An Ethnobotanical Study of Medicinal Plants Used By Palestinians … · 2018. 9. 19. · IV Abstract The study aimed at evaluating the current status of folk medicine treatment in

An Ethnobotanical Study of Medicinal Plants

Used By Palestinians in Gaza Strip

ؾؼث لث اطة ااغرخذح ف اطث١ح دساعح اثاذاخ

افغط١١١ ف لطاع

Samar Yousuf. Alghuff

Supervised by:

Dr. Tarek A. ElBashiti

Associate Prof. of biotechnology

Dr.Mohamad M. Abou Auda

Associate prof. of Botany

A thesis submitted in partial fulfillment

of the requirements for the degree of

Master of Science in biological science

July /2018

ضجــغ – حــلا١ــــــح الإعـــــــــاؼـاد

اثسث اؼ اذساعاخ اؼ١ا ػادج

اؼــــــــــــــــــــــح ــــــــــــــــــــ١ـو

اؼ اــــس١ــــاذــــ١ــــح ش ـــــــاخغر١

The Islamic University–Gaza

Deanship of Research and Graduate Studies

Faculty of Science

Master of Biological Science

II

إلــــــــــــــشاس

أا الغ أدا مذ اشعاح ار ذس اؼا:

An Ethnobotanical study of Medicinal Plants Used By

Palestinians In Gaza Strip

لث افغط١١١ ف لطاع اغرخذح ف اطة اؾؼث اطث١ح دساعح اثاذاخ

غضج

ألش تأ ا اؽرد ػ١ ز اشعاح إا راج خذ اخاؿ، تاعرثاء ا ذد الإؽاسج إ١ ز١ثا سد،

أ ز اشعاح وى أ أ خضء ا ٠مذ لث الاخش٠ ١ دسخح أ مة ػ أ تسث ذ أ

ؤعغح ذؼ١١ح أ تسث١ح أخش.

Declaration

I understand the nature of plagiarism, and I am aware of the University’s policy on

this.

The work provided in this thesis, unless otherwise referenced, is the researcher's own

work, and has not been submitted by others elsewhere for any other degree or

qualification.

:Student's name اع اطاة:

:Signature ارل١غ:

:Date 1/7/2018 اراس٠خ:

III

IV

Abstract

The study aimed at evaluating the current status of folk medicine treatment in

Gaza strip, identify medicinal plants still in use by therapists, and their importance in

health care. An ethnobotanical survey conducted investigated and recorded plant

species used in folk medicine. Information was collected by interview with 20

informants from different locations. The study documented 72 medical plant species

belonging to 33 families. The most frequently utilized plant parts were leaves 41%

followed by stems 18%, and seeds 14%. The majority of remedies were used to treat

gastrointestinal disorders, 37 species followed by circulatory system problems, 22

species, and Urinary system, 20 species.

The antimicrobial activity of eleven plant extracts and synergistic effect

between some plant extracts was evaluated against Escherichia coli and

Staphylococcus aureus.. The antibacterial activities of the extracts were evaluated

using the disk diffusion method, well diffusion method, minimum inhibitory

concentration (MIC) and minimum bacterial concentration (MBC). The results

showed that the E.coli was more susceptible to the employed plant extracts than

S.aureus. In most plant extracts the aqueous extract has greater effect in the

inhibition from ethanolic extract except Syzygium aromaticum which showed the

large inhibition zone on the bacteria and the convergence between the results of the

ethanol extract and aquatic extract in both disc and well diffusion method. On the

contrary of Syzygium aromaticum the Hibiscus sabdariffa shown the lowest MIC

value in aquatic extact Compared to ethanolic extract against two types of pathogenic

bacteria. The MBC results for the most of the plant species in each ethanolic and

aquatic extract against E.coli and S.aureus was >200. This current study detected the

effectiveness of synergistic effects to some extract with each other toward a

pathogenic bacteria some plant extracts showed weak synergistic effect using disc

and well diffusion method and low value of MIC.

Keywords : Ethnobotany, medicinal plant, antimicrobial, synergism, aquatic extract,

ethanolic extract, MIC, MBC.

V

Arabic abstract

إى رق اىػغ اىحبى ىيؼلاج اىشؼج ف قطبع غضح ، رحذذ اىجبربد اىطجخ اىز لا ذفذ اىذساعخ

أخشذ دساعخ اعزقظبئخ ثئخ ىزغدو الأاع رضاه رغزخذ قجو اىؼبىد ، أزب ف اىشػبخ اىظحخ.

ؼبىح صػ 02اىؼيبد خلاه اخشاء قبثيخ غ خغ حث راىجبرخ اىغزخذخ ف اىطت اىشؼج

صػذ ع جبر 20ػي ابم خزيفخ قطبع غضح. ثيغ ػذد اىجبربد الامثش اعزخذاب ف اىؼلاج اىشؼج

ػبئيخ جبرخ رغزخذ ف ػلاج اىؼذذ الاشاع. ثذ اىذساعخ ا امثش الاخضاء اىغزخذخ 33ػي

%. 41% يب اىجزس ثغجخ 41% احزيذ اىغقب اىشرجخ اىثبخ ثغجخ 14الاساق شنيذ غجخ اىجبد

00ي اىدبص اىذس الاػخ اىذخ ع 32مبذ اغيت اىجبربد رؼبىح اػطشاثبد اىدبص اىؼ

.ع جبر 02 يب اشاع اىدبص اىجىع

اىجبربد اىطجخ , ثبلإػبفخ اى اىزؤثش ع جبرزش لإحذ ػشش مب ر رق اىزؤثش اىؼذ ثن

ر رق اىشبؽ اىؼذ اىزآصس ىجؼغ ب ػذ الإششنخ اىقىخ اىجنزشب اىنسح اىؼقدخ اىزجخ .

حفش. قذسح ثنزش ىيغزخيظبد اىجبرخ, ثبعزخذا ؽشقخ الازشبس ف اىقشص مزىل ؽشقخ الازشبس ف اى

قخ أقو رشمض ثجؾ اىغزخيظبد اىجبرخ حذب ثبعزخذا ؽشقخ اىزخفف اىدضئ . اشبسد اىزبئح ذ

خ اىقىخ امثش فؼبىزب ػي اىجنزشب اىنسح شاىجبربد ػذ الإششاىغزخيظبد فؼبىخ ثؼغ ز

اىؼقدخ خبطخ ف اىغزخيظبد اىبئخ.

ف ؼظ اىغزخيظبد اىجبرخ مب رؤثش اىغزخيض اىبئ امجش رؤثش اىغزخيض الاثبى ف رثجؾ

اىز اظش اىغزخيض الاثبى زبئح افؼو Syzygium aromaticumثبعزثبء اه اى اىجنزش

Hibiscusش جبد اظ Syzygium aromaticumاىبئ ف رثجؾ اى اىجنزش. ػي اىؼنظ

sabdariffa قخ افؼو ف اىغزخيض اىبئ ثبىقبسخ غ اىغزخيض الإثبى ػذ ػ اىجنزشب اىغججخ

مب أظشد ثؼغ اىغزخيظبد اىجبرخ رؤثشا رآصسب ػؼفب ثبعزخذا ثبعزخذا ؽشقخ الازشبس ف ىلأشاع.

شقخ اىزخفف اىدضئ.اىقشص مزىل ؽشقخ الازشبس ف اىحفش ؽ

اىجبربد اىطجخ ، ؼبداد اىنشثبد، اىزآصس، غزخيظبد بئخ ، اىطت اىشؼج، اىنيبد اىفزبحخ3

غزخيض الإثبه، أقو رؤثش ثجؾ أقو رؤثش قبرو.

VI

Dedication

To the Spring that never stops giving, to who weaves my happiness with strings from

her merciful heart … my mother

To the big heart …. my father.

To my brothers and sisters who have been a great source of motivation and

inspiration .

To my daughter, my friend who suffered the difficulties with me Samar Abo Saffia

VII

Acknowledgments

My thanks first and foremost to Allah almighty, for his bounty, which enabled me to

accomplish this work. Praise be to Him first and foremost. who has been the source

of my energy throughout my study; without Allah none of this would be possible

All thanks to my supervisors Dr. Tarek Elbashiti and Dr. Mohamad Abou Auda for

their advice, guidance, and encouragement.

Also, I am very grateful for each of Farida Mousleh and Mahmoud Al Hindi from

Biological Control Unit and Samar Abo Saffia to help in the practical part of

antimicrobial methods of this research.

Finally, thanks are extended to everyone who has a hand in this work.

VIII

Table of content

Declaration ................................................................................................................... II

Abstract ...................................................................................................................... IV

Arabic abstract ............................................................................................................. V

Dedication .................................................................................................................. VI

Acknowledgments .................................................................................................... VII

Table of content ....................................................................................................... VIII

Table of Figures ........................................................................................................ XII

List of Table .......................................................................................................... XVII

List of Abbreviated Terms .......................................................................................XIX

Chapter 1: Introduction ............................................................................................. 1

1.1 Overview ................................................................................................................ 2

1.2 Main objective: ...................................................................................................... 5

1.3 Specific Objectives: ............................................................................................... 5

1.4 Significance : ......................................................................................................... 6

Chapter 2 : Literature Review .................................................................................. 7

2.1 The history of herbal treatment .............................................................................. 8

2.2 Ethnobotany .......................................................................................................... 9

2.2.1 Traditional ethnobotanical markets ............................................................. 9

2.3 Medicinal plants ..................................................................................................... 9

2.3.1 Future of Medicinal Plants .......................................................................... 9

2.4 Plants used in extraction ...................................................................................... 10

2.4.1 Ocimum basilicum (basil) : 11 ........................................................... اش٠سا

2.4.2 Hibiscus sabdariffa L. (Roselle) : 12 ................................................. اىشوذ٠ح

2.4.3 Trigonella foenum- graecum L. (Fenugreek seed) : 13 ........................ اسثح

IX

2.4.4 Coriandrum sativum (Coriander) : 14 ................................................. اىضتشج

2.4.5 Anethum graveolens (Dill) : 15 ....................................................... ػ١ خشادج

2.4.6 Syzygium aromaticum L. (Clove) : 16 ................................................. امشف

2.4.7 Glycyrrhiza glabra L. (Licorice) : 17 ........................................... اؼشق عط

2.4.8 Cuminum cyminum L. (Cumin) :18 ...................................................... اى

2.4.9 Cymbopogo schoenanthus.L (camel grass) :19 .................................. اسفا تش

2.4.10 Boswellia sacra Flueck (Olibanum or frankincense) : 20 .... زص اثا اذوش

2.4.11 Artemisia monosperma Delile (white wormwood) : 21 ....................... اؾ١ر

2.5 Microorganism ..................................................................................................... 22

2.5.1 Escherichia coli ........................................................................................ 22

2.5.2 Staphylococcus aureus ............................................................................. 23

2.6 Previous Studies:.................................................................................................. 24

Chapter 3: Materials and Methods ......................................................................... 27

3.1 Materials .............................................................................................................. 28

3.1.1 Study Area ................................................................................................. 28

3.1.2 Study population ........................................................................................ 29

3.1.3 Medicinal Plants Markets .......................................................................... 31

3.1.4 Plant Sample Collection: ........................................................................... 31

3.1.5 Culture media and chemicals ..................................................................... 33

3.1.6 Microorganisms ......................................................................................... 33

3.2 Methods: ........................................................................................................... 33

3.2.1 Ethnobotanical Methods: ........................................................................... 33

3.2.1.1 Survey and Identification: .................................................................. 33

3.2.2 Extraction methods : .................................................................................. 34

3.2.2.1 Preparation of plant extract: ............................................................... 34

X

3.2.2.2 Evaluation of antibacterial activity of plant extracts by disc diffusion

method: ........................................................................................................... 35

3.2.2.3 Evaluation of antibacterial activity of plant extracts by well diffusion

method: ........................................................................................................... 35

3.2.2.4 Determination of MIC and MBC of plant extracts by Microdilution

Method: ........................................................................................................... 35

3.2.2.5 Synergistic assay: ............................................................................... 36

Chapter (4) :Results .................................................................................................. 37

4.1 Taxonomic diversity of plants under investigation............................................. 38

4.2 Most cited plants and remedies ............................................................................ 40

4.3 Folk medical knowledge sources: ........................................................................ 57

4.4 Used parts ............................................................................................................ 57

4.5 Procurement methods of medicinal plants: .......................................................... 58

4.6 Evaluation of antibacterial activity of plant extracts by disc diffusion method .. 58

4.6.1 Against Escherichia coli ............................................................................ 58

4.6.1.1 The aquatic extracts ............................................................................ 58

4.6.1.2 The ethanolic extracts ......................................................................... 59

4.6.2 Against Staphylococcus aureus ................................................................ 59

4.6.2.1 The aquatic extracts: ........................................................................... 59

4.6.2.2 The ethanolic extracts: ........................................................................ 59

4.7 Evaluation of antibacterial activity of plant extracts by well diffusion method .. 60

4.7.1 Against Escherichia coli: ........................................................................... 60



4.7.2. Against Staphylococcus aureus: ............................................................... 61



XI

4.8 The minimum inhibitory concentration (MIC) & minimum bactericidal

concentrations (MBC) of plant extracts ..................................................................... 70

4.8.1 Against Escherichia coli: ........................................................................... 70

4.8.2 Against Staphylococcus aureus: ............................................................... 75

4.9 Evaluation the synergistic effect: ......................................................................... 80

4.9.1 The Synergistic effect between plant extract: ............................................ 80

4.9.1.1 Evaluation the synergistic effect by disc diffusion method: ............... 80

4.9.2 Evaluation the synergistic effect by well diffusion method ...................... 80

4.9.2.1 Against Escherichia coli: .................................................................... 80

4.9.2.2 Against S. aureus: ............................................................................... 80

4.9.3 The minimum inhibitory concentration (MIC) of mixed plant extracts

against isolated bacteria: .................................................................................... 82

4.9.3.1 Against Escherichia coli: .................................................................... 82

4.9.3.2 Against S. aureus: ............................................................................... 82

Chapter (5) Discussion and Conclusions ........................................................... 85

5.1 Study population: ................................................................................................. 86

5.2 Medicinal Plants Markets: ................................................................................... 86

5.3 Plants and parts used ............................................................................................ 87

5.4 Antibacterial Activity of the Plant Extracts ......................................................... 88

5.5 Conclusion: .......................................................................................................... 91

5.6 Recommendation: ................................................................................................ 91

References .................................................................................................................. 93

Appendix .................................................................................................................. 100

XII

Table of Figures

Figure (3.1) : Gaza strip map ( source : www.arij.org )............................................ 29

Figure (3.2): Distribution of study population by gender ......................................... 30

Figure (3.3): Distribution of study population by age .............................................. 30

Figure (3.4): Distribution of study population by educational level. ....................... 31

Figure (3.5) : Photograph shows interview with informants .................................... 34

Figure (4.1): Distribution of plant families by number of species ........................... 39

Figure (4.2): Sources of folk medical knowledge .................................................... 57

Figure (4.3): Percentage of used plant parts ............................................................. 57

Figure (4.4) Procurement methods of medicinal plants ............................................ 58

Figure (4.6) : The effect of aquatic extract to 6-Syzygium aromaticum, 7-Ocimum

basilicum,8-Artemisia monosperma, 9-Coriandrum sativum, 10-Cymbopogon

schoenanthus and 11- Glycyrrhiza glabra by disc diffusion method to against E. coli

................................................................................................................................... 64

Figure (4.5) : The effect of aquatic extract to 1- Anethum graveolens, 2- Hibiscus

sabdariffa, 3- Boswellia Carterii, 4- Cuminum cyminum, 5- Trigonella foenum

graecum by disc diffusion method to against E. coli ................................................. 64

Figure (4.8) : The effect of ethanolic extract to 6-Syzygium aromaticum, 7-Ocimum

basilicum,8-Artemisia monosperma, 9-Coriandrum sativum, and 10-Cymbopogon

schoenanthus by disc diffusion method to against E. coli ......................................... 64

Figure (4.7) : The effect of ethanolic extract to 1- Anethum graveolens, 2- Hibiscus


graecum by disc diffusion method to against E. coli ................................................. 64



graecum by disc diffusion method to against S. aureus ............................................ 65

Figure (4.9) : The effect of aquatic extract to 11- Glycyrrhiza glabra by disc

diffusion method to against E. coli ............................................................................ 65

XIII

Figure (4.12): The effect of aquatic extract to 11- Glycyrrhiza glabra by disc

diffusion method to against S. aureus ........................................................................ 65

Figure (4.11) : The effect of aquatic extract to 6-Syzygium aromaticum, 7-

Ocimumbasilicum,8-Artemisia monosperma, 9-Coriandrum sativum, and 10-

Cymbopogon schoenanthus by disc diffusion method to against S.aureus ............... 65

Figure (4.14) : The effect of ethanolic extract to 6-Syzygium aromaticum, 7-

Ocimum basilicum,8-Artemisia monosperma, 9-Coriandrum sativum, and 10-

Cymbopogon schoenanthus by disc diffusion method to against S.aureus. .............. 66

Figure (4.13) : The effect of ethanolic extract to 1- Anethum graveolens, 2-

Hibiscus sabdariffa, 3- Boswellia Carterii, 4- Cuminum cyminum, 5- Trigonella

foenum graecum by disc diffusion method to against S. aureus ............................... 66



graecum by well diffusion method to against E.coli ................................................. 66

Figure (4.15) : The effect of ethanolic extract to 11- Glycyrrhiza glabra by disc


Figure (4.18) : The effect of aquatic extract to 11- Glycyrrhiza glabra by well

diffusion method to against E.coli ............................................................................. 67

Figure (4.17) : The effect of aquatic extract to 6-Syzygium aromaticum,7-Ocimum


schoenanthus by well diffusion method to against E.coli. ......................................... 67

Figure (4.20) : The effect of ethanolic extract to 6-Syzygium aromaticum, 7-

Ocimum basilicum,8-Artemisia monosperma, 9-Coriandrum sativum, and 10-

Cymbopogon schoenanthus by well diffusion method to against E.coli. .................. 67

Figure (4.19) : The effect of ethanolic extract to 1- Anethum graveolens, 2-

Hibiscus sabdariffa, 3- Boswellia Carterii, 4- Cuminum cyminum, 5- Trigonella

foenum graecum by well diffusion method to against E.coli .................................... 67



graecum by well diffusion method to against S.aureus ............................................. 68

XIV

Figure (4.21) : The effect of ethanolic extract to 11- Glycyrrhiza glabra by well

diffusion method to against E.coli ............................................................................. 68

Figure (4.24) The effect of aquatic extract to 11- Glycyrrhiza glabra by well


Figure (4.23) The effect of aquatic extract to 6-Syzygium aromaticum, 7-Ocimum


schoenanthus by well diffusion method to against S.aureus ..................................... 68

Figure (4.26) The effect of ethanolic extract to 6-Syzygium aromaticum,7-Ocimum


schoenanthus by well diffusion method to against S.aureus ..................................... 69

Figure (4.25) The effect of ethanolic extract to 1- Anethum graveolens, 2- Hibiscus


graecum by well diffusion method to against S.aureus ............................................. 69

Figure (4.27) The effect of ethanolic extract to 11- Glycyrrhiza glabra by well


Figure (4.29): (MIC) of ethanolic extracts of 4-Cuminum cyminum 5-Syzygium

aromaticum 6-Trigonella foenum graecum against E. coli....................................... 72

Figure (4.28): (MIC) of ethanolic extracts of 1- Anethum graveolens 2- Hibiscus

sabdariffa 3- Boswellia Carterii against E. coli. ........................................................ 72

Figure (4.31): (MIC) of ethanolic extracts of 9- Coriandrumand 10-

sativumCymbopogon schoenanthus against E. coli. .................................................. 72

Figure (4.30): (MIC) of ethanolic extracts of 7-Ocimum basilicum 8-Artemisia

monosperma against E. coli. ...................................................................................... 72

Figure (4.32): (MIC) of ethanolic extracts of 11- Glycyrrhiza glabra against E. coli

................................................................................................................................... 73

Figure (4.33): (MIC) of aquatic extracts of 1- Anethum graveolens 2- Hibiscus

sabdariffa 11- Glycyrrhiza glabra against E. coli..................................................... 73

Figure (4.34): (MIC) of aquatic extracts of 3- Boswellia Carterii 4-Cuminum

cyminum against E. coli............................................................................................. 73

XV

Figure (4.35): (MIC) of aquatic extracts of 5-Syzygium aromaticum 6-Trigonella

foenum graecum against E. coli. ................................................................................ 73

Figure (4.36): (MIC) of aquatic extracts of 7-Ocimum basilicum 8-Artemisia

monosperma 9- Coriandrumand 10- sativumCymbopogon schoenanthus against E.

coli. ............................................................................................................................ 74

Figure (4.37): (MIC) of ethanolic extracts of 1- Anethum graveolens 2- Hibiscus

sabdariffa against S.aureus. ....................................................................................... 77

Figure (4.38): (MIC) of ethanolic extracts of 3- Boswellia Carterii 4-Cuminum

cyminum against S.aureus. ........................................................................................ 77

Figure (4.39): (MIC) of ethanolic extracts of 5-Syzygium aromaticum 6-Trigonella

foenum graecum against S.aureus. ............................................................................ 77

Figure (4.40): (MIC) of ethanolic extracts of 7-Ocimum basilicum 8-Artemisia

monosperma against S.aureus.................................................................................... 77

Figure (4.41): (MIC) of ethanolic extracts 9- Coriandrumand 10- sativum

Cymbopogon schoenanthus against S.aureus ............................................................ 78

Figure (4.42): (MIC) of ethanolic extracts of 11- Glycyrrhiza glabra against

S.aureus...................................................................................................................... 78

Figure (4.43): (MIC) of aquatic extracts of 1- Anethum graveolens 2- Hibiscus

sabdariffa and 11- Glycyrrhiza glabra against S.aureus. ......................................... 78

Figure (4.44): (MIC) of aquatic extracts of 3- Boswellia Carterii 4-Cuminum

cyminum against S.aureus. ........................................................................................ 78

Figure (4.45): (MIC) of aquatic extracts of 5-Syzygium aromaticum 6-Trigonella

foenum graecum against S.aureus. ............................................................................ 79

Figure (4.46): (MIC) of aquatic extracts of 7-Ocimum basilicum ......................... 79

8-Artemisia monosperma ........................................................................................... 79

9- Coriandruman sativum .......................................................................................... 79

10-Cymbopogon schoenanthus against S.aureus. ...................................................... 79

XVI

Figure (4.47): the MIC of aquatic extract of 12- Cuminum cyminum with Ocimum

basilicum and 13- Artemisia monosperma with Coriandrum sativum against S.

aureus......................................................................................................................... 83

Figure (4.48): the MIC of ethanolic extract of 12- Cuminum cyminum with

Ocimum basilicum and 13- Artemisia monosperma with Coriandrum sativum

against S. aureus. ....................................................................................................... 83

Figure (4.49): the MIC of aquatic extract of 12- Cuminum cyminum with Ocimum

basilicum and 13- Artemisia monosperma with Coriandrum sativum against E.coli 84

Figure (4.50): the MIC of ethanolic extract of 12- Cuminum cyminum with

Ocimum basilicum and 13- Artemisia monosperma with Coriandrum sativum

against E.coli .............................................................................................................. 84

XVII

List of Table

Table (2.1) : Classification of Ocimum basilicum .................................................... 11

Table (2.2) : Classification of Hibiscus sabdariffa L. ............................................... 12

Table (2.3) : Classification of Trigonella foenum- graecum L. ................................ 13

Table (2.4) : Classification of Coriandrum sativum L. ............................................. 14

Table (2.5) : Classification of Anethum graveolens L. ............................................. 15

Table (2.6) : Classification of Syzygium aromaticum. L ........................................... 16

Table (2.7) : Classification of Glycyrrhiza glabra L ................................................ 17

Table (2.8) : Classification of Cuminum cyminum L. ............................................... 18

Table (2.9) : Classification of Cymbopogo schoenanthus.L ..................................... 19

Table (2.10) : Classification of Boswellia sacra Flueck ........................................... 20

Table (2.11) : Classification of Artemisia monosperma Delile ................................ 21

Table (2.12) : Classification of Escherichia coli ...................................................... 22

Table (2.13) : Classification of Staphylococcus aureus........................................... 23

(Table 3.1) List of medicinal plants used in the antimicrobial & synergistic assay . 32

(Table 3.2) List of chemicals used in the study ........................................................ 33

Table (4.1) : Distribution of plant families by number of species ............................ 38

Table (4.2): Classification of the most used plants in popular treatments in the Gaza

Strip ............................................................................................................................ 41

Table (4.3) : Diversity of medicinal use- categories ................................................ 56

Table (4.4) : Antimicrobial Activity of Plant extracts against E. coli by well

diffusion method and disc diffusion method ............................................................. 62

Table (4.5): Antimicrobial Activity of Plant extracts on Staphylococcus aureus by

well diffusion method and disc diffusion method ..................................................... 63

Table (4.6) : The MICs & MBCs of plant extracts against E. coli .......................... 71

Table (4.7): The MICs & MBCs of extracts against S.aureus .................................. 75

XVIII

Table (4.8): Antimicrobial Activity between Plant extracts on E. coli by well


Table (4.9): Antimicrobial Activity between Plant extracts on S. aureus by well


Table (4.10): The minimum inhibitory concentration (MIC) between plant extracts

against isolated bacteria ............................................................................................. 82

XIX

List of Abbreviated Terms

MIC Minimum Inhibitory Concentration

MBC Minimum Bactericidal Concentration

DMSO Dimethyl Sulfoxide

TTC Triphenyl Tetra zolium Chloride

MHB Muller-Hinton Broth

MHA Muller-Hinton Agar

milligram/milliliter mg/ml

Millimeter mm

CFU Colony Forming Unit

1

Chapter 1

Introduction

2

1.1 Overview

Throughout history, human relied on various natural materials as a treatment

for several diseases. But it has been replaced by the most natural products with

synthetic drugs that are based on modern chemistry and biotechnology over the past

few decades (Jaradat, 2005).

We cannot be denied that the discovery of antibiotics is one of the most

important discoveries that have helped to combat bacterial infections that cause

various diseases but because of the erroneous and random use of antibiotic infectious

bacteria have formed resistance against these drugs, this is in addition to the side

effects that appear due to the use of antibiotics (Jouda et al., 2015) , for these reasons

there has been an increasing interest in the study of medicinal plants and their

traditional use in different parts of the world.

Although modern medicine may be available in many developing countries

but about two thirds of the population relies heavily on traditional practitioners and

medicinal plants to meet primary health care needs. Traditional herbal medicine is

often been used for historical, cultural, and ecological reasons, in particular this is

due to continued availability, better compatibility and high acceptance (Kunwar et

al., 2010).

Reliance on derivative treatments of medicinal plants, especially in

developing countries where the importance of modern drugs often absent or simply

too expensive. Low economic situation of developing countries leads to higher prices

for medicines and this makes medicinal plants and traditional medicine more

desirable by the local population (Randriamiharisoa et al., 2015).

Most of the world’s population still use herbs as a first choice to treat

diseases but besides the medicine field several mass-consumed eatables and

stimulants also have beneficial side-effects (Nemeth, 2012). The knowledge of

medicinal plants has been accumulated in the course of many centuries and the used

of these plant by indigenous cultures are useful not only to maintain their

biodiversity and cultural traditions but also for the health care community and the

development of drugs in the present and the future (Abou Auda, 2011).

3

Large population density in developing countries rely on plant resources for

health care. Allopathic medicine can cure a wide range of diseases, but rising prices

in some cases, side effects cause a lot of people to return to herbal medicines, which

tend to have fewer side effects (Mahmoud and Gairola, 2013)

WHO estimated in 2002 that 80% of the world's population in developing

countries rely mainly on medicinal plants and traditional medicine practitioners to

meet the primary health care needs (Ali-Shtayeh et al., 2011) and previous research

has shown that about 85% of traditional treatment involves in the use of plant

extracts (Quiroga et al., 2012).

However, Western countries have also seen a growing interest in natural

medicine. In the particular, it has spread in the herbal medicine market and become

rich in pharmacies and many stores (Jaradat, 2005).

Medicinal plants can be of great importance in the daily life of the people

who live near the places where they grow, not only for healing traditions, but as a

commodity out to urban areas where they are not found locally to be sold in the

market (Randriamiharisoa et al., 2015).

Several studies indicate that some plants contain compounds such as peptides,

aldehydes, alkaloidal constituents, some essential oils, phenols and water, ethanol,

chloroform, methanol. These plants shown therapeutic application against some

human diseases caused by bacteria, fungi and viruses (El Astal et al., 2005).

These natural products are still a major source of new drug discoveries: for

example, 65% of the drugs that were approved for marketing between the years 1983

and 1994 were based on natural sources (Jaradat, 2005). Several studies have shown

the possibility of using plant extracts in the therapy of some diseases caused by

bacterial infection and confirmed the effect of these extracts on bacterial growth,

these studies proved that some Palestinian plants have a significant impact on these

infectious bacteria (Ali-Shtayeh et al., 2014).

In the late 19th century scientific tests were conducted for the first time on

the properties of some plants in their resistance to certain microbes. The results were

documented today as a reliable source of modern medicine(Abu Shanab et al., 2015).

So medicinal plants have a promising future because there are about half million

4

plants around the world, and most of the medical activities of them have not

investigated yet, which could be decisive in the treatment of present or future studies

(Rasool Hassan, 2012).

In Palestine, like in many developing countries, medicinal plants play an

important role in primary health care and are widely used as complementary and

alternative medicine in the Traditional Arabic Palestinian Herbal Medicine, for the

maintenance of health, and for the treatment of various illnesses including chronic

diseases (Ali-Shtayeh et al., 2013). These herbal medicine considered an integral

part of the Palestinian culture and plays a pivotal and indispensable role in the

current folk healthcare (Jaradat, 2005).

On Palestine culture and economy the impact of local plant varieties is

significant. Many wild plant species are extensively used in Palestine cuisine and

many people, mostly in rural areas, still widely use medicinal plants for the treatment

of burns, disease and other (Hinnawi, 2010).

In Palestine, has a lot of ethnobotanical surveys were conducted to highlight

the importance of herbal medicine in the treatment of illnesses and diseases (Abd

Rabou et al., 2008). but there is a significant lack of information on the number and

type of medicinal plants used in Palestine, especially in Gaza Strip (Abou Auda,

2011).

The local market of medicinal plants in Gaza strip not documented so our

study focused on the Gaza strip and its medicinal plant markets. We focused also on

the varieties of medicinal plants used in medical treatments by traditional healers.

This study was the starting point for identifying the main groups of medicinal plants

used by folk medicine practitioners and the benefits derived from these plants.

Few studies have found combining antimicrobial agent with crude plant

extracts increase the efficiency of them against some kinds of pathogenic organism

(Elbashiti., et al 2011), so we focused also on the effect of extract of some plants on

S.aureus and E.coli bacteria and the synergistic effect of some plants on these

pathogenic bacteria.

5

1.2 Main objective:

The main objective of this study is to identify and classify the most important

medicinal plants that are used by folk therapists in Gaza Strip.

1.3 Specific Objectives:

1- To documenting and preserving traditional knowledge on the use of

medicinal plants among the local population in the Gaza Strip for the

management of various diseases.

2- To study the impact and effectiveness of certain medicinal plants used in the

treatment of some diseases

3- To ascertain the detailed information about the use of plants and healing

practices among citizens and therapists on traditional medicine.

4- To find out the efficiency of the most popular extracts against pathogenic

bacteria.

5- To determine the Minimum Inhibitory Concentration (MIC) and Minimum

Bactericidal Concentration (MBC) of the plant extracts.

6- To determine the synergistic effect of some plant extracts with each other on

pathogenic bacteria.

6

1.4 Significance :

This study is the current attempt to be the first of its kind in the Gaza Strip. It

will identify and give useful information on the classification and identification of

the most important medicinal plants in Gaza Strip used by locals and therapists on

traditional medicine in the treatment of some diseases. Moreover, the results of this

study should be a top priority for the people in terms of knowledge, management,

use, and research potential in the future.

7

Chapter 2

Literature Review

8

2.1 The history of herbal treatment

The concept of the use of plants by humans has changed since 1880s until the

present day. The method of use of some plants varies according to the culture of man

and his surrounding environment. Man depends on plants as a food, economic,

cultural and therapeutic source(Ari et al., 2015).

Herbal treatment is one of the oldest methods of treatment used by mankind

on the face of the earth, where the profession was passed from generation to

generation orally and without codification, where it became a traditional way among

the peoples and one of the most important habits passed through generations

(Revathi and Parimelazhagan, 2010)

The increasing demand for medicines manufactured from natural herbs led to

the exploitation of these plants in an uncontrolled and uncoordinated and this led to

the threat of natural plants in most countries (Kunwar et al., 2013).

Herbal treatment have not been well studied, tested, or documented in many

developing countries, and most of the information about these plants are still in the

hands of traditional healers and knowledge of healers is either lost or passed to next

generation by the word of mouth (Amiri and Joharchi, 2013). And because the

value of knowledge of treatments is eroding with the older generation, the

government should make efforts to preserve this knowledge, and promote the

traditional treatments and protection of natural resources of medicinal and aromatic

plants, and the initiation of research and development activities in this field. There is

a need to strengthen production and trade to get the commercial benefits of medicinal

and aromatic plants of the country's natural resources( Handa et al., 2006).

Over the long history of traditional use of medicinal plants in Palestine study

the use of ethnobotanical represents a unique opportunity (Ali-Shtayeh et al., 2015).

West Bank and Gaza Strip have lots of herbs and medicinal plants, which are widely

used for the treatment of various diseases and also in high demand (Abou Auda,

2012). But the constant and ongoing conflict with Zionist enemy has led to the

deterioration of health care services, natural resources and plant reduced the ability to

exploit these resources despite the popular awareness of these resources and their

uses in folk medicine (Handa et al., 2006).

9

2.2 Ethnobotany

The definition of ethnobotany can be summed up in four words: people,

plants, interactions, and uses, so its includes all types of relationships between

people and plants (Abbasi et al., 2011) and focuses on how and why local people use

medicinal herbal remedies and works to understand the relationship between humans

and local plants in their land (Hinnawi, 2010).

2.2.1 Traditional ethnobotanical markets

Around the world the traditional ethnobotanical markets has been recognized

as that which sell and trade medicinal herbs and their derivatives as it became a

regional trade between the countries of the world and express the culture of some

peoples and their traditions (Tinitana et al., 2016).

Traditional ethnobotanical markets includes two main branches: the first is

the market of traditional medicinal plants where the plants are sold raw, untreated

and dried. The second branch is the market for medicines manufactured from these

plants where the plants are sold individually or mixed with several other herbs to

treat a specific disease (Randriamiharisoa et al., 2015)

2.3 Medicinal plants

Medical plants are plants that have medicinal properties and are used for

therapeutic purposes because some or all of their parts contain an effective substance

for the treatment of diseases (Nemeth, 2012). It is also known as plants used to

maintain health or treat some diseases. The methods of using these plants vary from

one country to another around the world according to the traditions of this country

(Smith-Hall et al., 2012).

Any plant may be considered a medicinal plant if it contains an effective

substance whether this plant is wild or implanted and whether it is Vegetables , fruits

, or spices. Most of these plants were used mainly to save food or spices or to treat

gastric disorders because of their beneficial effects and taste (Nemeth, 2012).

2.3.1 Future of Medicinal Plants

Medicinal plants have a promising future, where all around the world about

half a million plant , and most of them their medical activities have not investigated

10

yet which could have been decisively in the treatment of many diseases (Rasool

Hassan, 2012). Recently, modern medicine has been adopted on the use of plant

extracts in order to reduce the use of chemical treatments and reduce their side

effects and with great technological advances, researchers can recognize the

importance of these large numbers of plants and use them to treat diseases as an

alternative to these chemical compounds (Yuan et al., 2016).

The effectiveness of some medical extracts has also been proven in the

treatment of certain incurable diseases such as cancer. The most important

characteristic of medicinal plants is therapeutic synergy where more than one plant

extract can be used simultaneously to treat a specific disease (Rasool Hassan, 2012).

Where popular healers famous for the creation of mixtures of a group of herbs

together to treat some diseases on the grounds that the plant cannot cure the disease

alone. Therefore, in this study we tested the synergistic effect of extracts to some of

these plants on some types of human pathogenic bacteria and their effectiveness in

eradicating the disease.

2.4 Plants used in extraction

The following are some of the plant extracts that have been studying its

antimicrobial activity against some pathogenic clinically isolated bacteria

11

2.4.1 Ocimum basilicum (basil) : اش٠سا

Basil or sweet basil (Ocimum basilicum L) is a plant that belongs to the

family Lamiaceae and has shown its potential to be therapeutic in averting several

diseases in various countries, some studies have shown that basil leaves contain

antioxidants, anti-bacterial and antiviruses, as well as cancer treatment, also basil oil

used in the prevention and treatment of cardiovascular disease (Sakr and Al-

Amoudi, 2012) (Table 2.1).

Table (2.1) : Classification of Ocimum basilicum

Kingdom Plantae

Ocimum basilicum L

Subkingdom Tracheobionta

Superdivision Spermatophyta

Division Magnoliophyta

Class Magnoliopsida

Subclass Asteridae

Order Lamiales

Family Lamiaceae

Genus Ocimum L.

Species Ocimum basilicum L

12

2.4.2 Hibiscus sabdariffa L. (Roselle) : اىشوذ٠ح

Roselle (Hibiscus sabdariffa L.) belongs to the family Malvaceae, is an important

annual crop grown successfully in tropical and sub-tropical climates

The whole plant can be used as a drink or dried sepals can be used only by soaking it

in water or boiling it and being taken as a hot drink, around the world roselle plant

have been used for its medicinal importance such as hypertension, pyrexia and liver

damage Recently the sepal extract has been used as an effective treatment against

leukemia (Mohamed et al., 2012 ). (Table 2.2).

Table (2.2) : Classification of Hibiscus sabdariffa L.

Kingdom Plantae

Hibiscus sabdariffa L




Class Magnoliopsida

Subclass Dilleniidae

Order Malvales

Family Malvaceae

Genus Hibiscus L.

Species Hibiscus sabdariffa L

13

2.4.3 Trigonella foenum- graecum L. (Fenugreek seed) : اسثح

Fenugreek (Trigonella foenum-graecum L.) is a plant that belongs to the

Fabacecae family. Fenugreek seeds have been known and valued as medicinal

material from very early times. The leaves and seeds of the fenugreek are used in

many countries in the treatment of some diseases such as diabetes, high cholesterol

and cancer as contain antibiotics for some types of bacteria and other diseases

(Moradi kor et al., 2013 ). (Table 2.3).

Table (2.3) : Classification of Trigonella foenum- graecum L.

Kingdom Plantae

Trigonella foenum -graecum L.




Class Magnoliopsida

Subclass Rosidae

Order Fabales

Family Fabaceae

Genus Trigonella L.

Species Trigonella foenum-graecum

L.

14

2.4.4 Coriandrum sativum (Coriander) : اىضتشج

Coriander (Coriandrum sativum L.) which belongs to the family Apiaceae is

one of miraculous herb that functions as both, spice as well as herbal medicine, it is

mainly cultivated from its seeds throughout the year.

Coriander is a seasonal herb that spreads throughout the mediterranean and is known

for its great medicinal value. This plant is used in folk medicine treatment because of

its highly aromatic and has multiple uses in food and in other industries (Bhat et al.,

2014). (Table 2.4).

Table (2.4) : Classification of Coriandrum sativum L.

Kingdom Plantae

Coriandrum sativum L.




Class Magnoliopsida

Subclass Rosidae

Order Apiales

Family Apiaceae

Genus Coriandrum L

Species Coriandrum sativum L.

15

2.4.5 Anethum graveolens (Dill) : ػ١ خشادج

Anethum graveolens L. commonly known as dill belonging to the family

Apiaceae, originated from mediterranean and West Asia, dill has a long history in

medicinal uses, where the leaves of this plant are used in food because it is rich in

minerals such as phosphorus, potassium and magnesium, as the seeds are used in the

treatment of cystitis, liver disease and insomnia (Chahal et al., 2017). (Table 2.5).

Table (2.5) : Classification of Anethum graveolens L.

Kingdom Plantae

Anethum graveolens L




Class Magnoliopsida

Subclass Rosidae

Order Apiales

Family Apiaceae

Genus Anethum L.

Species Anethum graveolens L

16

2.4.6 Syzygium aromaticum L. (Clove) : امشف

Syzygium aromaticum L. commonly known as Clove belonging to the family

from family Myrtaceae. The part used from the plant is the flower buds where they

are collected before flowering it is one of the most valuable spices that has been used

for centuries as food preservative and for many medicinal purpose. Clove is an

important medicinal plant due to the wide range of pharmacological effects

consolidated from traditional use for centuries. Clove oil is effective as a broad

spectrum antibiotic it is effective for the treatment of diarrhea and eradicating the

fungus that caused athlete's foot (Cortés-Rojasf et al., 2014). (Table 2.6).

Table (2.6) : Classification of Syzygium aromaticum. L

Kingdom Plantae

Syzygium aromaticum.L




Class Magnoliopsida

Subclass Rosidae

Order Myrtales

Family Myrtaceae

Genus Syzygium P

Species Syzygium aromaticum.L

17

2.4.7 Glycyrrhiza glabra L. (Licorice) : اؼشق عط

Glycyrrhiza glabra L., belonging to the family Fabaceae, is a plant which

grows in Egypt and other countries of the world, and has roots of great nutritional

and medical value (Zadeh et al,. 2013). Licorice is used in folk medicine as a

treatment for stomach and duodenal ulcers and is a good antidote to inflammation

and allergies (Damle, 2014 ). (Table 2.7).

Table (2.7) : Classification of Glycyrrhiza glabra L

Kingdom Plantae

Glycyrrhiza glabra L




Class Magnoliopsida

Subclass Rosidae

Order Fabales

Family Fabaceae

Genus Glycyrrhiza L

Species Glycyrrhiza glabra L

18

2.4.8 Cuminum cyminum L. (Cumin) : اى

Cuminum cyminum L., belonging to the family Apiaceae, it is considered as

one of the old medicinal herb in both Asia, Africa and Europe. Cumin was used in

the treatment of gastrointestinal diseases, gynecological diseases and also in the

treatment of dental pain and epilepsy, the seeds were also documented as stimulant,

carminative and astringent (Gohari, A. R., and Saeidnia, S., 2011). (Table 2.8).

Table (2.8) : Classification of Cuminum cyminum L.

Kingdom Plantae

Cuminum cyminum L




Class Magnoliopsida

Subclass Rosidae

Order Apiales

Family Apiaceae

Genus Cuminum L.

Species Cuminum cyminum L

19

2.4.9 Cymbopogo schoenanthus.L (camel grass) : اسفا تش

Cymbopogo schoenanthus.L belongs to the family Poaceae, it is characterized

by a distinctive and strong smell and is considered an important medicinal plant. It is

a powerful sedative for the digestive system and antiparasitic, Some studies have

shown the role of the camel grass in the treatment of heart disease and high

cholesterol (Sousa et al,. 2005 ). (Table 2.9).

Table (2.9) : Classification of Cymbopogo schoenanthus.L

ingdom Plantae

Cymbopogo schoenanthus.L




Class Liliopsida

Subclass Commelinidae

Order Cyperales

Family Poaceae

Genus Cymbopogon Spreng.

Species Cymbopogo schoenanthus.L

20

2.4.10 Boswellia sacra Flueck (Olibanum or frankincense) اذوش :زص اثا

Boswellia sacra Flueck or olibanum is a plant product. It is an oleo-gum-resin

produced by several species of tree belonging to the family Burseraceae. Since

ancient times, frankincense has been used in many countries, including Middle

Eastern countries, to treat many diseases, especially chronic inflammatory diseases.

Extensive studies on frankincense and its effect showed that this gum resin can

reduce the inflammation and pain in the body and relieve the related symptoms of

many diseases. The anti-inflammatory, anti-arthritic, anti-microbial and analgesic

effect, in addition to his role in the treatment of heart disease, asthma, skin, prostate

cancer and diabetes (Hamidpour et al., 2015). (Table 2.10).

Table (2.10) : Classification of Boswellia sacra Flueck

Kingdom Plantae

Boswellia sacra Flueck




Class Magnoliopsida

Subclass Rosidae

Order Sapindales

Family Burseraceae

Genus Boswellia

Species Boswellia sacra Flueck

21

2.4.11 Artemisia monosperma Delile (white wormwood) : اؾ١ر

Artemisia monosperma Delile commonly known as belong to the family

Asteraceae. The main medicinal use of this plant is that it works on the treatment of

diabetes, anti-convulsions, anti-microbial, and anti-malarial treatment, on the other

hand this plant is widely used in folk medicine as a treatment for hypertension

(Moufid, A., and Eddouks, M., 2012). (Table 2.11).

Table (2.11) : Classification of Artemisia monosperma Delile

Kingdom Plantae

Artemisia monosperma Delile




Class Magnoliopsida

Subclass Asteridae

Order Asterales

Family Asteraceae

Genus Artemisia L.

Species Artemisia monosperma

Delile

22

2.5 Microorganism

Clinical isolated bacteria used in the study are Escherichia coli and

Staphylococcus aureus.

2.5.1 Escherichia coli

Escherichia coli is a Gram negative rod non-sporing bacilli belongs to the

family Enterobacteriaceae, most E.coli are normal commensals found in the

intestinal tract, Pathogenic E.coli can be classified in to pathotypes by their

virulence factors, together with the type of disease, virulent strains causes

serious infections include meningeal, urinary tract, gastrointestinal (Welch,

2006). (Table 2.12).

Table (2.12) : Classification of Escherichia coli

Domain Bacteria

Escherichia coli

Phylum Proteobacteria

Class Gammaproteobacteria

Order Enterobacteriales

Family Enterobacteriaceae

Genus Escherichia

Species Escherichia coli

23

2.5.2 Staphylococcus aureus

Staphylococcus aureus (S.aureus) bacteria is a Gram-positive belongs to the

family Staphylococcaceae which characterized by individual cocci, which divide in

more than one plane to form grape-like clusters, it is are facultative anaerobic, non-

motile and do not form spores (Harris et al,. 2002).

S.aureus is a major pathogen that infects both hospitalized patients with

decreased immunity, and healthy people in the community (Harris et al,. 2002), this

bacteria cause many type of diseases where their proliferation in the blood leads to

secondary pneumonia, endocarditis, osteoarthritis and arthritis (Jouda, 2013). (Table

2.13).

Table (2.13) : Classification of Staphylococcus aureus

Domain Bacteria

Staphylococcus aureus

Phylum Firmicutes

Class Bacilli

Order Bacillales

Family Staphylococcaceae

Genus Staphylococcus

Species Staphylococcus aureus

24

2.6 Previous Studies:

In 2015 Randriamiharisoa et al identified and generated a list of medicinal

plants sold in the traditional markets of the capital of Madagascar, Antananarivo and

obtain an idea of the amount of medicinal plants sold on the large outdoor markets,

and estimates the economy of the medicinal plant markets.

In 2015 Ali-Shtayeh et al investigated a broad range of Palestine flora using

a methodology that enabled the discovery of plants with potential medicinal

characteristics. Some of the plants identified have a history of human use, while

others have never been used before for medicinal purposes. plant samples were

collected from 76 natural sites distributed in different geographical and climate

zones. Samples were evaluated for thirteen kinds of activity protected from disease

using bioassays field deployment based on the screen to technical nature of the

World Institute for Bio-Exploration. More than 1470 plant samples derived from 588

plant species belonging to 100 families were screened. Approximately 329 species

(56%) belonged to 12 families, notably the Papilionaseae, Asteraceae, Liliaceae,

Lamiaceae, Brassicaceae, and Apiaceae families.

In 2010 Revathi and Parimelazhagan observed that the tribal’s use 70 wild

valuable plant species belonging to 42 families which identified with pertinent

information with regard to their botanical and local name, family, parts used by the

local people for different human ailments. The common diseases treated by folk

therapies were asthma, digestive problems, paralyzes, skin diseases and diabetes. the

study provides an ethnobotanical data of the medicinal plants used by the tribal

people of Irulas to cure several diseases. and promoted a practical use of medical

plant which must be continued focusing on its pharmacological effectivness. Further

detailed exploration and collection of ethnobotanical information, chemical studies

and screening for medicinal properties will provide cost effective and reliable source

of medicine for the welfare of humanity.

In 2012 Abou Auda Performs an extensive survey on medicinal plant

diversity in the flora of Gaza Valley showed that a total of 183 species under 33

families are represented in the flora. The investigation for medicinal plant diversity

25

within these families revealed that out of these 183 species, 52 are medicinal and

many of them are mentioned at different literature.

In 2011 Abou Auda collected knowledge of plant species, which used by

the local inhabitants of villages of the middle area of Gaza Strip, Palestine. A total of

19 plant species belonging to 19 genera and 16 families have been listed which are

being utilized for treating 23 different diseases during 2008-2009. Herbs were found

to be the most consumed plants (10 species), followed by trees (5 species) and

shrubs (4 species)

In 2015 Ari et al indicated that 130 plants taxa belonging to 39 families were

recorded and collected. Hundred and seventy-eight different uses of these plants

were documented and used generally for medicinal (84), food (68), fodder (16),

household goods (3), dyes (3), handicrafts (3) and religious (1).

In 2012 Quiroga et al identified 258 uses traditional medicines, spanning a

total of 13 disease categories, including 91 species of local and foreign plants and

one unknown type of plant specimen. The most often treated digestive disorders

(55%) with medicinal plants, followed by the pain of musculoskeletal system (25%)

and skin disorders (24%). Hospital information indicates that the most common

diseases are acute respiratory infections (47%), diseases of acute diarrhea (37%).

And mostly used herbal remedies in the form of teas and decoctions. Informants are

used mainly native plant species, despite the fact that alien species have been

introduced to drugs.

In 2015 Rakotoarivelo et al identified the six most common diseases,

namely malaria, diarrhea and stomach pains, cough, bilharzia and dysentery. Among

the 209 species of plants that have been identified as the use of drugs, and used 83

species belonging to 49 families and 77 genera for the treatment of these diseases.

In 2014 Husein et al examined the ethanol extracts of six types of medicinal

plants that belong to three families on the basis of their uses in the Palestinian Arab

traditional herbal medicine for the treatment of various ailments as stomach acidity,

atherosclerosis, cancer, diabetes and food toxicity The extracts were tested against:

six bacterial strains including Staphylococcus aureus, Proteus vulgaris,

Pseudomonas aeruginosa, Salmonella typhi, Escherichia coli and Klebsiella

26

pneumoniae; five Candida albicans isolates, and two dermatophytes: Microsporum

canis, and Trichophyton rubrum

In 2015, Abu Shanab et al examined nine medicinal plants grown in

Palestine were examined in vitro for possible antibacterial activity against 6 bacterial

strains through partial dilution and propagation methods. Three plant extracts have

almost antibacterial activity against both gram-positive and gram-negative bacteria.

There are other three plant extracts were more active against gram-positive than

against gram-negative bacteria and these plant extracts were from the cycoproteum

spinosome, lunonia inermes and pseudium guagava. This study supports the use of

traditional medicine for various plant extracts in the treatment of various infections

caused by pathogenic bacteria in Palestine, either using single or combined extracts.

In 1998 Ali-Shtayeh et al evaluated ethanolic and aqueous extracts of 20

Palestinian plant species used in folk therapy were investigated for their

antimicrobial activities against five bacterial species (Staphylococcus aureus,

Escherichia coli, Klebsiella pneumonia, Proteus vulgaris, Pseudomonas aeruginosa)

and one yeast (Candida albicans). This study shown that most of the studied plants

are potentially a good source of antimicrobial agents and demonstrates the

importance of such plants in medicine and in assisting primary health care in this part

of the world.

In 2015 Elkichaoi et al detected the effectiveness of some fruit types extracts

against some types of pathogenic bacteria that known for their resistance to

antibiotics, sensitivity of each fruit extracts were tested by serial micro dilution

method and agar disc diffusion method, the efficacy of these extracts has been

demonstrated against Gram positive bacteria than Gram-negative bacteria due to the

different cell-wall shape of different types of bacteria.

In 2011 Elbashiti et al evaluate the antimicrobial activity of some local plant

extracts alone and synergism with antibiotics against E.coli and S.aurues. The results

showed that the plant extracts alone have rare effect against isolated bacteria, but the

extracts and there synergistic effect with antibiotics appear to be an inhibitor to

S.aurues (Gram positive bacteria) rather than E.coli (Gram-negative bacteria).

27

Chapter 3

Materials and Methods

28

3.1 Materials

3.1.1 Study Area

The Gaza strip

The total area of the historic Palestine is 27.009 km2. The area of west bank

is 5844 km2. The Gaza strip, an area of 365 km2 .The Gaza Strip is a narrow strip of

land on the Mediterranean coast. It borders the occupied lands to the east and north

and Egypt to the south. It is approximately 41 kilometers long, and between 6 and 12

kilometers wide (Environmental Assessment of the Gaza Strip, 2009).

The study area has a typical semi –arid Mediterranean climate, hot in summer

and cold in winter. The average daily mean temperature ranges from 25C˚ in summer

to 13C˚ in winter, with the average daily maximum temperature range from 29C˚ to

17C˚ and the minimum temperature range from 21C˚ to 9C˚ in summer and winter

respectively. The daily relative humidity fluctuates between 65% in daytime and

85% at night in summer and between 60% and 80% in winter (United Nations

Environmental Program – UNEP, 2003).

29

Figure (3.1) : Gaza strip map ( source : www.arij.org )

3.1.2 Study population

The interview was conducted with 20 informants from different areas of Gaza

Strip, including 2 females and 18 males, their ages ranged from 30 to 65 years, as

shown in Figure (3.2), (3.3),respectively.

http://www.arij.org/

30

Figure (3.2): Distribution of study population by gender

Figure (3.3): Distribution of study population by age

0

2

4

6

8

10

12

14

16

18

male

female

No. of informant

0

1

2

3

4

5

6

7

8

9

30-44 45-65 >65

No. of informant

31

Interviews were conducted with informants of different educational levels. Figure

(3.4) shows the level of education, (Illiterate, primary, secondary, diploma,

university).

Figure (3.4): Distribution of study population by educational level.

3.1.3 Medicinal Plants Markets

Medicinal plants sold in popular markets, which are special shops sell these

medicinal plants, mixtures or oils made from these plants. There are also centers use

these plants in alternative medicine treatment.

During this study, field work and survey to the vast majority of these markets

and centers where the most common plant species used by folk therapists in treating

some diseases. They were distributed throughout the Gaza Strip (7 in Gaza City, 2 in

Jabalya, 3 in Khan Younis, 3 in Nuseirat, 2 in Deir al-Balah, 1 in Rafah, 2 in

Maghazi).

3.1.4 Plant Sample Collection:

The Plants whose extracts were used in this study consisted of Ocimum

basilicum (basil), Hibiscus sabdariffa (Roselle), Trigonella foenum-graecum

(Fenugreek seed), Coriandrum sativum (Coriander), Anethum graveolens (Dill),

0

1

2

3

4

5

6

Illiterate primary Middle secondary university

No. of informant

32

Syzygium aromaticum (Clove), Glycyrrhiza glabra (Licorice), Cuminum cyminum

(Cumin), Cymbopogon schoenanthus (Camel grass), Boswellia sacra (Olibanum)

and Artemisia herba-alba (white wormwood) were bought from Al-Zawea popular

market.(Table 3.1).

(Table 3.1) List of medicinal plants used in the antimicrobial & synergistic assay

Tested

part

Family Local Arabic

Name

Scientific Name No.

Seeds Umbelliferae ػ١ ادشادج Anethum graveolens L 1

Sepals Malvaceae اىشوذ٠ Hibiscus sabdariffa L 2

Gum Burseraceae زص اثا

اذوش

Boswellia sacra Flueck 3

Dry

fruits

Apiaceae اى Cuminum cyminum L 4

Seeds Fabaceae اسثح Trigonellafoenum-

graecum L 5

Flower

buds

Myrtaceae امشف Syzygium aromaticum

(L.) Merrill & Perry 6

Leaves Lamiaceae اش٠سا Ocimum basilicum L. 7

Leaves Asteraceae اؾ١ر Artemisia monosperma 8

Seeds Apiaceae اىضتشج Coriandrum sativum L 9

leaves Poaceae اسفا تش Cymbopogon

schoenanthus Spreng 10

roots Fabaceae اؼشق عط Glycyrrhiza glabra L. 11

33

3.1.5 Culture media and chemicals

For carrying out this study we used Mueller Hinton Broth and Mueller-Hinton

agar. Also ethanol and distilled water used for extraction process, Triphenyl Tetra

zolium Chloride (TTC) were used as microbial growth indicator. Dimethyl sulfoxide

(DMSO) .(Table 3.2).

(Table 3.2) List of chemicals used in the study

3.1.6 Microorganisms

Two bacterial species, gram- positive Staphylococcus aureus and gram- negative

Escherichia coli were obtained from the biology department of the Islamic

University of Gaza (IUG) and the microbiology department of Al-Shifa hospital.

3.2 Methods:

3.2.1 Ethnobotanical Methods:

3.2.1.1 Survey and Identification:

Ethnobotanical information was obtained through interview which was

applied on People who use medicinal plants in folk remedies in popular markets of

Gaza city.

Questions addressed to the therapist were therefore, mainly focused on the

purpose of plant application, parts used, the manner of their preparation and

Chemicals Manufacturer

Mueller Hinton agar Liofilchem

Mueller Hinton Broth Liofilchem

Dimethyl Sulphoxide (DMSO) Applichem

Ethanol, Distilled Water CHEM Limited

Triphenyl Tetra zolium Chloride

(TTC)

Applichem

34

administration, forms of use, purchase method, method of storage and collection,

types of diseases which treated and how to prepare some popular prescriptions

Figure (3.5).

