FST 3201- 3 (BASIC FOOD MICROBIOLOGY)

EXPERIMENT TITLE :
MORPHOLOGY EXAMINATION OF YEAST AND MOULD

EXPERIMENT DATE:
14 JUNE 2023

LECTURER’S NAME:
PROF. MADYA. DR. FARINAZLEEN BINTI MOHAMAD GHAZALI
PROF. MADYA. DR YAYA RUKAYADI

MEMBERS OF SUBGROUP 6

NAME MATRIX NUMBER

NOR SYUHADA BINTI BAHARUDIN 210374

NURUL AISYAH BINTI ABDUL RAHMAN 209903

NOR SYAZANA BINTI ZAINUDDIN 210216

FARAH SOFEA BINTI SUHAIMEE 209799

ABSTRACT

Through the combination of macroscopic and microscopic analyses, a comprehensive

understanding of the macromorphology and micromorphology of yeast and mould was

achieved. This investigation contributes to the identification and differentiation of these

microorganisms, enabling better recognition and characterization in various fields such as food

microbiology, environmental monitoring, and pharmaceutical research.

Microscopic analysis involved observing the cellular structures and features of the organisms.

The results of the macroscopic examination revealed distinct differences in the appearance and

growth patterns of yeast and mould colonies. Yeast colonies appeared smooth, creamy, and

often exhibited a raised morphology. In contrast, mould colonies displayed a filamentous texture,

with a variety of colors and shapes observed. Microscopic examination provided further insights

into the cellular characteristics of yeast and mould. Yeast cells were observed to be oval or

spherical in shape, with a clear and distinct cell wall. Budding or vegetation, a process where

new cells form as small projections from the parent cell, was also evident in yeast samples.

Mould samples, on the other hand, displayed filamentous structures known as hyphae, which

consisted of elongated cells forming a network.

INTRODUCTION

Mycology is the study of fungi, their interactions with one another and other living things, and

the distinctive biochemistry that distinguishes them from other taxa. The eukaryotic organisms

known as fungi constitute a separate kingdom. Fungi were distinguished from both plants and

animal traits. Fungi are eukaryotic microorganisms which include yeasts, moulds, and

combinations of the two. Some fungi may cause autoimmune, allergic, cutaneous,

subcutaneous, or systemic conditions.

Moulds can be distinguished by the growth of hyphae that produce colony features.It is

characterised by long filaments that grow by apical extension. Hyphae may be irregularly or

sparsely septate, and their number of nuclei may vary. Apical elongation, the process by which

hyphae grow longer, calls for a delicate balance between the breakdown of existing cell walls

and the production of new ones. Conidiogenesis structure is frequently used to distinguish

moulds. The complete protoplasm of the peculiar sac-like cells termed sporangia, which are

produced by certain moulds, cleaves into spores known as sporangiospores. Moulds have

normal eukaryotic structures with a cell wall that is typically made of chitin, cellulose, or both on

rare occasions. Moulds may also pierce surfaces because they are obligate aerobes that

develop by lengthening the apical points of their hyphae.

Yeasts are microscopic, single-celled fungus that produce via budding. The existence or lack of

capsules, the size and structure of the yeast cells, the method of daughter cell production

(conidiogenesis), the creation of pseudohyphae and true hyphae, the presence of sexual

spores, and physiologic information are used to distinguish between the many yeast taxa. It has

a thin chitinous cell wall and its cell membrane that encloses the protoplasm contains all the

typical cell organelles, including ribosomes, mitochondria, ER, the nucleus, and other granules.
Macromorphology encompasses colony colour, size, shape, texture, and exudate production

(visible to the unaided eye). Conidiophore form, size, and texture, vesicle shape and size,

phialide shape and size, and conidiospore colour, size, shape, texture, and arrangement are all

included in micromorphology. Different species will have morphologies that are different from

one another. Thus, it will help in the identification of mould and yeast.

OBJECTIVE

The objective of the experiment is to examine the macro and micromorphology of yeast and
mould.
MATERIALS AND METHODS

A. MOULD EXAMINATION ORGANISMS

Pre-grown mould cultures on PDA ( Rhizopus oryzae, Aspergillus oryzae )

APPARATUS

Coverslip, Inoculation loop, Lactophenol cotton blue, Light microscope, Slide and cover slip,

