Kimbi JN PROTOCOL2 C.sinensis

EVALUATING THE ANTI-FUNGAL ACTIVITY OF Camellia sinensis (L) Kuntze EXTRACT ON ANTI-
FUNGI RESISTANT AND NON RESISTANT CANDIDASPECIES IN THE CITY OF DOUALA
Table of Content
INTRODUCTION.......................................................................................................................................2
I.RESEARCH QUESTION...........................................................................................................................4
II.RESEARCH OBJECTIVES........................................................................................................................5
II.1. General objectives ......................................................................................................................5
II.2. Specific objective........................................................................................................................5
III.LITERATURE REVIEW ..........................................................................................................................6
III.1. GENERALITIES ON THE PLANTS..........................................................................11
III.2 GENERALITIES OF CANDIDASPECIES.........................................................................................11
IV.MATERIAL AND METHOD .................................................................................................................16
V.EXPECTED RESULTS............................................................................................................................22
REFERENCES...........................................................................................................................................23
ANNEX....................................................................................................................................................28
1Presented by KIMBI JAEL NJINGI

INTRODUCTION
Fungal infections are one of the major threats to public health with and increasing frequency
of drug resistance (1) accounting for an excess of 1.7million deaths annually worldwide
(2)A major part of common localized infections are caused by Candidaspecies, they cause
major opportunistic and invasive infections. Candidaspecies that reside in healthy host
include Candidaalbicans, C. tropicalis, C. auris, C. parasitosis, and C. krusei.
Candidanormally resides inside the body such as the mouth, throat, gut and vagina without
causing problems. The current treatment regimens against candidaspecies are based on the
use of the following class of anti-fungi: echinocandines, polyenes, heterocyclic
benzofuranes, flucytosine, azoles just to name a few. Antifungal resistance is an increasing
problem with the fungus Candidarecording considerate percentage of resistance to
antifungal drugs, making them difficult to treat.(3)[3]
According to the studies carried out by CDC’s Antibiotic Resistance Threat (ART) in the
United States 2019, about 7% of all candidablood samples tested are resistant to fluconazole.
Although Candidaalbicans, is the most common cause of severe candidainfections, resistance
is most common to other species particularly candidaauris, candidaglabatra, and
candidaparapsilosis.(4) [4]
Finding alternative treatments which are effective, accessible and less toxic remains a major
challenge. Plant kingdom has always been a hub for many natural compounds with novel
structure, results of new researchers showed that plants are enriched of many bioactive
secondary metabolites such as saponins, alkaloids, terpenoids and phenolic acids which are
characterized by antifungal property[(5)]. Depending on that, these plants can be considered
as a potent future source for antifungal drugs. Epidemiological data suggest that the incidence
and prevalence of serious mycoses continues to be a public health problem, so the spread of
multidrug-resistant strain of fungus and reduced number of drugs available make it necessary
to discover new classes of antifungals from natural products. Some examples of plants with
antifungal properties include: Eugenia uniflora, Psidium guajava, Curcuma longa, Persea
Americana, Cassia alata just to name a few[(6)].
Camellia sinensis (tea plant) of the family theaceae are ever green, medium size wooden
shrubs growing to a height of 1.8m.[(7)] C. sinensis teas have attracted a great deal of

attention due to their numerous health benefits, including antioxidant, hypoglycemic,
anticarcinogentic, antimutagenic, hypocholesterolemic, antimicrobial [(8)]and antifungal
activities like other natural products[(9). Tea is the World’s most widely consumed non-
alcoholic beverage with essential economic and health benefits since it is an excellent source
of polyphenols, catechins, amino acids, flavonoids, carotenoids, vitamins, and
polysaccharides. The aim of this review is to summarize the main secondary metabolites in
tea plants, and the content in green or black tea. The application of these secondary
metabolites in the cosmetics and pharmaceutical industry is to be reviewed in this study.
Numerous functional genes and regulatory factors have been discovered, studied, and applied
to improve tea plants. Research advances, including secondary metabolites, applications,
omics research, and functional gene mining, are comprehensively reviewed here.
This study was designed to investigate the in-vitro anti-candidaeffects of tea extracts on
antifungal resistant and non-resistant strands
Tea biomolecules mainly consist of non-protein amino acids theanine, free sugars,
methylxanthine or purine alkaloids like caffeine, theobromine, theophylline and theacrine,
phenolic acids like gallic acid, and eight other catechins[ (10)]. However, the state of research
on tea regarding its pharmacological properties in relation to fungi is limited and the majority
of work has been conducted on green tea with very little on black and white tea. Various
studies have shown significant suppressive effects of green tea polyphenols against many
microorganisms. Black tea, a major source of phenols including theaflavins and thearubigins
has also shown to have antimicrobial properties both invivo and invitro.(11)
This study attempts to establish the potentiality of C. sinensis plant product as a novel
modalities in the line of new drug discoveries and as a remedy to the multiple antimicrobial
resistance.

