SCREENING SELECTED FLOWERS FOR

POTENTIAL ANTmACTERIAL

ACTIVITY

LING nNG JING

,npu$lUUII •• IUISITI .auYSla ·SaBAn

PLANT TECHNOLOGY PROGRAMME

SCHOOL OF SCIENCE AND TECHNOLOGY

UNIVERSITI MALAYSIA SABAH

2007

DECLARATION

I declared that this dissertation was the result of my own independent work, except where

otherwise stated.

to April .!007

"'PU$lUlU IIIUlSIlI IIlLAYiIA ·U ....

LING JING JING

HS2004-4380

11

PUMS99,1 UNlVERSITI MALAYSIA SABAH

. BORANG PENGESAHAN STATUS TESIS@

JUDUL: Seret,1!"!) S(lle(1t'cl fiolto'Jtn fa( po1fnt,,,J Allti~tHtefl'tiJ

A<f i"it~ .

r j nah :_-=.q.:.~.;,,,I'-,e,",Ic:._v_--,O-,F,-_.l'_',-i _f_"_"'-_____ __________ _ SF_~[ PENGAJlAN,_~_·_D__'4 __ _

S.y. L I~, JIN§ J I~~ (HURUF BESAR)

rncngd.-u membc:D4Ckan tesis ~SlSujllWOoktoc ~U.rl?)· ini.disimp.o di P.erpusw.:un Unl\'ccsiti Malaysia Sabah deng'JQ sYU<lt·syan.t kcgunun sc6erti bcrii..~t:

1. Tesu ad.lab bab1\ilik Univeniti Ml-bysi.aSab.ah. 2. . ~erpustakun Univcrsiti Mllaysi& SUaah dibclI&!b . .D membua[ $llinan untuk tujuao peng:aji<l.ll ubajl. 3 Perpust1k..aan dibcnar\:aa mcmbuat ulin.u. (esis inlscbagai bahan pcrtukaran antara institusi pcogajian

tinggi. 'UPUS1A KA Ali ! ··SiI. <and. lain ( I ) OHIYERSITI MIlAYSIHA!AH

o o

SULIT

TERHAD

TIDAK'ICRifAD

(TANOI\ TANGAN PENULtS)

latn.ltTc tap IltH J T!ft"tPt!ttt ~ 'toad . I.Ipptl · ~tI;l'I&1"'j

•

-,\TATAl"': • POlon, yanSlidubcrhl'lun.

(Mengitndungi. mlklu.rnat ~g berdujah keselamatu. atau kepco.tl!lJ;1.ll Mlli.ysia scperti yang termal..-rub di da.h.m. AKTA:RAHSlA R.ASMl1972)

(M::ngwu.ngi md:.1umat TERHAD yang tclah ditentubo oleb organisuilbadtn di ~DI peoydidikl1l dijd.nkan)

Diiahkan oleh

(TA . . N PUSTAKAWAN)

Nama rcnyc1ia

Tuikh:

•• Jikl leS;' ;ni SULIT at,u TERHAD, sil. lampirhn S\If1tt duipad& pih&k bcrl:ua..worpnisasl hcdi;cnUll dct\&llI mtftYitatc.n scltali scb.b dlll umpoh Inil ini pcflu dil::dubn lCb..gai SULIT dan TF.RilA.O.

@ Tesisdimlkludlw1 R:biI, .. i telis b.,i Iju .... h Doktor Fllu!ah. dan Satjana scc::ara ptnyelidihn. "!au dis-crus; b ... ~ f'CtICljitlll1Caofa kcrja kvmli dan flCI'lyc:iidiUn, atau Laponn Projck. Slljanl Muda (LPSM ).

-

AUTHENTICATION

Authenticated by Members of Dissertation Committee

1. SUPERVISOR

(MR. RUZAIDI AZLI BIN MOHO MOKHTAR)

2. EXAMINERI

(PROF. MADYA DR. MARKUS ATONG)

3. DEAN PfI"SmlU

UilnSlTIlUmll ·UBI'

(pROF. MADYA DR. SHARIFF A. K. OMANG)

, , :, ,;:t ' ,I, .....

