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( Urtica Dioica L.) LEAF, FRUIT and ROOT EXTACTS Msc. T

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( Urtica Dioica L.) LEAF, FRUIT and ROOT EXTACTS Msc. T PHYTOCHEMICAL SCREENING and ANTIMICROBIAL ACTIVITIES OF STINGING NETTLE (Urtica dioica L.) LEAF, FRUIT AND ROOT EXTACTS MSc. THESIS MEGERSA IDRIS AHMED FEBURARY 2021 HARAMAYA UNIVERSITY, HARAMAYA Phytochemical Screening and Antimicrobial Activities of stinging Nettle (Urtica dioica L.) Leaf, Fruit and Root Extracts A Thesis Submitted to the School of Biological Sciences and Biotechnology Postgraduate Programs Directorate Haramaya University In Partial Fulfillment of the Requirements for the Degree of Master of Science in Biotechnology Megersa Idris Ahmed January, 2021 Haramaya University, Haramaya APPROVAL SHEET HARAMAYA UNIVERSITY POSTGRADUATE PROGRAM DIRECTORATE As thesis Research advisors, we hereby certify that we have read and evaluated this Thesis, prepared, under our guidance by Megersa Idris entitled Phytochemical Screening and Antimicrobial Activities of stinging Nettle (Urtica dioica L.) Leaf, Fruit and Root Extracts.We recommend that it be submitted as fulfilling the thesis requirement. Zekeria Yusuf (PhD) ________________ _______________ Major Advisor Signature Date Ashebr Abraha (DVM, MSc, Assoc.Prof.) _______________ ________________ Co- Advisor Signature Date As member of the Board of Examiners of the M.Sc. Thesis Open Defense examination, we certify that we have read and evaluated the Thesis prepared by Megersa Idris and examined him. We recommend that the thesis be accepted as fulfilling the thesis requirements for the degree of Master of Science in field of Biological Sciences (Biotechnology). ________________ ________________ ________________ Chairperson Signature Date ________________ ________________ ________________ Internal examiner Signature Date ________________ ________________ _______________ External examiner Signature Date Final approval and acceptance of the Thesis is contingent upon the submission of its final copy to the council of Graduate Studies (CGS) through the candidate's department or school graduate committee (DGC or SGC). II DEDICATION This thesis work is dedicated to my mother Fatuma Ahmed and my wife Merkani Mohamed for their encouragement and support in different aspects. III STATEMENT OF THE AUTHOR By my signature below, I declare and affirm that this M.Sc Thesis is my own work. I have followed all ethical and technical principles of scholarship in conducting studies, data collection, data analysis, and compilation of this Thesis. Any scholarly matter that is included in the Thesis has been given recognition through citation. This Thesis has been submitted in partial fulfillment of the requirements for the Degree of Master of Science in Biological Science at Haramaya University. The Thesis is deposited in the Haramaya University Library and is made available to borrowers under rules of the Library. I solemnly declare that this Thesis has not been submitted to any other institution anywhere for the award of any academic degree, diploma, or certificate. Brief quotations from this Thesis may be made without special permission provided that accurate and complete acknowledgment of sources is made. Requests for permission for extended quotations from or reproduction of this Thesis in whole or in part may be granted by the Head of the School or Department when in his or her judgment the proposed use of the material is in the interest of scholarship. In all other instances; however, permission must be obtained from the author of the Thesis. Name: Megersa Idris Signature ------------------------ Place: Haramaya University Date of Submission: Feburary 2021. IV BIOGRAPHICAL SKETCH The Author Megersa Idris Ahmed was born on April 7, 1990 G.c in Haramaya District, East Hararghe, Oromia Regional State, Ethiopia.from his father Idris Ahmed and his mother Fatuma Ahmed in December, 1992. He attended his elementary school at Bate Elementary School from 1998 to 2004.Gc. He pursued Secondary Education at Haramaya Secondary School from 2005 to 2006. After successfully passing the Ethiopian General Secondary Education Certificate (EGSEC) examination, he joined Haramaya Preparatory School from 2007 to 2008, for higher education then, joined Ambo University on 2009 and graduated with BSc degree in Veternary Laboratory Technology on June 28, 2012.G.c. After graduation, he was employed by the Haramaya University College of Veternary Medicine, where he worked for three years. He then joined Haramaya University on October 2018 to pursue a study leading to the Degree of Master of Sciences in Biotechnology. V ACKNOWLEDGEMENTS Above all, I would like to praise and glorify the Almighty Allah who provided me with all the required resources, knowledge, strength and wisdom for the fulfillment of the task. First and foremost, it