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Thowiba Mohammed Eltayeb Mohammed Phytochemical Analysis and Antimicrobial Activities of Garad (Acacia nilotica L.) and Rumman (Punica granatum L.) Plants Thowiba Mohammed Eltayeb Mohammed B. Sc.( Honor) in Biology and Chemistry, Faculty of Education- Elhasahiesa, University of Gezira (2002) Post- graduate Diploma in Biosciences and Biotechnology, Center of Biosciences and Biotechnology, Faculty of Engineering and Technology (2004) M.sc in Biosciences and Biotechnology, Center of Biosciences and Biotechnology, Faculty of Engineering and Technology (Biosciences) (2009) A Thesis Submitted to the University of Gezira in Fulfillment of the Requirement for the Award of the Degree of Doctor of Philosophy in Biosciences and Biotechnology, (Microbiology) Center of Biosciences and Biotechnology Faculty of Engineering and Technology August 2015 1 Phytochemical Analysis and Antimicrobial Activities of Garad (Acacia nilotica L.) and Rumman (Punica granatum L.) Plants Thowiba Mohammed Eltayeb Mohammed Supervision Committee: Name Position Signature Prof / Awad Mohamed Abdel-Rahim Main Supervisor ………………. Dr. Mutaman Ali Abdelgadir Kehail Co – Supervisor ……………… Date: August, 2015 2 Phytochemical Analysis and Antimicrobial Activities of Garad (Acacia nilotica L) and Rumman (Punica granatum L.) Plants Thowiba Mohammed Eltayeb Mohammed Examination Committee: Name Position Signature Prof/ Awad Mohamed Abdel-Rahim Chairman --------------- Hassan Beshir ElAmin External Examiner -------------- Dr. Zakaria Ahmed Salih Internal Examiner --------------- Date of Examination 10 /08 /2015 3 بسم هللا الرحمن الرحيم قال تعالى: { وهو الذي أنزل من السماء ماء فأخرجنا به نبات كل شيء فأخرجنا منه خضرا خنرج منه حبا مرتاكبا ومن النخل من طلعها قنوان دانية وجنات من أعناب والزيتون والرمان مشتبها وغري متشابه انظروا إىل مثره إذا أمثر وينعه إن يف ذلكم ﻵيات لقوم يؤمنون } صدق هللا العظيم سورة اﻷنعام اﻵية 99 4 DeDICATION To my Parents Sister and her son (Mohammed) and Husband 5 ACKNOWLEDGEMENTS My grateful thanks to Allah who gave me the health or strength and the ability to achieve this humble study and with his will this achievement was completed. Also, I would like to express my gratitude and thanks to my main Supervisor: Prof / Awad Mohamed Abdel-Rahim, for his good guidance, my co- Supervisor Dr. Mutaman Ali A. Kehail Deepest thanks to my family, friends and colleagues for encouragement and support. Thanks and appreciation are due to the Staff of the Microbiology and staff of the food Laboratories, Department of Food Science and Technology, Faculty of Engineering and Technology University of Gezira. Especially Ustaz Hassan Ansari, and Mohammed Omer. Thanks to the staff of department of Chemistry &Biology faculty of Education- Elhasahiesa, University of Gezira. And to the staff of Chemistry &Biology Laboratories faculty of Education- University of Albutana. .. Especiall thanks to Dr. Salah Ahmed Moustafa , U. Ashwag Ahmed Abd-Elrahman, Elgid Esmaeel Mosa and his family (Gameela Elnour Omer and their sons & daughters) and U. Rehab Sideeq Ali Othman. 6 Phytochemical Analysis and Antimicrobial Activities of Garad (Acacia nilotica L.) and Rumman (Punica granatum L.) Plant Thowiba Mohammed Eltayeb Mohammed Ph.D. in Biosciences and Biotechnology (Microbiology), August, 2015 Center of Biosciences and Biotechnology, Faculty of Engineering and Technology (Biosciences) University of Gezira Abstract Sudan is rich in medicinal plants which synthesize substances that are useful to the maintenance of health in humans and animals. Garad and Rumman were reported to have some antimicrobial activity. The aim of the present study is to investigate and analyze the phytochemical compounds and the antimicrobial activities of the two plants. The extracts of the different parts of these plants were tested against mycelial growth and spore germination, using selective media and the cup plate Inhibition zone method was used for the solvents tested. The results showed that some parts of the two plants were containing flavonoids, tannins, saponine, sterole and alkaloids. The microbial tests indicated that the Garad seeds extract was highly effective against the radial growth of P. italicum. Leaves and bark extracts were effective against A. niger. Rumman Flower and peel extracts were better against P. italicum. However they were not effective against A. niger. All Rumman part extracts were effective against P. italicum. and only the flower extract was effective against the fresh and dry weights of A. niger. The spore germination tests showed that all the extracts of Garad plant parts were effective against P. italicum. Only the Rumman peels extracts were effective against P. italicum and A. niger. The inhibition zone tests showed that the Garad leaves and pods extracts were highly effective against