Kiliobas - Antimicrobial Activity Thesis

94 Pages·2014·1.33 MB·English

i

ANTIPLASMODIAL ACTIVITY OF AQUEOUS AND ETHANOLIC EXTRACTS OF
ANACARDIUM OCCIDENTALE AND CYMBOPOGON CITRATUS

KILIOBAS KIRI SHA’A
B.Sc., M.Sc. (MAIDUGURI)
PGNS/UJ/0081/06

A thesis in the Department of ZOOLOGY, Faculty of Natural Sciences,
Submitted to the School of Postgraduate Studies, University of Jos,
in partial fulfillment of the requirements for the award of the degree of
Doctor of Philosophy in Zoology (Parasitology) of the
UNIVERSITY OF JOS.

February 2014
ii

CERTIFICATION

This is to certify that this thesis has been examined and approved for the award of the degree of
DOCTOR OF PHILOSOPHY in Zoology (Parasitology) of the UNIVERSITY OF JOS.

----------------------------------------
Prof. (Mrs) O. O. Ajayi
Supervisor
Department of Zoology, University of Jos, Jos.

-----------------------------------------
Prof. (Mrs.) G. S. Mwansat
Head of Department
Department of Zoology, University of Jos, Jos
iii

DECLARATION
I hereby declare that this work is the product of my own research efforts, undertaken under
the supervision of Prof. (Mrs) O. O. Ajayi and has not been presented elsewhere for the award
of a degree or certificate. All sources have been duly distinguished and appropriately
acknowledged.

-------------------------------------- ………………..
Kiliobas Kiri Sha’a Date
PGNS/UJ/0081/06
iv

ACKNOWLEDGEMENTS
My sincere thanks and appreciation go to my supervisor, Prof. (Mrs) O. O. Ajayi for
her guidance, kindness and valuable contribution to this research. She has contributed in no
small way to the success of this work.
I also want to remember and thank my former supervisor, late Prof. J. A. Ajayi for his
immense contribution, his fatherly advice and encouragement.
My special thanks to the Head of Zoology Department, Prof. (Mrs) G. S. Mwansat for
her encouragement. I want to appreciate all staff members, academic and non-academic, of
Zoology Department for various contributions, especially the Postgraduate Seminar Co-
ordinator, Dr (Mrs) J.A. Yohanna and my internal assessors for their valuable contributions.
My gratitude to Prof. S. Oguche for his guidance, and for providing a study space for
me in Jos University Teaching Hospital. I remain grateful to Prof. Godwin Imade who agreed to
have some part of this study conducted in the Obstetrics and Gynaecology (O & G) Research
Laboratory, Jos University Teaching Hospital, Jos. I want to appreciate Mr Ayuba John Sendest,
also of O & G research laboratory for his friendliness and cooperation.
I want to thank Mr Yakubu Thomas Philip of Pharmacognosy Department, University
of Jos for his assistance during the extraction and phytochemical screening of extracts. The staff
of the Animal House, University of Jos assisted me so much in the conduct of research on
animal subjects. I specially thank Mr Luka Wazo and Mr Alfred A. Adolong for their kind
assistance.
I want to thank the Principal Investigator, Damboa Malaria Surveillance Sentinel site,
Dr. I. M. Watila for permiting me to conduct the in vitro assay in their Laboratory and for his
immense contribution to this work. My thanks also to Laboratory Staff of Damboa General
Hospital for their cooperation and assistance; of special mention are Mr Yahi Yanta Chiroma,
Mal. Baba Gana Mala, Mal. Bunu Hassan, and Dima M.
v

I want to appreciate the professional assistance of Pharmacist Amos Istifanus of the
Pharmacy Department, Jos University Teaching Hospital for technical assistance. Mr Emmanuel
Fyaktu of the Medical Microbiology Department, Faculty of Medicine, University of Jos
assisted in planning the in vitro assay. I appreciate him so much. My thanks and appreciation to
Mr Ahmed Danbello for his friendliness and assistance during the course of this research work. I
remain grateful.
I want to appreciate the kind contribution of my wife, Mrs Saratu K. Sha’a to this
research work. I remain grateful for her love, sacrifice and encouragement. My children have
been a source of encouragement during the course of this study. I thank them a lot.
My elder brother, late Hon. Daniel S. Kiri supported me financially, morally and
otherwise. I am very grateful to him for his brotherly care. I want to sincerely thank Hon. Iliya
D. Wanapiya for his encouragement and financial support. I appreciate his contribution so
much. My friend Pastor Ishaya Amos supported me prayerfully and financially all through the
course of this work. I am so thankful.
I thank my employers, Adamawa State Polytechnic, Yola for study fellowship.
Above all, I thank God for His goodness and for His wonderful works to me. To Him
be the glory, Amen.
vi

DEDICATION
This work is dedicated to my late father, Sha’a Kiri for his immeasurable contribution to my
education.
vii

TABLE OF CONTENTS
TITLE PAGE
DECLARATION …………...……………………………………………………………………ii
CERTIFICATION …………………………………………………………………....................iii
ACKNOWLEDGEMENT…………………………………………………..……...…………....iv
DEDICATION………………………………………………………………...…………………vi
TABLE OF CONTENTS……………………………………………………….........................vii
LIST OF TABLES ………………………………………………………………...…………….xi
LIST OF FIGURES……………………………………………………………………..............xii
LIST OF PLATES…………………………………………… ………………………………..xiii
ABSTRACT ……………………………………………………………………………………xiv
CHAPTER ONE
INTRODUCTION