Stereomicroscope, Immersion oil

METHODOLOGY

Pre-grown mould cultures on Potato Dextrose Agar (PDA) were given to the student. For

macromorphology, the mould colony size, colour, texture, shape, and exudate were observed

and recorded. For micromorphology, wet mount method (using lactophenol cotton blue as stain),

and hanging drop method can be used. For the wet mound method, a small amount of sample

from the edges of the mould colony was aseptically removed using a sterile inoculation loop

then was placed on a clean microscope slide. Then, a sufficient amount of lactophenol cotton

blue was dropped, and covered the slide slip carefully to avoid bubbles, and observed under

light microscope. The conidiophore shape, size, and texture, vesicle shape and size, phialide

shape and size, and conidiospore colour, size, shape, texture, and arrangement were observed

and recorded. For the hanging drop method, immersion oil was dropped on the edge of the

glass coverslip followed by sufficient mould culture in the middle of the coverslip. Then, the slide

was placed onto the glass coverslip invertedly and then observed under the microscope. The

conidiophore shape, size, and texture, vesicle shape and size, phialide shape and size, and

conidiospore colour, size, shape, texture, and arrangement were observed and recorded.
B. YEAST EXAMINATION ORGANISMS

Pre-grown yeast cultures on PDA

APPARATUS

Coverslip, Inoculation loop, Lactophenol cotton blue, Light microscope, Slide and cover slip,

Stereomicroscope, Immersion oil

METHODOLOGY

The yeast culture was given to the students. For macromorphology, the mould colony size,

colour, texture, shape, and exudate were observed and recorded. For micromorphology, wet

mount method (using lactophenol cotton blue as stain), and hanging drop method can be used.

For the wet mound method, a small amount of sample from the edges of the mould colony was

aseptically removed using a sterile inoculation loop then was placed on a clean microscope

slide. Then, a sufficient amount of lactophenol cotton blue was dropped, and covered the slide

slip carefully to avoid bubbles, and observed under light microscope. The conidiophore shape,

size, and texture, vesicle shape and size, phialide shape and size, and conidiospore colour,

size, shape, texture, and arrangement were observed and recorded. For the hanging drop

method, immersion oil was dropped on the edge of the glass coverslip followed by sufficient

mould culture in the middle of the coverslip. Then, the slide was placed onto the glass coverslip

invertedly and then observed under the microscope. The conidiophore shape, size, and texture,

vesicle shape and size, phialide shape and size, and conidiospore colour, size, shape, texture,

and arrangement were observed and recorded.

RESULTS

Table 1.0: Macromorphology

Name Aspergillus Oryzae Rhizopus Oryzae Yeast

Colony size Thick and Colonies up to 10mm Medium size colonies.

homogenous colonies

Colour Greenish-yellow spore Pale brown to brown Cream colony colour.

Texture Bumpy surface texture Cushiony and cottony Smooth and surface
texture glistening.

Shape Flask shape or Globose,subglobose or Spherical to egg-shaped to

cylindrical phialides oval in shape and filamentous.
root-shape rhizoids.

Exudate Filamentous Present root-like Produce filaments or hyphae

micro-fungus that structures called in their growth.
show round rhizoids, produce
vesicle,conidiophores elongated stolons with
and conidia that are branching hyphae
spherical and light which are coenocytic.
green in appearance.
MICROMORPHOLOGY

Table 2.0 - Mould

Test/microbes Aspergillus Oryzae Rhizopus Oryzae

Wet mound

Hanging drop

Table 3.0 - Yeast

Wet mound Hanging drop

yeast
DISCUSSION

In this experiment, we learn about morphological examination of yeast and mould which is the

study of fungi that covers fungal genetics, taxonomy and identification and advantages to

humans,animal and plants.Mold and yeast are members of the Fungi kingdom and are

classified as eukaryotes. Despite belonging to the same kingdom, they exhibit distinct

differences in their structure, function, color, and reproductive methods. Yeast is a unicellular

organism that can have a round, oval, or filamentous shape and in form and size very similar to

bacteria. The most prevalent form of asexual reproduction in yeasts is budding. In contrast,

mold is a multicellular organism that appears as clusters of mycelium or hyphae with diverse

colors. It can reproduce either sexually or asexually.

In addition, yeasts and moulds are very important to human as they utilized in the production of

various food items and ingredients, such as wine, beer, baked goods, and dairy products. For

example, molds are cultivated for use in both animal feed and human food production. They

play a crucial role in the creation of various food-related products, such as amylases and other

enzymes used in breadmaking, as well as citric acid found in soft drinks. Molds also play a

significant part in the maturation process of numerous oriental foods.therefore, Saccharomyces

cerevisiae and lactic acid bacteria, among other types of yeasts, have been traditionally

employed in the production of numerous fermented foods. Within the food industry, yeasts hold

a notable position in the manufacturing of alcoholic beverages, bioethanol, baker's yeast, and

various yeast-based products.

Soy sauce is a widely consumed Oriental fermented product that has gained popularity in

Western cuisine. It is a dark brown liquid with a salty taste and a pleasant aroma reminiscent of
meat extracts. Soy sauce serves as a seasoning agent and table condiment, enhancing the

flavor and color of various dishes such as meats, seafood, and vegetables.The manufacturing

process of soy sauce involves the fermentation of a mixture of soybeans, cereal (usually wheat),

and salt.