I. RESEARCH QUESTION
Does the ethanolic leaf extract of Camellia sinensis possess anti-fungal effect on
antifungal resistant Candidaspecies?

II. RESEARCH OBJECTIVES
II.1. General objectives
 Evaluate the antifungal activity of ethanolic leaf extract of Camellia sinensis

on resistant Candidaspecies as a remedy to Anti-microbial resistance (AMR).
II.2. Specific objective
 Identification and extraction of bioactive compounds found in green and black

tea.
 Collection and identification of antifungal resistant candidastrains
 To compare the antifungal activity of our extract on strains of resistant

Candidaspecies.

I. LITERATURE REVIEW
III.1. GENERALITIES ON THE PLANTS
Phytotherapy in the past and even now has been used to treat various ailments
even before the introduction of modern medicine. Herbal medicines are still
widely used in many parts of the world especially in areas where people have
limited or no access to modern health care (12). In most developing countries
where phyto-therapy is still very much used due to high cost in chemotherapeutics,
there is a need for scientific research to evaluate the biological activities of
medicinal plants. For results obtained may lead to the development and validation
of traditionally used medicinal plants.
Green tea is selected for this study because its consumption has its legendary
origins in China of more than 4,000 years ago. Green tea has been used as both a
beverage and a medicine in most of Asia (13). All the types of teas are
manufactured from the leaves of Camellia sinensis. Tea possesses significant anti-
oxidative, anti-inflammatory, anti-microbial, anti-carcinogenic, antihypertensive,
neuroprotective, cholesterol lowering and thermogenic properties(14)These due to
it rich concentration in bioactive polyphenols.
a) BOTANICAL STUDY
Table 1: Taxonomy of the genus Camellia sinensis
Superkingdom Eukaryote
Kingdom Plantae
Phylum Spermatophyta
Subphylum Angiospermae
Class Dicotyledonae
Order Theales
Family Theaceae
Genus Camellia
Species Camellia sinensis

i. BOTANICAL DESCRIPTION
Theaceae, the tea familyof plants in the order Theales. The

theaceae comprises about 40 genera of trees or shrubs native to
temperate and tropical regions.
Members of the family have evergreen leaves and flowers with

sepals and petals and numerous stamens inserted.(15)
 C.sinensis plants are evergreen, medium size woody shrubs growing into a
height of 1.8m.
 Leaves: oval and pointed at the tip, usually 5-10cm long, shiny, dark-green
above. Leaf margin finely dentate.
 Flowers: they are white, fragrant and up to 4cm diameter.
 Fruits: it’s a 3-angled capsule with seeds and is surrounded by a persistent

petal.(16)
Figure 1: Fresh tea leaves and flower
b) GEOGRAPHICAL STUDY
Camellia sinensis is a native plant from Asia-China, Tibet and

Northern india, these are the most prolific regions in the world.
Tea grows in worm but not exceedingly hot climate for proper
maturity. Fairly heavy rainfall of about 1500mm which is well
distributed over the growing period.
Deep, acidic and well drained fertile alluvial soil for the growing
of the crop(17)
In Cameroon, the following conditions are necessary for tea

production in the three tea growing regions of Cameroon (that is
the Ndu and ndawara tea estates in the North West, Tole tea
estate in the south west and in the west region).
 Annual rainfall of 2000 - 4000mm
 Relative humidity of 70 - 90%
 Sunshine hours of 5hrs/day
Culticated using the cutting method and a hectare of land can provide 2500
tones per year.
The type of tea produced in camerron is the brown tea.(18)

