ACKNOWLEDGMENTS I would like Thank Allah, the creator of the world for giving me strength and helping me to finish

this work. I would like to express my deep and sincere gratitude to my supervisor, Dr. Badria M. Abdel-Wasae, associate professor of Zoology

(Immunology and Parasitology) and Vice Dean for quality, Faculty of Applied Science, Taiz University for her excellent guidance, kind encouragement, scientific advice, and for her fruitful critical discussion and for reading and criticizing the thesis. I am deeply grateful to my supervisor Dr. Mohammad El- Ezzy assistant professor of Zoology (Cytology and Histology) and head of the Department of Biology, Faculty of Applied Science, Taiz University, for his detailed and constructive comments, and for his important support throughout this work. Special thank to Prof. Dr. Abdul Karim Nasher, professor of Parasitology, Faculty of Science, Sanaa University who provided me with the albino mice. I thank all the members and staff in the Biology Department, especially, Dr. Yasser Obady for his guidance in statistical analysis, Dr. Fouad Homran. I wish to extend my warmest thanks
to

Dr. Abdu Alwally Al-

Khulaidi for plant identification (Aloe vera) and I wish to thank Mr. Abdurhman Dabwan, the technician at Biology Department, Faculty of Education, Taiz University, for his assistance and support during the practical period. Finally, I thank my family, especially my mother, my brothers, for their loving support, my wife and my three daughters without their encouragement and understanding it would have been impossible for me to finish this work.
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ABSTRACT Visceral leishmaniasis is considered as a disease of serious medical importance. It infects humans, especially children and some domestic and wild animals. The disease is common in areas where the vector (sand fly) is present. The aims of this study can be summarized to: 1- Study the pathological effects of visceral leishmaniasis on experimental albino mice at different periods. 2- Study the healing effect of Aloe vera leaf extract on mice with visceral leishmaniasis. The present study consists of two parts. The first part of this study has three serological tests: (formol gel, Leishmania IgG/IgM Rapid TestCassette and Culture of parasites) were conducted for isolation of the visceral leishmaniasis parasite from patients in Taiz Governorate at April to December 2008. The second part of the study included infecting mice with visceral leishmaniasis parasites and treatment by the leaf extract of Aloe vera (15 mg / kg / 5 days). At 20, 40, 60, 80 and 100 day post infection 4 mice from each group were weighted as well as, study the morphological, histopathological, haematological according to international methods. The results of this study show that: * The albino mice are susceptible to infection with visceral leishmaniasis experimentally even 100 day from infection. * The mortality rate in albino mice due to visceral leishmaniasis reached to 11.11% and occurred in the first periods. * The decrease in weight of mice with visceral leishmaniasis, reached to 25% on day100 post infection compared to the control group on the same
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assays

and

biochemical

analysis

day. While, the treated mice with plant extract Aloe vera reached to 8 % at day 40. * The significant decrease in the values of serum albumin and glucose of mice with visceral leishmaniasis reaches to 32% and 42% at day 40 and 80 respectively, comparing to the control group in the same day, and the treated mice with plant extract Aloe vera (15 mg / kg / 5 days) changed the decline rate to 4% and 3.7 % at day 100. * There is a significant increase in the value of total protein and globulin in mice with visceral leishmaniasis to 87% and 331% at day 80, comparing with the control group in the same day. While, the treated group has an increases of total protein and globulin serum to 25% and 89 % respectively in the days 40 and 100, comparing with the control group in the same days. * A significant decrease in the values of haemoglobin, white the blood cells and platelets of mice with visceral leishmaniasis reach to 48, 33 and 42 % at day 60, 80 and 100, respectively, comparing with the control group. But the treated group with Aloe vera plant extract records a decrease in the values to 20%, 19% and 10% respectively at day 60, 80 and 100 respectively, comparing with the control group in the same day. * There is a significant increase in the concentration of GlutamicOxaloacetic Transaminase and Glutamic-Pyruvic Transaminase in the mice with visceral leishmaniasis. The percentage reaches to 626 and 470% respectively at day 20, comparing with the control group. While, the treated group recorded the increase value of those enzymes to 66% and 45% respectively at day 80, comparing with the control group. * Increase in the values of urea and the creatinine in mice with visceral leishmaniasis reach to 128% and 40% respectively in day 20 comparing with the control group. While, the increase in the same values of urea and

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the creatinine in treated group reach to 23% and 3.2 % respectively at day 60 and 80 comparing with the control group. * Histopathological results include widespread damage in the liver, spleen, kidneys and small intestine of the mice with visceral leishmaniasis. These effects have appeared in first periods. The injury disorders and destroys the general organization of the tissues and the abundance of inflammatory and infiltrators cells. While, the cells swollen, and high damages in nucleus are seen. The emergence of haemorrhage, especially among renal tubules and hepatic lobules of the cells has manifested necrosis as well. * Administration Aloe vera plant crude to mice with visceral leishmaniasis has stimulated tissues to heal and get repaired gradually. Nevertheless some of the pathological changes stilled limited. *On the other hand, the study confirms the hazard of these parasites on human and animal health as well as shows the successful role of medical plants Aloe vera leaf extract on the treatment of this disease, in a time was called to use natural therapy.

