Antimicrobial Activity of some Sudanese Plants against Mycetoma Causing Agents with emphasis ...

140 Pages·2011·1.35 MB·English

ﻢﻴﺣﺮﻟا ﻦﻤﺣﺮﻟا ﷲا ﻢﺴﺑ

University of Khartoum
Gradute College
Medical & Health Studies Board

Antimicrobial Activity of some Sudanese Plants against Mycetoma Causing
Agents with emphasis on Prosopis chilensis plant

By

Fatima Abdalla Mohammed Ali

BSc. In chemistry & Zoology U of K

MSc. Pharmacognosy U of K

A thesis submitted in fulfillment for the requirement of the Degree of PhD in
Chemistry. January 2010

Supervisor

Professor KamalEldin Eltayeb Ibrahim

MD. PhD Chemistry

Professor of Pharmaceutical Chemistry

U of K

2010
Dedication

To my family
To those who have given me their time and energy to supply
valuable facts and opinions
To my mother soul
ACKNOWLEDGMENT

Before everything and after everything I am thankful to the most Merciful,
Omnipotent, Omnipresent and Omniscient Almighty Allah. With his help and
blessing I was able to achieve this mission. My deepest thanks to my supervisors
Professor KamalEdin Etayeb Ibrahim Department of Pharmaceutical Chemistry
Faculty of Pharmacy U of K and Professor Ahmed Hassan Fahal Department of
Surgery Faculty of Medicine U of K for their assistance, interest and guidance
throughout this study. Deep gratitude and many thanks to all members of the staff
of Mycetoma Research Center at Soba teaching hospital for their help in collection
and culture of the pathogens.
I am also indebted to Dr Wail Alsadig Medicinal and Aromatic Plant Institute for
their help in the taxonomical identification of the plant species.
I would like to express my thanks and gratitude to Juba University Vice chancellor
and dean of college of Education Ministry of Higher Education for the financial
support for the short fellowship to Pakistan.
It is indeed a pleasure for me to express my indebtedness and gratitude to Prof. Dr.
M. Iqbal Choudhary, the acting Director of the International Center for Chemical
Sciences (ICCS), University of Karachi, Karachi, Pakistan for his kind offer of a
chance to do the isolation and spectroscopic analysis. My sincere appreciation to
Dr. Mohamed A. Mesaik, Dr. Panjwani Centre for Molecular Medicine and Drug
Research (PCMD) for their distinct and thorough guidance in conducting the
immunomodulating test.
I wish to acknowledge the kind cooperation of the technical staff members in the
Department of Pharmaceutical Chemistry Faculty of Pharmacy U of K. Finally I am
greatly in debated to my father, sister and brothers for their patience understanding
and moral support.
  i
ABSTRACT