Tea catechins are affected by plucking seasons and plucking
positions(19). The secondary metabolites in tea leaves vary from
different areas since the climate, soil composition, variety, and
other environmental factors are different. However, more details
for geographical environments such as altitude, slope, and
aspect, which would influence the chemical compositions in tea
leaves has not been reported.
Our case study the Ndu Tea Estate, located at the Northeast edge
of the bamenda grasfield. Ndu is the highest elavation town in
Cameroon with an average temperature of 27℃
b) PHYTOCHEMICAL AND PHARMACOLOGICAL STUDY OF Camellia

sinensis
 PHYTOCHEMICAL STUDIES
Various production technics produces different types of teas with

different concentrations in secondary metabolites (SMs), for the
non-fermented tea we can get the green and white tea, semi-
fermented (red tea), and fermented (black tea) all with relatively
different levels of secondary metabolites. Tea biomolecules mainly
consist of non-protein amino acids, theanine, free sugars, methyl
xanthine (caffeine, theobromine, theophylline and theacrine),
phenolic acids like gallic acid and eight other cathechin, with
epigallocatechin-3-gallate as main catechin of green tea
cumulatively they are called polyphenol
According to Mohammed HY et al study on the antifungal effects

of tea leaf polyphenopls on Candidaalbicans, catechins showed
stronger antifungal activity against C. albicans PTCC-5027.
Catechin’s MIC90 (that is the concentration of catechins causing
90% growth inhibition of tested strain of C. albicans) and MFC (the
minimum antifungal susceptibility of catechin) were determined by
Macro dilution test and calculation after 24 and 48hours.(9)
Tea polyphenols includes catechins, flavonoids, anthocyanins and

phenolic acids
Table 2:
Bioactive Components Uses

compounds
Catechins Catechin©, Anti-cancers, anti-
epicatechin(ec), viral or anti-
epigallocatechin(EGC) microbial and anti-
epicatechin oxidant effects.
gallate(ECG)
epigallocatechin
gallate(EGCG)
Flavonoids Flavonol glycosides, Anti-inflammatory
Myricetin glycosides, and anti-oxidants
quercetin glycosides and properties.
Behenyl glycosides
anthocyanines Anti-inflamatory,
anti-oxidant and
protects against
type 2 diabetes
Phenolic acids Gallic acid, chlorogenic Anti-oxidant,
acid, P-coumarinic acid, antimicrobial and
ellagic acid, quinic acid anti-inflammatory.
and tea gallate
The table above is a summary of the bioactive compounds found in

tea.(20)
 THE PHARMACOLOGYCAL STUDY
Teas can be classified depending on the degree of fermentation such

as green tea (unfermented tea), white and yellow tea (lightly
fermented), oolong tea (semi-fermented tea), black tea (fermented
tea) and pu-erh tea (post –fermented tea). Flavonols (primary
catechins such as epigallocatechin gallate), glycosyl derivatives
(that is apigenin, mtricetin,quercetine, rutin), teaflavins and
thearubigins have been identified as the main bioactive compounds
in the leaves of Camellia sinensis. According to Kokate CK.
Antifungal activity decreases when the extent of tea fermentation is
increased, implying stronger activity.
b) STEPS INVOLED IN PLANT COL LECTION
- Ethnobotanic study
- Identification of the plant
- Harvesting
- Washing
- Drying
- Crushing
- Extraction

III.2 GENERALITIES OF CANDIDASPECIES
a) DEFINITION
Candidarepresents a large genus of Ascomycetous yeast, consisting of

about 150 species and more than 20 species are of clinical importance.
They are common inhabitants of the skin, mucuous membranes of
tracheal, gastrointestinal and genitourinary tracts. The most
predominant species isolated from patients is C. albicans. Among the
non-albican candidaspecies, C. glabrata,is the most frequent detected,
followed by C. parasilopsis, C.tropical and C. krusei (21)
b) CLASSIFICATION
Table 2: Taxonomy of the genus Candida
Superkingdom Eukaryote
Kingdom Fungi
Subkingdom Dikarya
Phylum Ascomycota
Subphylum Saccharomycotina
Class Saccharomycetes
Subclass Saccharomycetidae
Order Saccharomycetales
Family Debaryomycetaceae
Genus Candida
c) PHYSIOPATHOLOGY
Candidaspecies live as commensals on skin, gastrointestinal and

genitourinary tracts. Repeated and lengthly episodes of neutropenia
provides favourable conditions for candidagrowth especially in
critically ill patients facing AIDS, blood cancer, bone marrow or solid
organ transplants. Increased use of immunosuppressive agents, broad
spectrum antibiotics, anticancer therapies, the use of indwelling
catheters and long-term corticoid therapy are also predisposing factors
for acquiring Candidainfections.(22)

