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Antimicrobial Effects of Gomphrena and Vernonia Extracts Against Staphylococcus Species and Candida Species from Women with Vulvovaginitis in Zaria, Nigeria

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Antimicrobial Effects of Gomphrena and Vernonia Extracts Against Staphylococcus Species and Candida Species from Women with Vulvovaginitis in Zaria, Nigeria ANTIMICROBIAL EFFECTS OF GOMPHRENA AND VERNONIA EXTRACTS AGAINST STAPHYLOCOCCUS SPECIES AND CANDIDA SPECIES FROM WOMEN WITH VULVOVAGINITIS IN ZARIA, NIGERIA. BY Esther Iyadunni, OGUNDANA, B.Sc BOTANY (KSU, 2007) M.Sc/SCIE/8486/2010-2011 A THESIS SUBMITTED TO THE SCHOOL OF POSTGRADUATE STUDIES, AHMADU BELLO UNIVERSITY, ZARIA IN PARTIAL FULFILMENT OF THE REQUIREMENTS FOR THE AWARD OF MASTER OF SCIENCE (M.Sc) DEGREE IN BOTANY DEPARTMENT OF BIOLOGICAL SCIENCES, FACULTY OF SCIENCE, AHMADU BELLO UNIVERSITY, ZARIA, KADUNA STATE, NIGERIA MARCH, 2015 i DEDICATION This thesis is dedicated to the Almighty God, the Maker and the Finisher, the beginning and the end and to all women, both singles and pregnant. ii DECLARATION I declare that the work in this thesis titled „‟ Antimicrobial Effects of Gomphrena And Vernonia Extracts Against Staphylococcus Species and Candida species from Women with Vulvovaginitis in Zaria, Nigeria‟‟ has been performed by me in the Department of Biological Science, Faculty of Sciences, Ahmadu Bello University, Zaria, Nigeria under the supervision of Dr. M. A. Adelanwa, Prof. A.K Adamu and Dr. E. E. Ella. The information derived form the literature has been duly acknowledged in the text and a list of references provided. No part of this thesis was previously presented for another degree or diploma at this or any other Institution. Esther Iyadunni OGUNDANA _____________________ _________________ Signature Date iii CERTIFICATION This thesis entitled „‟ ANTIMICROBIAL EFFECTS OF GOMPHRENA AND VERNONIA EXTRACTS AGAINST STAPHYLOCOCCUS SPECIES AND CANDIDA SPECIES FROM WOMEN WITH VULVOVAGINITIS IN ZARIA, NIGERIA‟‟ by Esther Iyadunni OGUNDANA (M.Sc/SCIE/8486,2010-2011) meets the regulations governing the award of degree of Master of Science in Botany of Ahmadu Bello University, Zaria and is approved for its contribution to knowledge and literary presentation. Dr.M.A Adelanwa __________________ ________________ Chairman, Supervisory Committee Signature Date Department of Biological Sciences Ahmadu Bello University, Zaria Prof. A. K. Adamu __________________ ________________ Member, Supervisory Committee Signature Date Department of Biological Sciences Ahmadu Bello University, Zaria Dr.E.E. Ella __________________ ________________ Member, Supervisory Committee Signature Date Department of Microbiology, Ahmadu Bello University, Zaria Prof. A.K. Adamu __________________ ________________ Head of Department, Signature Date Department of Biological Sciences Faculty of Science, A.B.U, Zaria Prof A.Z. Hassan __________________ ________________ Dean, School of PostgraduateStudy Signature Date Ahmadu Bello University, Zaria iv ACKNOWLEDGEMENTS I am grateful to God Almighty for His mercies throughout this research. You made it possible, Lord. My gratitude goes to my supervisors: Dr. M.A. Adelanwa, Prof. A.K. Adamu and Dr. E.E. Ella, for their time, support, efforts and corrections throughtout the period of this research. May God crown your efforts worthy and your children will excel by His grace. Remain blessed. My sincere appreciation to my parents, Mr. and Mrs. A.A. Ogundana for their parental love, words of encouragement, support and prayers. My sister and her husband, Solicitor and Barrister and Mrs Babajide Aminu, for their financial support and words of encouragement. My brother, flying Sergent Olarenwaju Ogundana, for his financial contribution and for bieng there for me. Also to my brother, Emmanuel Durojaiye Ogundana, my niece Kolawole Folakemi Faith, Akinyele Tunrayo and my nephew, Emmanuel Tope Ogundana, for their lovely support. My gratitude goes to the school, Ahmadu Bello University and the Departments, of Biological Sciences and Microbiology for giving me the opportunity to pursue my dreams. To the Heads of the Departments, Biological Sciences and Microbiology, lecturers and Staff that have helped in one way or the other, God bless you. My thanks to all laboratory technicians for their guidance and time during the course of this work. God will always be there for you all. Special thanks to Dr. Uchendu Chidiebene for his contributions and the knowledge shared towards this research. The wisdom God has given you, the devil will not steal it from you. Thanks for being there. How can I forget my fiancé for his love, support and word of encouragement. May God take our love beyond this level, amen. I can not afford to forget my friends and colleaques. My stay on campus would have been borring without you guys. God bless you all. v ABSTRACT A total of 300 samples of High Vaginal swabs (HVS) were collected from women of reproductive