MLT422: CLINICAL MICROBIOLOGY I

BACTERIAL CULTURE MEDIA:

TYPES AND APPLICATIONS

By:
NURAINI SOFEA BINTI AZAMI
2019488802
HS2412A
 Table of Contents

Contents Pages

Culture Media Types and Applications

Liquid Media…………………………………………………………………………………………….…2-3

Basal Media………………………………………………………………………………………………….4

Enrichment Media………………………………………………………………………………………..5

Selective Media…………………………………………………………………………………………..6-7

Differential Media……………………………………………………………………………………….8-9

Transport Media……………………………………………………………………………………..…10-11

References……………………………………………………………………………………………………………….11

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 1. Liquid media

- Liquid media contain specific amounts of nutrients but do not have a trace of gelling
agents such as gelatin or agar.
- Sometimes refered as ‘broth’.
- Liquid medium serves various purposes such as propagation of a large number of
organisms, fermentation studies, and various other tests.
- Bacteria grow uniformly producing general turbidity.

Examples Applications
1. Tryptic Soy Broth (TSB) - Widely used for the isolation of bacteria from clinical
specimens, supporting the growth of the majority of
pathogenic bacteria.
- May be used for preparing dilutions of organism for colony
counts, preparation of standard inoculum and antibiotic
sensitivity testing for both fastidious and non-fastidious
microorganisms.

2. Peptone water - Used for the growth of the organism and a base for
determining carbohydrate fermentation patterns of non-
fastidious organisms.
- Also used for the detection of indole production by the
organism.

3. Nutrient broth - Used for cultivating a broad variety of fastidious and non-
fastidious microorganisms.

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4. Selenite-F broth - Used for isolating Salmonella and Shigella from stool.

5. Glucose broth - Used for blood culture.

- Used to culture fastidious organism such as Streptococci.

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 2. Basal media

- Media that will support the growth of microorganisms that do not have special nutritional
requirements.
- Often used in the preparation of enriched media to maintain stock cultures.

Examples Applications
1. Nutrient broth - Used for cultivating a broad variety of fastidious and
non-fastidious microorganisms.

2. Nutrient agar - Used for the cultivation of microbes.

- Supporting growth of a wide range of non-fastidious
organisms.

3. Peptone water - Used for the growth of the organism and a base for
determining carbohydrate fermentation patterns of
non-fastidious organisms.
- Also used for the detection of indole production by
the organism.

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 3. Enrichment media

- Media that are enriched (added) with whole blood, lyzed blood, serum, special extracts
or vitamin to support the growth of pathogens that require additional nutrients.
- Commonly used to grow as many different types of microbes as are present in the
specimen.

Examples Applications
1. Blood agar - Used to grow fastidious organisms and to
differentiate bacteria based on their hemolytic
properties.

2. Chocolate agar - Also known as heated blood agar or lysed blood agar.
- Used to isolate and grow fastidious respiratory
bacteria, such as Haemophilus influenzae and Neisseria
meningitides.

3. