ISOLATION AND
IDENTIFICATION OF MOULD FROM FELINE
FUR
NIK NOOR SYAMIMI ISMAIL
1207-2101
DIPLOMA IN BIOTECHNOLOGY
PRESENTED : BY
SUPERVISOR :
MRS. NORASNIDA KAMARULZAMAN
INTRODUCTION Scientific name for cat is Felis catus Felidae is the biological family of the cats. The
most familiar felid is the domestic cat, but the family includes
all other wild cats,
including the big cats. Hundreds of millions of cats are kept as pets around the world. Cats are also used in the fur trade, as food and to control pests.
OBJECTIVES
1. To identify the mould present on cat
fur
2. To study the differ group of mould of
indoor and outdoor cat
ISOLATION & IDENTIFICATION Isolation• The term ‘isolate’ can be applied to any
form or species for separation.• The method is to isolate individual cells
and produce pure culture is by prepare a streak plate and let it grow in incubator
Identification• By taking the specimen to glass slide
that together with stain to see the structure of specimen under microscope
ONE OF FUNGUS DISEASE ON CAT ulcerative dermatitis;
cryptococcal rhinitis through the facial bones
Cryptococcosis is common for where frequent at upper respiratory, and occasional pulmonary involvement; often swelling of the bridge of the nose
DERMATITIS TO HUMAN A fungal dermatitis which
caused by a fungus named Trichophyton Verrucosum not a worm. It gets it's name from ringworm because of the shape of the skin disorder.
Human skin infected with ringworm
Cat skin infected with ringworm
INDOOR & OUTDOOR CATS Indoor Catoften called housecat because they are kept indoor (in house) and valued by humans for companionship, and their ability to hunt household pests.
Outdoor CatCat that not owned by any one individual, but is generally friendly to people and may be fed by several households
METHODOLOGY Preparation of Agar
Weight the agar
powder
Calculate the
standard method
Mix with distilled water
Heat and stir on
hotplate and stirrer
Autoclave
Rinse with water to let
it cool
Pour plate Agar hardened and ready to be
use
SABOURAUD’S DEXTROSE AGAR (SDA)
Selective medium is formulated to allow growth of fungi, cultivate dermatophytes and inhibit the growth of bacteria
Dermatophytes is a fungus that commonly causes skin disease in animals and humans.
YEAST EXTRACT PEPTONE DEXTROSE (YPD/YEPD)
YPD, is a complete medium for yeast growth and not a selective medium.
Nutritionally rich medium for the growth of
Saccharomyces cerevisiae which is a species of yeast used in baking, winemaking & brewing of beer making
LACTOPHENOL COTTON BLUE STAINS The cotton blue is the one that gives the
colour and permit to visualize the fungi under microscope. The stain adheres (stick or absorb) to the chitin of the fungi walls of hyphae and conidia
Hyphae
METHODOLOGY Staining of Fungus by using Lactophenol
cotton blue
Add a drop of stain
Flame the needle
Pick mycelia Spread to the stain
Flame the needle
Place coverslipView under microscope
AREA SPECIMENS
1. NOSE2. CHIN
3. CHEEK (LEFT)4. UPPER HEAD
3
2
14
METHODOLOGY Taking the specimen
Sterilized transport medium
was open
Cotton swab taken &
swabbed on the places
(nose, head, cheek &
chin)
Cotton swab was
inserted to the
transport medium
and labelled
Streak to agar using cotton swab
that was swabbed
Lid of agar plate was
closed and
inverted
Agar plates
were put in
incubator at 25oC &
for 5-7 days
RESULTS
CANDIDA ALBICANS1
From experimentLight microscope40x magnifier
From referenceLight microscope150x magnifier
ASPERGILLUS NIGER2
From experimentLight microscope40x magnifier
From referenceLight microscope100x magnifier
CLADOPHIALOPHORA CARRIONII
3
From experimentLight microscope40x magnifier