Background
Mycetoma is a chronic granulomatous disease of subcutaneous and deep tissue.
Progressive destruction of tissue leads to loss of function of the affected site. The
pathogens are found in the soil as saprophytes, inoculated through the skin by minor
cuts or scratches by thorn or splinter wounds. Mycetoma infection is caused by true
fungi (eumycetoma) or by higher bacteria of the class actinomycetes
(actinomycetoma). The incidence of mycetoma in Sudan has not changed; around
400 new cases are seen in hospitals and outpatient clinics every year. Treatment of
Eumycetoma infection is difficult and the rate of recurrence after surgery is high in
spite of medical treatment in combination with surgery. The quest for effective
medical treatment was therefore actively persued.
Objective
1-To test the antimicrobial activity of 45 indigenous plant extracts (leaves, bark,
roots, pods, seeds and fruits), against mycetoma pathogens, using agar diffusion
method.
2-To achieve NCCLS modified in vitro test against Madurella mycetomatis and
immunomodulation property for Prosopis chilensis (leaves, bark and root), the most
active plant as well as the chemical constituents.
Methodology
70 plant extracts were prepared by small scale extraction procedure with 70%
methanol. The mycetoma pathogens were isolated from patients and cultured using
different media. The antimicrobial activity was assayed in vitro using different
methanolic extracts. Large scale extraction procedure was done with different
solvent fractions for Prosopis chilensis three parts. In vitro susceptibility for
Madurella mycetomatis hyphal suspension as well as immunomodulation property
  ii
was tested using different human blood cells Prosopis chilensis three parts extract
and fraction.
Results
Twenty four plant extracts from 70 extracts(from 45 plant species) exhibited
various degrees of antimicrobial activity. MIC range between 0.2-4 mg/5ml against
the three organisms. Crude methanolic extracts from the leaves and bark of P.
chilensis showed the most potent activity against M. mycetomatis of MIC value
0.26 mg/5ml medium and 0.5 mg/5ml medium respectively. While S. nigrum fruits
showed MIC against M.mycetomatis of 0.8 mg/5ml and M. olifera seeds 0.95
mg/5ml and L. sativum seeds of 1mg/5ml. P. chilensis leaves showed 0.35 mg/5ml
medium against A. pelletieri and 0.3 mg/5ml medium against S. somaliensis . P.
chilensis leave and the bark methanol crude extract exhibited antifungal activity
against M.mycetomatis at concentration range from0.02 to 1mg/ml. The leaves and
the bark of P. chilensis crude extract inhibit the growth completely at both
concentrations 1mg and 0.5 mg/2ml medium. While PcR-1 {5-hydroxy4’-
hydroxy7-Orhamnopyrnoside flavone (57)}, isolated from the root is the most
potent compound at 1mg/2ml in compared to the control (ketoconazol 1mg/2ml).
PcL-7 Piperidine-2-methyl -3 acetyl-6-dodcyl- 25-methyl –indolizin-ol (59),
isolated from the leaves, showed fungal growth inhibition at 1mg/2ml medium.
The leaves extracts, showed a dose- dependent effect with > 90% inhibition at the
highest concentration (100 µg/ml), have effect on the cell response.
The methanol extract of the leave is most active with inhibition 99.8 ± 0.1% at 12.5
µg/ml while the bark and root show high activity at 100µg/ml and they revealed
variable inhibition % and LC < 12.5. Immunosuppressive activity were found
50
significantly (P < 0.01) suppressing the proliferation of T-cells.

  iii
Conclusion
Out of 70 methanolic extracts of different plants P. chilensis leaves and barks were
found to be the most potent antimicrobial agents against mycetoma causing
pathogens. The alkaloidal compounds isolated from the leaves extract were the
most potent antifungal among other examined compounds.
These studies clearly demonstrate that P. chilensis plant is rich source of
antimicrobial and immunomodulating substances. The majority of the isolated
compounds were found to possess significant antifungal activity and good
candidates for developing immunosuppressive or antimicrobial agents.