Figure (3.5) : Photograph shows interview with informants

3.2.2 Extraction methods :

3.2.2.1 Preparation of plant extract:

Ten grams of plant material were placed in a flask with 150 ml of solvent

(Ethanol or distilled water) and placed in the microwave extractor for 60s (1min) to

be exposed. After being taken and cooled in room temperature for one min, they

were placed again in the microwave extractor and returned 12 times. The previous

steps were repeated with the plant samples eleven times using 80% ethanol and again

using distilled water (Jin-Yu Hao et al., 2002). The extracts were transported into

glass watch and by oven at 45ºC overnight to remove the solvent.

One gram of each extract was dissolved in 5 ml of dimethylsulfoxide (DMSO).

Thus 200 mg/ml of stock was obtained as a standard concentration of extracts

(Jouda, 2013). The dried extracts were stored in sterile glass bottles at -20 ºC until

use.

35

3.2.2.2 Evaluation of antibacterial activity of plant extracts by disc diffusion

method:

Agar disc-diffusion method was followed to determine the antimicrobial activity

of the selected plant extracts against test microorganism, sterile filter paper discs

approximately 6mm in diameter were impregnated with stock extracts and placed on

the surface of a MHA that has been inoculated with test microorganism. After 24

hours, the antimicrobial activity was evaluated by measuring zones of inhibition of

microbial growth surrounding the plant extracts (Sharma, 2011). The assay was

repeated trice.

3.2.2.3 Evaluation of antibacterial activity of plant extracts by well diffusion

method:

Well diffusion method was followed to determine the antimicrobial activity of

the selected plant extracts against test microorganism. After solidification wells

(6mm diameter) were cut into the agar and test plant extracts were introduced into

the wells, 50µl of the plant extracts were tested in a concentration of 100 mg/ml

After 24 hours, the antimicrobial activity was evaluated by measuring the diameter of

the inhibition zone formed around the plant extracts well. (Abu-Shanab et al.,

2004). The assay was repeated trice.

3.2.2.4 Determination of MIC and MBC of plant extracts by Microdilution

Method:

Minimum inhibitory concentration (MIC) which means the lowest

concentration of the plant extract at which the bacteria does not show visible

growth was determined by the micro-dilution method (96- well plates). Plant

extracts were diluted a number of times through a sterile diluent (MHB). A final

concentration from 200 to 0.1953 mg/ml was used for each plant sample Then

added 10 μl of inocula of overnight growth microorganisms to each well except a

positive control. Inoculum with media was used as a negative control and extract

with media was used as a positive control (Cheraif et al, 2007) After incubation

at 37°C for 24 h. 50μl of a 0.01% triphenyl tetrazolium chloride (TTC) solution

added to the well as indicator and the plate was incubated for one hour. Since the

colorless tetrazolium salt is reduced to red colored product by biological active

36

bacteria, the inhibition of growth can be detected when the solution in the well

remains clear after incubation with (TTC) (Abu-Shanab et al., 2004).

To determine MBC (lowest concentration of the plant extract at which

inoculated bacteria was totally killed) MHA inoculated with broth which taken

from each well and incubated for 24hr at 37ºC the lowest concentration that

revealed no visible bacterial growth after sub-culturing was taken as MBC 10%

DMSO solution served as negative controls(Kang et al., 2011).

3.2.2.5 Synergistic assay:

A modification of previously described procedures (Elbashiti et al, 2011)

was followed to evaluate of the synergistic effect. The antimicrobial activities of

the selected plant extracts a gainst the tested microorganism were compared with

each other, after mixed 50gm of Coriandrum sativum extract with 50gm of

Artemisia monosperma extract and mixed 50gm of Cuminum cyminum extract

with 50gm of Ocimum basilicum, the antimicrobial activity was evaluated by

well diffusion method, disc diffusion method and MIC.

37

Chapter (4)

Results

38

4.1 Taxonomic diversity of plants under investigation

The study documented 72 medical plant species belonging to 33 families that

were the most used in Traditional Arabic Palestinian Herbal Medicine in Gaza

strip (Table 4.1). The most represented families were: Lamiaceae (8 species),

Apiaceae (8 species) and Asteraceae (7 species) (Table 4.1). Figure (4.1).

Table (4.1) : Distribution of plant families by number of species

No. of Species اؼائح اثاذ١ح Family Name NO.

Lamiaceae 1 اؾف٠ح 9

Apiaceae 2 اخ١١ح 8

Asteraceae 3 اد١ح 7

Fabaceae 4 اثم١ح 5

Brassicaceae 5 اص١ث١ح 4

Lauraceae 6 اغاس٠ح 3

Malvaceae 7 اخثاص٠ح 3

Ranunculacea 8 اؾم١م١ح 2

Zingiberaceae 9 اضدث١١ح 2

Euphorbiaceae 10 اغعث١ح 2

Poaceae 11 اد١١ح 2

Apocynaceae 12 اذف١ح 2

Rosaceae 13 اسد٠ح 2

Myrtaceae 14 ا٢ع١ح 2

Lythraceae 15 اسائ١اخ 1

Hypericaceae 16 اا٠ثشو١ح 1

valerianaceae 17 ااسد١٠ح 1

Arecaceae 18 اخ١١ح 1

Urticaceae 19 امش٠ص١ح 1

Theaceae 20 ػائح اؾا 1

39

No. of Species اؼائح اثاذ١ح Family Name NO.

Equisetaceae 21 اىثاث١ح 1

Polygonaceae 22 اثطثاط١ح 1

Burseraceae 23 اثخس٠اخ 1

Asparagaceae 24 ا١١ح 1

Ginkgoaceae 25 ادى١اخ 1

Caryophyllaceae 26 امشف١ح 1

Sapotaceae 27 اغثذ١ح 1

Cyperaceae 28 اغؼذ٠ح 1

Plantaginaceae 29 اس١ح 1

Papaveraceae 30 اخؾخاؽ١ح 1

Portulacaceae 31 اشخ١ح 1

Rutaceae 32 اغزات١ح 1

Araliaceae 33 الأسا١ح 1

Figure (4.1): Distribution of plant families by number of species

0

1

2

3

4

5

6

7

8

9

No. of Species

40

4.2 Most cited plants and remedies

Based on the study survey, it was found that there were many plants that are

used in folk remedies. The method of preparation of these plants varies according to

the type of disease and the part used. Table (4.2) shows the classification of the most

used plants in popular treatments in the Gaza Strip. Also the plant is classified as

medical plants if mentioned by more than two therapists.

The remedies were administered orally or used externally according to the

disease and preparation method. Where the informants administer their remedies in

various forms including standard decoction prepared by boiling plant parts in hot

water, volatile compound and oils, use as ointment and use as paste. The remedies

were administered orally or used externally according to the disease and preparation

method.

41

Table (4.2): Classification of the most used plants in popular treatments in the Gaza Strip

Mode of

preparation and

use

Medicinal use Part used Arabic

name

Common name Family Scientific name No

Boiled leaves Urinary tract infections and

hormonal regulator

Above-

ground

part

تشدلػ ا

شدلػ

Marjoram Lamiaceae Origanum majorana L. 1

Boiled seeds Diuretic - digestion disorder

- asthma

Leaves and

Seed

اؾش ا

اؾش

Fennel Apiaceae Foeniculum vulgare Mill. 2

Boiled seeds

And oil as an

external treatment

Calming the nerves - colic -

cough and asthma

Seed ا١غ Anise Apiaceae Pimpinella anisum L. 3

Boiled leaves and

flowers

Cough - headache - skin -

colic - wounds

Leaves and

flowers

Golden اثاتح

cotula

Asteraceae Matricaria aurea (Loefl)

Sch.Bip

4

Raw fruit Boiled

peel

Anemia - stomach ulcer -

heart

All part of

fruit

Pomegranate Lythraceae Punica granatum L. 5 اشا

https://en.wikipedia.org/wiki/Lythraceae

42

Mode of

preparation and

use


name


Boiled bark Nausea - Diabetes bark امشفح cinnamon Lauraceae Cinnamomum verum J.Presl. 6

Drenched flowers

Oils as an external

cream

Depression - the effects of

addiction

Above-

ground

part

Saint John's wort Hypericaceae Hypericum perforatum L. 7 لذ٠ظ ٠زا

Boiled flowers or

leaves

Respiratory Allergy - Cough

- Calming the stomach

Above-

ground

part

Little-leaf Linden اض٠ضف

or greenspire

Malvaceae Tilia cordata Mill. 8

Oil Extract

Boiled roots

Indigestion - Epilepsy Root اغث spikenard valerianaceae Nardostachys grandiflora

DC.

9

Boiled dried fruits Prostate - urinary tract

infection

Fruits saw palmetto Arecaceae serenoa repens (w. bartram) ت١ظ ؾاس

small

10

Boiled seeds Increase breast milk -

diabetes

Seed اسثح Fenugreek Fabaceae Trigonella foenum-graecum

L.

11

43

Mode of

preparation and

use


name


Boiled leaves or

roots or eaten raw

Hemorrhoids - diabetes -

inflammation of the nerves

Whole

plant

امش٠ـ ا

اسش٠ك

Nettle Urticaceae Urtica urens L. 12

Soaked crushed

seeds

Diabetes - diuretic -

constipation - high

cholesterol

Seed ارشظ اش White Lupine Fabaceae Lupinus albus L. 13

Soaked leaves Hypertension - Indigestion -

Immunodeficiency

Leaves الاخضش Green tea Theaceae camellia sinensis (L.) Kuntze 14 اؾا

Soaked leaves Kidney stones - Arthritis -

Pulmonary tuberculosis

Leaves ر٠ اسصا Horsetail field Equisetaceae Equisetum arvense L. 15

Extract Oil

Boiled leaves

Diarrhea - headache -

sunburn - urinary tract

infection

Above-

ground

Part

اش٠سا ا

اسثك

basil Lamiaceae Ocimum basilicum L. 16

44

Mode of

preparation and

use


name


Boiled roots Stomach disorder -

constipation - tonic for the

liver

Root اشاذ Rhubarb Polygonaceae Rheum Palmatum L. 17

Extract oil and raw

seeds

Rheumatism - Hypertension

- diabetes - colon

Seed زثح اثشوح ا

اسثح اغداء

Black cumin Ranunculacea Nigella sativa L. 18

Soaked leaves and

branches

Gastrointestinal disorders -

Elimination of intestinal

worms

Leaves اؾ١ر White Wormwood Asteraceae Artemisia monosperma

Delile

19

Boiled Rhizomes Hypertension - stomach pain

- vertigo - memory

activation - high lipids

Rhizome اضدث١ Ginger Zingiberaceae Zingiber officinale Roscoe

20

Extract Oil Gastrointestinal tract - hair

loss - skin

Fruits and

Seed

Castor beans Euphorbiaceae Ricinus communis L. 21 اخشع

45

Mode of

preparation and

use


name


Dried fruits or fresh

leaves

Dysentery - vomiting -

indigestion - colon disorder

Leaves and

Fruits

Coriander Apiaceae Coriandrum sativum L. 22 اىضتشج

Boiled roots Diabetes - sexual impotence Roots اضع Zallouh Apiaceae Ferula hermonis Boiss 23

Boiled or Soaked

Flowers

Hypertension - Circulatory

system

flowers اىشوذ٠ Rosella Malvaceae Hibiscus sabdariffa L. 24

Boiled leaves Gastrointestinal pain - tonic

for memory – colds

Above-

ground

Part

Rosemary Lamiaceae Rosmarinus officinalis L. 25 او١ ادث

Soaked gum Colon - intestinal

disinfectant - activates

memory - diabetes

Gum

زص اثا

اذوش

Olibanum or

Frankincense

Burseraceae Boswellia sacra Flueck 26

46

Mode of

preparation and

use


name


Boiled leaves and

extract oil from

seeds

Overweight - Asthma - High

cholesterol

Whole

plant

Geranium grass Poaceae Cymbopogon schoenanthus اسفا تش

Spreng

27

Extract gum Gastrointestinal tract -

urinary system -gingivitis-

hair

Barks and

Fruits

الاخ٠د dragon blood tree Asparagaceae Dracaena cinnabari Balf.f. 28

Boiled leaves Asthma - activates memory

– Hypertension- Diabetes

Leaves ادىح ginkgo Ginkgoaceae Ginkgo biloba L. 29

Boiled leaves and

extract oil

Respiratory System -

Digestive System -

Circulatory System

Above-

ground

Part

Wild اضػرش اثش

thyme

Lamiaceae Thymus sepyllum L. 30

Boiled leaves Urinary tract infection -

anemia - rheumatism

Above-

ground

Part

Silvery سخ اساح

Whitlow wart or

silver nailroot

Caryophyllaceae Paronychia argentea Lam. 31

47

Mode of

preparation and

use


name


Soaked seeds or

added to milk or

bread

Skin - Diabetes -

Overweight

Seed اؾ١ا shea butter tree Sapotaceae Vitellaria paradoxa

C.F.Gaertn.

32

Boil the leaves and

twigs

Digestive system -

Hypertension

Above-

ground

Part

Sage Lamiaceae Salvia officinalis L. 33 ا١ش١ح

Dry Rhizomes

powder

Antibacterial - gallstones Rhizome اىشو Turmeric Zingiberaceae Curcuma longa L. 34

Boiled powder

leaves

Intestinal disinfectant -

Constipation

Leaves اغا ىح coffee senna Fabaceae Senna occidentalis (L.) Link. 35

Cooked or boiled

leaves

Laxative for the stomach -

Calming cough - Diabetes

Leaves اخث١ضج mallow Malvaceae Malva parviflora L. 36

48

Mode of

preparation and

use


name


Boiled seeds or eat

raw leaves

Gastrointestinal tract -

urinary system

Seeds and

leaves

ادشادجػ١ Dill Apiaceae Anethum graveolens L. 37

Soaked leaves Diuretic - laxative - joint

pain

leaves اسشخ Ghalisum Apocynaceae Cynanchum argel Delile 38

Boiled , dipped or

emulsified

Heart tonic - blood vessels Above-

ground

Part

Hawthorn Rosaceae Crataegus aronia L. 39 صػشس ؽائه

Boiled powder

twigs

Gonorrhea - Thyroid -

analgesic

stalk امغظ اذ Costus Asteraceae Saussurea costus (Falc.)

Lipsch

40

Boiled dried leaves Disinfectant of toxins -

reduces cholesterol - skin -

indigestion

leaves اغاس Laurel Lauraceae Laurus nobilis L. 41

49

Mode of

preparation and

use


name


Emulsion or

immersed

Vaginitis - diarrhea -

stomach analgesia

Above-

ground

Part

lady's mantle Rosaceae Alchemilla Vulgaris L. 42 سخ الاعذ

raw leaves or

boiled seeds

Prostate enlargement -

diabetes - hepatitis

Seed and

leaves

خشف١ؼ اد

ا ؽن

اداي

milk thistle Asteraceae Silybum marianum (L.)

Gaertn.

43

Boiled leaves Urinary system - menstrual

pain - sores

leaves و١ظ اشاػ shepherd's purse Brassicaceae Capsella bursa-pastoris (L.)

Medik.

44

Boiled leaves or

extract oil

Headache - cough leaves اىافس camphor tree Lauraceae Cinnamomum camphora (L.)

J.Presl.

45

Boiled leaves Respiratory System -

Digestive System - Allergies

leaves اى١١ا Red River

Gum

Myrtaceae Eucalyptus camaldulensis

Dehn.

46

50

Mode of

preparation and

use


name


Dry Roots boiled

for 10 minutes

Kidney failure - rheumatism

- hemorrhoids

Roots اؼالي Alhagi Manna Fabaceae Alhagi graecorum Medik. 47

Emulsion of

flowers

De-worming - antifungal -

anti-inflammatory - gout

Flowers الألسا ا

اثغ

Chamomile. Asteraceae Chrysanthemum coronarium

L.

48

Boiled flowers Arteriosclerosis - skin -

psoriasis - urinary system

Flowers تمح اه ا

ؽارشج

fumitory Papaveraceae Fumaria officinalis L. 49

Eat fresh or boiled

seeds and leaves

Urinary system - digestive

system - antibacterial

Above-

ground

Part and

seeds

اشخح ا

اثمح

Verdolaga or

Purslane

Portulacaceae Portulaca oleracea L. 50

Boiled flowers or

boiled herb as a

whole

Cough - nervous tension Flowers

and leaves

ؽمائك اؼا

ا اس

Windflower or

crown anemone

Ranunculaceae Anemone coronaria L. 51

51

Mode of

preparation and

use


name


Grind the herb and

mix with honey or

Soaked herb

Menstrual pain - Postpartum

bleeding - Prostate

enlargement

Whole

plant

Chaste tree Lamiaceae Vitex agnus-castus L. 52 وف ش٠

raw or boiled

leaves

diabetes -bacterial infection

and rheumatism

leaves اغش٠ظ Endive or escarole Asteraceae Cichorium endivia L. 53

Boiled seeds or

extract oil from

buds

Analgesic - Fever -

Allergies

Flower

buds and

seeds

Clove Myrtaceae Syzygium aromaticum (L.) امشف

Merrill & Perry

54

Soaked dry roots Anemia - Digestive System

- Headache

Roots زؾ١ؾح الان Dong Quai Apiaceae Angelica sinensis (Oliv.)

Diels

55

Raw leaves or

boiled seeds

Obesity - diabetes - arthritis

- digestive system

Above-

ground

Part and

seeds

celery Apiaceae Apium graveolens L. 56 اىشفظ

52

Mode of

preparation and

use


name


Raw or boiled

leaves and extract

oil

Digestive System -

Headaches - Colds

Above-

ground

Part

Spearmint Lamiaceae Mentha spicata L. 57 اؼغ

Soaked roots or as

bandages

Rheumatism - Colds -

urinary system

Roots ا ػالش لشزا

ػد امشذ

Akarkara or

Pellitory

Asteraceae Anacyclus pyrethrum (L.)

Link

58

Soaked Crushed

fruits

Gout - urinary system -

infertility - liver - skin

Fruits زة اؼض٠ض ا

عؼذ از٠ز

Yellow nutsedge or

chufa flatsedge

Cyperaceae Cyperus esculentus L. 59

Soaked or boiled

roots

Gastric ulcers - weakened

immunity

Roots اؼشق عط Liquorice Fabaceae Glycyrrhiza glabra L. 60

Oil extract as

external ointment

Scabies - psoriasis Latex اذفح oleander Apocynaceae Nerium oleander L. 61

53

Mode of

preparation and

use


name


Boiled leaves or

seeds , raw seeds or

extract oil

Rheumatism - sciatica -

anemia

Leaves and

Seeds

Gardn Cress Brassicaceae Lepidium sativum L. 62 زة اشؽاد

Soaked of whole

plant

Diabetes - Irritable Bowel -

Hemorrhoids - Laxative of

the intestine

Whole

plant

fleaworts Plantaginaceae Plantago lagopus L. 63 غا اس

Soaked leaves Diabetes - Cancer - skin

diseases

Leaves اسثب annual mercury Euphorbiaceae Mercurialis annua L. 64

Raw leaves or

extract oil

Hair loss - Anemia -

Hypertension - Cholesterol -

Diabetes

Above-

ground

Part

ادشخ١ش ا

اشوا

Garden rocket Brassicaceae Eruca sativa Miller 65

Boiled dry fruits Gastrointestinal tract -

respiratory system - anemia

Dry fruits اى Cumin Apiaceae Cuminum cyminum L. 66

54

Mode of

preparation and

use


name


Soaked leaves Digestive System -

Menstrual disorders

circulatory system

Leaves عزاب/ف١د Rue Rutaceae Ruta chalepensis L. 67

Soaked leaves Respiratory system -

Irritable colon - a sedative

of the nerves

Leaves ا١غح Lemon Balm Lamiaceae Mellisa officinalis L. 68

Boiled or soaked

seeds

Urinary system - increases

immunity - circulatory

system - diabetes

Seeds اؾؼ١ش Barley Poaceae Hordeum vulgare L. 69

Soaked roots Impotence - Immune

Deficiency - Memory

Activation

Roots ادغه Ginseng Araliaceae Panax Ginseng Baill. 70

55

Mode of

preparation and

use


name


Soaked leaves Stomachache, colic,

Diabetes,

leaves ادؼذج felty germander Lamiaceae Teucrium polium L. 71

Soaked seeds or Oil

extract

Diabetes – Cancer-

Rheumatism- Gout-

Circulatory System

Seeds اخشدي mustard Brassicaceae Brassica alba L. 72

56

Medicinal plants were reported to be used for the treatment of several

ailments as shown in Table (4.3). The largest number of remedies was used to treat

gastrointestinal disorders were represented by 37 plants. The second commonly used

remedies were for the treatment of circulatory system and were represented by 22

plants. As for remedies for the Urinary system were represented by 20 plants.

Table (4.3) : Diversity of medicinal use- categories

Disease No. of Plants

Digestive System 37

Circulatory system 22

Urinary system 20

Diabetes 18

Respiratory system 14

Nervous system 13

Skin, burns, and hair 12

Weight loss 10

Skeletal and muscular system 7

Immunity system 7

Reproductive system 7

Cancer 3

Others 11

57

4.3 Folk medical knowledge sources:

With respect to sources there is no processor qualified academically for this

profession, parents and grandparents seem to be the main source of traditional

knowledge as 75% of the study population reported to gather their knowledge from

this sources. Other information sources were obtained from old medical books and

others. Figure (4.2).

Figure (4.2): Sources of folk medical knowledge

4.4 Used parts

The most repeatedly utilized plant parts were leaves, stems and seeds which were

represented by 40%, 18% and 14%, respectively. Other plant parts were also reported

to be used in folk remedies as shown in Figure (4.3).

Figure (4.3): Percentage of used plant parts

0%

20%

40%

60%

80%

parents andgrandparents

old medicalbooks

others

Percentage ofinformants %

Leaves 41%

Stem 18%

Seeds 14%

Flower 8%

Fruits 7%

Roots 7%

Others 5%

58

4.5 Procurement methods of medicinal plants:

The findings of the current study showed that 35% of informants imported

medicinal plants from abroad, 30% purchased plants from local market, 20%

collected plants from nature and 15% obtained the plants by the three previous

methods (Figure 4.4). These plants are often stored in special bags or tightly sealed

plastic containers. However, there are difficulties in obtaining some plant species due

to the scarcity of some of these plants or the high prices.

Figure (4.4) Procurement methods of medicinal plants

4.6 Evaluation of antibacterial activity of plant extracts by disc diffusion

method

The results of antibacterial activity of aquatic and ethanolic extracts of all the

eleven plants tested individually against the two isolated bacterial species, which are

known to cause infection in humans. Each experiment replicated three time at least.

4.6.1 Against E. coli

The disc diameters of zone of inhibition of plants extracts against E.coli are

shown in Table (4.4).

4.6.1.1 The aquatic extracts

The aquatic extracts of all the plants screened showed various inhibitory effects

ranged between (8-17mm) in diameter, highest effect towards E.coli showed by

(Syzygium aromaticum) (with a 17 mm zone of inhibition) followed by (Hibiscus

0%

5%

10%

15%

20%

25%

30%

35%

Imported Localmarket

Collectfrom

nature

all ofthem

Percentage ofinformants%

59

sabdariffa, Artemisia monosperma and Coriandrum sativum (with a 8 mm zone of

inhibition). The lowest activity was the Anethum graveolens, Boswellia Carterii,

Cuminum cyminum, Trigonella foenum graecum, Ocimum basilicum, (with a 7 mm

zone of inhibition). No effect was shown for either Cymbopogon schoenanthus and

Glycyrrhiza glabra (with a zero mm zone of inhibition). Shown in (Table 4.4) and

(Figure 4.5 and 4.6)

4.6.1.2 The ethanolic extracts

The ethanolic extract of Syzygium aromaticum.L were the most effective

extract against E.coli which it is showing the highest antibacterial activity against

this bacteria with inhibition zone diameter of 17 mm followed strictly were extracts

of Hibiscus sabdariffa with inhibition zone diameter of 8 mm, no effect was shown

for Ocimum basilicum, Boswellia Carterii, Cuminum cyminum, Trigonella foenum

graecum, Anethum graveolens, Artemisia monosperma, Coriandrum sativum,

Glycyrrhiza glabra, and Cymbopogon schoenanthus. Shown in (Table 4.4) and

(Figure 4.7, 4.8 and 4.9)

4.6.2 Against Staphylococcus aureus

The disc diameters of zone of inhibition of plants extracts against S.aureus are

shown in Table (4.5).

4.6.2.1 The aquatic extracts:

The Aquatic extract of Hibiscus sabdariffa were the most effective extract

showing the highest antibacterial activity against S. aureus bacteria with inhibition

zone diameter of 10mm followed by Syzygium aromaticum.L with inhibition zone

diameter of 9mm, but there are no antibacterial activity shown for Ocimum

basilicum, Boswellia Carterii, Cuminum cyminum, Trigonella foenum graecum,

Anethum graveolens, Artemisia monosperma, Coriandrum sativum, Glycyrrhiza

glabra, and Cymbopogon schoenanthus. Shown in (Table 4.5) and (Figure 4.10,

4.11 and 4.12)

4.6.2.2 The ethanolic extracts:

Only Syzygium aromaticum.L with inhibition zone diameter of 12mm and

Hibiscus sabdariffa with inhibition zone diameter of 7mm showing antibacterial

60

activity against S. aureus bacteria, but there are no antibacterial activity shown for

Ocimum basilicum, Boswellia Carterii, Cuminum cyminum, Trigonella foenum


Glycyrrhiza glabra, and Cymbopogon schoenanthus Shown in (Table 4.5) and

(Figure 4.13, 4.14and 4.15)

4.7 Evaluation of antibacterial activity of plant extracts by well diffusion

method

Antibacterial activity of aquatic and ethanolic extracts of all the eleven plants

where tested against each E.coli and S.aureus by well diffusion method. Each

experiment replicated three time at least.

4.7.1 Against E. coli:

The inhibition zone of plants extracts against E.coli are shown in Table (4.4).


Various inhibitory effects ranged between 11- 25 mm in diameter showed by

aquatic extracts of all the plants against E.coli. The largest zone of inhibition were

observed from the Syzygium aromaticum.L with inhibition zone diameter of 25mm,

followed by Hibiscus sabdariffa, Boswellia Carterii, Anethum graveolens, Cuminum

cyminum, Ocimum basilicum, Trigonella foenum graecum, Glycyrrhiza glabra, with

inhibition zone diameter of 16, 16, 13, 14, 12, 11and 11mm respectively. There are

no antibacterial activity shown for Coriandrum sativum and Cymbopogon

schoenanthus. Shown in (Table 4.4) and (Figure 4.16, 4.17, 4.18).


The ethanolic extract of Syzygium aromaticum were the most effective extract

against E.coli which is showing the highest antibacterial activity against this bacteria

with inhibition zone diameter of 25 mm followed strictly were extracts of Hibiscus

sabdariffa with inhibition zone diameter of 13 mm. There was no effect shown for

Ocimum basilicum, Boswellia Carterii, Cuminum cyminum, Trigonella foenum


Glycyrrhiza glabra, and Cymbopogon schoenanthus Shown in (Table 4.4) and

(Figure 4.19, 4.20 and 4.21).

61

4.7.2. Against Staphylococcus aureus:

The inhibition zone of plant extracts against S.aureus are shown in Table(4.5)


The aquatic extracts of the tested plants give various degree of activity (11-

24mm) inhibition zone against S.aureus. The extract of Syzygium aromaticum

presented the highest antibacterial activity with inhibition zone diameter of 24mm

followed by Ocimum basilicum with inhibition zone diameter of 23mm followed by

Hibiscus sabdariffa, Boswellia Carterii, Cuminum cyminum, Trigonella foenum

graecum, which have an intermediate activity against S.aureus with inhibition zone

diameter of 16, 14, 13and 13 mm, respectively. The lowest activity was the Anethum

graveolens, Artemisia monosperma and Coriandrum sativum with inhibition zone

diameter of 11mm. Cymbopogon schoenanthus and Glycyrrhiza glabra have no any

antibacterial activity against S.aureus. Shown in (Table 4.5) and (Figure 4.22, 4.23

and 4.24).


The ethanolic extract of Syzygium aromaticum were the most effective extract

against S.aureus which is showing the highest antibacterial activity with inhibition

zone diameter of 22 mm, followed by Hibiscus sabdariffa, Anethum graveolens,

Cymbopogon schoenanthus, Coriandrum sativum and Artemisia monosperma which

have an intermediate activity against S.aureus with inhibition zone diameter of 17,

16, 14, 12 and 9 mm respectively. Boswellia Carterii, Glycyrrhiza glabra, Trigonella

foenum graecum, Ocimum basilicum and Cuminum cyminum have no any

antibacterial activity against S.aureus. Shown in (Table 4.5) and (Figure 4.25, 4.26,

4.27)

62

Table (4.4): Antimicrobial Activity of Plant extracts against E. coli by well diffusion

method and disc diffusion method.

Well diffusion method (mm) Disc diffusion method (mm) A*A*A

Average of inhibition zone.(± SD)

Control Water Ethanol Control Water Ethanol Plant extract

- 14 (0) - - 7 (0) - Anethum graveolens

- 16(0.82) 13 (0.82) - 7 (0.47) 8 (0.82) Hibiscus sabdariffa

- 16(0.82) - - 7 (0.47) - Boswellia Carterii

- 13(0.47) - - 7 (0.47) - Cuminum cyminum

- 11(0) - - 7 (0.47) - Trigonella foenum

graecum

- 25(0.47) 25(0.47) - 17 (0.47) 17 (0.82) Syzygium

aromaticum

- 12(0.82) - - 7 (0.47) - Ocimum basilicum

- 13(0) - - 8 (0.47) - Artemisia

monosperma

- - - - 8 (0.47) - Coriandrum sativum

- - - - - - Cymbopogon

schoenanthus

- 11(0) - - - - Glycyrrhiza glabra

* Antimicrobial Activity Assays.