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CONTENTS
Chapter 1 subchapter Title INTRODUCTION Aim of The Work 2 2.1 2.2 2.3 2.4 2.5 2.6 2.7 2.8 3 3.1 3.2 3.3 3.4 3.5 3.6 3.7 3.8 3.9 4 4.1 4.2 4.3 4.4 5 LITERATURE REVIEW Morphology And Life Cycle Modes of Transmission Distribution of Leishmania Classification The Symptoms of Leishmaniasis Control of Leishmaniasis The Pathological Studies Aloe vera Plant MATERIALS AND METHODS Isolation of The Parasite Experimental Animals And Animals Grouping Experiment design Plant Material And Extraction Pathological Studies Morphological Studies Biochemical Studies Haematological Studies Statistical Analysis RESULTS Gross Pathology Histopathological Observation Morphological Studies Biochemical And Haematological Analysis DISCUSSION Conclusion And Recommendations REFERENCES APPENDICES
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page 1 2 3 4 5 7 10 11 12 17 18 24 24 28 29 29 29 30 30 33 33 34 34 34 75 77 99 11 112 138

LISTS OF FIGURES
Chapter 2 2 3 3 3 3 4 Figure 1 2 1 2 3 4 1-6 TITLE Life cycle of Leishmania parasites Distribution of visceral Leishmania in world Serum samples of patients Formol gel test Leishmania IgG/IgM test Leishmania culture Gross Pathological pictures of infected mice with visceral leishmaniasis Photomicrographs of sections of the liver of infected mice and blood smear Photomicrographs of sections of the liver of control mice Photomicrographs of sections in the liver of infected mice Photomicrograph of sections in liver of infected and treated mice Photomicrographs of sections of the kidney of control mice Photomicrographs of section in the kidney of infected mice Photomicrograph of sections in kidney of infected and treated mice Photomicrographs of sections the spleen of control mice Photomicrographs of sections in the spleen of infected mice Photomicrograph of sections in spleen of infected and treated mice Photomicrographs of sections the small intestines of control mice Photomicrographs of sections in the small intestines of infected mice Photomicrographs of sections small intestines of infected and treated mice Comparing the values of weight (g/dl) in three experimental groups Comparing the values of biochemical tests in the experimental groups Comparing the values of haematological tests in the experimental groups page 5 9 25 25 25 26 44

4 4 4 4 4 4 4 4 4 4 4 4 4 4 4

7-14 15-16 17-26 27-34 35-36 37-46 47-54 55-56 57-66 67-74 75-76 77-86 87-94 95 96-103

46,48 48 48,50,52 52,54 54 56,58 58,60 62 62,64 66,68 68 68,70,72 72,74 75 77-91

104-106

93-97

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LIST OF ABBREVIATIONS
AIDS ANOVA AS ASTD AVL BALB BC BCG BS CL CV DCL DEG DNA EDTA ELISA FCS FIG. G GC GOD GP63 H H&E HA HG HIV IFCC IFN IFN-L Acquired Immunity Deficiency Syndrome Analysis of Variances Absorbance of Sample Absorbance of Standard Aloe vera Leaf Bagg Albino Mice Bowmans Capsule Bromocresol Green Blood Sinusoids Cutaneous Leishmaniasis Central Vein Diffuse Cutaneous Leishmaniasis Degeneration Desoxyribonucleic Acid Dipotassium Ethylenediaminetetraacetate Enzyme-Linked Immunosorbant Assay Fetal Calf Serum Figure Glomerulus Goblet Cell Glucose Oxidase Glycoprotein (Promastigote Surface Protease) Hepatocytes Haematoxylin And Eosin Haemorrhage Hepatic Granulomatosis Human Immunodeficiency Virus International Federation Of Clinical Chemistry And Laboratory Medicine Gamma Interferon Interferon-L

VII

IgG IgM IL INF K L LP LPG M MCL MW NB NEC NK NNN O.D PN POD P-Y S SAG SGOT SGPT SM TH1 TH2 TLC V VI VL WHO

Immunoglobulin G Immunoglobulin M Interleukin Infiltration Kupffer Cell Leishmania Lamina Propria Lipophosphoglycan Mucosa Mucocutaneous Leishmaniasis Molecular Weight Nutrient Broth Necrosis Natural Killer Novy-Macneal-Nicolle Optical Density Pyknotic Nuclei Peroxidase Peptone Yeast Spaces Sodium Antimony Gluconate Serum Glutamic-Oxaloacetic Transaminase Serum Glutamic-Pyruvic Transaminase

Submucosa
T-Helper 1 Lymphocytes T-Helper 2 Lymphocytes Thin Layer Chromatography

Vacuolation Villi
Visceral Leishmaniasis World Health Organization

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