1/10/23, 2:39 PM Culture Media: Classification, Types, and Relevance - Conduct Science

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1/10/23, 2:39 PM Culture Media: Classification, Types, and Relevance - Conduct Science

What Is Culture Media?

Culture media are mediums that provide essential nutrients and minerals to support the
growth of microorganisms in the laboratory.

Microorganisms have varying nature, characteristics, habitat, and even nutritional

requirements, thus it is impossible to culture them with one type of culture media. However,
there are also microorganisms that can’t grow on a culture media at all in any condition –

these are called obligate parasites.[1]

Culturing microorganisms is essential for diagnosing infectious diseases, obtaining antigens,

developing serological assays for vaccines, genetic studies, and identification of microbial
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Furthermore,
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essential for isolating pure cultures, storing culture stock, studying
biochemical reactions,
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preservatives effect, testing viable count, and testing antibiotic sensitivity.[2]
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This article will focus on the composition, classification, and types of culture media used in
microbiology labs to study a spectrum of microbial forms.

Classification and Types of Culture Media

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1/10/23, 2:39 PM Culture Media: Classification, Types, and Relevance - Conduct Science

Growing microorganisms in the lab involve mimicking the organisms’ natural habitat or
environment, and this is possible in the laboratory by formulating culture media that meets
their requirements. Therefore, many culture media were developed by scientists according to
the microbial species to be cultured.

The basic media contains a source of carbon & energy, nitrogen source, growth factors, and

some trace elements.[1] Some commonly used media components include peptone, agar,
water, casein hydrolysate, malt extract, meat extract, and yeast extract. In addition, the pH of

the medium should be set accordingly.[3]

However, some additional components or nutrients are added to the media when growing
specific microorganisms.

Culture media can be classified in three ways: based on their consistency, nutritional

component, and applications.[1]

A. Classification of culture media based on consistency

1. Solid media: In these media, the agar which is an unbranched long chain of
polysaccharides is added in the concentration of 1.5-2.0%. Most commonly, 1.3% agar is

used to prepare solid media in labs. The agar-containing media solidifies at 37 ºC.[1]
Sometimes, in the place of agar, some other inert solidifying agents are used, such as
gellan gum.
Solid media are used to grow microorganisms in their full physical form, prepare bacterial

pure cultures, or isolate bacteria to study colony characteristics.[1] The bacterial growth
on solid media varies in appearance as mucoid, round, smooth, rough, filamentous,
irregular, and punctiform. The media is not hydrolyzed by microorganisms and is free

from growth-inhibiting substances.[3] Examples of solid media are blood agar, nutrient
agar, McConkey agar, and chocolate agar.

2. Semisolid media: This media has 0.2-0.5% agar concentration, and due to the reduced
agar concentration, it appears as a soft, jelly-like substance. It’s mainly used to study the
motility of microorganisms, distinguish between motile and non-motile bacterial strains
(through U-tube and Cragie’s tube), and cultivate microaerophilic bacteria – bacteria on
this media appear as a thick line. Examples of semi-solid media are: Hugh and Leifson’s
oxidation fermentation medium, Stuart’s and Amies media, and Mannitol motility media.
[1]

3. Liquid media: These media do not contain any traces of solidifying agents, such as agar
or gelatin, and large growth of bacterial colonies can be observed in the media. Liquid
media uses
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Other than these, there are also biphasic media, which consist of both solid and liquid media.
And sometimes in the place of agar, egg yolk and serum are added to the media as a

solidifying agent.[3] Learn more on how to make agar plates here.

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1/10/23, 2:39 PM Culture Media: Classification, Types, and Relevance - Conduct Science

While naturally, these substances are liquid, they are solidified by using heat, and the
prepared media is sterilized using the inspissation technique. Examples are Lowenstein
Jensen medium and Dorset egg medium, which contain egg yolk, and Loeffler’s serum slope,

which contain serum.[3]

B. Classification based on the nutritional component

1. Simple media: It’s a general-purpose media that supports the growth of non-fastidious
microbes, and it is primarily used for the isolation of microorganisms. Examples are
nutrient broth, peptone water, and nutrient agar.

2. Complex media: These are media containing nutrients in unknown quantities that are
added to bring about a particular characteristic of a microbial strain. Examples are
tryptic soy broth, blood agar, and nutrient broth.