  iv
ﻢﻴﺣﺮﻟا ﻦﻤﺣﺮﻟا ﷲ ا ﻢﺴﺑ
ﺚﺤﺒﻟا ﺔﺻﻼﺧ
ﺔﻴﻔﻠﺨﻟا
ﺪﻨﻋ . ﻪﻘﻴﻤﻌﻟا ﻪﺠﺴﻧﻻاو ﺪﻠﺠﻟا ﺖﺤﺗ ﻰﻣﻮﺒﻴﺒﺣ ﻰﻌﺿﻮﻣ مرﻮﺗ ﻮهو ,ﻪﻨﻣﺰﻤﻟا ضاﺮﻣﻻا ﻦﻣ ﺎﻣﻮﺘﺴﻳﺎﻤﻟا ﺮﺒﻨﻌﺗ
بوﺮﻜﻴﻣ ﺪﺟﻮﻳ .ﺮﺘﺒﻠﻟ بﺎﺼﻤﻟا ﻮﻀﻌﻟا ضﺮﻌﻳ ﺎﻤﻣ ﻎﻟﺎﺑﺮﻴﻣﺪﺗ ﻰﻟا ﻪﺠﺴﻧﻻا ضﺮﻌﺘﺗ ﺔﻠﺤﻔﺘﺴﻤﻟا ﻪﺑﺎﺻﻻا
ﺔﻨﻌﻃ وا ﺮﻴﻐﺻ حﺮﺟ وا شﺪﺨﻟا ﻖﻳﺮﻃ ﻦﻋ بﺎﺼﻤﻟا ﻢﺴﺟ ﻰﻟا ﻞﺧﺪﻳو ﻪﺑﺮﺘﻟا ﻰﻓ ﻞﺻﻮﺤﺘﻣ ﺎﻣﻮﺘﺴﻳﺎﻤﻟا
. ﺔآﻮﺷ
eumycetoma ﻰﻓ ﺎﻤآ ﻪﻴﻘﻴﻘﺣ تﺎﻳﺮﻄﻓ ﺎﻣﻮﺘﺴﻳﺎﻤﻟا ضﺮﻣ ﺐﺒﺴﺗ
Actinomycetes ﺔﻠﻴﺼﻓ ﻦﻣ ﺎﻴﻠﻋ ﺎﻳﺮﺘآﺎﺑو
تﺎﻴﻔﺸﺘﺴﻤﻟا ﻰﻓ ﺎﻳﻮﻨﺳ ﻩﺪﻳﺪﺟ ﻪﺑﺎﺻا ﻪﻟﺎﺣ 400 ﻰﻟا ﻞﺼﺗ ﺚﻴﺣ ﻪﻳراﺮﻜﺘﻟا ﻪﺑﺎﺻﻻا تﻻﺪﻌﻣ دادﺬﺗ
ﻰﻟا ﻞﺼﺗ ﺚﻴﺣ ﻪﺣاﺮﺠﻟا ﺪﻌﺑ ﻰﺿﺮﻤﻟا سﺎﻜﺘﻧﻻا تﻻﺎﺤآ ﻩﺮﻴﺒآ تﻻﺪﻌﻣ ﺮﻬﻈﺗ ﺎﻤآ .ﻪﻴﺟرﺎﺨﻟا تادﺎﻴﻌﻟاو
ﻩﺪﻴﺟ ﻪﻴﻟﺎﻌﻓ تاذ ىﺮﺧا تادﺎﻀﻣ ﻦﻋ ﺚﺤﺒﻠﻟ ىدا ﺎﻤﻣ ﻪﻴﺋﺎﻴﻤﻴﻜﻟا ﻪﻳودﻼﻟ ﻪﻴﺒﻧﺎﺠﻟارﺎﺛﻼﻟ ﺔﻏﺎﺿﻻﺎﺑ %80
. ﻪﻴﺗﺎﺒﻧ ردﺎﺼﻣ ﻦﻣ
ﺚﺤﺒﻟا ﻦﻣ ضﺮﻐﻟا
. تﺎﻳﺮﻄﻔﻟاو ﺎﻳﺮﺘﻜﺒﻟا ﺪﺿ ﻪﻳﻮﻴﺣ ﻪﻴﻟﺎﻌﻓ ﺎﻬﻟ ﻪﻴﻧادﻮﺳ لﻮﺻا ﻦﻣ ﻪﻠﻔﻠﺘﺨﻣ عاﻮﻧا تاذ تﺎﺒﻧ 45 ﻰﻟاﻮﺣ رﺎﻴﺘﺧا
. تﺎﺗﺎﺒﻨﻟا ﻩﺬﻬﻟ ﻪﻔﻠﺘﺨﻣ ءاﺰﺟﻻ ﺺﻠﺨﺘﺴﻣ 70 ﺎﻬﻨﻣ
