PHBIOSC4 | Pharmaceutical Microbiology & Parasitology

Culture Media

LEARNING OBJECTIVES DEFINITION

1. Describe the different types of culture A culture medium is anything that supports
media the growth and physiologic requirements of
2. Classify the different types of culture microorganisms. It is basically an aqueous
media solution to which all the necessary nutrients
3. Describe the preparation of culture have been added. Depending on the type,
media physical properties, and combination of
nutrients, different classifications of culture
TOPIC OUTLINE media can be made.

1. Definition of culture media CLASSIFICATION

2. Classification of culture media
3. Preparation of culture media Culture media can be classified according
to three primary levels:
1. Physical State
OVERVIEW 2. Chemical Composition
3. Functional Type
This lesson for online distance learning
provides Pharmacy students the essential 1. Physical State
information to prepare culture media in
the laboratory and utilize culture media Liquid Media – these are water-
appropriately for the growth and study of based solutions that do not solidify
microorganisms. at temperatures above freezing
point. These media are commonly
termed broths, milk or infusions.
ACTIVATING PRIOR KNOWLEDGE

How are microorganisms cultured in the Semisolid Media – these exhibit a

laboratory? What medium are used to clotlike consistency at ordinary
facilitate their growth? room temperature. These media
contain an amount of solidifying
agent (agar or gelatin) which
INTRODUCTION thickens them but does not produce
a firm substance.
Microorganisms need nutrients and
certain environmental conditions in order Solid Media – these provide a firm
to grow and reproduce. In the surface on which cells can form
environment, these microorganisms have discrete colonies and are
adapted to the habitats most suitable to advantageous for culturing and
their needs. In the laboratory, however, isolating bacteria and fungi. Solid
these requirements must be met by a media come in two form: liquefiable
culture medium. or reversible solid media and
nonliquefiable or non-reversible
solid media.

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Culture Media

CLASSIFICATION

2. Chemical Composition Enrichment Media – these are

designed to increase the number of
Synthetic – these compositions are desired microorganisms to a
chemically defined. Synthetic media detectable level without stimulating
are composed of pure organic and the rest of the bacterial population.
inorganic compounds which have It contains complex organic
molecular contents specified by substances such as blood, serum,
means of an exact formula. hemoglobin or special growth
factors.
Non-synthetic – these complex
media contain at least one Examples:
ingredient that is not chemically Blood Agar – contains general
defined, not a simple compound, nutrients with 5% blood added. It
pure compound and not is used both as an enriched
representable by an exact chemical medium and as a differential
formula. Most of these substances medium. Exotoxins called
are extracts of animals, plants or hemolysins cause lysis of the
yeasts. red blood cells. The degree of
hemolysis is an especially useful
3. Functional Types tool for identification of many of
the Gram positive cocci. These
General-purpose Media – these organisms are grouped into
contain mixture of nutrients that three categories based on
could support the growth of hemolytic reactions:
pathogens and nonpathogens alike. Beta hemolysis completely
They are designed to grow as broad lyses the red blood cells and
a spectrum of microbes as possible. hemoglobin; this results in
complete clearing around
Examples: colonies.
Nutrient Agar – contains beef Alpha hemolysis refers to
extract and peptone, as well as the partial lysis of RBCs and
1.5% agar by weight hemoglobin and produces a
Nutrient Broth – contains beef greenish discoloration of the
extract and peptone dissolved in blood agar around the
water colonies.
Trypticase Soy Agar – contains No hemolysis, sometimes
partially digested milk protein called gamma hemolysis
(casein), soybean digest, NaCl, results in no change of the
and agar medium.

