Antibacterial Activity of Leaves and Fruits Extracts Of the Gumbil Tree (Cordia africana L.) Fadia SaifAldin Edrees Mohmmed B. Sc. In Biology, Faculty of Education, University of Holy Quran and Islamic Sciences, 2011. Postgraduate Diploma in Biosciences and Biotechnology, Faculty of Engineering and Technology, University of Gezira, 2013 A Dissertation Submitted to the University of Gezira, in Partial Fulfillment of the Requirement for the Award of the Degree of Master of science in Biosciences and Biotechnology (Biosciences) Center of Biosciences and Biotechnology Faculty of Engineering and Technology, 6/ 1/ 2015 Antibacterial Activity of Leaves and Fruits Extracts Of the Gumbil Tree (Cordia africana L.) Fadia SaifAldin Edrees Mohmmed 1 Supervision Committee: Name Position Signature Prof. Awad Mohamed Abdel-Rahim Main Supervisor …………... Dr. Yasir Mohamed Abdelrahim Co-supervisor …………... Date of Examination: 6/ January /2015 Antibacterial Activity of the Extracts of the Leaves and Fruits Of the Gumbil Tree (Cordia africana L.) Fadia SaifAldin Edrees Mohmmed Examination Committee: 2 Name Position Signature Prof. Awad Mohamed Abdel-Rahim Chairperson …………... Prof. Ahmed Elawad Elfaki External Examiner …………... Dr. Mai Abdalla Ali Abdalla Internal Examiner ………... Date of Examination: 6/ January /2015 Dedication To my Dear father Beloved mother Brothers and sisters (my grandmother) Uncles and my aunts Teachers & Friends 3 ACKNOWLEDGEMENTS My grateful thanks to ALLAH who gave me the health and ability to achieve this humble study, and with his will this achievement was properly completed. I would like also to express my thanks to all those who helped me to carry out this work especially Professor Awad Mohamed Abdel- Rahim, the main supervisor and Dr. Yasir Mohammed Abdel-Rahim, co- supervisor and Dr. Mutaman Ali for assistance. Thanks are also due the Technical Staff of the Microbiology Laboratory, Faculty of Engineering and Technology, University of Gezira. Great thanks are due to my family and friends for support and encouragement. 4 Antibacterial Activity of Leaves and Fruits Extracts of the Gumbil Tree (Cordia africana L.) Fadia Saif Aldean Edrees Mohmmed M. Sc. In Biosciences and Biotechnology (Biosciences) Center of Biosciences and Biotechnology Faculty of Engineering and Technology University of Gezira ABSTRACT Plant extracts were used to treat different diseases because they are cheap and effective, but pharmacists are often reluctant to prescribe them because of knowledge deficiency, real concerns about the product safety, concerns about liability, and the presence of pathogens contamination. The use of plants and their extracts as remedies for curing many diseases have stimulated studies for investigating the presence of effective antimicrobial substances in them. The present study was investigating the antibacterial properties of the tree Cordia africana (Gumbil) leaves and fruits extracts. The inhibition zone and the total plate count methods were used in this study. The results of the study showed that the extracts were highly effective against the two tested bacteria. It inhibited growth of the gram- negative (E. coli) bacterium. The extracts of leaves with methanol, hexane, ethanol and water were effective giving; 10, 10, 10 and 12 mm diameter, respectively. However, the extracts of the fruit with ethanol