Effects of Khat (Catha Edulis) on Some Blood Contents and the Tissues of the Digestive System of Rabbits
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Effects Of Khat (Catha edulis) On Some Blood Contents And The Tissues Of The Digestive System Of Rabbits By Mohammad Abdulwali Saeed Ali Al-Hegami B. Sc. A THESIS Presented in Partial Fulfillment of the Requirements for the Degree of MASTER OF SCIENCE In Zoology Biology Department Faculty of Science Sana’a University Yemen February 2001 Effects Of Khat (Catha edulis) On Some Blood Contents And The Tissues Of The Digestive System Of Rabbits BY Mohammad Abdulwali Saeed Ali Al-Hegami Approved A - Supervision Committee Members: 1 - Prof. Dr. Abdul Karim A. Nasher 2 - Dr. Abdulgalil S. Griuaty B - Examination Committee Members: 1 - Prof. Dr. Abdul Karim A. Nasher 2 - Prof. Dr. Abubakr A. Al-Qirbi 3 - Prof. Dr. Foad Al-Khelli Date Thesis is presented, February, 2001 Supervisors Prof. Dr. Abdul Karim A. Nasher Professor of Zoology Biology Department - Faculty of Science Sana’a University - Yemen Dr. Abdulgalil S. Griuaty Assistant Professor of Zoology Biology Department - Faculty of Science Sana’a University - Yemen DEDICATION With lots of Affection to: My Dear Parents My Brothers and My Sisters My Wife and My Children The Everlasting Source of Love and Sacrifice. ﺑﺴﻢ ﺍﻪﻠﻟ ﺍﻟﺮﲪﻦ ﺍﻟﺮﺣﻴﻢ ﺻﺪق اﷲ اﻟﻌﻈﻴﻢ ﺳﻮرة اﻟﺒﻘﺮة ﺁیﺔ 32 ACKNOWLEDGMENTS First and foremost, thanks are due to god, the most beneficent and merciful. I would like to acknowledge with gratitude the sincere supervision of Dr. Abdul Karim A. Nasher, Professor of Zoology, Biology Dept., Faculty of Science, Sana’a University for his guidance, continuous encouragement, helpful comments, valuable advice and support during the course of this study. I would also like to thank Dr. Abdulgalil S. Griuaty, Assistant Professor of Zoology, Biology Dept., Faculty of Science, Sana’a University for his role in preparing the study plan. Thanks are also extended to Prof. Imad M. Al-Ani, Faculty of Medicine, Sana’a University for his help in the examination and interpretation of digestive system sections. I would also like to express my deep gratitude to my family for their continuous encouragement and support during my work. I am indebted to the authorities of Sana’a University and in particular the Deanship of the Faculty of Science, and the staff of the Department of Biology for their help, support and provision of research facilities. Thanks are also extended to the staff of Higher Studies and Scientific Research, Sana’a University for their support. I would like to express my sincere gratitude to all my friends, for their helpful assistance during this study. i SUMMARY The present study is planned to elucidate the possible effects of Khat on the blood contents and tissues of the digestive system in experimental rabbits. Thirty-two adult rabbits of both sexes were used in the present study. Rabbits were chosen for this study because they can consume Khat as well as for their availability. The animals were divided into two groups: 1. The experimental group consisted of 24 rabbits (12 males and 12 females). Each animal received daily 600 g fresh alfalfa supplemented by 50 g of commercial concentrated fodder and 40 g fresh leaves of Sawtty Khat per one kg body weight for 4.5 successive months. 2. The control group consisted of 8 rabbits (4 males and 4 females) and each of them received the fresh alfalfa diet and the supplement fodder only. Venous blood samples were collected from all rabbits every 1.5 months during the study period and analyzed to assess blood parameters (CBC and serum). Determinations of CBC parameters were performed by routine clinical haematology methods. Serum parameters (blood sugar, total protein, cholesterol and triglycerides) were also determined by clinical chemistry methods. For histological part of this study, about 10 cu. mm tissue pieces of the stomach, small intestine, caecum and liver, were taken rapidly from each ii killed animal, and put immediately in a fixative solution (10% Neutral buffered formaline), and left for 24 hours. The fixed tissues were washed in running tap water and with 70 % ethanol several times. Finally, the paraffin technique was used in preparing microscopical sections. All the data were statistically analyzed by using student t -test. The results of this study can be summarized as follows: In the hematological part of this study, all the results of complete blood count (CBC) parameters and serum analysis were statistically significant (P < 0. 01). Details of these results are: 1. The total of erythrocyte count (RBCs), haemoglobin (Hb %) and haematocrit (Hct) decreased significantly by an average of (15. 91%, P < 0.01), (14.43%, P < 0.01), and (14.50%, P < 0.01) respectively in Khat fed rabbits as compared to their equivalent controls at the end of the experiment. 