11\

IV

ACKNOWLEDGEMENTS

First of all, I wish to convey my sincere thanks to my supervisor, Mr. Ruzaidi Azli Bin

Mohd. Mokhtar for his guidance, patience and concern throughout the study. Besides, I would like

to express my deepest gratitude to my ex-supervisor, Dr. Jaya Vejayan Palliah for his valuable

advice ofthi, study. I want to appreciate Mr. Zainal from Malaysian Cocoa Board (Inanam branch)

for supplying me the bacterial strains. I am also most grateful to Mr. Patrick Chee for supplying

some of the flowers in Orchid de villa. Finally, I would like to thank my family and also my

friends for the love and support.

~ UMS ."< UNIVERSlTI MALAYSIA SASAH

v

ABSTRACT

The objectives of this study were to extract the selected flowers with methanol, evaluate

their antibacterial activity against four standard bacteria by using disc diffusion method and

compare the antibacterial activity of the flowers, The ten species of flowers used in this study were

Yellow allamanda (Allamanda cathartica). Madagascar periwinkle (Catharanthus roseus).

Crown-of-Thoms (Euphorbia millt), Bachelor's button (Gomphrena gJobosa), Hibiscus (Hibiscus

rosa sinensis), Spider lily (Hymenoca/lis Iit/oratis), Mile-A-Minute Vine (Ipomoea eairica),

Lantana (Lantana camara), Toothache plant (Spilanthes acmel/a) and Crepe jasmine

(Tabernaemonrana divaricata). The methanolic extracts of the flowers were screened for

antibacteriaJ activity against Escherichia coli, Salmonella thyphimurium, Staphylococcus aureus

and Pseudomonas aeruginosa. The screening on the flowers revealed. varying degree of

antibacterial activity against various pathogenic bacteria. All the flower samples except Bachelor's

button showed clear inhibition zone as a result of antibacterial activity towards S. aureus. The

flower sample that was identified showing the significant antibacterial activity against S. allreus

was Crown-of-Thoms, with the inhibition diameter of 4.33 mm. Yellow allwnanda, Madagascar

periwinkle, Mile-A-Minute Vine, Toothache plant and Crepe jasmine showed the same diameter of

2.00 mm. Spider lily showed inhibition of 1.80 mm. Hibiscus exerted 1.00 mm diameter of

inhibition while Lantana showed the smallest inhibition zone of 0.50 mm. However, the entire 10

sample flowers did not show any inhibition zone as a result of antibacterial activity towards

Escherichia coli. Six flower samples exhibited inhibition zones as a result of antibacterial activity

towards Salmonella Ihyphimurillm. From the result, Spider lily, Toothache plant and Crepe jasmine

showed the same inhibition zone with diameter of 1.50 mm. Madagascar periwinkle, Hibiscus and

Toothache plant were less resistant to S. lhyphimllrium as it exened 1.33 mrn, 0.67 mm and

1.00 mm diameter of inhibition zone respectively. The remaining flower samples did not exert any

clear zone. There were 5 out of 10 flower samples that inhibited the positive results with the

Pseudomonas aeruginosa tested for the antibacterial activity. There were no obvious differences

between the tested plant samples of Hibiscus and Toothache plan~ showing the same inhibition

diameter of 1.00 mm. Madagascar periwinkle and Spider lily both showed 1.17 rom of inhibition

UMS UNIVERSITI MALAYSIA SABAH

VI

diameter. Crown-of-Thoms showed the largest inhibition zone of 1.50 mm. The remaining flower

sample revealed negative results. In conclusion, the antibacterial test showed the most significant

result in S. aureus with the flower named Crown-of-Thoms, with the mean diameter of 4.33 nun.

