MIC500

EXPERIMENT 1

MOLD: MICROSCOPIC IDENTIFICATION AND

IDENTIFICATION OF MOLD FROM FOOD

NAME : NURHUSNA NABILAH BINTI CHE RUSSLEE

STUDENT NUMBER :

GROUP :

DATE OF EXPERIMENT : 25th SEPTEMBER 2018

DATE OF SUBMISSION : 2nd OCTOBER 2018

1. INTRODUCTION

Moulds are categorised as fungi that develop in the form of multi-cellular filaments, namely
hyphae. Moulds are responsible for biodegradation in foods by breaking down the complex
organic matter into simpler substances which can result in food spoilage. The undesirable decay
of the foods progressively develops after the ripening stage or when the food is physically
damaged (such as bruised fruits as a result of poor handling/delivery). The growth of moulds is
detected when distinctive blue/green "fur" or orange spots develop on the affected foods.

Molds are widely distributed in nature, and are common contaminants of agricultural
commodities, foods, beverages, and feed. These products constitute a rich habitat for
microorganisms because they contain sufficient nutrients such as carbohydrates, proteins, lipids,
and other growth factors as well as moisture. (Markhali, 2008)

Fungi is a taxonomic Kingdom that is composed of well over 99,000 species including yeast,
molds, smuts and rusts among others. Being a very diverse category, Fungi may be single celled
or multicellular organisms that can be found virtually anywhere (particularly moist places).
(Moore D, 2011)

Also referred to as Mould, mold is a type of fungi that is genetically similar to yeast. Like
yeast, molds are also eukaryotic and can be found in moist places. They are also decomposers
and use enzymes to breakdown dead organic materials (leaves, wood, plants etc) which are
sources of energy. Unlike yeast, molds are more plant like with long filaments that are commonly
known as hyphae, which tends to grow on the surface on inside the substance being broken
down. According to Moore, Robson and Trinci (2011) Given that mold grows by extension of the
hyphae, a colony of mold can grow to cover a surface of several square feet.
2. OBJECTIVES
a) To allow the students to make pure cultures studies of some of the common moulds found in
food.
b) To make isolates from naturally contaminated foods.
c) To become familiar with staining and observing mold morphology

3. MATERIALS
a) Pure cultures (on Patato Dextrose Agar plates) of the following genera of mouldsare
provided
i) Rhizopus spp.
ii) Mucor spp.
iii) Penicillium spp.
iv) Aspergillus spp.
v) Neurospora spp.
vi) Fusarium spp.
b) Metheylene blue staining
c) Cellophane tape
d) Microscope
e) 10% glcerol
f) Food sample (tempe and bread)
g) PDA plates

4. METHODS
1. Macroscopic examination (Table 1.1)
The cultures was examined by using the low power of the compound microscope.
2. Microscopic examination (Table 1.1)
About 3 cm of cellophane tape was cut and the sticky surface was faced down a
gently touched against the mould growth. The strip then was transfered to a drop of
methylene blue stain on a glass slide. The cultures were futher examined for more
detailed structures.
a) Spores (type and arrangements)
b) Hypae and septation
c) Color of mycelium
d) Any special structures
3. Some of the mould food provided was studied and the mould was identified by
focusing the low power lens on the food specimens. At the same time, isolates was
made by streaking a small sample of the food on the PDA plates. The sample was
incubated at 27°C for 24 to 48 hours. The macroscopic and microscopic structure of
mold was examined and described. (Table 1.2)
5. RESULTS
1. Macroscopic an Microscopic examination culture provided

Table 1.1: Structures observed macroscopically and microscopically

Mold Color of Hypae and Spores (types/ Any special Illustration

mycelium septation arrangements) structures and labeled
Rhizopus White Nonseptate Sporangia at the Fluffy
spp. mycelia branching tips of the appearence.
hypae sporangiophores White to
are rounded gray-brown
on surface
Mucor spp. Grey to Nonseptate Sporangiophores Fluffy
brown hypae located at the appearance
end of the resembles
hyphae, globose cotton candy
sporangium.
Penicellium Yellowish Profusely The Dense brush-
spp. green branced with conidiospores like spore-
septate hypae are arranged in bearing
chain from the structure
tips of the
phialides
Aspergillus Bluish Glass like Basipetal (ball Fluffy
spp. green hypae and and chains) appearance
septate resembles
cotton candy

Neurospora Yellowish Septate hypae Branching Powdery

spp. brown conidiosphore form

Fusarium The filaments Form Long

spp. are hyaline chlamydospores filamentous
and septate in chains structure
2. Isolation and identification of mold from food sample.

My food sample is : bread and tempe

Table 1.2: Structure observed macroscopically and microscopically from food sample.

My mold Spores (types/ Hypae and Colour of Any special Illustration

arrangements) septation mycelium structures and labeled

Bread Conidia extend Nonseptate White Have a very

from and glass- mycelium dark conidia
circumferentially like hypae
from and
obscure vesicle,
biserate

Tempe Sporangiophores Nonseptate Greenish- Produces

located at the brown upward
end of the sweeping
hyphae, globose appearence
sporangium.
DISCUSSION

This experiment allow students to study the macroscopic and microscopic

morphology of different cultures. I was able to observe the different structure of mold.
Rhizopus appears as short strands with oval-shaped heads, looking like a balloon on a
string. The head is where the spores of this type of mold are contained. The rhizopus spp
are characterized by a body of branching mycelia composed of three types of hyphae
which are stolons, rhizoids, and usually unbranching sporangiophores. It is a very-fast-
growing, spreading type of mold which has white mycelia and black sporangia (Petuzello,
2018). Rhizopus species form rhizoids at the base of the sporangiophores. The black
sporangia at the tips of the sporangiophores are rounded. It have fluffy appearence
which is like cotton candy structure. Probably the most common of the Rhizopus species
is R. stolonifer, the so-called bread mold.