d) ANTIMICROBIAL RESISTANCE
Antimicrobial Resistance (AMR) occurs when bacteria, viruses, fungi

and parasites change over time and no longer respond to medicinnes
making infections harder to treat and increasing the risk of disease
spread, severe illness and death.(23)the main drivers of AMR include
misuse and over use of antimicrobials; lack of access to clean water,
sanitation and hygiene, poor infection and disease prevention and
control in health care facilities.
We have primary and secondary resistance
 Primary resistance also known as intrinsic resistance, is a

predictable trait that refers to the organism’s natural susceptibility
to an antimicrobial. For example, C. krusei is less susceptible to
fluconazole than C.albicans. Hence if C.keusei is isolated
fluconazole will generally not be selected for treatment.[(24)
 Secondary or acquired resistance is much less predictable and

potentially more problematic than primary resistance under
conditions like environmental exposure to antifungal agents, a
population of initially susceptible fungi may begin to express
resistance. This may result from an expression of newly acquired
genetic alterations, translation and expression of previously
repressed metabolic pathways, or unmasking of an existing less
susceptible fungal subpopulation.

Photo 2: Exploring the mechanism of intrinsic and extrinsic

mechanism of antifungal resistance.(25)
b) IDENTIFICATION
Test and kits available for Candidaspecies(26)
a) Conventional methods
Germ tube test Presence of hypha in C.albicans 24-36
Chlamysdospore Sporulation 24-72
formation
Biochemical test Assimilation/fermentation of carbohydtrates 48
API Candida Assimilation/fermentation of carbohydrates 18/24
ID YST/VITEK 2 Biochemical tests 15
API 20C AUX Assimilation tests
Pagno-levin Agar Color change/reduction of triphenyltetrazolium to 48
triphenylformazin
CandidaID Agar Hydrolysis indolyl glucosaminide by C.albicans 48
Albican ID A chromogenic substrate hydrolysed by the hexosaminidase >24
of C.albicans
Fluoroplate A fluorogenic substrate hydrolysed by hexosaminidase of >24
C.albicans
CHROM Agar β-glucosaminidase metabolized to produce colored colonies 48
of different Candidaspp
BiGGY Agar Reduction of bismuth sulfate to bismuth sulphide 48
Corn Meal Agar Stimulates sporulation in Candida >48
CandidaDiagnosti Glucosaminidase hydrolysis by different species of Candidato 48
c Agar produce varying color
AlbiQuick Detection of the enzymes β–galactosaminidase and ւ–proline >24
aminopeptidase
ChromID Candida Hydrolysis of hexosaminidase chromogenic substrate 24

producing different colours
CandidaSelect 4 Candidaidentification based on specific enzymatic activity 48
resulting in the formation of colored colonies
b) Molecular methods
Serological >24
FTIR Immunological assay based antigen-antibody reaction against >24
manna, manoprotein, glucan, HSP90, enolase…etc
MALDI-TOF-MS Spectrum difference of protein among Candidaspecies 24
PNA FISH Spectrum difference of protein among Candidaspecies >24
RFLP Restriction digestion and hybridization with complementary 48-72
DNA targets
Microsattelite Based on PCR amplification 24
typing
Multilocus typing Based on PCR amplification 24
RAPD Based on PCR amplification <12
Macrospically colonies of candida, on the routinely used sabouraud destose agar (SDA) and
are cream to yellow in colour. Depending on the species colony texture they may be smooth,
glistering, dry, wrinkled or dull.(27)

Figure 3: is an image of candidainfections in the skin and

mouth(28)

i. MATERIAL AND METHOD
1. Type of study
It will be an experimental study
2. Place of study
 Our plant will be harvested at the Ndu tea estate in the Donga-
mantung Division by factory manager.
 Then it will be sent to the Nation HEBARIUM in Yaounde of

identification.
 Extraction and identification of phytochemical compounds in the

inorganic laboratory of the Faculty of medicine and pharmaceutical
sciences in the University of Douala (FMPS-UD).
 Finally the evaluation of the anti-fungal effect on sorted out

resistant candidastrains in the Douala gyneco-obstetric and
pediatric hospital (DGOPH).
1. Period of study
The study will be done for a period of six months, ranging from the 14 th
November 2022 to the 14 of May 2023.
2. Study population
Our study will be done on Candidaspp isolates obtained from different

samplings done in the DGOPH.
 Inclusion criteria: Candidaspp isolates
 Exclusion criteria: non-resistant Candidaspp
1. Materials
Table : various materials to be used according to objectives and their uses
Activity Material Uses