(ىﺮﻄﻓ 1 ﺎﻳﺮﺘﻜﺒﻟا ﻦﻣ 2) ﺎﻣﻮﺘﺴﻳﺎﻤﻟا تﺎﺑوﺮﻜﻴﻣ ﻦﻣ ثﻼﺛ ﻰﻠﻋ تﺎﺼﻠﺨﺘﺴﻤﻟا ﻩﺬه ﺔﻴﻟﺎﻌﻓ رﺎﺒﺘﺧا
. رﺎﺟﻻا ﻂﺳو قاﺮﺘﺧا ﺔﻘﻳﺮﻃ ماﺪﺨﺘﺳﺎﺑ
Prosopis chilensis ﺖﻴﻜﺴﻤﻟا تﺎﺒﻧ ﺔﻴﻟﺎﻌﻓ رﺎﺒﺘﺧا ﻚﻟﺬآ
ﻞﻜﻟ ﻲﻋﺎﻨﻤﻟا ﺮﻳﻮﺤﺘﻟا ﺔﻴﺻﺎﺧ رﺎﺒﺘﺧا ﻚﻟﺬآو ﻪﻴﻄﻴﺨﻟا تﺎﻳﺮﻄﻔﻟا ﺔﻋارﺰﻟ ﻪﺛﺪﺤﺘﺴﻤﻟا ﺔﻘﻳﺮﻄﻟا ماﺪﺨﺘﺳﺎﺑ
. تﺎﺒﻨﻟا اﺬﻬﻟروﺬﺠﻟاو ءﺎﺤﻟاو قاروﻻا ﻦﻣ
ﺚﺤﺒﻟا داﻮﻣو قﺮﻃ
لﺰﻋ . صﻼﺨﺘﺳﻼﻟ ﺮﻐﺼﻤﻟا راﺪﻘﻤﻟا ماﺪﺨﺘﺳ ﺎﺑ %70 ﻰﻠﻴﺜﻤﻟا لﻮﺤﻜﻟا ﻦﻣ ﺺﻠﺨﺘﺴﻣ 70 ﺮﻴﻀﺤﺗ
ﻢﺴﺠﻟا جرﺎﺧ ﻲﺑوﺮﻜﻴﻤﻟا ﻂﻴﺒﺜﺘﻟا رﺎﻴﺘﺧا . ﻪﻔﻠﺘﺨﻣ طﺎﺳوا ﻰﻓ ﺎﻬﻋرزو ﻰﺿﺮﻤﻟا ﻦﻣ ﺎﻣﻮﺘﺴﻳﺎﻤﻟا تﺎﺑوﺮﻜﻴﻣ
.رﺎﺟﻻا ﻂﺳو قاﺮﺘﺧا ﺔﻘﻳﺮﻄﺑ تﺎﺻﻼﺨﺘﺴﻤﻟا ﻞﻜﻟ
ماﺪﺨﺘﺳا ﻢﺛ ﻦﻣو , ﺮﺒﻜﻤﻟاراﺪﻘﻤﻟا ماﺪﺨﺘﺳﺎﺑ ﺖﻴﻜﺴﻤﻟا تﺎﺒﻧ روﺬﺟو ءﺎﺤﻟ , قاروﻻ ﺔﻔﻠﺘﺨﻣ تﺎﺼﻠﺨﺘﺴﻣ ﺮﻴﻀﺤﺗ
ﺮﻄﻔﻟ ﻢﺴﺠﻟا جرﺎﺧ ىﻮﻴﺤﻟا ﻂﻴﺒﺜﺘﻟا ﺔﻴﻠﺑﺎﻗ رﺎﺒﺘﺧا ﻢﺛ ﻦﻣو ﻪﻴﻄﻴﺨﻟا تﺎﻳﺮﻄﻔﻟا ﺔﻋارﺰﻟ ﻪﺛﺪﺤﺘﺴﻤﻟا ﺔﻘﻳﺮﻄﻟا
Madurella mycetomatis
. ﺖﻴﻜﺴﻤﻟا تﺎﺒﻨﻟروﺬﺠﻟاو ءﺎﺤﻠﻟاو قاروﻻا ﻦﻣ ﻞﻜﻟ ﻲﻋﺎﻨﻤﻟا ﺮﻳﻮﺤﺘﻟا ﺔﻴﺻﺎﺧ رﺎﺒﺘﺧا ﻚﻟﺬآو