Figure 1: the flow chart of production of soy sauce

To create koji, steamed soybeans and roasted wheat are inoculated with either Aspergillus

oryzae or Aspergillus sojae. This mixture is then combined with water and salt and undergoes
fermentation for a period of 6 to 12 months. Following fermentation, the resulting product is

pressed to extract the fermented juice known as shoyu.

Other than that, fungi play a significant role in the production of two types of cheeses:

blue-veined cheeses and Camembert/Brie. Examples of blue-veined cheeses include

Roquefort, Gorgonzola, and Stilton, which rely on the mold Penicillium roqueforti and the

bacterium Streptococcus lactis. On the other hand, Camembert and Brie cheeses require

Penicillium camemberti and lactic acid-producing streptococci for their production. Additionally,

the mold Oospora lactis and the bacterium Bacterium linens may also contribute to the

manufacturing process of these cheeses.

CONCLUSION

To conclude, at the end of the experiment we can investigate the macro and micromorphology

of yeast and mould. Micromorphology is lower fungi while macromorphology is higher fungi like

mushrooms.
QUESTIONS

Table 1: Common foodborne moulds, their micromorphology, and their advantages/

disadvantages.
MOULD 1 MORPHOLOGY ADVANTAGE

Produces round greywhite Responsible for the main

colonies on agar. Colony size flavour and odour of
considered small (25-35 mm Camembert cheese, and
after 7 d incubation; Petri dish sausages.
is 90 mm). Colony texture is
floccose (cottony).

Penicillium camemberti

MOULD 2 MORPHOLOGY ADVANTAGE

Aspergillus molds appear as Some Aspergillus species are

fluffy colonies with a green used in food production
color. They produce processes, such as the
conidiophores (aerial hyphae) production of soy sauce and
bearing conidia (asexual certain types of cheese. They
spores) in chains. also contribute to the food
fermentation of products such
Aspergillus flavus as tempeh.

MOULD 3 MORPHOLOGY ADVANTAGE

Fusarium molds produce Some Fusarium species have

woolly colonies with colors industrial usage, such as the
ranging from white to pink. production of enzymes and
They produce both organic acids which can be
macroconidia and used in food processing.
microconidia.

Fusarium oxysporum
Table 2: Common foodborne yeasts, and their micromorphology, and their advantages /
disadvantages
MOULD 1 MORPHOLOGY DISADVANTAGE

Aspergillus molds appear as Primary contaminant of corn and

fluffy colonies with a green color. peanut. Produces carcinogenic
They produce conidiophores mycotoxin (aflatoxins).
(aerial hyphae) bearing conidia Certain species of Aspergillus can
(asexual spores) in chains. produce mycotoxins, such as
aflatoxins, which are potent
carcinogens and can contaminate
Aspergillus flavus food crops, particularly nuts,
grains, and dried fruits.

MOULD 2 MORPHOLOGY DISADVANTAGE

Produces round greywhite Penicillium molds can produce

colonies on agar. Colony size mycotoxins, such as ochratoxin,
considered small (25-35 mm which can contaminate grains,
after 7 d incubation; Petri dish is coffee beans, and other food
90 mm). Colony texture is commodities. Additionally, certain
floccose (cottony). species can cause spoilage of
fruits, vegetables, and bakery
Penicillium camemberti products.

MOULD 3 MORPHOLOGY DISADVANTAGE

Fusarium molds produce woolly Certain species of Fusarium can

colonies with colors ranging produce mycotoxins, such as
from white to pink. They fumonisins and trichothecenes,
produce both macroconidia and which can contaminate grains,
microconidia. especially corn and wheat. These
mycotoxins pose health risks to
humans and animals.

Fusarium oxysproum
 Table 2: Common foodborne yeasts, their micromorphology, and their advantages/
disadvantages.

YEAST 1 MORPHOLOGY ADVANTAGE

Saccharomyces Saccharomyces cerevisiae appears Saccharomyces cerevisiae is widely

cerevisiae as oval-shaped or ellipsoidal cells, used in baking and brewing. It plays a
typically 5-10 micrometers in crucial role in the leavening of bread,
diameter. Under favorable conditions, where it ferments sugars and
they reproduce by budding, forming produces carbon dioxide, causing
daughter cells. dough to rise. In brewing, S.
cerevisiae is responsible for alcohol
fermentation.

YEAST 2 MORPHOLOGY ADVANTAGE

Candida albicans Candida yeasts are oval or elongated Certain Candida species, such as
cells, usually around 4-8 micrometers Candida milleri, contribute to the
in diameter. They can form both flavor development of some cheese
budding and pseudohyphae varieties, including Roquefort cheese.
(elongated chains of cells).