3. Synthetic media: Synthetic media is a type of chemically defined media and is produced
from pure chemical substances. A defined media refers to a medium having a known
concentration of ingredients, like sugar (glucose or glycerol) and nitrogen source (such
as ammonium salt or nitrate as inorganic nitrogen). It is generally used in scientific

research, and an example is Czapek Dox Medium.[1]

C. Classification of culture media based on

application/chemical composition

Basal media: These are routinely used simple media having carbon and nitrogen sources
that boost the growth of many microorganisms. They are also known as general-purpose
media and are considered non-selective media.

The basal media do not require enrichment sources for the growth of non-fastidious

bacteria and are suitable for growing Staphylococcus and Enterobacteriaceae.[1]

They are generally used to isolate microorganisms in labs or in sub-culturing processes.

Examples are nutrient broth, nutrient agar, and peptone water.

Enriched media: This media is prepared by adding additional substances like blood,
serum, or egg yolk in the basal medium. It’s used to grow fastidious microorganisms as
they require additional nutrients and growth-promoting substances.

Examples are chocolate agar, blood agar, and Loeffler’s serum slope. Chocolate media is
used to grow N. gonorrhea while blood agar (which is prepared by adding 5-10% blood by

volume to a blood agar base) is used to identify hemolytic bacteria.[2]

Selective
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growth
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The selective growth of microbes is decided by adding substances like antibiotics, dyes,
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salts, or by pH adjustments.

Below is a list of common selective media and the bacteria they’re used to culture:[2]

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1/10/23, 2:39 PM Culture Media: Classification, Types, and Relevance - Conduct Science

S.
Culture media Inhibiting substances Bacteria
No

1 Thayer Martin Agar Contains antibiotics; vancomycin, Used for Neisseria gonorrhoeae
colistin, and nystatin
2 MacConkey’s Agar Contains bile salts Used for Enterobacteriaceae members
3 Lowenstein Jensen Medium Addition of malachite green Used for M.tuberculosis
4 Mannitol Salt Agar Contains 10% NaCl Used to recover S.aureus
5 Crystal Violet Blood Agar Contains 0.0002% crystal violet Used for Streptococcus pyogenes

6 Thiosulfate citrate bile salts Have elevated pH of about 8.5-8.6 Used for isolating Vibrio cholerae
sucrose (TCBS) agar
7 Wilson and Blair’s Agar Addition of dye brilliant green Used for recovering S. typhi
8 Potassium tellurite medium Contains 0.04% Potassium tellurite Used to recover C.diphtheriae
9 Pseudosel Agar (cetrimide agar) Contains cetrimide (antiseptic agent) Used to recover Pseudomonas aeruginosa
Contains bile salts, brilliant green, and Used for the isolation of Salmonella,
10Enrichment media: It’sAgara liquidsodium
medium,
citrate used to increase the relative concentration of
Salmonella-Shigella
which causes typhoid
certain microbes before culturing them on a solid medium plate. It’s used as a broth
medium and inhibits the growth of commensal species of microorganisms (those who
live in close association with each other) in the clinical specimen.
It’s also used in isolating fecal and soil microorganisms. Examples are selenite F broth
which is used to isolate Salmonella typhi from a fecal sample, tetrathionate broth, and

alkaline peptone water.[1]

Differential or indicator media: It contains certain indicators like dyes or metabolic

substrates in the medium composition which gives different colors to colonies of different
microbial species when they utilize or react with these components.
It allows the growth of more than one microorganism, however, the bacterial colonies are
differentiated based on their color when a chemical change occurs in the indicator, such

as neutral red, phenol red, methylene blue. Examples are:[1]

Blood agar: In blood agar, three types of blood cell lysis or hemolysis are observed:

alpha, beta, and gamma hemolysis.[5] It allows the growth of many microorganisms,
however, their ability to lyse blood cells differs, and this helps to distinguish the
bacterial colonies.
For example, S. pyogenes completely lyse blood cells (beta hemolysis), thus causing
total clearing of the media around its colonies. S. pneumoniae partially lyse red blood
cells, resulting in a greenish-colored medium, while gamma hemolytic
microorganisms like Enterococcus faecalis, Staphylococcus saprophyticus, and
Staphylococcus epidermidis, can’t lyse red blood cells, thus causing no color change

in the medium.[6]

Mannitol salts agar: The fermentation of mannitol by Staphylococcus aureus causes

the media to change to yellow, however, coagulase-negative staphylococci that
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cause to
fermentation to appear in pink.[7]
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MacConkey agar: It differentiates the gram-negative bacteria based on their lactose
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Citrobacter, and Enterobacter forms pink-red colonies, while lactose non-fermenters,
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like Salmonella, Shigella, Proteus, Providencia, Pseudomonas, and Morganella form

pale or colorless colonies.[8]

Thiosulfate citrate bile salts sucrose (TCBS) agar: The media contain sucrose, which
is utilized by ferment microbes and helps to distinguish them from non-ferment
microorganisms. Based on this characteristic, different colored bacterial colonies are
formed on the media that help to identify and distinguish them from each other.