. ﺔﻔﻠﺘﺨﻤﻟا ﻞﻴﻠﺤﺘﻟا قﺮﻃ ماﺪﺨﺘﺳﺎﺑ ﺔﻴﺋﺎﻴﻤﻴآ تﺎﺒآﺮﻣ ةﺪﻋ ﻞﺼﻓ ﻢﺛ ﻦﻣو
ﺚﺤﺒﻟا ﺞﺋﺎﺘﻧ ﺔﺸﻗﺎﻨﻣ
ﻂﻴﺒﺜﺗ ﺔﺟرد ﻞﻗﺎﺑ ﺎﻣﻮﺘﺴﻳﺎﻤﻟا تﺎﺒﺒﺴﻣ ﻦﻣ ثﻼﺛ ﺪﺿ ﻰﺑوﺮﻜﻴﻤﻟا ﻂﻴﺒﺜﺘﻟا ﻦﻣ ﺔﺟرد ﺎﻬﻟ ﻰﺗﺎﺒﻧ ﺺﻠﺨﺘﺴﻣ 24
ﻂﺳﻮﻟا ﻦﻣ ﻞﻣ 5 ﻞﻜﻟ ماﺮﺠﻠﻣ 4-0.2 ﻦﻴﺑ جواﺮﺘﺗ
ﺎﻣﻮﺘﺴﻳﺎﻤﻟا تﺎﺑوﺮﻜﻴﻣ ﻂﻴﺒﺜﺘﻟ ﺎﻌﺟﺎﻧ ﺮﺜآا ﺖﻴﻜﺴﻤﻟا تﺎﺒﻧ قارواو ءﺎﺠﻟ ﻦﻣ ﻞﻜﻟ ﻞﺜﻴﻤﻟا تﺎﺼﻠﺨﺘﺴﻣ تﺪﺑا ﺚﻴﺣ
ﻞﻣ 5 ﻞﻜﻟ ماﺮﺠﻠﻣ 0.5و ﻞﻣ 5 ﻞﻜﻟ ماﺮﺠﻠﻣ 0.26 قاروﻻا ﺺﻠﺨﺘﺴﻤﻟ ﺖﻧﺎآ ﻻرﻮﻳدﺎﻤﻟا ﺮﻄﻔﻟ ﻂﻴﺒﺜﺗ ﺔﺟرد ﻞﻗا
  v
0.8 ﻂﻴﺒﺜﺗ ﺔﺟرد ﻞﻗا ﻻرﻮﻳدﺎﻤﻟا ﺮﻄﻓ ﻩﺎﺠﺗ ﺔﻴﻟﺎﻌﻓ ﺐﻳﺪﻟا ﺐﻨﻋ ةﺮﻤﺜﻟ ﻞﻴﺜﻤﻟا ﺺﻠﺨﺘﺴﻣ ىﺪﺑا . ءﺎﺤﻟا ﺺﻠﺨﺘﺴﻤﻟ
1 دﺎﺷﺮﻟا تﺎﺒﻧ روﺬﺑو ماﺮﺠﻠﻣ 0.95 ﻂﺴﻴﺜﺗ ﺔﺟرد ﻞﻗا قاوﺮﻟا تﺎﺒﻧ روﺬﺑ ﺺﻠﺨﺘﺴﻣ ﻚﻟﺬآ ﻞﻣ 5 ﻞﻜﻟ ماﺮﺠﻠﻣ
ﺔﺟرد ﻞﻗا ﺎﻣﻮﺘﺴﻳﺎﻤﻠﻟ ﻪﺒﺒﺴﻤﻟا ﺎﻳﺮﺘﻜﺒﻠﻟ ﺔﺤﺿاو ﺔﻴﻟﺎﻌﻓ ﺖﻴﻜﺴﻤﻟا تﺎﺒﻧ قاروا ﺺﻠﺨﺘﺴﻣ ىﺪﺑا ﺎﻤآ . ماﺮﺠﻠﻣ
S.somalensis ﺎﻳﺮﺘﻜﺒﻟ ماﺮﺠﻠﻣ 0.3و A.pelleteri ﺎﻳﺮﻨﻜﺒﻟ ماﺮﺠﻠﻣ 0.35 ﻂﻴﺒﺜﺗ
ﺎﻄﻴﺒﺜﺗ ﻰﻠﺜﻴﻤﻟا ءﺎﺤﻟاو قاروﻻا ﺎﺼﻠﺨﺘﺴﻣ ىﺪﺑا ﺔﻴﻄﻴﺨﺒﻟ تﺎﻳﺮﻄﻔﻟا ﻂﻴﺒﺜﺘﻟ ﺔﺛﺪﺤﺘﺴﻤﻟا ﺔﻘﻳﺮﻄﻟا رﺎﺒﺘﺧا ﺪﻨﻋ