UMS UNIVERsm MAlAYSIA SASAI

vii

ABSTRAK

Objektif kajian ini adaJab untuk mengekstrak bunga-bunga pilihan dengan metanol, mengkaji

aktiviti antibakteria terhadap 4 spesies bakteria dengan menggunakan kaedah penyerapan piring!

dan membandingkan keputusan aktiviti anti-bakteria di antara jenis-jenis bunga dan jenis-jenis

bakteria. Sepuluh ekstrak bunga yang digunakan daJam eksperimen ini adalab Yellow allamanda

(A I/amanda cathartica), Madagascar periwinkle (Catharanthus roseus), Crown-of-Thorns

(Euphorbia milll), Bachelor'S bulton (Gomphrena globosa), Hibiscus (Hibiscus rosa sinensis),

Spider lily (Hymenocal/is Jiltoralis), Mile-A-Minute Vine (Ipomoea cairica), Lantana (Lantana

camara), Toothache plant (Spilanthes acmeI/o) dan Crepe jasmine (Tabernaemontana divaricata),

Sernua ekstrak bunga tersebut diuji ke atas bakteria Escherichia coli, Salmonella thyphimurium,

Staphylococcus aUreus dan Pseudomonas aeruginosa. Kajian terhadap bunga tersebut telah

memherikan hasil keputusan yang herbeza disebabkan oleh darjab perhezaan antibakteria. Semua

bunga kecuali Gomphrena globosa telab menghasilkan zon perencatan terhadap S. aurells. Sampel

bunga yang memherikan keputusan yang ketara terhadap aktiviti aotibakteria S. aureus adaJab

Euphorbia mill;, dengan diameter zon perencatan sebanyak 4.33 mm. Allamanda ca/hartiea,

Cotharan/hus roseus, Ipomoea eairica, Spilanthes aemella dan Tabernaemomana divaricala

menunjukkan keputusan yang sarna iaitu, 2.00 mm. Hymenocallis Iiltoralis memberikan diameter

sebanyak 1.80 mm. Hibiscus rosa sinensis menunjukkan keputusan 1.00 mm. Lamana camara

pula mernherikan diameter sebanyak 0.50 mm. Namun begitu, semua sampel bunga tersebut tidak

menunjukkan sebarang aktiviti anti-bakteria terhadap Escherichia coli. Selain itu, sampeJ bunga

tersebut juga memberikan aktiviti zon perencatan anti-bakteria yang lemah terhadap Salmonella

Ihyphimur;um. Daripada keputusan ini, Hymenocallis lilloralis, Spilan/hes aemella dan

Tabernoemontana divaricala memberikan keputusan yang sarna dengan zon perencatan 1.50 mm.

Cotharanlhw· roseus, Hibiscus rosa sinensiS dan Spilanthes acmello adalah kurang resisten

terhadap S. Ihyphimurium kerana masing-masing menghasilkan diameter 1.33 mm, 0.67 mm dan

1.00 mm. Sampel bunga yang lain tidak menunjukkan sebaraog tindak baJas antibakteria. Terdapat

lima daripada sepuluh sampel bunga yang memherikao keputusan positif terhadap Pseudomonas

aerllginosa. Tidak terdapat perbezaan yang ketara di antara Hibiscus rosa sinensis dan Spilanlhes

~ ,..< UMS UNIVERSITI MALAYSIA SABA

VIU

Cicmella yang dikaji iaitu menujukkan diameter sebanyak 1.00 mm. Ca,haranthus rosells dan

Jiymenocallis /il/oralis kedua-duanya memberikan 1.17 rom diameter zan perencatan. Euphorbia

»Ii/Ii menunjukkan zon perencatan 1.50 mm. Sampel yang lain memberikan keputusan neg.tif.

l<.esimpulanny~ ujian antibakteria menunjukkan keputusan yang paling ketara dengan S. DUfeus

diuji terhadap Euphorbia milli, dengan purata diameter zon perencatan 4.33 rnm.

@) !JIY.lS

V11l

acmella yang dikaji iaitu menujukkan diameter sebanyak 1.00 mm. Catharamhus rosellS dan

Hymenocallis lil/oralis kedua-duanya memberikan 1.17 nun diameter zon perencatan. Euphorbia

mill; menunjukkan zon perencatan 1.50 mm. Sampel yang lain memberikan keputusan negatif.