From referenceLight microscope40x magnifier
ASPERGILLUS FLAVUS4
From experimentLight microscope40x magnifier
From referenceLight microscope40x magnifier
5 SACCHAROMYCES CEREVISIAE
From experimentLight microscope40x magnifier
From referenceLight microscope100x magnifier
TRICHOPHYTON RUBRUM6
From experimentLight microscope40x magnifier
From referenceLight microscope40x magnifier
CUNNINGHAMELLA BERTHOLLETIAE
7
From experimentLight microscope40x magnifier
From referenceLight microscope40x magnifier
BLACK
OUTDOOR CATHOSTEL TPM COLLEGE2-4 YEARS OLDFEMALE
SDA agar YPD agar
HEAD
4. Aspergillius flavus 7. Cunninghamella bertholletiae
5. Saccharomyces cerevisiae7. Cunninghamella bertholletiae
4
7
5
7
SDA agar YPD agar
CHEEK
4. Aspergillius flavus 2. Aspergillus niger
2
4
SDA agar YPD agar
CHIN
2. Aspergillus niger4. Aspergillus flavus
Contaminated
4
2
SDA agar YPD agar
NOSE
3. Cladophialophora carrionii4. Aspergillus flavus7. Cunninghamella bertholletiae
Contaminated
4
73
OREO
OUTDOOR CATHOSTEL TPM COLLEGE1.5-2 YEARSMALE
SDA agar YPD agar
HEAD
2. Aspergillus niger3. Cladophialophora carrionii6. Trichophyton rubrum
5. Saccharomyces cerevisiae
2
6 3
5
SDA agar YPD agar
CHEEK
2. Aspergillus niger5. Saccharomyces cerevisiae
5. Saccharomyces cerevisiae
2
5 5
SDA agar YPD agar
CHIN
4. Aspergillus flavus6. Trichophyton rubrum
Contaminated
46
SDA agar YPD agar
NOSE
5. Saccharomyces cerevisiae
5. Saccharomyces cerevisiae
55
OYEN
INDOOR CATFLAT, WEST PORT,KLANG2-3 YEARSMALE
SDA agar YPD agar
HEAD
2. Aspergillus niger4. Aspergillus flavus7. Cunninghamella bertholletiae
Contaminated
4
2
7
SDA agar YPD agar
CHEEK
1. Candida albicans6. Trichophyton rubrum
5. Saccharomyces cerevisiae7. Cunninghamella bertholletiae
1
7
6
SDA agar YPD agar
CHIN
2. Aspergillus niger4. Aspergillus flavus6. Trichophyton rubrum
2. Aspergillus niger3. Cladophialophora carrionii
4
2
6
3
2
SDA agar YPD agar
NOSE
2. Aspergillus niger4. Aspergillus flavus3. Cladophialophora carrionii
5. Saccharomyces cerevisiae7. Cunninghamella bertholletiae
4 2 73 5
DISCUSSION The fungus growth on SDA agar and YDP agar
are Aspergillus niger, Aspergillus flavus, Candida albicans and yeasts
Overall, Aspergillus sp. is have the most growth of fungus that have on agar plates (KSJang, 2007) For the fungus growth; indoor cat have more fungus species numbers than outdoor cat KSJang, 2007
This is because indoor cat lives in a house is humid, limited ventilation and space (flat house).
The air also mixed with chemicals and skin debris from the people live in the house
DISCUSSION Contaminated agar plates were due to
unsterilized condition of incubator and laboratory where moths and ants can get in to the incubator
It also because the agar plates were not sealed as the Parafilm were absent
CONCLUSION Through this experiment, all the
objectives have accomplished. The species of mould that both indoor and outdoor cat have on cat fur is Aspergillus sp. The group of moulds or fungus present on fur of indoor have more mould than outdoor cat
REFERENCES Kye-Seung Jang, Yeo-Hong Yun, Hun-Dal Yoo, &
Seong Hwan Kim (2007). Molds Isolated from Pet Dogs. doi:10.4489/MYCO.2007.35.2.100
Tournas, V., Stack, M. E., Mislivec, P. B., Koch, H. A., & Bandler, R. (2001, April). BAM: Yeasts, Molds and Mycotoxins. BAM: Yeasts, Molds and Mycotoxins.
Hall, L., Wohlfiel, S., & Roberts, G. D. (2003). Experience with the MicroSeq D2 Large-Subunit Ribosomal DNA Sequencing Kit for Identification of Commonly Encountered, Clinically Important Yeast Species. Introduction to Clinical Mycology, Part 4.
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