Extraction -Beaker -Used as vessel for
extraction
-Stirrer -Used to homogenize
the mixture of the
dried powder and
-Filter paper solvents
-Used to separate
-Funnel crude extracts from
solvents
-Used to channel liquid
or fine grained
substances into a
-Rotatory evaporator container with a small
opening
-Used for efficient and
gentle removal of
-Ethanol 70 degrees solvents from samples
by evaporation.
-Used for the
extraction of plants
constituents.
Phytochemical -HCL -Test for flavonoids
screening -H2SO4 and NaOH -Test for phenols
-Dragendroff’s -Test for alkaloids
reagent(potassium -Test for proteins
bismuth iodide solution)
-Biurets test -Test for proteins
-Distilled water -Dissolve extracts
Evaluation of anti- -Electric balance -For measuring weight
fungal activity -Petri dish 90mm -To smear culture
media
-Sabouraud dextrose -Growth of yeast
agar(SDA)
-Normal saline -For preparation of the
inoculum
-Incubator -To provide adequate
temperature for fungi
growth.
-Refrigerator -For conservation of
samples and prepared
culture media
-Bunsen burner
-To provide and

maintain a sterile
perimeter
-Autoclave -To sterilize culture
media and other
equipments.
-Test tubes -Preparation of
inoculum and test
extracts
-Sterile swaps -For sample collection
Material for optic -Glass slides
microscopy -Cover slides
-loops
-Optic microscope
-Pipette 100µl
-Pipette tips 200-1000µl
Material for data -A pen
collection and -A laboratory manual
tracability -Computer
2. Methods
a) Administrative procedure
Apply for an authorization which permits this work to take place. These
authorization letter is addressed to;
- The ethic clearance committee of the FMPS-UD
- The director of the DGOPH
a) Recruitment procedure
All candidaspp strains identified in these hospital using the different

conventional methods of identification are to be recruited in this study
b) Extraction
 Collection of plant: after identification at the National Hebarium,

the leaves are washed under a running tap to eliminate dust and
other foreign particles then dried for about 6days
 500gm of fresh C. sinensis leaves will be dried at a temperature of

32 - 35℃.(29) 45gms of the powdered leaves will be extracted in a 500ml
cornical flask with 70 degrees ethanol (ehanolic extraction) and slightly
warm water (aqueous extraction).(30)
 The extracts will be separately filtered using sterile Whatman no.1

filter paper.
 Concentration by rotary evaporate apparatus
 Extracts will be stored in the refrigerator for further use.

Percentage yield (%) =( mass of extract(g) / mass of ground powder

(g)) X 100
a) Phytochemical screening
Medicinal plants contain natural compounds called phytoconstituents

which works together with nutrients and fibre to form defense against
disease and stress conditions. Primary constituents consist of common
sugars, amino-acids, chlorophyll and proteins. Secondary constituents
consist of alkaloids, terpenoids, steroids and flavonoids (31)
 Test for alkaloids
- Name of reagent: Mayer’s reagent
- Procedure : in each tube containing 2ml of extract, will be added 5 drops

of HCL (2%), then 2drops of Mayer’s reagent
- Expected results: whitish precipitate.

 Test for flavonoids
- Name of test: shinoda test
- Procedure: 1ml of methanol will be added to 2ml of crude extract and later
0.5ml of NaOH 10%
- Expected results: formation of a yellow precipitate.
 Test for poly phenols and phenols
- Name of test: Liebermann’s test
- Procedure: to 2ml of the crude extract, 1ml of Fe(III)chloride and 5 drops

of Potassium ferric cyanide will be added and homogenized
- Expected results: a deep red or greenish precipitate or blue color will

indicate the presence of phenols and polyphenols respectively.
a) Identification of resistant Candidastrains
 Sample collection
Patients after being consulted and sent to the clinical laboratory by any physician, will be
escorted to the collection room by the personnel in charge. The collection procedure will be
done according to the type of sample needed; High vaginal specimens, Blood specimen,
Sputum specimen, Feces specimen, Nail clippings, Pus and scares.
 Culture will be done using the SDA
Identification of resistant species will be analyzed using the following

methods;
- Api candidagallery
- Germ-tube test
- Disc diffusion method
- vitek®
- concentration gradient strip (E-test)
- Mass spectrophotometry

a) Evaluation of the anti-fungi activity
Antifungal susceptibility test are grouped into two categories: quantitative

methods that permits the determination of the presence or absence of
antifungal activity by sensitively testing on fungi stains and qualitative
method that permits the determination of inhibitory parameters such as
MIC and MFC of the fungi.