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1/10/23, 2:39 PM Culture Media: Classification, Types, and Relevance - Conduct Science

For example, V. cholerae ferment the sucrose and form slightly flattened yellow
colonies having opaque centers and translucent peripheries. Whereas, V.
parahaemolyticus can’t ferment the sucrose and forms green to blue-green colonies.
[9]

Transport media: Transport media are useful for clinical specimens which are required to
be transferred immediately to labs to maintain the viability of potential pathogens and to
prevent overgrowth of commensals or contaminating microorganisms. Some of them are
semi-solid in consistency, and examples include:
Sach’s buffered glycerol saline: It’s used to transport feces from patients suspected
to be suffering from bacillary dysentery.

Cary Blair transport and Venkatraman Ramakrishnan media: Fecal samples

collected from suspected cholera patients are transported using these media.

Pike’s medium: A throat specimen containing Streptococci is transported using this

medium.[2]

Anaerobic media: This media is for anaerobic bacteria which require low oxygen levels,
extra nutrients, and reduced oxidation-reduction potential. It is supplemented with hemin
and vitamin K nutrients and oxygen is removed by boiling it in a water bath and sealing it

with paraffin film.[2]

Examples are: Thioglycollate broth and Robertson Cooked Meat (RCM) medium which is

commonly used to grow Clostridium spp.[2]

Assay media: It’s used for amino acids, vitamins, and antibiotics assays. For example,
antibiotic assay media is used to determine the antibiotic potency of microorganisms.

Storage media: It’s used to store microorganisms for a longer period, examples are chalk

cooked meat broth and egg saline medium.[2]

Conclusion

Culture media is a source of nutrients and growth factors required for the growth of
microorganisms and even plants in laboratory conditions. Every organism has different
nutritional requirements based on its habitat or living conditions. Therefore, a single
formulation of culture media can’t be used to grow all organisms in labs.

Many types of culture media have been developed by scientists to grow selective or desired
microorganisms. These are classified based on their nutrient composition, consistency, and
application or use in life science laboratories.

This website
Culture usesserve
media cookies to purposes in labs like isolating specific strains of microorganisms,
several
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Thus, before deciding which culture media to use, it is critical to determine the purpose of
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your study morein some cases the type of microorganism you want to study. This narrows
down your choices and helps you to choose which media is best for your experiment, without
wasting your time and effort.

References:

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1/10/23, 2:39 PM Culture Media: Classification, Types, and Relevance - Conduct Science

1. Tankeshwar Acharya (2021). Bacterial Culture Media: Classification, Types, Uses. Retrieved
from https://microbeonline.com/types-of-bacteriological-culture-medium/.

2. Fatima Aiman (2022). Microbial Culture Media- Definition, Types, Examples, Uses.
Retrieved from https://microbenotes.com/types-of-culture-media/#application-of-
culture-media.

3. Rao Sridhar. Bacterial Culture Media. Retrieved from

https://www.microrao.com/micronotes/pg/culture_media.pdf.

4. Aryal Sagar (2022). Salmonella Shigella (SS) Agar- Composition, Principle, Preparation,
Results, Uses. Retrieved from https://microbenotes.com/salmonella-shigella-ss-agar/.

5. Tankeshwar Acharya (2021). Blood Agar and Types of Hemolysis. Retrieved from
http://microbeonline.com/blood-agar-composition-preparation-uses-and-types-of-
hemolysis/.

Anjali Singh
Anjali Singh is a freelance writer. Following her passion for science and research, she did her
Master’s in Plant Biology and Biotechnology from the University of Hyderabad, India. She has a
strong research background in Plant Sciences with expertise in Molecular techniques, Tissue
culture, and Biochemical Assays. In her free time outside work, she likes to read fictional books,
sketch, or write poems. In the future, she aspires to pursue a doctorate in Cancer Biology while
continuing her excellence as a scientific writer.

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