KesimpuJannya, ujian antibakteria menunjukkan keputusan yang paling ketara dengan S. aureus

diuji terhadap Euphorbia mi/li, dengan purata diameter zon perencatan 4.33 mm.

UMS , "

DECLARATION

AUTUENTICATION

AKNOWLEGDEMENT

ABSTRACT

ABSTRAK

LIST OF CONTENTS

LIST OF TABLE

LIST OF FIGURE

LIST OF UNlT

LIST OF ABBREVIATION

CUAPTER I INTRODUCTION

1.1 Introduction

1.2 Objectives

LIST OF CONTENTS

CUAPTER 2 LITERATURE REVIEW

2.1 AI/amanda cotharrieD L.

2.2 Cafharanfhus roseus G.Don

2.3 Euphorbia milii

2.4 Gomphrena g/obosa L.

2.5 Hibiscus rosa sinensis

2.6 Hymenocallis litloralis

2.7 Ipomoea eairiea L.

2.8 Lamono camara

2.9 Spilanlhes acmella

2.10 Tabernaemontana diVOT/cola

Page

II

"' 'V v

VII

'X

XI'

xiii

XV

XVI

3

4

6

8

JO

12

13

14

17

18

20

'X

2.11 Bacteria used in this study

2.11.1 Escherichia coli

2.11.2 Siaphylococcus QureUj'

2.11.3 Salmonella Iyphimurium

2.11.4 Pseudomonas aeruginosa

CHAPTER 3 MATERIALS AND METHODS

3.1 Materials

3.1.1 General materials

3.1.2 I nstrurnentation

3.1.3 Bacterial strains

3.1.4 Plant materials

3.2 Methods

3.2. 1 Overall procedure

3.2.2 Determination of antibacterial activity

a. Preparation of culture media

h. Plant extraction

c. Antibacterial assay

3.2.3 Statistical analysis

CHAPTER 4 RESULTS

4.1

4.2

4.3

Screening for antibacterial activity

Experimental results

Data analysis

CHAPTER 5 DISCUSSION

5.1 Different observation between Gram-positive and Gram-negative bacteria

5,2 Consideration of methanol used as extraction solvent

5.3 Considerations of sample discs preparation

22

22

23

24

25

25

26

26

28

29

31

33

34

35

35

46

51

54

55

x

5.4 Antimicrobial agent

5.5 Gentamicin antibiotic discs as reference discs

5.6 The Disc-Diffusion Method

CHAPTER 6 CONCLUSION

6.1

6.2

Conclusion

Suggestion for further study

REFERENCE

APPENDIX

55

56

56

58

58

60

64

Xl

XlI

LIST OF TABLE

No.ofTitle Page

3.1 List ofbacteri. will be used for this study. 26

3.2 List of flowers will be used in this study 27

4.1 Antibacterial activities of sample discs after incubation 24 hours 36

~ UMS ."< UNIVERSlTI MALAYSIA SASAH

LIST OF FIGURE

No.ofTitie Page

2.1 AI/amanda cQthartica L. 5

2.2 Catharanthus roseus G. Don. 6

2.3 Euphorbia mili. 9

2.4 Gomphrena globosa L. 10

2.5 Hibiscus rosa sinensis 12

2.6 Hymenocallis lilloralis 14

2.7 Ipomoea eairiea L. 15

2.8 Lantana camara 17

2.9 Spi/anthes acmella 19

2.10 Tabernaemontana divaricata 20

3.3 Schematic representation of overall procedure 28

3.4 Schematic representation of agar and broth preparation 30

3.5 Schematic representation of flower extraction procedures 32

4.1 Growth inhibition of E. coli by different samples. (I, AI/amanda cathartiea; 2. Calharanlhus roseus; 3, Euphorbia milii; 4, Gomphrena globosa) 37

4.2 Growth inhibition of E. coli by different samples. (5, Hibiscus rosa sinensis; 6, Hymenoeallis lillora/is; 7, Lantana camara; 8, Ipomoea eairiea) 38

4.3 Growth inhibition of E. coli by different samples. (9, Spilanlhes aemel/a; 10, Tabernaemonrana divarieQfa; C. Blank; G, IO)Jg Gentamicin disc) 38