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Culture Media

CLASSIFICATION

Selective Media – these contain one Mannitol Salt Agar – was

or more agents that inhibit the developed by microbiologists
growth of a certain microbe or seeking a method for isolating
microbes and therby encourage only Staphylococcus aureus, a
certain microbes to grow. Selective pathogen frequently transmitted
media are very important in primary by contaminated food. This
isolation of a specific type of medium contains 7.5% salt,
microorganism from samples which is inhibitory to most
containing many different species bacteria other than
like feces, saliva, skin, water and Staphylococcus.
soil. Thayer-Martin Agar - is a
chocolate agar designed to
Examples: isolate Neisseria gonorrhoeae.
MacConkey Agar (MAC) – is a Saboraud’s Dextrose Agar (SDA)
widely used culture medium – is used for fungi. It contains
which is both selective and gentamicin and has a low pH
differential. The medium is that will kill most bacteria.
primarily used to differentiate Hektoen Enteric Agar – is a
between Gram negative bacteria selective medium with bile salts
while inhibiting the growth of added to inhibit Gram positive
most Gram positive bacteria. organisms. The comparatively
Addition to the nutrient agar high concentration of bile salts
base of bile salts and crystal inhibits not only Gram positive,
violet will inhibit the growth of but also some Gram negative
most Gram positive bacteria, organisms, but not Salmonella
making MacConkey agar and Shigella species.
selective. Lowenstein-Jensen Medium
Eosin Methylene Blue (EMB) – (LJ) – is used to isolate
is a selective and differential Mycobacterium.
medium used for isolation and Mueller-Hinton Agar (MHA) – is
differentiation among members used to isolate Neisseria
of the Enterobacteriaceae. Eosin gonorrhea and N. meningitides
methylene blue agar selects for and also for antimicrobial
Gram negative bacteria. It sensitivity testing.
contains methylene blue and Cetrimide Agar – is used to
eosin dyes to inhibit the growth isolate Pseudomonas
of Gram positive bacteria. aeruginosa.
Salmonella-Shigella Agar (SSA)
– is used to isolate Salmonella
and Shigella species.

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Culture Media

CLASSIFICATION

Differential Media – these allow resulting in a characteristic

growth of several types of greenish, metallic sheen.
microorganisms and are designed to Organisms which do not ferment
display visible differences among lactose will be colorless, taking
those microorganisms. on the color of the medium. This
Differentiation shows up as medium has been widely used in
variations in colony size or in color, the past to screen for coliforms
in media color changes or in the in the water.
formation of gas bubbles and Triple Sugar Iron Agar –
precipitates. differentiates bacteria based on
their ability to ferment glucose,
Examples: lactose and/or sucrose, and to
MacConkey Agar – reduce sulfur to hydrogen
differentiates between lactose- sulfide. It is used primarily to
fermenting coliforms and distinguish the morphologically
lactose nonfermenters, which similar bacteria of
include potential pathogens. Enterobacteriaceae, all of which
Lactose, a fermentable ferment glucose to an acid end
carbohydrate, and neutral red, a product.
pH indicator, are added to Mannitol Salt Agar – is used to
differentiate the lactose positive differentiate pathogenic
coliforms from the potentially Staphylococcus species from
pathogenic lactose nonpathogenic members of the
nonfermenters. When lactose is genus Micrococcus. If the
fermented, acid products lower organism does not use mannitol,
the pH below 6.8, with the the medium will remain red (no
resulting colonial growth turning change). If the organism does
pinkish-red. If an organism is ferment mannitol, it will create
unable to ferment lactose, the metabolic by-products which are
colonies will be colorless. acidic – and the surrounding
Eosin Methylene Blue – medium will be yellow.
differentiates those with ferment Salmonella-Shigella Agar (SSA)
lactose (the coliforms) from the – is used to differentiate
coliforms which do not ferment Salmonella and Shigella species
lactose. It contains lactose, whether they are lactose (red
Small amounts of acid colonies) and non-lactose
production result in a pink fermenters (colorless colonies)
colored growth, while large as well as hydrogen sulfide
amounts of acid cause the acid producers (black color).
to precipitate on the colony,

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Culture Media
PREPARATION REFERENCES

A complete set of instructions for the Ho, H., Roldan-Gan, R., & Verbo, V.
preparation of culture media is given on the (2006). Microbiology and Parasitology
label of each bottle. Laboratory Manual for the Health
Sciences. C&E Publishing Inc.
During the preparation of culture media, it is Kreislur, J.E. (2011). Microbiology for
important to take note if the medium that is Nursing Students. Educational
being prepared is a tube or plated medium. Publishing House.
In preparing tube medium, it is necessary to Basit, S.A. & Juayang, A.C. (2018). OBE
dispense first the medium before sterilizing Manual in Microbiology and Parasitology
while for the preparation of plated medium, for Health Sciences. APD Educational
it is important to sterilize first before Publishing House.
dispensing in Petri dishes.

Watch the step by step procedure in

preparing culture media thru this link
https://bit.ly/3coatAS.

SUMMARY

Culture media contains the necessary

nutrients required for the culture of
microorganisms in the laboratory hence is
essential in the study of microbiology. The
utilization of an appropriate type of culture
media allows the growth, selection or
differentiation of microorganisms being
studied. Proper preparation of culture media
should be observed to prevent
contamination from unwanted
microorganisms.

LEARNING ACTIVITY

Answer the following questions:

1. Why should culture media be sterilized
before and after use?
2. Why is McConkey Agar selective as well
as differential? Explain.
3. What is chocolate agar used for? What
is its difference with blood agar?
4. Outline the steps in culture media
preparation.

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