Control= DMSO

(-) No inhibition zone.

Standard deviation = SD

63

Table (4.5): Antimicrobial Activity of Plant extracts on Staphylococcus aureus by

well diffusion method and disc diffusion method


Average of inhibition zone.(± SD.)


- 11(0.94) 16(0.47) - - - Anethum graveolens

- 16(0.47) 17(0.47) - 10(0.82) 7 (0) Hibiscus sabdariffa

- 14(0) - - - - Boswellia Carterii

- 13(0.47) - - - - Cuminum cyminum

- 13(0.47) - - - - Trigonella foenum

graecum

- 24 (0) 22 (0.82) - 9 (0.94) 12 (0.47) Syzygium aromaticum

- 23(0.47) - - - - Ocimum basilicum

- 11(0.47) 9(0.47) - - - Artemisia

monosperma

- 11(0.82) 12(0.47) - - - Coriandrum sativum

- - 14(0.82) - - - Cymbopogon

schoenanthus

- - - - - - Glycyrrhiza glabra

* Antimicrobial Activity Assays.

Control= DMSO

(-) No inhibition zone.

Standard deviation = SD

.

64

Figure (4.6) : The effect of aquatic

extract to 6-Syzygium aromaticum,

7-Ocimum basilicum,8-Artemisia

monosperma, 9-Coriandrum sativum,

10-Cymbopogon schoenanthus and

11- Glycyrrhiza glabra by disc

diffusion method to against E. coli


extract to 1- Anethum graveolens,

2- Hibiscus sabdariffa, 3- Boswellia

Carterii, 4- Cuminum cyminum,

5- Trigonella foenum graecum by disc


Figure (4.8) : The effect of ethanolic




and 10-Cymbopogon schoenanthus by

disc diffusion method to against E.

coli







65






diffusion method to against S. aureus


extract to 11- Glycyrrhiza glabra by disc


Figure (4.12): The effect of aquatic

extract to 11- Glycyrrhiza glabra by disc

diffusion method to against S. aureus



7-Ocimumbasilicum,8-Artemisia



disc diffusion method to against

S.aureus

66






disc diffusion method to against

S.aureus.



2- Hibiscus sabdariffa,

3- Boswellia Carterii, 4- Cuminum

cyminum, 5- Trigonella foenum

graecum by disc diffusion method to

against S. aureus





5- Trigonella foenum graecum by well

diffusion method to against E.coli


extract to 11- Glycyrrhiza glabra by

disc diffusion method to against S.

aureus

67


extract to 11- Glycyrrhiza glabra by well



extract to 6-Syzygium aromaticum,7-

Ocimum basilicum,8-Artemisia



well diffusion method to against E.coli.






well diffusion method to against E.coli.







68






diffusion method to against S.aureus



well diffusion method to against E.coli

Figure (4.24) The effect of aquatic


well diffusion method to against S.

aureus

Figure (4.23) The effect of aquatic





well diffusion method to against

S.aureus

69

Figure (4.26) The effect of ethanolic

extract to 6-Syzygium aromaticum,7-

Ocimum basilicum,8-Artemisia



well diffusion method to against

S.aureus






diffusion method to against S.aureus



well diffusion method to against S.

aureus

70

4.8 The minimum inhibitory concentration (MIC) & minimum bactericidal

concentrations (MBC) of plant extracts

Extracts were tested against the bacterial isolates for their inhibitory activity,

using a common broth microdilution method in 96 multiwell microtiter plates in two

fold dilution series of these extracts. It was prepared as 200, 100, 50, 25, 12.5. 6.25,

3.125, 1.562, 0.781 & 0.390 mg/ml for the aquatic and ethanolic extracts in

triplicated. The results showed antibacterial activity against E.coli and S.aureus, the

average of the obtained minimum inhibitory concentrations (MICs) & minimum

bactericidal concentrations (MBCs) is listed in Tables (4.6),(4.7), Respectively.

4.8.1 Against E.coli:

As shown in Table (4.6) the MIC values of all tested plant extracts against

E.coli. The MIC values of the ethanolic extracts of Anethum graveolens and

Cuminum cyminum was 12.5mg/ml, for each of Ocimum basilicum, Artemisia

monosperma, Glycyrrhiza glabra and Cymbopogon schoenanthus was 6.25 mg/ml.

While for Boswellia Carterii and Coriandrum sativum was 1.56mg/ml. The MIC for

Hibiscus sabdariffa, Syzygium aromaticum and Trigonella foenum graecum against

E. coli was the least one 0.78 mg/ml as shown in (Figures 4.28 to Figure 4.32).

The MIC results of the aquatic extracts of Anethum graveolens, Coriandrum

sativum and Cymbopogon schoenanthus was 25mg/ml, and the MIC value of

Artemisia monosperma and Glycyrrhiza glabra was 12.5 mg/ml. The MIC for

Ocimum basilicum, Trigonella foenum graecum, Cuminum cyminum, Syzygium

aromaticum and Boswellia Carterii against E. coli were 6.25, 3.125, 1.56, 1.56 and

0.78 mg/ml, respectively. The least value was 0.39 mg/ml for Hibiscus sabdariffa.

(Figure 4.33 to Figure 4.36).

As shown in table (4.6) the MBC of the ethanolic extracts of Glycyrrhiza

glabra 100mg/ml and Syzygium aromaticum 200mg/ml. The MBC value was

>200mg/ml for Anethum graveolens, Hibiscus sabdariffa, Boswellia Carterii,

Cuminum cyminum, Trigonella foenum graecum, Coriandrum sativum, Cymbopogon

schoenanthus, Artemisia monosperma and Ocimum basilicum. The MBC values of

aquatic extracts for all eleven sample was >200mg/ml.

71

Table (4.6) : The MICs & MBCs of plant extracts against E. coli

MIC conc. MBC conc.

Solvent

Plant extract

Ethanol Water Ethanol Water

Anethum graveolens 12.5 25 >200 >200

Hibiscus sabdariffa 0.78 0.39 >200 >200

Boswellia Carterii 1.56 0.78 >200 >200

Cuminum cyminum 12.5 1.56 >200 >200

Trigonella foenum graecum 0.78 3.125 >200 >200

Syzygium aromaticum 0.78 1.56 200 >200

Ocimum basilicum 6.25 6.25 >200 >200

Artemisia monosperma 6.25 12.5 >200 >200

Coriandrum sativum 1.56 25 >200 >200

Cymbopogon schoenanthus 6.25 25 >200 >200

Glycyrrhiza glabra 6.25 12.5 100 >200

72

Figure (4.29): (MIC) of ethanolic

extracts of 4-Cuminum cyminum

5-Syzygium aromaticum

6-Trigonella foenum graecum

against E. coli.


extracts of 1- Anethum graveolens

2- Hibiscus sabdariffa 3- Boswellia

Carterii against E. coli.


extracts of 9- Coriandrumand 10-

sativumCymbopogon schoenanthus

against E. coli.


extracts of 7-Ocimum basilicum

8-Artemisia monosperma against E.

coli.

73


extracts of 11- Glycyrrhiza glabra

against E. coli

Figure (4.33): (MIC) of aquatic

extracts of 1- Anethum graveolens 2-

Hibiscus sabdariffa 11- Glycyrrhiza

glabra against E. coli


extracts of 3- Boswellia Carterii

4-Cuminum cyminum against E. coli


extracts of 5-Syzygium aromaticum

6-Trigonella foenum graecum against

E. coli.

74

Figure (4.36): (MIC) of aquatic extracts

of 7-Ocimum basilicum 8-Artemisia

monosperma 9- Coriandrumand 10-

sativumCymbopogon schoenanthus

against E. coli.

75

4.8.2 Against Staphylococcus aureus:

The MIC values of the ethanolic extracts against S. aureus of Glycyrrhiza

glabra was 50mg/ml. The MIC of Boswellia Carterii, Cuminum cyminum and

Trigonella foenum graecum was 25mg/ml and for each of Anethum graveolens,

Hibiscus sabdariffa, Ocimum basilicum, Artemisia monosperma, Coriandrum

sativum, Cymbopogon schoenanthus was 12.5mg/ml. The least value was 0.39

mg/ml for Syzygium aromaticum. (Table 4.7) and (Figure 4.37 to Figure 4.42).

The MIC of the aquatic extracts of Cuminum cyminum was 50mg/ml, for

Anethum graveolens, Boswellia Carterii, Trigonella foenum graecum, Ocimum

basilicum, Cymbopogon schoenanthus, Coriandrum sativum, Artemisia monosperma

was 25mg/ml, and for Glycyrrhiza glabra was 12.5mg/ml. The least value was

6.25mg/ml for Hibiscus sabdariffa and Syzygium aromaticum (Table 4.7) and

(Figure 4.43 to Figure 4.46)

The MBC value was >200mg/ml for aquatic and ethanolic for all eleven plant

extracts.

Table (4.7): The MICs & MBCs of extracts against S.aureus

MIC Conc. MBC Conc.

Solvent

Ethanol

Water

Ethanol

Water Plant extract

Anethum graveolens 12.5 25 >200 >200

Hibiscus sabdariffa 12.5 6.25 >200 >200

Boswellia Carterii 25 25 >200 >200

Cuminum cyminum 25 50 >200 >200

Trigonella foenum graecum 25 25 >200 >200

Syzygium aromaticum 0.39 6.25 >200 >200

76

Ocimum basilicum 12.5 25 >200 >200

Artemisia monosperma 12.5 25 >200 >200

Coriandrum sativum 12.5 25 >200 >200

Cymbopogon schoenanthus 12.5 25 >200 >200

Glycyrrhiza glabra 50 12.5 >200 >200

77



Hibiscus sabdariffa against S.aureus.


extracts of 3- Boswellia Carterii 4-

Cuminum cyminum against S.aureus.




S.aureus.


extracts of 7-Ocimum basilicum 8-

Artemisia monosperma against

S.aureus.

78


extracts 9- Coriandrumand 10-

sativum Cymbopogon schoenanthus

against S.aureus


extracts of 11- Glycyrrhiza glabra

against S.aureus



Hibiscus sabdariffa and 11-

Glycyrrhiza glabra against S.aureus.


extracts of 3- Boswellia Carterii 4-

Cuminum cyminum against S.aureus.

79




S.aureus.


extracts of 7-Ocimum basilicum

8-Artemisia monosperma

9- Coriandruman sativum

10-Cymbopogon schoenanthus

against S.aureus.

80

4.9 Evaluation the synergistic effect:

4.9.1 The Synergistic effect between plant extract:

We estimated in vitro synergism between extracts of Cuminum cyminum with

Ocimum basilicum, and Artemisia monosperma with Coriandrum sativum against S.

aureus and E. coli

4.9.1.1 Evaluation the synergistic effect by disc diffusion method:

4.9.1.1.1 Against E. coli:

The aquatic extract of Cuminum cyminum with Ocimum basilicum shown

inhibition zone diameter of 7mm but there was no effect of ethanolic extract from

them against E. coli. The ethanolic extract of Artemisia monosperma with

Coriandrum sativum shown inhibition zone diameter of 8mm and no effect was

shown for aquatic extract Table (4.8).

4.8.1.1.2 Against S. aureus:

As shown in Table (4.9) there was no inhibition zone for both Cuminum

cyminum with Ocimum basilicum and Artemisia monosperma with Coriandrum

sativum in each of aquatic and ethanolic extract.

4.9.2 Evaluation the synergistic effect by well diffusion method

4.9.2.1 Against E. coli:

As shown in Table (4.8) the aquatic extract of Cuminum cyminum with

Ocimum basilicum shown inhibition zone diameter of 7mm but there was no effect of

ethanolic extract from them against E. coli. The aquatic and ethanolic extract of

Artemisia monosperma with Coriandrum sativum shown inhibition zone diameter

of 8mm.

4.9.2.2 Against S. aureus:

The table (4.9) shown that there was no any effect for both Cuminum

cyminum with Ocimum basilicum and Artemisia monosperma with Coriandrum

sativum in each of aquatic and ethanolic extract against S. aureus

81

Table (4.8): Antimicrobial Activity between Plant extracts on E. coli by well

diffusion method and disc diffusion method


Average of inhibition zone. (± SD.)


- 7(0) - - 8(0) - Cuminum cyminum

with Ocimum

basilicum

- 8(0) 8(0) - - 7(0) Artemisia

monosperma with

Coriandrum sativum

* Antimicrobial Activity Assays. Control= DMSO

(-) No inhibition zone. Standard deviation = SD

Table (4.9): Antimicrobial Activity between Plant extracts on S. aureus by well

diffusion method and disc diffusion method

Well diffusion method(mm) Disc diffusion method(mm) A*A*A

Average of inhibition zone. (± SD.)


- - - - - - Cuminum cyminum

with Ocimum

basilicum

- - - - - - Artemisia

monosperma with

Coriandrum sativum

* Antimicrobial Activity Assays. Control= DMSO

(-) No inhibition zone. Standard deviation = SD

82

4.9.3 The minimum inhibitory concentration (MIC) of mixed plant extracts

against isolated bacteria:

4.9.3.1 Against E. coli:

As shown in table 4.10 and figure 4.49 and 4.50, the MIC of the aquatic

extract of Cuminum cyminum with Ocimum basilicum was 6.25 mg/ml and the

ethanolic extract was 25 mg/ml. The MIC for Artemisia monosperma with

Coriandrum sativum of aquatic extract was 12.5 mg/ml and the ethanolic extract was

25 mg/ml

4.9.3.2 Against S. aureus:

The MIC results of the aquatic extract of Cuminum cyminum with Ocimum

basilicum was 25mg/ml and in ethanolic extract was 50mg/ml. The MIC for

Artemisia monosperma with Coriandrum sativum of aquatic and ethanolic extract

was 25 mg/ml against S.aureus (Table 4.10) and (figure 4.47 and 4.48).

Table (4.10): The minimum inhibitory concentration (MIC) between plant extracts

against isolated bacteria

E.coli MIC

Conc.

S. aureus. MIC

Conc.

Solvent

Plant extract

Ethanol Water Ethanol Water

Cuminum cyminum with Ocimum basilicum 25 6.25 50 25

Artemisia monosperma with Coriandrum

sativum

25 12.5 25 25

83

Figure (4.47): the MIC of aquatic

extract of 12- Cuminum cyminum

with Ocimum basilicum and 13-

Artemisia monosperma with

Coriandrum sativum against S.

aureus.