4.4 Growth inhibition of S. Ihyphimurium by different samples. (I, AI/amanda 39 calhartiea; 2, Calharanthus roseus; 3, Euphorbia milii; 4, Gomphrena globosa)

xiii

4.5 Growth inhibition of S. thyphimurium by different samples. (5, Hibiscus rosa sinensis; 6, Hymenocallis lilloralis; 7, lAntana camara; 8, Ipomoea cairica)

4.6 Growth inhibition of S. thyphimurium by different samples. (9, Spilanthes

40

acmella; la, Tabernaemontana divaricata; C, Blank; G, 10).lg Gentamicin disc) 40

4.7 Growth inhibitions ofS. aureus by different samples. (I , Allamanda cathartica; 2, Catharal1lhus roseus; 3, Euphorbia milii; 4, Gomphrena globosa) 41

4.8 Growth inhibitions of S. aureus by different samples. (5, Hibiscus rosa sinensis; 6, Hymenoeallis lil/aratis; 7. Lantana camara; 8.1pomoea eairico)

4.9 Growth inhibitions of S. aureus by different samples. (9, Spilan/hes

42

oemello; 10, Tabernaemontona divarieala; C, Blank; G, 10).lg Gentamicin disc) 42

4.10 Growth inhibition of P. aeruginosa by different samples. (1, Allamanda calharlica; 2, Carharanthus roseus; 3, Euphorbia milii; 4, Gomphrena globosa) 43

4.11 Growth inhibition of P. aeruginosa by different samples. (5, Hibisclis rosa sinensis; 6, Hymenocal/is Iil/araUs; 7. Lamana camara; 8, Ipomoea cairica) 44

4.12 Growth inhibition of P. aeruginosa by different samples. (9, Spilanthes acmel/a; 10, Tahemaemontana divaricata; C, Blank; G, 10 ~g Gentamicin disc) 44

4.13 Inhibition zone of different sample discs tested against Staphylococcus allrellS 46 after 24 hours incubation. (Values were mean ± SO)

4. J 4 Inhibition zones of different sample discs tested against Escherichia coli 47 after 24 hours incubation. (Values were mean ± SD)

4.15 Inhibition zone of different sample discs tested against Salmonella 48 thyphimurillm after 24 hours incubation. (Values were mean ± SD)

4.16 Inhibition zones of different sample discs tested against Pseudomonas 49 aerug;nosa after 24 hours incubation. (Values were mean ± SD)

xiv

xv

LIST OF UNIT

m meter

em centimeter

mm mil limeter

run nanometer

ml milliliter

111 microliter

g gram

ppm parts per million

'C degree Celsius

~ UMS . ..< UNIVERSlTI MALAYSIA SASAH

LIST OF ABBREVIATION

E. coli

S. ryphimurium

P. aerllginosa

S. Dureus

Escherichia coli

Salmonella typhimllr;um

Pseudomonas aerugillosa

SlaphyiococclIs Qllreus

xv,

CHAPTER I

INTRODUCTION

1.1 Introduction

According to an estimation of World Health Organization (WHO), nearly 80 per<:ent of

the populations in developing countries rely on traditional medicine especially plant drugs

for their priroary health care needs (Yang, 2005). Medicinal plants are important for

pharmacological research and drug development such as thernpeutic agents and as models

for pharmacologieally active compounds. The advantages of using plant derived

medicines are that they are relatively safer than synthetic alternatives, offering profound

therapeutic benefits and more affordable treatment.

Many of the exploration and utilization of natural products as antimicrobials arise

from microbial sources. It was the discovery of penicillin that led to later discoveries of

antibiotics such as streptomycin, aureomycin and chloromycetin. Though most of the

clinically used antibiotics are produced by soil micro-organisms or fungi, higher plants

have also been a so""", of antibiotics. Examples of these are the antibiotic action of

allinine in Allium salivum (garlic) and the antimicrobiaJ action berberines in goldenseal

2

(Hydras/is canadensis). Plant based antimicrobials represent a vast untapped source for

medicines. Continued and further exploration of plant antimicrobials needs to occur.

Plants based antimicrobials have enormous therapeutic potential. They are

effective in the treatment of infectious diseases while simultaneously mitigating many of

the side effects that are often associated with synthetic antimicrobials. They are effective

and gentle. Many plants have tropisms to specific organs or systems in the body.

PhytOmedicines usually have multiple effects on the body. Their actions often act beyond

the symptomatic treatment of disease. An example of this is Hydrastis canadensis.

Hydrastis canadensis not only has antimicrobial activity, but also increases blood supply

to the spleen promoting optimal activity of the spleen to release mediating compounds.

Other examples like morphine from opium use as narcotic and analgesic, latex exuded

from the poppy seed for gout, cocaine from cocoa leaf as potential local anesthetic, garlic

for blood and heart remedies, and so on (Sharma, 2004).

Antibiotics have changed our life in the sense that it led to new standards of health

for billions of people. Many of the life-threatening infections of previous centuries are

now conveniently cured by medicine. Antibiotics also have applications as feed additives.

growth stimulants, pesticides and wider agricultural uses. Nowadays, many bacterial

strains are resistant to antibiotics treabnent due to the indiscriminate usage of antibiotics.

The phenomenon of antibiotic resistance by bacteria bas becoming a medical disaster as

we may be entering a post-antibiotic era where antibiotics are no longer effective.

3

Over the past few years, researches had shown an increase in antibiotic-resistant

bacteria such as gentamicin-resistant Staphylococcus aureus and methicillin-resistant

Staphylococcus aureus (MRSA). The diseases caused by MRSA have exceeded 30

percent in some countries such as southern Europe and the United States. Moreover,

reports on staphylococcal infections with reduced susceptibility or resistance to

vancomycin are emerging (Faber et al., 2005). As a result of the worldwide increase of

gram-positive antimicrobial-resistant pathogens, morc studies on the search for novel­

infective agents are carrying on for resolutions (Detighanyar et 01., 2005).

In this study, certain pJant extracts were used to screen their antimicrobial

activities towards standard bacterial strains. Some plants were selected based on their

traditional utilization in the treaUnenl of microbial diseases such as skin disease, acne,

abscesses and SO on.

1.2 Objectiv ..

There were three objectives formed from this current study. First, to extract the selected

flowers with methanol. Second, to evaluate the antibacterial activity of selected flowers

against four standard bacteria [Staphylococcus aureus (ATCC 25923), Escherichia coli

(ATCC 25922). Pseudomonas aeruginosa (ATCC 27853) and Salmonella typhimurium

(ATCC 14028)] by using disc-diffusion technique. Third, to compare the antibacterial

activity (inhibition zone) of selected flowers.

CHAPTER 2

LITERATURE REVIEW

2.1 AI/amanda calharlica L.

Allamanda calhartica L., originated from America, is a common garden and street

tropical omamentallatieiferous shrub that can grow up to four meters. It is also known as

yellow a1lamanda (English), bunga lelur raja (Malay), jaharsonlalcka (Sanskrit), and

jasmine d'amarille, orelie de 10 guyane. a/amande (French). Allamanda cathartica L. has

poisonous white latex. Its leaves are simple, verticillate, sessile, glabrous beneath and

shining. Blade is oblong with 8 em to 15 em long, 4 em to 5 em wide, laneeolate, base

acute and apex acuminate. Flowers are showy, bright yellow, large with 7 em long, tube

with 3 em long and inflated with 2 em diameter. Lobes are ovate oblong contorted in buds.

This plant is illustrated in Figure 2.1.

5

f---l 2cm

Figure 2.1 AI/amanda ca/hartiea L.

In various parts of Southeast Asia.. the leaves of AI/amanda calharlica L. are used

in small quantity as laxative and emetic because high doses of this plant are toxic. Besides,

its leaves are used as laxatives, the barks are hydragogue in ascite and roots are given in

cases of snakebites in India. AI/amanda catharlica L. shows antimicrobial activity,

cytotoxic and molluscicidal properties. Extracts of this plant inhlbited in vitro Klebsiella

sp., Staphylococcus sp. and fungal growth. Its derivatives which known as Allamandine

were reported having anti leukaemic, tumor inhibiting and molluscicidal properties.

