University of Southern Mindanao

College of Veterinary Medicine

Vet Micro 311
General Veterinary Microbiology

Name: RANGAS, RIZA MAE D. Date: December 14, 2023

Year/Course/Section: 3DVM-A Score: ______________

FUNGAL ISOLATION
Introduction
Fungi are eukaryotic microorganisms. Fungi can occur as yeasts, molds, or as a
combination of both forms. Fungi are everywhere in very large numbers—in the soil and
the air, in lakes, rivers, and seas, on and within plants and animals, in food and clothing,
and in the human body. Together with bacteria, fungi are responsible for breaking down
organic matter and releasing carbon, oxygen, nitrogen, and phosphorus into the soil and
the atmosphere. Fungi are essential to many household and industrial processes, notably
the making of bread, wine, beer, and certain cheeses. Fungi are also used as food; for
example, some mushrooms, morels, and truffles. Fungi plays an important role in
medicine by yielding antibiotics such as penicillin the first of a series of antibiotics—many
of them derived from fungi—that have revolutionized medical practice.
Materials
• Bottled glass • Inoculating loop
• Cotton plug • Alcohol lamp
• Moldy bread • Frosted glass slides
• Forceps and cover slip
• Lactophenol blue dye
Procedure
Inoculation
1. Prepare medium for fungal Isolation (e.g., Sabouraud Dextrose Agar, Potato
Dextrose agar, Malt Agar).
2. Dispense in the bottle; slant (tip of agar should not be near the bottle neck)
3. Allow to harden; refrigerate if not used immediately.
4. Obtain inoculum (e.g., contaminated/ moldy feeds, grains, moldy bread, and
other environmental sources).
5. Using a forceps, obtain a small portion of the sample and “plant” on the
surface of the agar.
6. Observe daily and note the rate of growth. Describe the colonial
characteristic.
7. Prepare two slides of fungal mount and stain with lactophenol blue dye and
take note the morphology of the fungus under the microscope.
 Staining
1. Add one drop of the lactophenol cotton blue stain on the glass slide.
2. Scoop an adequate amount of sample from the colony by the use of sterile
inoculating loop.
3. Gently mix the sample with the LBCP stain.
4. Holding the coverslip between forefinger and thumb, touch one edge of the
drop of LPCB stain with the coverslip edge, and lower gently, avoiding air
bubbles.

Results and Observation

Figure 1. Fungal growth day 1 Figure 2. Fungal growth day 3

Bread mold grew after 24h of An increase of cloudy undulate-lobate

incubation. White-green cloudy greenish-white appearance was
appearance growth is observed. observed. Filamentous growth is also
observed in the day of incubation.

Figure 3. Fungal growth day 5

Growth continues to increase in span days
of incubation. An irregular-formed,
scalloped-margin, and circular growth
were observed.
 Day of Shape Size Surface Color Opacity Elevation Margin
incubation
Day 1 Irregular Mediu Dull White - Opaque Convex- Lobate
m green umbonate,
flat
Day 3 Irregular, Large Dull, Greenish- Opaque, Convex- Even,
filamentous wrinkle white & translucent umbonate, wavy,
, Bluish- flat lobate
punctiform white
spore
Day 5 Irregular, Large Dull, Green- Opaque, Convex, Even,
filamentous wrinkle yellow, translucent umbonate, wavy,
yellowish- flat scalloped
blue,
white

Figure 4. Fungal mount slide 1. Figure 5. Fungal mount slide 2.

Sporangium, sporangiophore, hyphae Similar classifications were
were prominent classification observed in observed in slide 2.
slide 1.

Conclusion
After a performing all the procedures and observation, the morphological traits were
collected and determined. The fungus development in the incubated bottled glass
demonstrates how the morphology and traits of the isolated colony grow and alter with
time. The colonies' macroscopic and microscopic morphologic features were clearly
visible, as shown in the figure and table with the data above. Furthermore, the two mount
slides' comparable observed classifications show that they originate from the same
organism.
Documentation

References

Fungus | Definition, Characteristics, Types, & Facts. (2023, December 13).

Encyclopedia Britannica. https://www.britannica.com/science/fungus/Importance-of-
fungi

McGinnis, M. R. (1996). Introduction to Mycology. Medical Microbiology - NCBI

Bookshelf. https://www.ncbi.nlm.nih.gov/books/NBK8125/