ﺐآﺮﻣ ﻞﺼﻓ ﻢﺗ . ﻞﻣ 2 ﻞﻜﻟ ماﺮﺠﻠﻣ 1-0.02 ﻦﻴﺑ حواﺮﺘﺗ ﻮﻤﻧ ﻂﻴﺒﺜﺗ ﺔﺟرد ﻞﻗا ﻻرﻮﻳدﺎﻤﻟا ﺮﻄﻓ ﻦﻣ ﻞﻜﻟ ﺢﺿاو
PcR-1 {5-hydroxy4’-hydroxy7-Orhamnopyrnoside flavone (57)}
ﺐآﺮﻣ ﻚﻟﺬآو لوﺰﻧﺎآﻮﺘﻴﻜﻟا ءاود ﻢﻜﺤﻤﻟا ﺔﻴﻟﺎﻌﻔآ ﻊﺟﺎﻧ ﻂﻴﺒﺜﺗ وذ ﺖﻴﻜﺴﻤﻟا تﺎﺒﻧ روﺬﺟ ﻦﻣ
PcL-7 Piperidine-2-methyl -3 acetyl-6-dodcyl- 25-methyl –indolizin-ol (59)
ﺮﻳﻮﺤﺘﻟا ﺔﻴﺻﺎﺨﻟ ﺢﺿاو ﺎﻄﻴﺒﺜﺗ قاروﻼﻟ ﺺﻠﺨﺘﺴﻣ ىﺪﺑا . ﻞﻣ 2 ﻞﻜﺗ ماﺮﺠﻠﻣ 1 ﻂﻴﺒﺜﻧ ﺔﺟرد ﻞﻗا قاروﻻا ﻦﻣ
ﺎﻳﻼﺨﻠﻟ ﺢﺿاو ﻂﻴﺒﺜﺗ ﺎﻀﻳا ﻪﻟو ﺔﻴﻋﺎﻨﻤﻟا ﺎﻳﻼﺨﻠﻟ 90%< ﻂﻴﺒﺜﺘﻟ ماﺮﺟوﺮﻜﻴﻣ 100 ﻩﺮﺛﺆﻤﻟا ﺔﻋﺮﺠﻟا ﻰﻋﺎﻨﻤﻟا
. ماﺮﺟوﺮﻜﻴﻣ 12.5> ةﺮﺛﺆﻤﻟا ﺔﻋﺮﺠﻟا ﻪﻳوﺎﻔﻤﻠﻟا
جﺎﺘﻨﺘﺳﻻا
ﺎﻣﻮﺘﺴﻳﺎﻤﻟا تﺎﺑوﺮﻜﻴﻤﻟ ﻰﻄﻴﺒﺜﺗ ﺮﺛا تاذ داﻮﻣ ﻰﻠﻋ ىﻮﺘﺤﺗ تﺎﺗﺎﺒﻨﻟا نا ﺔﺳارﺪﻟا ﻩﺬه ﻦﻣ ﺖﻄﻠﻨﺘﺳا
ﺮﺒﺘﻌﻳ اﺬﻟو . ﺰﻴآﺮﺗ ﻞﻗﺎﺑ ﺮﻄﻔﻟاو ﺎﻳﺮﺘﻜﺒﻟا ﻦﻣ ﻞﻜﻟ ﺢﺿاو ﺎﻄﻴﺒﺜﺗ ﺮﻬﻇا ىﺬﻟا ﺖﻴﻜﺴﻤﻟا تﺎﺒﻨآ
ﻰﻋﺎﻨﻤﻟا ﺮﻳﻮﺤﻨﻟا ﻂﻴﺒﺜﺗو ﺎﻣﻮﺘﺴﻳﺎﻤﻟا مرﻮﺗ جﻼﻌﻟ رﺎﺒﺘﺧا ﻊﺿﻮﻣ ءﺎﺤﻟاو قاروﻻا ﻦﻣ ﻞآ ﺺﻠﺨﺘﺴﻣ
. ضﺮﻤﻟا اﺬﻬﻟ