2. The mean of total count of leucocytes (WBCs) decreased significantly by an average of (16.18%, P< 0.01), in Khat fed rabbits as compared to their equivalent controls at the end of the experiment. This decrease of WBCs was observed in the lymphocytes by an average of (30.15%, P < 0.01), while the number of granulocytes and MID increased by an average of (11.16%, P < 0.01) and (13.02%, P < 0.01), respectively in Khat fed rabbits compared to their equivalent controls at the end of experiment. 3. The mean of total count of Platelets significantly increased by an average of (47.07 %, P. < 0.01), in Khat fed rabbits compared to their equivalent controls at the end of the experiment. iii 4. Mean blood sugar increased significantly by an average of (13. 29%, P < 0.01) in Khat fed rabbits compared to their equivalent controls at the end of the experiment. 5. The mean of total Serum Proteins decreased significantly by an average of (17. 89 %, P < 0.01) in Khat fed rabbits compared to their equivalent controls at the end of the experiment. 6. Total Cholesterol decreased significantly by an average of (7. 92 %, P < 0.01) in Khat fed rabbits compared to their equivalent controls at the end of the experiment. 7. Triglycerides decreased significantly by an average of (13. 24%, P < 0.01) in Khat fed rabbits compared to their equivalent controls at the end of the experiment. Results on the effects of Khat on the tissues of the digestive system in rabbits showed that daily Khat administration in experimental rabbits caused very little structural changes. These changes were observed in the stomach, duodenum, jejunum and liver tissues. No changes were observed in the ileum and caecum tissues in Khat fed rabbits as compared to these organs in the control animals. These changes are: A. Stomach: Histological examination of the Sections in the stomach of the Khat fed rabbits showed a slight necrosis of the epithelial cells passing down into the gastric pits while no effect was seen in the control rabbits. iv B. Small Intestine: Histological examination of the Sections in the small intestine of the Khat fed rabbits showed that the affected organs were the duodenum, and jejunum while there was no change in the ileum tissues in Khat fed rabbits as compared to their respective controls. The effect observed in the duodenum was shown as necrotic villi with degenerative epithelial cells and dilated blood vessels congested with blood. In the jejunum severe necrosis was observed on the lining of this organ. This necrosis severely affected the villi and extended into their lamina propria. C. Caecum: There was no detectable change in caecum tissues in Khat fed rabbits as compared to their respective controls. D. Liver: Histological examination of the liver Sections of Khat fed rabbits showed that the central vein was congested with blood. Moreover, there was a proliferation of Kupffer cells in the livers of Khat fed rabbits, which was not observed in the control rabbits. Results on the effects of Khat on the body weight of rabbits showed that daily Khat administration in experimental rabbits caused significant decrease in the means of body weight by an average of (24. 34 %, P. < 0.01), in Khat fed rabbits as compared to their equivalent controls. v It was also noticed that Khat had some effect on the behavior of Khat fed rabbits as evidenced by loss of appetite and aggressive fights among three male rabbits, which resulted in severe injuries of these rabbits. Moreover, increased movements was observed in the Khat fed rabbits 2-3 hours after Khat consumption, and returned to normal behavior thereafter. vi Table of Contents Contents Page No. ACKNOWLEDGEMENTS i SUMMARY ii TABLE OF CONTENTS vii LIST OF TABLES ix LIST OF FIGURES x ABBREVIATIONS xii LIST OF APPENDICES xiii CHAPTER 1 1. INTRODUCTION 1.1. General considerations (1) 1.2. Literature review (5) 1.3. Morphology of Khat plant (8) 1.4. Cultivation of Khat plant (9) 1.5. Chemical constituents of Khat (13) 1.5.1. Alkaloids (13) 1.5.2. Amino acids (19) 1.5.3. Tannins (19) 1.5.4. Vitamins, minerals, etc. (19) 1.6. Khat chewing habit in Yemen (21) 1.7. Aim of the work (23) vii CHAPTER 2 2. MATERIALS AND METHODS 2.1. General Layout of the experiment (24) 2.1.1. Khat leaves (24) 2.1.2. The rabbits (24) 2.2. Methods (25) 2.3. Chemicals and instruments used in this study (32) 2.4. Statistical analysis of the results (35) CHAPTER 3 3. RESULTS AND DISCUSSION 3.1. Effects of Khat on blood contents (37) 3.1.1. Complete blood count (37) 3.1.2. Serum contents (41) 3.2. Effects of Khat on the tissues of the digestive system (50) 3.3. Effect of Khat on the body weight (58) 3.4. Effect of Khat on the behavior of rabbits (61) CONCLUSION AND RECOMMENDATIONS (62) REFERENCES (63) APPENDICES (74) ARABIC SUMMARY (86) viii List of Tables Table Page No.