Name: GALINATO, Jewel R.

Date of Experiment: May 10, 2021

Year and Section: BSN 1A Date of Submission: __________

Experiment No 5 Cultural Characteristic of Microorganisms

I. INTRODUCTION

I. LEARNING OBJECTIVES
At the end of the exercise, the students should be able to:
1. Examine the growth of bacteria in agar plates according to size of the colony, margin or edge,
surface texture, elevation, consistency, optical features, pigmentation and odor.

The cultural characteristics of organisms like bacteria pertain to their microscopic appearance on
different kind of culture media. Culture media can be prepared in various forms depending on the desired
use. Microbes are cultured or grown into culture media to increase their number during the incubation
period. After a suitable period, some of the dispersed cells develop into colonies. A colony is a population
of cells arising from a single bacterial cell in a solid medium. It is visible to the naked eye as discrete
entities.
Bacterial growth in different culture media is characterized by its appearance. Colonial
characteristics and the amount of growth after incubation are essential information in the identification of
bacteria. The morphology of colonies can be described by size, shape, color, edge, or margin. These
features are observed by looking at the top of the colony itself. Colony elevation is apparent when viewed
from the sides at eye level. The growth of bacteria in a broth is described based on the turbidity of the
broth after incubation.
II. MATERIALS
 Inoculated agar plates
 Inoculated agar slants
 Inoculated broth media
 Inoculated agar deep or butts
IV. PROCEDURES
A. Examine the characteristics of bacterial colonies in agar plates and determine the following
features:
1. Size. Only isolated or well-separated colonies should be measured. The time at which the
measurement is made must be noted since young colonies are smaller than older ones.
a. Pinpoint – extremely small colony, measuring only a fraction of millimeter in
diameter or less than 1 mm
b. Large – colonies measuring 5 to 10 mm in diameter
2. Margin or edge and forms. The periphery of bacterial colonies may have different
patterns depending on the species.
a. Entire or circular edge
b. Irregular projections such as undulate, filamentous, lobate, erose or curled
c. Forms such as punctiform, circular, filamentous, irregular, rhizoid, or spindle
3. Elevation. Bacterial colonies may be thin or thick, and their surfaces may be flat or
raised and may be varying degrees of convexity.
4. Surface texture. A bacteria colony’s texture may be described as:
a. Smooth – shiny and glistening
b. Rough – dull, granular and matte
c. Mucoid – slimy, gummy
d. Wrinkled – crumpling, folding
5. Consistency. This growth feature can be determined by touching the colony using an
inoculating needle.
a. Butyrous – the colony has a butter-like consistency
b. Viscous or stringly – a portion of the colony may come off the agar surface
c. Rubbery – the whole colony comes off the agar surface
d. Dry, brittle, powdery – the colony breaks when touched by a needle
6. Optical features. Degree of opacity may be expressed as:
a. Opaque – impenetrable to light
b. Transparent – a clear image is seen and as if there is no intervening material
c. Translucent – permits the passage of light
7. Chromogenesis or pigmentation. There are bacterial cultures that produce and retain
water-insoluble pigments intracellularly, hence causing the colonies to become colored
(pigmented). Some bacterial strains with pigmentation are as follows:
a. Staphylococcus aureus – gold
 b. Serratia marcescens – red
c. Micrococcus luteus – yellow
d. Chromobacterium violaceum – violet
8. Odor. This characteristics is only noted if the smell emitted distinctive (i.e., sweet,
putrefactive, or fruity).
B. Examine the characteristics growth of the bacterial colonies in agar slants according to the
following features:
1. Amount of growth. The abundance of growth may be described as none, scanty,
moderate, or abundant.
2. Color. Pigmentation is observed in the bacterial colony that is visible within the medium.
Most bacteria that lack chromogenesis exhibit a white growth.
3. Opacity. Degree of opacity may be expressed as opaque, transparent, or translucent.
4. Forms. The following illustrations of the different forms help in differentiation:
C. Examine the growth characteristics of bacteria in broth culture media and take note of the
following features:
1. Amount of growth. To determine the amount of growth, it is necessary to shake the tube
to disperse the organism and describe whether it is scanty, moderate, or abundant.
2. Distribution and type of growth
a. Uniformly distributed throughout the medium or evenly turbid
b. Scum or film (pellicle) – growth is confined to the surface of the broth
c. Sedimentary – accumulation may be described as granular, flocculent, or viscous
d. Ring around the top rim of the medium
D. Examine the growth characteristics of bacteria in agar deep or butt culture medium according
to the following features:
1. Growth within the line of inoculation (non-motile)
2. Growth spread or not within the line inoculation (motile)
V. ILLUSTRATIONS
Sketch the bacterial growth in the following:
Streak Plate Streak Slant
 Broth Butt/Slant