  vi
CONTENTS
Acknowledgment ................................................................................. i
Abstract ................................................................................................ ii
Arabic abstract ..................................................................................... v
List of Content ................................................................................... vii
List of Tables ..................................................................................... xi
List of Figures .................................................................................. xiii
List of Compounds ............................................................................ xv

1. INTRODUCTION . .………...........…………………………… 1
1.1. Naturally Occurring Products with Antimicrobial activity …........... 1
1.1.2 Antimicrobial Activity of Phenols and Phenolic Acids …............…. 2
1.1.3 Antimicrobial Activity of Quinones …..............………………...….. 3
1.1.4 Antimicrobial Activity of Flavones, Flavonoids and Flavonols .......... 4
1.1.5 Antimicrobial Activity of isoflavonoids ….........……………….…… 5
1.1.6 Antimicrobial Activity of Tannin ……………...........…………….... 6
1.1.7 Antimicrobial Activity of Coumarins……………………...............… 7
1.1.8 Antimicrobial Activity of Essential Oils And Terpenoids ..........……. 8
1.1.9 Antimicrobial Activity of Alkaloid ………..........……………....…... 8
1.1.10 Antimicrobial Activity of Polypeptides and Lectins …….........….…. 9
1.1.11 Plant Products as Antimicrobial Agents …….…………................... 10
1.2.1 Mycetoma Disease ………………………………………................. 12
1.2.2 Mycetoma In Sudan …………….,,,…....………………...............… 13
1.2.3 Mycetoma Treatment…………………………………………............ 15
1.2.3 InVitro Susceptibility of M. Mycetomatis to treatments by a
Modified NCCLS Method ……….………...................………........ 16
1.3 Immune Defense ……………..................…………………….......... 17
1.3.1 Immunomodulation Property ……….........……………………..…... 18
1.3.2 Oxidative Burst Activity …………...........……………………….…. 18
1.3.3 Lymphocyte Cells ………………………...........………………..….. 19
1.3.3.1 T-Cells Proliferation .……............……………………………….… 20
1.3.4 Immunological Changes in Myctoma Disease …............………….... 21
1.4 Some Sudanese Plant used in the present Study .................................. 23
1.4.1 Acacia nilotica (L.) Wild .................................................................... 23
1.4.2 Calotropis procera (Ait.) ................................................................... 23
1.4.3 Capparis decidua (Forsk.) .................................................................. 24
1.4.4 Cassia alata Linn. .............................................................................. 24
1.4.5 Cassia occidentalis Linn. ................................................................... 24
  vii
1.4.6 Citrulus colocynthis L. Schard ....................................................... 24
1.4.7 Cordia sinensis lam. ....................................................................... 25
1.4.8 Datura stramonium Linn. ............................................................... 25
1.4.9 Euphorbia heterophylla Linn ......................................................... 25
1.4.10 Grewia villosa Wild. ........................................................................ 26
1.4.11 Lawsonia inermis L. ........................................................................ 26
1.4.12 Lepidium sativum Husk. .................................................................. 26
1.4.13 Moringa oleifera Lam. .................................................................... 27
1.4.14 Nymphaea lotus L. ........................................................................... 27
1.4.15 Ocimum citrodorumL. .................................................................... 27
1.4.16 Polygonum glabrum Wild. .............................................................. 27
1.4.17 Solanum nigrum Linn. ..................................................................... 28
1.5 Family Fabaceae ………………………………………….....…… 28
1.5.1 Sub Family Mimosoideae ………………………………..…......… 29
1.5.2 Genus Prosopis ……………………………………………........... 29
1.5.3 Prosopis Chilensis (Molino) Stuntz Species ……………...........…. 30
1.5.4 Habitat of Mesquite …………………………………………......... 31
1.5.5 Medicinal Uses in Genus Prosopis …………………………......… 32
1.6 The Biological and Antimicrobial Activity In Genus Prosopis ....… 33
1.7 Chemical Constituents in Genus Prosopis ……………………...... 38
1.7.1 Alkaloids ……………………………………….……………….... 38
1.7.1.1 Piperidine Alkaloids ………………………………………......…. 39
1.7.2 Flavonoids …………………………………………………........... 42
1.7.3 Cyclic Hydrocarbons ……………………………………….......... . 46
1.7.4 Lipids And Sterols ……………………………………….….......... 47
1.7.5 Taninns ………………………..………………………….…........... 47
1.7 Chemical Constituents Isolated From P. Chilensis ………...........… 49
1.8 Study Objectives ………….………………........………………….... 50

2. MATERIAL AND METHODS ……………..........……….......... 51
2.1 Plant Materials ………….………………………….......……....… 51
2.2 Chemicals and Materials Used ...................... …….…….......…… 52
2.3 Media, Reagents and Equipments Used in the Biological Assay ..... 53
2.4.1 Preparation of Reagents and Media Used In Biological Assay ....... 54
2.4.1.1 Borate Buffer Preparation ................................................................ 54
2.4.1.2 Hanks Balanced Salt Solution (HBSS--) .………………….............. 54
2.4.1.3 Hanks Balanced Salt Solution (HBSS++) ……...….......................... 54
2.4.1.4 Preparation of Luminol …………………………………...........…. 55
2.4.1.5 Blood Agar ………………………...........……............................… 55
2.4.1.6 Lowenstein Jensen Acid Medium ………...........………..….......... 55
  viii