and hexane were highly effective both giving 12 mm compared methanol, chloroform and petroleum 5 ether which gave 10 mm. The water extract was the least (8 mm). However, the effect of the fruit extracts was more serious against Staph. aureus than on E. coli. Hexane and petroleum ether and ethanol were highly effective (12, 11, 11 mm, diameter, respectively) while methanol and chloroform gave intermediate effects (10 mm) and the water was the least effective (7 mm).The inhibitory effect of the water extract against the tested bacteria was more effective at the higher concentrations of the extracts (75 and 100%,). The effect of the water extract on the number of bacterial colonies was highly pronounced. The number of E. coli was decreased from 226 colonies at the control treatment to 29 colonies only at the higher concentration. A similar was recorded for the Staph. aureus. It could be recommended that, further tests should be carried on other microorganisms, and the active antibacterial components need to be verified. 6 (.Cordia africana Lانُشاؽ انؼًاد نبكخيشيا نًسخخهظاث أوساق ورًاسشجشة انقًبيم ) فاديت سيف انذيٍ ادسيس يحًذ انًاجسخيش في انعهىو وانخقُيت انبيىنىجيت )عهىو أحيائيت( يشكز انعهىو وانخقُيت انبيىنىجيت كهيت انهُذست وانخكُىنىجيا جايعت انجزيشة انًهخض حسخخذو يسخخهظاث انُباحاث في يعانجت انعذيذ يٍ اﻷيشاع ورنك ﻷَها سخيظت وفعانت، غيش أٌ انًخخظظيٍ في يجال انظيذنت غيش يخحًسيٍ ﻻسخخذايها ورنك نعذو انًعشفت انخايت بها ونعذو انخأكذ يٍ سﻻيخها وقذسحها في انًعانجت ووجىد انخهىد بانكائُاث انًًشػت بها. اسخخذاو انُباحاث ويسخخهظاحها نًعانجت بعغ اﻻيشاع حفزث انباحزيٍ نذساست انًىاد انفعانت انًىجىدة بها كؼًاداث نهًكشوباث. حى في هزا Cordia)انبحذ دساست انخىاص انًزبـت نهبكخيشيا بًسخخهظاث أوساق ورًاس شجشة انقًبيم . حى اسخخذاو ؿشيقخي انًُـقت انًزبـت وانعذد انكهي نهًسخعًشاث. أظهشث انُخائج (africana ًَى انبكخيشيا أٌ نخهك انًسخخهظاث فعانيت عانيت ػذ انبكخيشيا انخي حًج دساسخها. فقذ ربــج . كاَج يسخخهظاث اﻻوساق بانًيزاَىل وانهكسيٍ وانًاء أكزش (E. coli)انسانبت نظبغت جشاو فعانيت حيذ أعـج 01,01,01,01 يهى قـش بانخىاني . بيًُا كاَج يسخخهظاث انزًاس باﻻيزاَىل وانهكسيٍ هًا اﻻكزش فعانيت حيذ اعـً كﻻهًا 01 يهى يقاسَت بًسخخهظاث انًيزاَىل وانكهىسوفىسو واﻻيزش انبخشوني انخي اعـج 01 يهى قـشاً. وكاٌ انًسخخهض انًائي هى اﻻقم ( Staph. aureus)فعانيت )4يهى(. هزا وكاَج يسخخهظاث انزًاس اكزشفعانيت عهي انبكخيشيا . وكاَج يسخخهظاث انهكسيٍ واﻻيزش انبخشوني واﻻيزاَىل أكزش E. coli يقاسَت بانبكخيشيا )00.00.01 يهى عهي انخىاني(. بيًُا أعـج Staph. aureusفعانيت ػذ انبكخيشيا يسخخهظاث انًيزاَىل وانكهىسوفىسو حأريشاث يخىسـت )01يهى( وكاٌ انًسخخهض انًائي هى اﻷقم فعانيت )3يهى(. كاٌ انخاريش انًائي انًزبؾ نهًسخخهض انًائي أكزش فعانيت عهً انخشكيزاث انعانيت )011,31%( . كاٌ حأريش انًسخخهض انًائي عهي حعذاد انًسخعًشاث انبكخيشيت أكزش وػىحاً. حيذ اَخفغ انخعذاد يٍ 112 يسخعًشة في حجشبت انًقاسَت اني 15 يسخعًشة فقؾ . حىطي انذساست Staph. aureusعهً انخشكيز انعاني. انُخائج يشابهت في حسجيهها نبكخيشيا 7 بإجشاء انًزيذ يٍ اﻻخخباساث عهً كائُاث حيت دقيقت أخشي وكزنك نهخعشف عهً انًكىَاث انفعانت ػذ انبكخيشيا. List of Contents Topic Page ------ ------------------------------------------------------------------ No. - ----------- I Main page II Supervision committee III Examination committee Dedication Iv Acknowledgements V Abstract (English) VI Abstract (Arabic) VII List of Contents VIII 8 List of Tables X List of Figures Xi CHAPTER ONE INTRODUCTION Introduction 1 Objectives 4 CHAPTER TWO LITERATURE REVIEW 