YEAST 3 MORPHOLOGY ADVANTAGE

Zygosaccharomyces Zygosaccharomyces yeasts are Certain Zygosaccharomyces species

rouxii oval-shaped cells, typically larger are involved in food fermentation
than Saccharomyces cerevisiae, processes, such as the production of
ranging from 10-15 micrometers in traditional fermented soybean
diameter. They reproduce through products like soy sauce and miso.
budding.
 YEAST 1 MORPHOLOGY DISADVANTAGE

Saccharomyces Saccharomyces cerevisiae appears While S. cerevisiae is generally

cerevisiae as oval-shaped or ellipsoidal cells, beneficial, excessive growth or
typically 5-10 micrometers in contamination with specific strains
diameter. Under favorable conditions, can lead to spoilage of certain food
they reproduce by budding, forming products, such as fruit juices or wines.
daughter cells. Additionally, in rare cases, it can
cause infections in individuals with
compromised immune systems.

YEAST 2 MORPHOLOGY DISADVANTAGE

Candida albicans Candida yeasts are oval or Candida yeasts can also cause
elongated cells, usually around 4-8 spoilage of food, especially in acidic
micrometers in diameter. They can or sugary environments. Additionally,
form both budding and some Candida species can cause
pseudohyphae (elongated chains of infections in humans, particularly in
cells). individuals with weakened immune
systems.

YEAST 3 MORPHOLOGY DISADVANTAGE

Zygosaccharomyces Zygosaccharomyces yeasts are Zygosaccharomyces yeasts can also

rouxii oval-shaped cells, typically larger cause spoilage in some food
than Saccharomyces cerevisiae, products, including fruit juices and
ranging from 10-15 micrometers in wines. They can produce off-flavors
diameter. They reproduce through and cloudiness, rendering the product
budding. unappealing.
REFERENCES

Admin. (2022, November 8). The difference between mold and yeast - a detailed overview.
BYJUS.https://byjus.com/biology/difference-between-molds-and-yeasts/#:~:text=Mould%20
is%20multicellular.,be%20round%2C%20oval%20or%20filamentous.

Babel, F.J. The role of fungi in cheese ripening. Econ Bot 7, 27–42 (1953).
https://doi.org/10.1007/BF02984987

Lorenzo, J. M., Munekata, P. E., Dominguez, R., Pateiro, M., Saraiva, J. A., & Franco, D. (2019,
September 29). Main groups of microorganisms of relevance for food safety and stability:
General aspects and overall description. Innovative Technologies for Food Preservation.
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7150063/#:~:text=Microbiology%20is%20im
portant%20to%20food,%2C%20bakery%2C%20and%20dairy%20products

Libretexts. (2021a, March 6). 8.15: Human uses of fungi. Biology LibreTexts.
https://bio.libretexts.org/Bookshelves/Introductory_and_General_Biology/Book%3A_Introdu
ctory_Biology_(CK-12)/08%3A_Protists_and_Fungi/8.15%3A_Human_Uses_of_Fungi#:~:t
ext=Yeasts%20are%20used%20in%20bread,sauce%2C%20tempeh%2C%20and%20chee
ses

Monika, b. (2018, October 29). How to make soy sauce? (with production process and Flow
Chart): Industrial Microbiology. Biology Discussion.
https://www.biologydiscussion.com/industrial-microbiology-2/how-to-make-soy-sauce/how-t
o-make-soy-sauce-with-production-process-and-flow-chart-industrial-microbiology/86711

Michael R. McGinnis & Stephen K.Introduction to mycology - medical microbiology - NCBI

bookshelf. (n.d.). https://www.ncbi.nlm.nih.gov/books/NBK8125/

Nayak, S., Kavitha, B., Sriram, G., Saraswathi, T. R., Sivapathasundharam, B., & Dorothy, A. L.
(2012). Comparative study of Candida by conventional and CHROMagar method in
non-denture and denture wearers by oral rinse technique. Indian Journal of Dental
Research, 23(4), 490. DOI:10.4103/0970-9290.104956
Escott, C., Del Fresno, J. M., Loira, I., Morata, A., & Suárez-Lepe, J. A. (2018).
Zygosaccharomyces rouxii: Control strategies and applications in food and winemaking.
Fermentation, 4(3), 69.DOI:10.3390/fermentation4030069

Gordon, T. R., & Martyn, R. D. (1997). The evolutionary biology of Fusarium oxysporum. Annual
review of phytopathology, 35(1), 111-128.
http://www.extento.hawaii.edu/kbase/crop/Type/f_oxys.htm

Parapouli, M., Vasileiadis, A., Afendra, A. S., & Hatziloukas, E. (2020). Saccharomyces
cerevisiae and its industrial applications. AIMS microbiology, 6(1), 1.doi:
10.3934/microbiol.2020001