HES 032 BSN Lab Activity 6

HES 032-Microbiology and Parasitology
College of Nursing
RAD Learning (AY:__________)
Group Leader: ___________________________________
Group Members: _______________________________________

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ACTIVITY NO. 6: Culture Media
LEARNING OBJECTIVE: At the end of the laboratory period, the student should be able to:
1. Identify the different classification of culture media

2. Differentiate culture and culture media
3. Illustrate and differentiate each according to its function or use, preparation, composition
and physical state.
DISCUSSION:
A nutrient material prepared for the growth of microorganisms in a laboratory is called a culture
medium. Some bacteria can grow well on just about any culture medium; others require special
media, and still others cannot grow on any nonliving medium yet developed. Microbes that are
introduced into a culture medium to initiate growth are called an inoculum. The microbes that
grow and multiply in or on a culture medium are referred to as a culture.
The development of microorganisms upon the culture media is dependent upon several
important factors:
1. The proper food elements must be available.

2. Oxygen must be available as required.
3. A certain degree of moisture is necessary.
4. A medium must be of the proper pH reaction.
5. Proper temperature relations must prevail.
6. The medium must be sterile.
7. Contamination must be prevented.
Compiled by: Claire Mae Caval, RMT

Microbiology and Parasitology – Instructor
Southwestern University PHINMA
CLASSIFICATION OF CULTURE MEDIA
Culture media can be classified according to three primary levels:
1. Physical State
2. Chemical Composition
3. Functional State
A. According to their Physical State
1. Solid Media – consist mainly of substances mixed in a solidified agar, gelatine o albumin
matrix. Agar is usually added in concentration of 2-3%. Such preparations
G i i i rtions of 1-1.5% and such media solidify at temperature
i i gar or gelatine coagulates with h , g , L ff ’ c g
serum medium and most of the solid media for the cultivation of Mycobacterium
tuberculosis.
2. Semi-solid Media – are especially used for keeping stocks of the bacteria for long
periods of time. Agar is usually added in amounts of 0.5-1.0%.
3. Liquid Media – are prepared without such substances as gelatine or agar.
B. According to their Chemical Composition
1. Synthetic Media – one in which the chemical composition of the ingredients is known.
Ex : Ri g ’ i L c ’ i
2. Non-synthetic Media – one in which the precise composition of some or all the nutritive
elements used is not definitely known. Example: Meat extract broth
3. Living tissue – one in which living tissue cells are present. These are especially used for the
cultivation of rickettsiae, Chlamydia and viruses. Examples: HeLa cell lines

C. According to their Use/Functional Type:
Most media used in the clinical bacteriology laboratory belong to one of the following types:
1. Simple Media – are ordinary media which are not enriched and in which non- fastidious
organisms are easily cultivated. Examples: Nutrient broth and Nutrient agar
2. Specific Media – are special media which are used for the cultivation of a particular
organism. Ex : L ff ’ g S dium, Lowenstein-Jensen medium,
S ’ D x g F ch ’ i
3. Enriched Media – include 5% sheep blood agar, enriched chocolate agar and several
infusion-based media, all of which may be further enriched by additives. These media must be
carefully tested for sterility before they are used since even fastidious organisms are likely to
grow. Examples: Blood Agar, Chocolate Agar
4. Differential Media – i c g h M c y’ and EMB agars, Triple Sugar Iron

and Mannitol Salt Agar, which contain dyes, sugars and indicators designed to elicit a
characteristic biochemical response (usually a color) and are used to differentiate groups of
organisms such as lactose fermenters and non- lactose fermenters.
5. Selective Media – contain, in addition to components found in differential media, agents

designed to further inhibit most Enterobacteriaceae and to select from the specimen those
strains resistant to the higher concentrations of the selective components. Some enteric media
combine both selective and differential characteristics. Hektoen enteric agar and xylose lysine
desoxycholate agar may be classed as selective because they are somewhat more inhibitory
than MacConkey selective media also are capable of detecting H2S production. Examples:
MacConkey Agar, Eosin Methylene Blue, Mannitol Salt Agar, Thayer Martin Medium, Saboraud
Agar, Hektoen Agar
6. Transport Media – are devised for prolonging the survival of microorganisms when a
significant delay occurs between collection and culturing. Examples: Cary Blair Transport
Medium, Amies Transport Medium.
If culture media is prepared in a plate (Plate Media), weigh, autoclave, then dispense the culture
media but if prepared in a tube (Tube Media), weigh, dispense, then autoclave to sterilize the
culture media.

References:
Engelkirk, P., & Engelkirk, J. (2015). Burton's Microbiology for the Health and Sciences. 10th ed., Philadelphia: Lippincott Williams &
Wilkins
Tortora G., F. B. (2016). Microbiology an Introduction, 12th Edition. United States of America: Pearson Education
Southwestern University College of Nursing (2021). HES 032-Microbiology and Parasitology Laboratory Compilation. Cebu City.
Bregente, C. (Unpublished)
OBSERVATIONS:
A. Plate Media
Blood Agar McConkey Agar Chocolate Agar
Eosin Methylene Blue Agar Nutrient Agar Salmonella Shigella Agar

B. Tube Media
TSI LIA SCA SIM NA Broth
C. Tabulate and classify the different types of culture media above and determine
their uses:

STUDY GUIDE QUESTIONS:
1. Enumerate the steps in preparing a plate and tube culture media.
2. Why should the Petri dish be inverted during incubation and storage in the
refrigerator?


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HES 032-Microbiology and Parasitology

Group Leader: ___________________________________

Group Members: _______________________________________

ACTIVITY NO. 6: Culture Media

1. Identify the different classification of culture media

1. The proper food elements must be available.

Compiled by: Claire Mae Caval, RMT

Culture media can be classified according to three primary levels:

A. According to their Physical State

3. Liquid Media – are prepared without such substances as gelatine or agar.

B. According to their Chemical Composition

Compiled by: Claire Mae Caval, RMT

4. Differential Media – i c g h M c y’ and EMB agars, Triple Sugar Iron

5. Selective Media – contain, in addition to components found in differential media, agents

Compiled by: Claire Mae Caval, RMT

Blood Agar McConkey Agar Chocolate Agar

Eosin Methylene Blue Agar Nutrient Agar Salmonella Shigella Agar

Compiled by: Claire Mae Caval, RMT

TSI LIA SCA SIM NA Broth

Compiled by: Claire Mae Caval, RMT

1. Enumerate the steps in preparing a plate and tube culture media.

Compiled by: Claire Mae Caval, RMT

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