2.1. Introduction 5 2.2. Plant investigated 8 2.2.1 Local names 8 2.2.2 Geo graphic distribution 8 2.2.3 Tree management 9 2.2.4 Taxonomy and nomenclature 9 2.2.5 Botanical Description 9 2.2.6 Products and services 10 2.2.7 Biology 10 2.2.8 Ecology 11 2.3. Antimicrobial activity of plant extracts 11 2.4 Bacteria 13 2.4.1 Bacteria under test 13 2.4.1.1 E. coli 12 2.4.1.2 Staphylococcus aureus CHAPTER THREE MATERIALS AND METHODS 3.1. Sample collection 14 3.2 Source of cultures 14 3.3. Media used 14 9 3.3.1 Bacteriological media 14 3.3.1.1 Nutrient Agar 14 3.3.1.2 Nutrient Broth 14 3.4. Methods Used 15 3.4.1 The Cup-Plate Agar Diffusion Method 15 3.4.2 The Dilution Plate Method 15 3.5. Statistical Analysis 15 CHAPTER FOUR RESULTS 4.1. Effect of Different Extracting Solvents 16 4.2. Effect of Different Concentrations of the Aqueous 21 Extracts on the Inhibition Zone 4.3 Effect of Different Concentrations of the Extracts 21 on the Number of Colonies CHAPTER FIVE 30 DISCUSSION CHAPTOR SIX CONCLUSIONS AND RECOMMENDATIONS 5.1 Conclusions 33 5.2 Recommendations 33 REFERENCES 34 11 List of Tables Table Title Page No. No. 4.1 Effect of different plant parts extracts obtained by 17 different solvents on the inhibition zone (mm) of E. coli 4.2. Effect of different plant parts extracts obtained by 19 different solvents on the inhibition zone (mm) of Staph. aureus 4.3. Effect of different concentrations of Cordia africana 22 extracts on the inhibition zone of (mm) E. coli 4.4. Effect of different concentrations of Cordia africana 24 11 extracts on the inhibition zone (mm) of Staph. aureus 4.5. Effect of different concentrations of Cordia africana 26 (leaves) extracts on the number of colonies of E. coli 4.6. Effect of different concentrations of Cordia africana 28 (leaves) extracts on the number of colonies of Staph. aureus List of Figures Figure Title Page No. No 4.1 Effect of different plant parts extracts obtained by 18 different solvents on the inhibition zone of (mm) E. coli 4.2. Effect of different plant parts extracts obtained by 20 different solvents on the inhibition zone of (mm) Staph. aureus 4.3. Effect of different concentrations of Cordia africana 23 12 extracts on the inhibition zone (mm) of E. coli 4.4. Effect of different concentrations of Cordia africana 25 extracts on the inhibition zone of (mm) Staph. aureus 4.5. Effect of different concentrations of Cordia africana 27 leaves extracts on the number of colonies of E. coli 4.6. Effect of different concentrations of Cordia africana 29 leaves extracts on the number of colonies of Staph. aureus INTRODUCTION Natural products from plants have played major sustaining roles in the life of humans, especially for food sources and for medicinal products. Nature has provided mankind with folk medicines for centuries and continues to be the richest source of bioactive chemicals for the development of modern drugs. Herbalism is a traditional medicinal or folk medicine practice, based on the use of plants and plant extracts. Herbalism is also known as botanical medicine, medical herbalism, herbal medicine, herbolog, and phytotherapy. The scope of herbal medicine is sometimes extended to include fungal and bee products, as well as minerals, shells and certain animal parts. Pharmacognosy is the study of medicines derived from natural sources (Acharya and Shrivastava, 2008).