age (15-50 years) for suspected vaginitis at Ahmadu Bello University, Health Care Center and Samaru Clinic, Zaria, Kaduna State, Nigeria. The specimens were investigated for the presence of Candida species and Staphylococcus species and the effect of Gomphrena celosioides and Vernonia perrottetii extracts on the isolates. The antimicrobial activity of the extracts was performed against Candida spp and Staphylococcus spp isolated using agar well diffusion method. Total number of 153(51%) patients were found to be infected, with 79(26.3%) having Candidiasis and 74(24.7%) having Staphylococcal vaginal infection. Organisms were found to be more prevalent among the age group of 21-30 years followed by 31-40 years old. There was a significant association in the use of birth control pills, tight underwear, tight clothing, pregnancy and vaginitis (χ2= 82.78, p<0.001 OR =20.07 CI = 9.192 to 43.79, χ 2= 23.06 p<0.001 OR = 4.836 CI = 2.440 to 9.587, χ 2= 8.292 p<0.004 OR = 2.385 CI = 1.306 to 4.354, χ2 = 30.95 p<=0.0001 OR = 4.338 CI = 2.541 to 7.405, respectively). There was no significant (p>0.05) association between age and vaginitis, marital status, use of vagina cream by the respondents and the use of antibiotics and vaginitis infections (χ2 = 4.560, p = 0.2070, df = 3; χ2 = 0.06609, p = 0.7971, df = 1; χ2 = 2.796, p = 0.0945, OR = 1.679, CI = 0.9108 to 3.096, and χ2 = 0.05774, p = 0.4473, OR = 0.8367, CI=0.5281 to 1.326 respectively). Biochemical characterization of the 50 randomly selected Candida spp using API 20 AUXC showed that Candida tropicalis were isolated from 13(4.33/26%) followed by C. glabrata and C. albicans 3(1%/6%) and (1%/6%), C. parasiplosis and C. lambica 1(2%/0.33%) and 1(2%/0.33%), respectively. The results of the ten randomly selected Staphylococcus spp using Microgen kit also showed that S. vi xylosus, S. aureus and Staphylococcus spp were present in 3 samples 3(1%) and 3(1%) respectively and, S. warneri in one sample 1(0.3%). Antibiotic sensitivity testing was performed for both Candida spp and Staphylococcus spp using agar well diffusion method. The results showed that the organisms exhibited multiple resistances against ketoconazole among the 5 antifungal drugs used. The plant extracts sensitivity showed that almost all the Candida spp isolated were resistant to the plant extracts both singly and in combination, except for only 7 isolates that were susceptible at 2000mg/ml to V. perrottetii extracts. The results of the aqueous plant extracts sensitivity against Staphylococcus species at 500mg/ml and 1000mg/ml showed that V. perrottetii produced a zone of inhibition greater than the one produced by the antibiotics used for S. xylosus (a). The methanol extracts of Vernonia perrottetii (100mg/ml) showed that S. xylosus (a) had the same zone of inhibition as the antibiotic used (amoxycillin). Both aqueous and methanolic extracts of the two plant also did better for S. aureus where the antibiotics did not produce any zone of inhibition. The results of the minimum inhibitory concentration of Vernonia perrottetii (aq) MIC500/MBC>1000 showed bacteriostatic activity against only one organism. V. perrottetii (Me) tested against all the organisms exhibited bacteriostatic activity MIC500/MBC>1000 respectively. Aqueous combination of the extracts MIC500/MBC>1000 exhibited bacteriostatic activity to S. aureus and methanol combination of the extracts was bactericidal to S. xylosus. The present study showed that there was a high prevalence of Candida spp and Staphylococcus spp as well as strong association with some of the risk factors for vaginitis, and this suggest that, public enlightening should be carried out among women. It can be deduced from this study that the combination of the plant extracts did not perform better against both Candida spp and Staphylococcus spp isolated than when applied singly or in combination, implying that the two selected plant extracts might not vii be combine together. It is therefore suggested that the individual plants be combined with other plants to see if a better result could be obtained. viii TABLE OF CONTENTS Title page .................................................................................................................. ….i DEDICATION .............................................................................................................ii DECLARATION ....................................................................................................... iii CERTIFICATION ..................................................................................................... iv ACKNOWLEDGEMENTS ........................................................................................ v ABSTRACT ................................................................................................................ vi TABLE OF CONTENTS ........................................................................................... ix LIST OF TABLES
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