Figure (4.48): the MIC of ethanolic




Coriandrum sativum against S.

aureus.

84

Figure (4.49): the MIC of aquatic




Coriandrum sativum against E.coli

Figure (4.50): the MIC of

ethanolic extract of 12- Cuminum

cyminum with Ocimum basilicum

and 13- Artemisia monosperma with

Coriandrum sativum against E.coli

85

Chapter (5)

Discussion and

Conclusions

86

Plant kingdom is one of the most important sources of the natural product due

to both medicinal and economical values (Kharjul et al., 2012). Many of medicinal

plants are known for decades as essential resources to human health.

The traditional cultural use of plant recipes and their protective and curative

significance is most likely associated with low cost, limited side effects, easy access

and the popular relationship with them.

People continue to use traditional remedy by consuming special medicinal

plant species sold at local markets and due to the limited number of studies in

Palestine in this field (Ali- Shtayeh and Jamous, 2006). The current study aimed to

evaluate the traditional use of medicinal plants by therapists in Gaza strip, as a part

of a series of ethno-botanic studies for estimation of the status of traditional herbal

medicine in the Palestinian communities.

5.1 Study population:

The current study shows that men were predominantly represented in the

samples where the folk therapy with medicinal plants considered more common

among men more than women with age group between 45–65 years, and this is not

consistent with study (Idu et al., 2010) which reported that the majority of medicinal

plant traders in Nigeria were women (64.29 %) between the age group of 40 – 60

years. The main source of traditional knowledge was also mainly rely on the

information and experiences gained from parents and grandparents and although

there is a similarity in some of the therapeutic recipes, each healer has its own

knowledge and tries to keep it secret and not exchange with everyone.

These findings clearly show that herbal medication still holds an important

status in folkloric medicine in Gaza strip, and these agree with Ali-Shtayeh and

Jamous (2006) which note that a people in Gaza strip and West Bank depend on a

monthly average visit to folk therapies at least 2-6 time on month.

5.2 Medicinal Plants Markets:

Most of the plant materials used in folk medicine is sold in herbal shops and

mostly being imported from other countries. Most of the plant materials were in

dried form and sold either singularly or as a mix with other plants for a particular

treatment (Idu et al., 2010). So to collect information about the markets of medicinal

87

plants, a series of semi-structured interviews were conducted with 20 informant of

therapist and sellers of medicinal plants in Gaza Strip. Seventy two species of

ethnomedicinal plants belonging to 33 families were considered the most widely

used by therapist in the Gaza Strip. The most important family was Lamiaceae with 9

species, followed by Apiaceae with 8 species, Asteraceae with 7 species and

Fabaceae with 5 species other families with low numbers. (Table 4.1).

5.3 Plants and parts used

In the current study leaves were reported to be the most commonly used plant

parts for treatment (Figure 4.3). This might indicates that leaves are the major site of

active ingredients, and these agree with studies which observed that aerial parts are

the most frequently used parts in medicinal therapies (Amiri and Joharchi, 2013;

Arellanes et al., 2013), but these disagree with previous study which observed that

the underground parts were most used by therapies, while the whole plant and leaves

were second and third respectively (Maneenoon et al., 2015).

Each plant has a different method of use according to the part from the plant

used and the method of preparation of the prescription depending on the type of

disease. Most therapeutic prescriptions were prepared by simply boiling or soaking

the plant and were administered as tea. Table (4.2).

Plant recipes may include one or more plants, and one plant can be used to

treat many diseases. This is why it is difficult to identify certain plants for certain

diseases (Ali-Shtayeh et al., 2000)

These study showed that plants sold at the traditional markets were mostly

used for gastrointestinal disorders and these agree with previous study of (Amiri and

Joharchi, 2013; Maneenoon et al., 2015). However, our results indicate that the

number of plant species that treating the gastrointestinal system was (37) species,

followed by circulatory system (22), urinary system (20), diabetes (18), respiratory

system (14), nervous system (13), skin, burns, and hair (12), weight loss (10),

skeletal and muscular system (7), immunity system (7), reproductive system (7),

cancer (3), and (11) for other diseases.

88

5.4 Antibacterial Activity of the Plant Extracts

Many naturally occurring compounds found in plants have been served as a

source of antimicrobial agents against pathogens. This study evaluates the ability of

the aquatic and ethanolic extract of 11 plants species from the 72 species to inhibit

the growth of two type of pathogenic bacterial isolates E. coli and S. aureus.

Both disc diffusion method and well diffusion method were used to assess the

activity of plant extracts. It is noted that obtained results from well diffusion method

much better than that of disc diffusion method against the two types of bacterial

isolate and this finding in agreement with the previous study of (Al-Talib et al.,

2016). These results may be due to that the plant extracts in well diffused method

diffused in the plate more than that of disc diffusion method. Although, the low

recorded values of some plant extracts can be attributed to the fact that the extracts

are in crude form, containing very small amounts of biologically active compounds

(Karmegam et al., 2008). Probably part of prevention or inhibitor material for the

growth of microorganisms in plant extracts may lose their inhibitory power during

extraction methods (Elkichaoi., et al 2015).

The most results of disc and well diffusion method of the aqueous plant

extracts provided more strong antimicrobial activity compared to ethanolic extracts

against the isolate bacterial. This may be due to the aquatic extract found to be richer

in polar phenol than in ethanolic extracts (Triantaphyllou et al., 2001) which in

consistence with traditional therapies who use primarily water as the solvent. Also it

is found that the ethanolic plant extracts of Syzygium aromaticum.L have large

inhibition zone against E. coli, but gave a close result against S. aureus in both

ethanolic and aquatic extracts in both disc and well diffusion method. This may be

related to different solubility of the active constituents in the different solutions

(Parekh and Chanda., 2007).

It is shown that excellent effect of Syzygium aromaticum.L and Hibiscus

sabdariffa.L against both E.coli and S.aureus that infect the gastrointestinal tract and

these agree with (Tolulope, M., 2007 ; Cortés-Rojasf et al., 2014). Although these

two plant species have not been mentioned by therapists for treatment of

gastrointestinal disorder as shown in Table (4.2).

89

Microdilution method was used to determine the lowest plant extracts

concentration that inhibiting the growth of the bacteria isolates. As shown in

Tables(4.7), (4.8), the MIC value of Syzygium aromaticum.L of ethanolic extract

against S.aureus was found as the lowest 0.39mg/ml. On the other hand, the aquatic

extract of this plant species shown 6.25mg/ml against S.aureus. It also showed least

concentration of ethanolic extracts against E.coli with value 0.78mg/ml. On the

contrary of Syzygium aromaticum.L the Hibiscus sabdariffa.L shown the least MIC

value in aquatic extact. Compared to ethanolic extract against the two pathogenic

bacterial isolates. Where the MIC value of Hibiscus sabdariffa.L in aquatic extract

against S.aureus was 6.25mg/ml but the ethanolic extract value was 12.5mg/ml, and

the MIC value of this plant species in aquatic extract against E.coli was 0.39mg/ml

and the ethanolic extract was 0.78mg/ml.

Some researchers however, have reported that the activity in the disc

diffusion assay does not necessarily correlate to low MIC & MBC values in the

microtitre plate method (Ncube et al., 2008). This finding were agreed with our

results where the result of MBC of the ethanolic extracts of Glycyrrhiza glabra was

100mg/ml against E.coli. In contrast, the MBC results for the rest of the plant species

in both ethanolic and aquatic extract against E.coli was > 200 mg/ml.

The difference in antimicrobial activity of the plant extract can be due to the

freshness of the plant material, the physical factors (temperature, water) or the drying

method used prior to the extraction process and the extraction method (Okigbo and

Mmeka., 2008). As well, the successive isolation of plant compounds from plant

materials depends largely on the type of solvent used in the extraction process. We

have seen this clearly in the results of Syzygium aromaticum.L extracts which gave

high results against E.coli and S.aureus in the ethanolic extracts than in the aquatic

extract.

On the basis of the antibacterial assay of this study E. coli (Gram negative) as

found more susceptible to the employed plant extracts than S. aureus (Gram positive)

and this disagree with the previous study of Elbashiti which show that there was no

antibacterial activity of plant extracts against E.coli (Elbashiti., et al 2011) and this

due to the different cell-wall shape of different types of bacteria (Elkichaoi., et al

2015).

90

This current study detected the effectiveness of the synergistic effects of

some plant extract when mixed with each other toward a pathogenic bacterial

isolates. Plant extracts mixing gave weak results compared to the use of plant

extracts alone, this result disagree with (Karmegam., et al 2012) which showed that

the antimicrobial activity of aqueous and ethanolic leaf extracts of some plants gave

lower inhibition zones when used alone than that of the extract combinations.

One of the prescriptions from a popular therapist was tested and the

synergistic effect was tested of Cuminum cyminum with Ocimum basilicum, and of

Coriandrum sativum with Artemisia monosperma against each of E.coli and

S.aurues.

As shown in Table (4.9) there was antagonistic effect in the disc diffusion

method of ethanolic and aquatic extracts of mixed Cuminum cyminum with Ocimum

basilicum and mixed of Coriandrum sativum with Artemisia monosperma against

S.aurues. On the other hand, these mixed plant extracts showed weak synergistic

effect against E.coli with inhibition zone 7-8mm. (Table 4.8).

As for well diffusion method, the results were different. The Table (4.10)

showed that the effect of the aquatic extract of each plant separately on a bacterium

gave average results between 13-23 mm inhibition zone but when mixed the extracts,

there was no synergistic effect against S.aureus. These extracts showed 7-8 inhibition

zone against E.coli in aquatic extracts which was lower than that of each extract

alone.

Therefore, the results of this prescription were weak compared with the use of

each extract alone, and this is clearly shown in the results of MIC, where all extracts

showed a decrease in MIC to test synergistic effect. This finding was in agreement

with the previous study of Adwan & Mhanna (2008) which explained this as these

crude extracts have many different phytochemicals which might inhibit bacteria and

fungi by different mechanisms.

91

5.5 Conclusion:

The folk treatment and the use of medicinal plants in therapy is still transferred

from grandfathers and parents to younger generation and not depend on academic

study.

This study indicates that a large number of ethnomedicinal plant (72) species are

still the most useful in the folk treatment in Gaza strip. Majority of these plants

imported from other countries and some of them considered as local.

our results indicate that the number of plant species that treating the

gastrointestinal system was (37) species, followed by circulatory system (22), urinary

system (20), diabetes (18), respiratory system (14), nervous system (13), skin, burns,

and hair (12), weight loss (10), skeletal and muscular system (7), immunity system

(7), reproductive system (7), and cancer (3).

The most results of disc and well diffusion method of the aqueous plant extracts

provided more strong antimicrobial activity compared to ethanolic extracts against

the isolate bacterial. The strongest effect against E. coli was recorded in MIC in

aquatic extract of Hibiscus sabdariffa.L with value 0.39mg/ml. But the Syzygium

aromaticum.L observed strong effect in each S. aureus and E. coli in ethanolic

extract by all types of evaluation methods.

5.6 Recommendation:

1- The need to maintain knowledge through documentation and to encourage people

working in the field (therapists) with a control by the Ministry of Health.

2- The need to conduct awareness seminars by the Ministry of Health on the harms

and benefits of plant extracts and how to use them properly.

3-Toxicity studies of active plants should be conducted to determine safety indicators

for extracts.

4- Determine the interaction of these crude extracts with each other to determine the

validity of the recipes based on mixing the medicinal plants together by therapists.

5- It is essential to make the complete inventory of the medicinal component of the

flora of Palestine for conservation and sustainable use.

92

Chapter (6)

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100

Appendix

101

Appendix: A

Interview Question

ؼيبد شخظخ3

اىدظ رمش ] [ اث ] [

] [ 44] [ امجش 44 -14 ] [ 14-32] [ 02-41اىؼش

] [ اثزذائ ] [ زعؾ ] [ اىغز اىزؼي ا

ثب ] [ خبؼ ] [

ــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــ

ز ز ربسط ز اىخ؟ .4

ا رحظو ػي اىؼيبد اىزؼيقخ ثبىؼلاج ثبلأػشبة؟ .0

و بسط احذ غشك ف اىؼبئيخ ز اىخ؟ .3

و رق ثزؼي أ احذ ز اىخ حز لا رذثش؟ .1

رؼبىد ف الاعجع؟؟ و ىل صثبئ دائ؟؟م ػذد اىشػ اىز .4

و خذ اشاع حذدح زؼبىح ب اغيت اىبط ثبلأػشبة؟؟ ارمش ثؼغ ز .4

الاشاع؟

ا رحظو ػي الاػشبة؟؟ .2

و راخ طؼثبد ف اىحظه ػي اىجبربد اىطجخ؟ ب ز اىظؼثبد؟؟ .1

طؼت اىحظه ػيب؟؟و خذ جبربد رغزخذب ف اىؼلاج بدسح .2

ارمش اعبء ثؼغ ز اىجبربد؟؟

و اخذ حبلاد رؤثشد ثشنو عيج اىؼلاج ثبلأػشبة؟؟ .42

و رق ثئػذاد اىزشمجبد اىذائخ ثفغل؟؟ ا مبذ الاخبثخ ؼ... ارمش ثؼغ .44

اىطفبد اىزقيذخ اىز رق ثئػذادب ثفغل ا أن ؟

ز رذخو ف اغيت اىؼلاخبد اىز رق ثئػذادب ؟؟ارمش اعبء ثؼغ اىجبربد اى .40

غبىجب أ خضء اىجبد ز اعزخذا ف اػذاد اىطفبد اىطجخ ؟؟ .43

و رؤذ خد شامض اىؼلاج ثبلأػشبة ؼزذح صاسح اىظحخ اىفيغطخ؟؟ .41

ا مبذ اخبثزل لا... ارمش الاعجبة؟؟

بطو ػي شبدح زخظظخ ثزا اىدبه؟؟و رؤذ ا ن اىؼبىح ثبلأػشبة ح .44

و ىذل أ اقزشاحبد ا لاحظبد حه ػع اىذساعخ ) اعزخذا اىجبربد اىطجخ .44

ف اىؼلاج اىشؼج(

102

ثؼذ الاخبثخ ػ الاعئيخ اىغبثقخ و نل رؼجئخ زا اىدذه؟؟ .42

لازظاخ الاػذادطش٠مح ا الاشاض ار ٠ؼادا ادضء اغرخذ اع اثاخ اشل

103

Appendix: B

Photograph of survey

104

105

Documents

An Ethnobotanical Study of Medicinal Plants Used By Palestinians … · 2018. 9. 19. · IV Abstract The study aimed at evaluating the current status of folk medicine treatment in