VI. RESULTS AND DISCUSSION

1. Write the cultural characteristics of the bacterial culture in the agar plate.

Cultural characteristics Bacterial culture 1 Bacterial culture 2

in the agar plate (Normal Body (Common
Flora:_Candida Albicans_) Item :_______________)

Size of colony Approx. 1 mm

Margin or edge Circular
Surface texture Mucoid
Elevation Convex
Consistency Viscous
Opacity Translucent
Chromogenesis Micrococcus luteus
Odor

2. Write the cultural characteristics of the bacterial culture in the agar slant.

Cultural characteristics Bacterial culture 1 Bacterial culture 2

in the agar slant (Normal Body (Common
Flora:_Escherichia Coli_) Item:_______________)

Amount of growth Moderate

Pigmentation Milky
 Opacity Translucent
Forms Spindle

3. Write the cultural characteristics of bacterial culture in the broth tube

Cultural characteristics in the Bacterial culture 1 Bacterial culture 2

broth tube (Normal Body (Common
Flora:_Proteus Item:_______________)
Mirabilis_)
Amount of growth Abundant
Distribution and type of growth Scum/film

4. Write the cultural characteristics of bacterial culture in the agar deep/butt.

Cultural characteristics in the Bacterial culture 1 Bacterial culture 2

agar deep/butt (Normal Body (Common
Flora:_Shigella Item:_______________)
dysenteriae_)
Type of growth Non-motile
Forms Irregular

VII. GUIDE QUESTIONS

1. What is the significance of describing the cultural or colonial characteristics of bacteria?

 Features of the colonies may help to pinpoint the identity of the bacterium. Different species of
bacteria can produce very different colonies. In the above picture of a mixed culture, an agar plate
that has been exposed to the air and many different colony morphologies can be identified.

2. How would you determine whether a colony is a contaminant or a real bacterial culture?
 First, perform gram staining and look at the morphology of the bacterial cells, if contaminated more
than one cell type shall be visible. Then, streak the culture on a suitable agar based medium and
observe color and type of cfus. If Bacillus sp. you are using, heat the culture at 80 C for 10 min and
culture on a suitable medium. Most contaminating non sporulating cells will be killed and bacilli
survive.
 3. How can you determine whether or not there is bacterial culture growth in nutrient broth?
 Bacterial growth in broths is indicated by the development of a cloudy appearance. If the newly
inoculated broth looks cloudy at the start, you will have no way to determine if this is due to bacterial
growth during the incubation period.

4. What are the advantages of using a solid culture medium over a liquid form?
 Advantages of solid media: (a) Bacteria may be identified by studying the colony character, (b)
Mixed bacteria can be separated. Solid media is used for the isolation of bacteria as pure culture.
'Agar' is most commonly used to prepare solid media. It is used for profuse growth, e.g. blood culture
in liquid media

VIII. CONCLUSIONS
How do you tell the difference between a tiger and a wolf? By studying the animal's features.
Similarly, each species of bacteria has properties that, when combined, can be used to distinguish the
bacterial species in question from other species of bacteria.

Microorganisms' cultural characteristics will aid in the differentiation of microorganisms based

on their physical appearance. To study an organism's cultural characteristics (also known as colonial
morphology), you would require a plate with isolated colonies of the bacteria to be observed. Keep in
mind that various types of media, each with its own set of nutrients, can have an effect on the cultural
characteristics of bacteria. Some media are much more nutritive and will promote vigorous growth. Some
forms of media can stifle development. The temperature at which the bacteria is incubated can also have
an impact on colonial morphology. Any bacteria grow faster at body temperature than at room
temperature, and vice versa. Some bacteria express certain characteristics, such as pigment production,
more intensely at some temperatures than others. The amount of time the bacteria is incubated can also
have an impact on colonial characteristics including colony size. Bacterial development may be hampered
by the presence of other species. Colonial morphology may also be impaired if the original inoculum is
too dense or too diluted.