INTRODUCTIO Fungi are heterotrophic eukaryotic organisms.

The kingdom fungi contain both

N multicellular organisms, or molds and unicellular organisms knows as yeasts. Fungi may
be plant like and possess a cell wall, they do not contain chlorophyll nor do they
photosynthesize. Yeast are unicellular, avoid shapes organisms that are usually larger
than bacteria. Yeast possess a nucleus and reproduce by budding or binary fussion.

Fungi can be classified by the type of sexual spores they produce. Based on the type of
spore, fungi can be classified into four main division; Zygomocota, Ascomycota,
Basidiomycota and Deuteromycota.

OBJECTIVE To identify the presence of fungus.

PRINCIPLE A slide culture is a rapid method of preparing fungal colonies for the examination and
identifications. Permits fungi to be studied virtually in situ with as little disturbance as
possible. A little amount of nutritional substance is given for fungi and they are allowed
to grow in a suitable condition. In slide culture we are growing the fungi directly on the
slide on a thin film of agar, after a good growth they are treated with Lactophenol
Cotton Blue stain and observed under the microscope.
EQUIPMENT 1. Sabouraud Dextrose Agar (SDA)
AND MATERIAL 2. Sterile petri dish
3. U-shaped glass rod
4. Filter paper
5. Sterile distilled water 4ml
6. 95% Ethyl alcohol
7. Lactophenol cotton blue stain
8. Sterile straight wire
9. Sterile forcep
10. Isolated colonies
11. Safety cabinet
12. Gloves
13. Lab coat
14. Incubator
15. Light microscope
16. Glass slide
PROCEDURE Culture procedure
(ACTIVITY 1) 1. Isolated colonies cultured in Sabouraud Dextrose Agar.
2. The equipment such as petri dish, glass rod, glass slide sterile in autoclave in
121oC for 15 minutes.
3. Placed filter paper on petri dish.
4. Cut a 5mm square block from Sabouraud Dextrose Agar (SDA) small enough to fit
under a coverslip by using sterile wire loop.
5. Put the block of Sabouraud Dextrose Agar transferred on to the glass slide.
6. Inoculate the fungal isolated by stabbing into the medium. Colonies took out
from the pentone water and placed on the agar block at 4 sides of the square
shape by using the sterile wire loop.
7. Put the coverslip on top surface of the agar block.
8. 4ml distilled water added on the filter paper.
 9. Sealed the petri dish with cellophane tape.
10. The petri dish incubated at 25oC in incubator for 1 – 2 weeks.
11. After 2 weeks, proceed with staining procedure.

Staining procedure
1. A few drops of 95% ethyl alcohol added on the slide after incubation for
disinfection.
2. Removed the cover slip, Lactophenol Cotton Blue stain as mounting medium
added on the slide.
3. Observed the glass slide under light microscope.
4. Record the results.
RESULT Aspergillus

Vesicle

Hyphae

Phialide
Stipe

Conidia

Metulae

DISCUSSION Fungi are heterotrophic eukaryotic organisms to the kingdom fungi. It contains both
multicellular organisms, or molds and unicellular organisms known as yeast. Although
fungi may be plantlike and possess a cell wall, they do not contain chlorophyll nor do
they photosynthesize. Yeast are unicellular, ovoid shape organisms that are usually
larger than bacteria. Yeast possess a nucleus and reproduce by budding, or binary
fission. A bud begins as a tiny protrusion of the parent cell that eventually enlarges in
size and pinches off. Molds grow as multicellular hair like filaments called hyphae into
tangled masses called mycelium. If the hypae contain internal cross walls (septa), they
are considered septate, and those without the septa are considered nonseptate or
aseptate.

A fungal culture help to diagnose fungal infections, a health problem related by

exposure to fungi. The test may help identify specific fungi, guide treatment, or
determine if a fungal infection treatment is working. A slide culture is a rapid method of
preparing fungal colonies for the examination and identifications. Permits fungi to be
studied virtually in situ with as little disturbance as possible. A little amount of
 nutritional substance is given for fungi and they are allowed to grow in a suitable
condition. In slide culture we are growing the fungi directly on the slide on a thin film of
agar, after a good growth they are treated with Lactophenol Cotton Blue stain and
observed under the microscope. The Lactophenol Cotton Blue stain is a special stain to
examine fungal elements following either a tape preparation or scraping. The
Lactophenal Cotton Blue stain contains phenol, which will kill the organisms, lactic acid
which preserves fungal structures, and cotton blue which stains the chitin found in the
fungal cell walls.

In this experiment, the media that used to supports rapid growth of pathogenic and
non-pathogenic fungi is Sabaraud Dextrose Agar (SDA). Sabaraud Dextrose Agar is a
universal primary medium for fungal culture, to prevent the growth of Actinomyces
which others grows well on Sabouraud dextrose agar (SDA). This agar to isolate
dermatophytes from non-sterile specimens and was a low pH 5.6 medium. Agar is
solidifying agent and easy to cut in block.

CONCLUSION In conclusion, the rapid slide culture is rapid method of preparing fungal colonies for
examination and identification. Fungi are identified mostly by close examination of its
morphology and the characteristics it possess. In slide culture, the fungi are growing
directly on the slide on a thin film of agar. By doing this there is no need to remove a
portion of the fungus from a culture plate and transfer it on the slide.

REFERENCES https://ecommons.aku.edu/cgi/viewcontent.cgi?article=1069&context=books
http://faculty.fiu.edu/~gantarm/Experiment%203;%20Identification%20of%20Fungi.pdf
https://www.slideshare.net/MrSSenthilPrabhu/identification-of-fungi
https://www.youtube.com/watch?v=wcNnfW5eI1o