6 Average (mean ± s.d.) WBCs differential count of experimental 42 rats fed Detarium senegalense fruit flour for 6 weeks. 7 Average (mean ± s.d.) serum metabolites values of experimental 42 rats fed Detarium senegalense fruit flour for 6 weeks. 8 Average (mean ± s.d.) serum enzymes values of experimental rats 43 fed Detarium senegalense fruit flour for 6 week. 9 Average (mean ± s.d.) body weight changes of experimental rats 47 injected subcutaneously with Detarium senegalense fruit flour water extract for 6 weeks. 10 Average (mean ± s.d.) blood values of experimental rats injected 48 subcutaneously with D. senegalense fruit flour water extract for 6 weeks. 11 Average (mean ± s.d.) WBCs differential count values of 48 experimental rats injected subcutaneously Detarium Senegalense fruit flour water extract for 6 weeks. 12 Average (mean ± s.d.) serum metabolites values of experimental 49 rats injected subcutaneously Detarium senegalense fruit flour water extract for 6 weeks. 13 Average (mean ± s.d.) serum enzymes values of experimental rats 49 injected subcutaneously Detarium senegalense fruit flour water extract for 6 weeks. vii Gibreil, Hanan M. N.,Ph.D. Thesis,2014 14 Average (mean ± s.d.) body weight changes of experimental rats 52 injected intraperitoneally with Detarium senegalense fruit flour water extract for 6 weeks. 15 Average (mean ± s.d.) blood values of experimental rats injected 53 intraperitoneally with D. senegalense fruit flour water extract for 6 weeks. 16 Average (mean ± s.d.) WBCs differential count values of 53 experimental rats injected intraperitoneally Detarium Senegalense fruit flour water extract for 6 weeks. 17 Average (mean ± s.d.) serum metabolites values of experimental 54 rats injected intraperitoneally Detarium senegalense fruit flour water extract for 6 weeks. 18 Average (mean ± s.d.) serum enzymes values of experimental rats 54 injected intrperitoneally Detarium senegalense fruit flour water extract for 6 weeks. viii Gibreil, Hanan M. N.,Ph.D. Thesis,2014 List of Figures Figure Page No. Title 1 Distribution of test plant in Africa. 17 2 Detarium senegalense fruit. 19 Graphical representation of the concentrations of trace minerals using 3 38 XRF-Spectrometer system. 4 Average growth curves of treatment groups. 41 5 Average growth curves of treatment groups. 47 6 Average growth curves of treatment groups. 52 ix Gibreil, Hanan M. N.,Ph.D. Thesis,2014 List of Plates Plate No. Title Page 1 The Detarium senegalense savanna type tree. 18 2 Detarium senegalense fruit. 18 3 Presser machine at 15 ton/cm2 38 4 The XRF Spectrometer system with liquid nitrogen. 38 5 Thickening of alveolar walls with slight interstitial proliferation of type 44 II cells. Peribronchial mononuclear cells infiltrate in the lung of rat in group C. H&E x100. 6 Thickening of alveolar walls with severe interstitial proliferation of type 44 II cells in the lung of rat in group D. H&E x100. 7 Thickening of alveolar walls with slight interstitial proliferation of type 45 II cells in the lung of rat in group D. H&E x100. 8 Peribronchial mononuclear cells infiltrate in the lung of rat in group D. 45 H&E x100. 9 Focal infiltration of mononuclear cells and neutrophils around blood 46 vessels in the lung of rat in group D.