C. albicans, and bark extract was highly effective against C. valida. The bark and the seeds extracts were effective against C. tropicalis, while, the pods extract was more effective against Pichia sp., E. coli and S. paratyphi. However, the pods, bark and leaves extracts were highly effective against Staphylococcus sp. Rumman peels extracts were more effective against C. tropicalis and all of it's parts were effective against the three bacteria (E. coli, S. paratyphi and Staphylococcus sp.). The solvent tests showed that the methanolic pod extracts of Garad were effective against the tested fungi, as well as Pichia sp. and C. tropicalis. The leaves extracts were effective against C. albicans, and Staphylococcus sp. , while, the bark extracts were effective against C. tropicalis and C. valida. However, the seeds extracts were effective against C. albicans, C. valida, and C. tropicalis. The methanolic Rumman peels extracts were effective against the two fungi, and C. valida. The ethanolic bark, seeds and pods extracts of Garad were effective against E. coli and S. paratyphi. C. valida was the only sensitive yeast against the bark and the peel ethanolic extracts of Rumman. Peels extracts were more effective against the three tested bacteria. The bark and flower effective against E. coli. All the petroleum ether and hexane extracts of the two plants parts were not effective against all the tested organisms. Both Garad and Rumman plants were containing inhibitory compounds against different microorganisms. It could be suggested that Garad and Rumman extracts could be used for treating some skin diseases. 7 . تحليل المركبات النباتية والنشاط الميكروبي لنباتي القرض والرمان ثويبة محمد الطيب محمد الدكتوراه في العلوم والتقنية البيولوجية )أحياء دقيقة( مركز العلوم والتقنية البيولوجية- كلية الهندسة والتكنولوجيا جامعة الجزيرة ملخص الدراسة السودان غني بالنباتات التي تنتج مواد مفيدة لصحة اﻹنسان والحيوان. نباتات القرض والرمان ثبت أن بها بعض المواد المثبطة للكائنات. هدفت هذه الدراسة لبحث وتحليل المركبات الكيميائية والنشاط المضاد للميكروبات لنباتي القرض والرمان. المستخلصات المختلفة ﻷجزاء النباتين تم اختبارها ضد النمو الميسيليومي وانبات الجراثيم, باستخدام أوساط غذائية انتقائية. وتم استخدام طريقة المنطقة المثبطة ﻻختبار المذيبات المختلفة., أثبتت النتائج أن بعض أجزاء النباتين تحتوي على مركبات مثل الفﻻفونويدات, تانين, سابونين,استيرول وألكالويدات. أشارت نتائج اﻻختبارات الميكروبية إلى أن مستخلص بذور القرض كان عالي الفعالية ضد النمو القطري للفطر P.italicum . هذا وكانت مستخلصات اﻷوراق واللحاء فعالة ضد الفطر .A niger . مستخلصات أزهار وقشرة الرمان أكثر فعالية ضد الفطر P.italicum , ولكن لم تكن فعالة ضد الفطر A. niger . كل مستخلصات أجزاء نبات الرمان كانت أكثر فعالية ضد الفطر P.italicum بينما كانت مستخلصات اﻷزهار هي الوحيدة الفعالة ضد الوزن الجاف والرطب للفطر A. niger. أظهرت إختبارات إنبات الجراثيم إلى أن كل مستخلصات نبات القرض كانت فعالة ضد الفطر P.italicum . مستخلصات قشرة الرمان كانت الوحيدة اﻷكثر فعالية ضد نمو الفطرين P.italicum و A. niger . اختبارات المنطقة المثبطة أظهرت أن مستخلصات أوراق وحوافظ القرض هي اﻷكثر فعالية ضد الخميرة C. albicans و مستخلصات اللحاء كانت اﻷكثر فعالية ضد الخميرة C.valida. و مستخلصات اللحاء والبذور كانت أكثر فعالية ضد الخميرة C.tropicalis وفي حين كانت مستخلصات الحوافظ هي اﻷكثر فعالية ضد .E. coli Pichia sp و و S.paratyphi ومع ذلك كانت مستخلصات الحوافظ واللحاء واﻷوراق أكثر فعالية ضد البكتيريا Staphylococcus sp. مستخلصات قشرة الرمان كانت أكثر فعالية على الخميرة C.tropicalis, وكل أجزاء النبات فعالة ضد أنواع البكتيريا الثﻻث ) E. coli و S. paratyphi و Staphylococcus sp)(. أظهرت اختبارات المذيبات المختلفة إلى أن المستخلصات الميثانولية لحوافظ القرض كانت هي اﻷكثر فعالية ضد الفطريات المختبرة, باﻹضافة للخمائر .Pichia sp و C.tropicalis. وفي حين كانت مستخلصات اﻷوراق هي اﻷكثر فعالية ضد الخميرة C.albicans والبكتيريا Staphylococcus sp بينما كانت مستخلصات اللحاء أكثر فاعلية ضد الخميرة C.tropicalis و C.valida. مستخلصات قشرة الرمان الميثانولية كانت أكثر فعالية ضد الفطرين باﻹضافة إلى الخميرة C.valida. و كانت مستخلصات لحاء وبذور و حوافظ القرض اﻹيثانولية أكثر فعالية ضد البكتيريا E.coli والبكتيريا S.paratyphi. علما بأن الخميرة C.valida هي الوحيدة التي كانت أكثر حساسية من بين الخمائر لمستخلصات اللحاء والقشرة اﻹيثانولية لنبات الرمان. مستخلصات القشرة هي اﻷكثر فعالية ضد أنواع البكتيريا الثﻻث المختبرة, أما مستخلصات اللحاء واﻷزهار هي الفعالة ضد البكتيريا E. coli . كل مستخلصات القرض والرمان التي تم استخﻻصها بواسطة إيثر البترول والهكسان كانت غير فعالة ضد كل الكائنات الحية التي تم اختبارها. اتضح أن نباتي القرض والرمان بهما مواد مثبطة لكائنات حية دقيقة مختلفة. يمكن التوصية باستخدام مستخلصات تلك النباتات لمعالجة بعض اﻷمراض الجلدية. 8 TABLE OF CONTENTS Subject Page Dedication Iii ACKNOWLEDGEMENTS Iv ENGLISH ABSTRACT V ARABIC ABSTRACT Vi TABLE OF CONTENTS Vii LIST OF TABLES Ix LIST OF FIGURES Xii CHAPTER ONE: INTRODUCTION 1 CHAPTER TWO: LITERATURE REVIEW 3 2.1 INTRODUCTION
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