i. EXPECTED RESULTS
 Get to fine bioactive components in the ethanolic extracts of

C.sinensis.
 Obtain inhibitory effects of the extract on resistant Candidastrains.
 Proof its efficiency as a remedy to multiple drug resistance.

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EVALUATING THE ANTI-FUNGAL ACTIVITY OF Camellia sinensis (L) Kuntze EXTRACT ON ANTI-

FUNGI RESISTANT AND NON RESISTANT CANDIDASPECIES IN THE CITY OF DOUALA

1Presented by KIMBI JAEL NJINGI

2Presented by KIMBI JAEL NJINGI

3Presented by KIMBI JAEL NJINGI

4Presented by KIMBI JAEL NJINGI

II. RESEARCH OBJECTIVES

II.1. General objectives

 Evaluate the antifungal activity of ethanolic leaf extract of Camellia sinensis

II.2. Specific objective

 Identification and extraction of bioactive compounds found in green and black

 Collection and identification of antifungal resistant candidastrains

 To compare the antifungal activity of our extract on strains of resistant

5Presented by KIMBI JAEL NJINGI

III.1. GENERALITIES ON THE PLANTS

Table 1: Taxonomy of the genus Camellia sinensis

6Presented by KIMBI JAEL NJINGI

Theaceae, the tea familyof plants in the order Theales. The

Members of the family have evergreen leaves and flowers with

 Flowers: they are white, fragrant and up to 4cm diameter.

 Fruits: it’s a 3-angled capsule with seeds and is surrounded by a persistent

Figure 1: Fresh tea leaves and flower

Camellia sinensis is a native plant from Asia-China, Tibet and

In Cameroon, the following conditions are necessary for tea

 Annual rainfall of 2000 - 4000mm

 Relative humidity of 70 - 90%

 Sunshine hours of 5hrs/day

The type of tea produced in camerron is the brown tea.(18)

b) PHYTOCHEMICAL AND PHARMACOLOGICAL STUDY OF Camellia

Various production technics produces different types of teas with

According to Mohammed HY et al study on the antifungal effects

Tea polyphenols includes catechins, flavonoids, anthocyanins and

Bioactive Components Uses

The table above is a summary of the bioactive compounds found in

 THE PHARMACOLOGYCAL STUDY

Teas can be classified depending on the degree of fermentation such

b) STEPS INVOLED IN PLANT COL LECTION

- Identification of the plant

10Presented by KIMBI JAEL NJINGI

III.2 GENERALITIES OF CANDIDASPECIES

Candidarepresents a large genus of Ascomycetous yeast, consisting of

Table 2: Taxonomy of the genus Candida

Candidaspecies live as commensals on skin, gastrointestinal and

11Presented by KIMBI JAEL NJINGI

Antimicrobial Resistance (AMR) occurs when bacteria, viruses, fungi

We have primary and secondary resistance

 Primary resistance also known as intrinsic resistance, is a

 Secondary or acquired resistance is much less predictable and

12Presented by KIMBI JAEL NJINGI

Photo 2: Exploring the mechanism of intrinsic and extrinsic

Test and kits available for Candidaspecies(26)

13Presented by KIMBI JAEL NJINGI

14Presented by KIMBI JAEL NJINGI

Figure 3: is an image of candidainfections in the skin and

15Presented by KIMBI JAEL NJINGI

It will be an experimental study

 Then it will be sent to the Nation HEBARIUM in Yaounde of

 Extraction and identification of phytochemical compounds in the

 Finally the evaluation of the anti-fungal effect on sorted out

Our study will be done on Candidaspp isolates obtained from different

 Inclusion criteria: Candidaspp isolates

 Exclusion criteria: non-resistant Candidaspp

Table : various materials to be used according to objectives and their uses