is my great pleasure to wholeheartedly extend my most profound and sincere gratitude to my major advisor Dr Zekeria Yusuf and co-advisor Dr. Ashebr Abraha (DVM) for their valuable comments, suggestions, assistance and critical guidance in the preparation of the proposal and thesis write up. Their ever readiness to provide assistance, guidance and advice greatly helped me to carry out the study. VI ACRONYMS AND ABBREVIATIONS ALP Alkaline Phosphatase ALT Alanine Transaminase AST Aspartate Aminotransferase BHA Butylated Hydroxyanisole BHT Butylated Hydroxytoluene CLSI Clinical and Laboratory Standard Institute COX-1 Cyclo Oxygenases DPPH Diphenyl-2-Picrylhydrazyl radical EGF Epidermal Groth Factor FIRI Fasting Insulin Resistance Index LD50 Median Lethal Dose LDL Low Density Lipoprotein MBC Minimum Bactericidal Concentration MFC Minimum Fungicidal Concentration MHA Mueller-Hinton Agar MIC Minimum Inhibitory Concentration MRSA Methicillin-Resistant Staphylococcus Aureus NA Nutrient Agar OGTT Oral Glucose Tolerance Test PDA Potato Dextrose Agar PSA Prostate-Specific Antigen PSA Prostate-Specific Antigen RSV Respiratory Syncytial Virus SHBG Sex Hormone Binding Globulin SHBG Human Sex Hormone Binding Globulin TNF-α Tumor Necrosis Factor UDA Urtica Dioica Agglutinin VLDL Very Low Density Lipoprotein VII TABLE OF CONTENTS APPROVAL SHEET II DEDICATION III STATEMENT OF THE AUTHOR IV BIOGRAPHICAL SKETCH V ACKNOWLEDGEMENTS VI ACRONYMS AND ABBREVIATIONS VII TABLE OF CONTENTS VIII LIST OF TABLE XI LIST OF TABLE IN THE APPENDIX XII 1. INTRODUCTION 1 2. LITERATURE REVIEW 4 2.1. Botanical Description of Nettle 4 2.2. Traditional Medicinal Properties 4 2.3. Phytochemical Composition 5 2.4. Antimicrobial Properties 6 2.4.1. Root aqueous extract 6 2.4.2. Root non-aqueous extract 7 2.4.3. Root hydroalcoholic extract 7 2.4.4. Leave aqueous extract 8 2.4.5. Leave non-aqueous extract 8 2.4.6. Leave hydroalcoholic extract 9 2.4.7. Seed non-aqueous extract 11 2.4.8. Seed hydroalcoholic extract 11 VIII 2.4.9. Aerial aqueous extract 11 2.4.10. Aerial non-aqueous extract 12 2.4.11. Aerial hydroalcoholic extract 12 2.5. Pharmacological Properties 12 2.5.1. Antiproliferative activity 13 2.5.2. Anti-inflammatory activity 13 2.5.3. Antioxidant activity 14 2.5.4. Analgesic, antinociceptive and antiulcer properties 14 2.5.5. Antidiabetic and antihypertensive action 15 2.5.6. Effect on platelet aggregation, hyperlipidemia, atherosclerosis, and anti allergic activity 16 2.6. Toxicity 16 2.7. Modes of use and Precautions 17 2.8. Test methods used to Investigate the Antimicrobial Effect of Nettle 17 3. MATERIALS AND METHODS 20 3.1. Description of Study Area 20 3.2. Collection of Plant Material and Extract Preparation 20 3.2.1 Crude extraction 20 3.2.2. Preparation of different concentrations of the crude extracts 21 3.4. Phytochemical Screening of the Plant Materials 21 3.5. Media Preparation and Standardization of Inoculum 23 3.6. Disk diffusion method 23 3.6.1. Inoculation of mueller hinton agar plates 24 3.6.2. Measuring zones of inhibition 24 3.6.3. Determination of minimum inhibitory concentration 24 IX 3.6.4. Determination of minimum bactericidal concentration and minimum fungicidal concentrations 25 3.7. Data Analysis 26 4. RESULTS AND DISCUSSION 27 4.1. Qualitative Analysis of Phytochemical Composition of U. dioica L.Leaf, Fruit and Root Crude Methanolic Extracts 27 4.2. Antimicrobial Activities of Stinging Nettle Leaf, Fruit and Root Methanolic Extracts 27 4.3. Minimum Inhibitory Concentration , Minimum Bactericidal Concentration and Minimum Fungicidal Concentration of Stinging nettle Leaf, Fruit and Root Methanolic Extracts 31 5. SUMMARY, CONCLUSION AND RECOMMENDATION 34 5.1. Summary 34 5.2. Conclusion 35 5.3. Recommendation 36 6. REFERENCES 37 7. APPENDICIES 50 X LIST OF TABLE Table page 1. Preliminary phytochemical screening of crude extracts from stinging nettle (U. dioicaL.) leaf, fruit and root 27 2. Antibacterial Activity of the methanolic extracts of stinging nettle leaf, fruit and root as mean of inhibition diameter zone against Gram-Positive and gram-Negative Pathogenic Bacteria . 29 3. Antifungal Activity of the methanolic extracts of stinging nettle (Urtica dioica L.) leaf, fruit and root as mean of inhibition diameter zone against fungal spp. 30 4. Minimum inhibitory concentration and minimum bactericidal concentration of methanolic leaf, fruit and root extracts of Urtica dioica L.) 32 5. Minimum inhibitory concentration and minimum fungicidal concentration of methanolic leaf, fruit and root extracts of Urtica dioica (L.) 33 XI LIST OF TABLE IN THE APPENDIX Table page 1. Antibacteria activity based on diameter of zone of inhiontion 51 2. Data for antifungal activity 52 XII Phytochemical Screening and Antimicrobial Activities of stinging Nettle (Urtica dioica L.) Leaf, Fruit and Root Extracts ABSTRACT Medicinal
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