The Mucor fungi cause the group of infections referred to as zygomycosis. Mucor is a
filamentous fungus found in soil, plants, and decaying fruits. Colonies of Mucor grow rapidly
at 25-30°C it also have fluffy appearence which is like cotton candy structure. From the front, the
color is white initially and becomes grayish brown in time. Sporangia located at the end of the
hyphae which extend from a thin sporangiosphores forming a ball-shaped clusters of spores and
the hypae appeared to be nonseptate. The sporangia was contain in a vesicle where columella
was present. The columella functions for nutrient exchange between the active protoplasm
below and the developing spores inside the upper portion of the sporangial head.

Penicillum spp. are filamentous fungi they are recognized by their dense brush-like spore-
bearing structures. They have branched conidiospores. The spores are produces in chain from
the tips of the phialides, with the youngest spore at the base of the chain. It is iniatially white and
becone yellowish with time. The hypae cointain septa that divide the hypae into separate cells.
Penicillium spp exert desirable effects on the ripening of specific cheeses. The enzymes of the
moulds are able to break down the fats and proteins which results in generating characteristic
flavours of the cheese (Markhali, 2008).

The colonies of Aspergillus spp are fast growing white to bluish green which consists
of a dense felt of erect conidiosphores. The hypae are nonseptate and hyaline which
make up a mycelium. The conidiophores originate from the basal foot cell located on the
supporting hyphae and terminate in a vesicle at the apex. Covering the surface of the
vesicle entirely or partially on the upper surface are the flask-shaped phialides. Over the
phialides are the round conidia forming radial chains. Aspergillus species are common
contaminants of starchy foods such as bread and potatoes, and grow in or on many
plants and trees.

Neurospora are recognized by the orange color of the vegetative spores. It grows on
dough forming a lot of conidia which cause a bad contamination. Neurospora species are
molds with broadly spreading colonies, with abundant production of ascomata. The
hyaline appears to be yellowish brown and becoming dark and opaque with time.
Neurospora is widely used in genetics as a model organism because it is quickly
reproducing and can survive on minimal media. It have branching conidiosphores which
oval in shape.

Fusarium is a filamentous fungus that is widely distributed in plants and soils which
can cause crop disease in a wide range of commodity crops, from cereals to melon,
pepper, potato and tomato (Early, 2009). Their colonies are usually fast growing which
have pale color with a fluffy cotton candy-like aerial mycellium. Microscopically, the
hyphae of Fusarium in tissue resemble those of Aspergillus spp where the filaments are
hyaline, septate and formed chlamydospores in chains

While for food sample we are using bread and tempe as our isolates. On the bread,
we found a Rhizopus characteristic of mold which it has a very dark conidia. The conidia
extend from circumferentially from and obscure vesicle, biserate looking like a balloon
on a string. It also have nonseptate, glass-like hypae and have a fluffy macroscopical
appearence.

Whilst, the tempe sample shows a Mucor spp. structure where sporangiophores
located at the end of the hyphae, globose sporangium. Sporangia located at the end of
the hyphae which extend from a thin sporangiosphores forming a ball-shaped clusters of
spores and the hypae also appeared to be nonseptate. The sporangia was contain in a
vesicle where columella was present. Its also have a fluffy macroscopical appearence
resemble those as Mucor spp.

The amount of spreading colonies can be reduce by adding dichloran (2,6-dichloro-4-

nitroaniline) which previously identified as a mold-spreading inhibitor. Colony diameters
decreased as the dichloran concentration increased. While dichloran-glcerol (DG18) Agar
Base is based on the formulation described by Hocking and Pitt and is recommended for
the enumeration and isolation of xerophilic moulds from dried and semi-dried foods
which help to recover molds from food that have lower water activity. Examples of these
are dried fruits, spices, confectionery, cereals, nuts and dried meat and fish products.

From this activity we can identify the distinctive differences between sporangium and
conidiosphores which is sporangium are specialized aerial hypae that bear specialized
sacs called sporangia which contain non-motile sporangiosphores. While cinidiosphores
are erect, septate hypae which bear non-motile spores calleed conidia at its tip. Other
than that sporangiosphores are enclosed in a specialized enclosure called sporangia
whilst conidia are not enclosed in an enclosure.
6. CONCLUSION
As a conclusion, from this experiment we can identify the type of mold based on their
different macroscopic and microscopic characteristics. It grows in the form of
multicellular filament called hypae which can easily recognized by it’s cottony
appearence and sometimes a powdery appearence. Most mold grow well at ordinary
temperature. It also gives unpleasent smell like in a moist room. Each mold have
different sets of spores arrangement, hypae and septation, and even color. Thus we can
identify the type of mold we are handling with.

7. REFERENCES

Works Cited
Anderson, H. (2010). Mold Under Microscope (The Fungi Kingdom).

Bullerman, L. (2003). Encyclopedia of Food Sciences and Nutrition (2nd Edition).

Early, R. (2009). Foodborne Pathogens (Second Edition).

Markhali, F. S. (2008). Microscopic and Macroscopic Identification of Moulds in foods. Australia.

McDonald, W. (2001). Aspergillus Species.

Moore D, R. G. (2011). 21st Century Guidebook to Fungi.

Petuzello, M. (2018). Rhizopus Fungus Genus.

Ryan KJ, R. C. (2004). Sherris Medical Microbiology.