1.1 BACKGROUND TO THE STUDY…………………………………………………………1
1.2 MALARIA HIGH RISK GROUP ……………………………………………….................2
1.3 SOCIO-ECONOMIC IMPLICATIONS OF MALARIA…...………….................................3
1.4 SCREENING OF TRADITIONAL MEDICINAL PLANTS FOR
ANTIMALARIAL ACTIVITIES………………...................................................................5
1.5 STATEMENT OF RESEARCH PROBLEM ……………………………………................5
1.6 JUSTIFICATION …………………………………………………….……………………..7
1.7 AIM OF THE STUDY………………………………………………………………………8
1.8 THE OBJECTIVES OF THE STUDY..……………………………………………………..8
1.9 RESEARCH HYPOTHESES ……………………………………………………………….8
CHAPTER TWO
LITERATURE REVIEW

2.1 MALARIA TREATMENT: A HISTORICAL BACKGROUND…………………………..9
viii

2.2 RESISTANCE TO ANTIMALARIAL DRUGS AND THE USE OF
ARTEMISININ COMBINATION THERAPY (ACT) ……….………………….............10
2.3 MEDICAL PLANTS AS ALTERNATIVE SOURCES OF NEW
ANTIMALARIAL COMPOUNDS……………………………………………………….12
2.4 IN VITRO AND IN VIVO ANTIMALARIAL ACTIVITIES OF
MEDICINAL PLANTS REPUTED TO TREAT MALARIA…………………….............12
2.5 ACUTE TOXICITY TESTING…………………………………………………...............15
2.6 ANTI-INFLAMMATORY AND ANALGESIC PROPERTIES OF
MEDICINAL PLANTS…………………………………………………………………..16

CHAPTER THREE
MATERIALS AND METHODS

3.1 EXPERIMENTAL PLANTS……………………………………….…………………….19
3.1.1 Anacardium occidentale………………………………………………………………....19
3.1.2 Cymbopogon citratus ………………………………………………................................19
3.2 COLLECTION OF EXPERIMENTAL PLANTS’ PARTS ……………………………..20
3.3 PREPARATION OF EXTRACTS …………………………………................................20
3.3.1 Extraction of Plant Materials ……………………………………....................................20
3.4 PHYTOCHEMICAL SCREENING OF CRUDE EXTRACTS ………………...……….20
3.4.1 Test for Flavonoids…………………………………………………………………...…..21
3.4.2 Test for Saponins……………………………………………………................................21
3.4.3 Test for Tannins…………………………………………………………………..............21
3.4.4 Test for Anthraquinones……………………………………………………………...…..21
3.4.5 Test for Glycosides…………………………………………………………………...…..21
3.4.6 Test for Cardiac Glycosides (Keller Kiliani test)………………..….................................22
3.4.7 Lieberman’s Test-Steroids and Terpenes…………………………………………………22
ix

3.4.8 Test for Flavonoids…………………………………………………………………….....22
3.4.8.1 Lead Acetate Test for Flavonoids………………………………...……………………..22
3.4.8.2 Sodium Hydroxide Test for Flavonoids…………………………………………………22
3.4.9 Test for Carbohydrates……………………………………………....................................23
3.5 BLOOD SAMPLES COLLECTION AND IN VITRO STUDY……...............................23
3.5.1 Inclusion Criteria……………………………………………….......................................23
3.5.2 Exclusion Criteria…………………………………………………...................................23
3.5.3 Screening for Plasmodium falciparum……………………………..................................23
3.5.4 Collection of Plasmodium falciparum Isolates ………………….....................................24
3.5.5 Preparation of Culture Medium………………………………….....................................24
3.5.6 Preparation of Extracts’ Solution…………………………………...................................25
3.5.7 In vitro Assay……………………………………………………….................................25
3.5.8 Thresholds for In vitro Antimalarial Activity……………………..................................26
3.6 COLLECTION OF EXPERIMENTAL ANIMALS……………………………………..26
3.7 ACUTE TOXICITY TEST……………………………………….....................................26
3.8 ANTI-INFLAMMATORY TEST …………………………………………………….…27
3.8.1 Topical Oedema: Xylene Induced Oedema………………………...................................27
3.9 TEST FOR ANALGESIC PROPERTIES…………………………………………............27
3.9.1 Acetic Acid Induced Writhing……………………………………...……………..............27
3.10 STATISTICAL ANALYSIS …………………………………………………..............28
x

CHAPTER FOUR
RESULTS

4.1 EXTRACTION AND PHYTOCHEMICAL SCREENING ……......................................31
4.1.1 Yield of Extracts…………………………………………………………………..………31
4.2 IN VITRO ASSAY ………………………………………………………………….……31
4.3 ACUTE TOXICITY TEST ………………………………………....................................48
4.4 ANTI-INFLAMMATORY TESY: TOPICAL OEDEMA ………....................................48
4.5 ANALGESIC PROPERTIES OF PLANTS’ EXTRACTS……………………………….51
4.5.1 Acetic Acid Induced Writhing Test……………………………………………..………...51
CHAPTER FIVE
DISCUSSION

5.1 ANTIMALARIAL ACTIVITIES OF ANACARDIUM OCCIDENTALE AND
CYMBOPOGON CITRATUS……………………………………….....................................54
5.2 ACUTE TOXICITY IN MICE………………………………………………………..……57
5.3 ANTI-INFLAMMATORY ACTIVITY……………………………....................................57
5.4 ANALGESIC PROPERTIES OF PLANTS’ EXTRACTS………………………………...58
5.5 CONCLUSION………………………………………………………………….………….60
5.6 CONTRIBUTION TO KNOWLEDGE…………………………….....................................60
5.7 RECOMMENDATIONS FOR FURTHER WORK………………………………………..61
REFERENCES …………………………………………………………………………….63
APPENDIX……………………………………………………………………….………...79