Further report showed that cytotoxic-.guided fractionation of ethylacetate leaves using Sc-

(7) yeast strain resulted in the isolation of isoplumericin and plumericin iridoids. This was

previously reported as antimicrobial and moUuscicidal principles of Plumieria spp. (Wiart,

2000).

• , • , ,

!I: -. "" -' to: ~.

E~ • ! • •

6

2.2 Calharanthus roseus G. DOD

Catharanthus roseus G. Don or Madagascar periwinkle (EngLish)~ kemunting cina

(Malay); Ihenbanmahnyoban (Burma); pervenche de Madagascar (French); is an

herbaceous subshrub initially indigenous to Madagascar and now widely distributed

throughout warm regions where it is cultivated as ornamental . Catharanlhlls roseus G.

Don is an erect, perennial herb that has high pinky-red stem, much branched. Its leaves

simple, thinly fleshy, opposite, glabrous on both sides, dark shining and petiolate. The

inflorescence is racemose, with the flowers in axillary pairs. The flowers are rose-purple

or white or white with a rose-purple spot in the centre. This plant is illustrated in

Figure 2.2.

1------1 2cm

Figure 2.2 Catharanthus raseus G. Don.

®!J,MA~

60

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Aqil. F., Khan, M. S. A., Owais, M. & Ahmad, I., 2005. Effect of certain bioactive plant

extracts on clinical isolates of I}-Iactamase producing methicillin resistant

Staphylococcus aureus. Journal Basic Microbial 45 (2), 106-114.

Bauer, A. W. , Kirby, W. M. M., Sherris, J. C. & Turch, M. 1966. Antibiotic susceptibility

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Brock, T. D., Madigan, M. T. , Martinko, J. M. & Parker, J. 1994. Biology 0/ Aficroorganisms. 7th Ed. Prentice-Hall International, Inc. pp. 344-352.

Chin, H. F. 1998. The Hibisclls. Qlleen a/Tropical Flowers. Tropical Press Sdn. Bhd,

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Detighanyar. P .. Burger, C., Zeitlinger, M. , Islinger, F., Kovar. F., Muller, M., KIoft, C.

& Joukhadar. C. 2005. Penetration of cinezolid into soft tissues of healthy

volunteers after single and multiple doses. Antimicrobial agents and

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Faber, C., Stallmann, H. P., Lyaruu, D. M., Joosten, U., Eiff, C. Y., Amerongen, A. Y.

N., & Wuisman, P. I.. J. M. 2005. Comparable efficacies of the antimicrobial

peptide human Lactoferrin 1-11 and Gentamicin in a chronic Methicillin­

Resistant. Staphylococcus aureus Osteomyelitis model. Antimicrobial agents and

chemotherapy 49 (6): pp. 2438-2444.

61

Ferreira. A. A., Amaral, F. A., Duarte, I. D., Oliveira, P. M., Alves, R. B., Silveira. D.,

Azevedo, A. 0 ., Raslan, D. S. & Castro, M. S. 2006. Antinociceptive effect from

Ipomoea cairica extract. Journal ofEthnopharmacology 105 (1-2): pp. 148-153.

George, D. P. R. 2001. Encyclopedia of Medicinal Planls. Editorial Safeliz, Spain.

Goh, K. L. 2001. Malaysian Herbs. Volume. I. Percetakan Advanco Sdn. Bhd., Kuala

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Heinerman, J. 1998. Miracle Healing Herbs. Prentice Hall, New Jersey.

Idso, S. B., Kimball , B. A., Pettit, G. R., Gamer, L. C. & Backhaus, R. A. 2000. Effects

of atmospheric C~ enrichment on the growth and development of Hymenocallis

lil/oraUs (Amaryllidaceae) and the concentrations of several antineoplastic and

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~UMS ~ UNIVERSlTI MALAYSIA SABA