ﻢﻴﺣﺮﻟﺍ ﻦﻤﺣﺮﻟﺍ ﷲﺍ ﻢﺴﺑ University of Khartoum The Graduate College Medical & Health Studies Board Faculty of Public and Environmental Health Department Food Hygiene and safety Antibacterial Activity of Trigonella foenum greacum linn (Fenugreek) Seed - Extracts Against Some Species of Enterobacter. By Nafisa Eltigani Hamid Kroum B.Sc. (Honours) (U of K) 2003 A thesis submitted in partial fulfillment for the requirements of the Degree of Master in Public and Environmental Health (Food Hygiene and Safety) Supervisor Dr. Amira Abdel Azeem Behairy (Janeuary- Septemper 2009) University of Khartoum Dr. El Bushra El Sheikh El Nur(October 2009 – Janeuary 2010) University of Khartoum 2009 1 ﺔﹶﻳﻵَ ﺍ :ﻰﻟﺎﻌﺘ لﺎﻗ ﻪﻨﻤ ﻓﹶ ﺎﺒﺣ ﺎﻬ ﻨﻣ ﺎﻨﺟ ﺮ ﺧ ﺃﹶﻭ ﺎﻫ ﺎﻨﻴﻴﺣ ﺃﹶ ﹸﺔﺘﻴﻤ ﻟﹾﺍ ﺽ ﺭ ﺄﹶﻟﹾﺍ ﻢﻬﻟﹶ ﹲﺔﻳﺁَﻭ ) ( ﻥﻮﹸﻠﹸﻛﺄﹾﻳ (33ﺔﻳﻻا ﺲﻳ ةرﻮﺳ) 2 To my Mother, my Father and my Brothers For their encouragement and appreciated help To my teachers, colleagues and To everyone who encouraged me to complete this research. And all whom I love Chapter One Introduction and Literature Review 3 Table of Content Items Page Number Table of contents I- II Acknowledgement III Abbreviations IV English Abstract V- VI Arabic Abstract VII -VIII List of Figures IX-X List of Tables X I Chapter One Introduction 1 -3 Literature Review 1.1 Fenugreek (Trigonella foenum graecum Linn) 3 1.1.1 Traditional and Medicinal Uses of Fenugreek 4 -5 1.1.2 Chemical constituent 6 -7 1. 1.3 Edible Uses 7 1. 1.4 Fenugreek's Possible Side Effects 7 -8 1. 3 Enterobacteriace 9-10 1 .3.1 Escherichia coli 10-11 1 .3.2 Klebsiella pneumonia 11-14 1.3.3 Salmonella typhi 13-14 1.4 Pseudomonas aeruginosa 14-16 4 1.5 Staphylococcus aureus 17-18 1.6Antibiotic 18-20 Chapter Two Material and methods 2.1. Materials: 21 2.1.1. Tested organisms 21 2.1.2. Plant material 21 2.2Methods: 22 2.2.1. Preparation of plant extracts 2.2.2. Preparation of standard bacterial suspension 22-23 2.2. 3. Preparation of culture media 23 2.2.3.In vitro testing of extracts for antibacterial activity 23 2.2.4. Antibiotic used for comparison 24 Chapter Three Results 25 Chapter Four Discussion 43-45 Conclusion 46 Recommendations 47 References 48-54 5 All praise and thanks to Almighty Allah, the most Merciful and Beneficent who gave me life and strength to finish this research. I am especially grateful to my supervisors Dr. Amira Abdel Azeem Behairy and Dr. El Bushra El Sheikh El Nur for their helps and advices that gave me through my work. I would like to acknowledge the staff of Microbiology laboratory of Medicinal and Aromatic Plants Research Institute Finally I would like to thank everybody who collaborated by his opinions, advices or efforts in preparing this research 6 Abbreviation ATCC American Type Culture Collection E.c Esherichia coli K.p Klebsiella Pnemoniae P.a Pseudomonas aeruginosa Sl.t Salmonella typhi S.a Staphylococcus aureus WHO World Health Organization 7 Abstract Back ground Trigonella foenum graecum L. (fenugreek) is considered to have along history in medicine. It was used by Chinese for improvement of metabolic activities and the general heath. It is also used as spice and a cure in many parts of the world. In Sudan, it is used in folkrolic medicine for the ttreatment of boils, diarrhea and abdominal pain. Study Design Experimental study Setting This study was conducted in The Microbiology laboratory of Medicinal and Aromatic Plants Research Institute - National Center for Research - Khartoum. Sudan Objective To examine the antibacterial activity of Trigonella foenum greacum (fenugreek) crude seed extracts against of Esherichia coli, Klebsiella Pnemoniae, Salmonella typhi, Pseudomonas aeruginosa and Staphylococcus aureus. Methods The plant tissues were extracted with water and two organic solvents, namely methanol and chloroform. . The cup-plate agar diffusion method was used to determine the antibacterial activity of three extracts (water, chloroformic and methanolic 8 extracts) of Trigonella foenum-graecum against five standards bacterial strains Esherichia coli ATCC 25922, Klebsiella Pnemoniae ATCC 53657, Pseudomonas aeruginosa ATCC 27853, Salmonella typhi ATCC 0650 as Gram negetive bacteriaum and Staphylococcus aureus ATCC 25923 as Gram positive bacteriaum. Results Most of the standard bacteria were tested resistant to extracts of Trigonella foenum-graecum, except Salmonella typhi and Staphylococcus aureus which revealed sensitivity to chloroformic extract at 20% concentration with inhibition zone dimiter of 15mm for Salmonella typhi and 13mm for Staphylococcus aureus. Conclusion -Chloroformic extract of Trigonella foenum-graecum seeds is effecttive againist on Salmonella typhi and Staphylococcus aureus bacteria. -Methanolic and water extracts of Trigonella foenum-graecum seeds are very poor antibacterial agents. Recommendation: Based on the results obtained, it is recommended that, - Traditional using of Trigonella foenum-graecum seed may help in protecting from bacterial diseases casued by Salmonella typhi and Staphylococcus aureus and may assist in controling bacterial growth in food. - Chemical investigation of the chloformic extract components of Trigonella foenum-graecum seed is to beconducted in order to determine the active ingredients - Uses of Trigonella foenum-graecum as food and a cure because it effect on Salmonella typhi it cause typhoid fever, and also effect on Staphylococcus aureus bacteria can cause a range of illnesses from minorskin infections, to life-threatening diseases such as pneumonia, 9 meningitis, osteomyelitis, endocarditis, Toxic shock syndrome (TSS), and septicemia. ﺔﺴﺍﺭﺩﻟﺍ ﺹﻠﺨﺘﺴﻤ 10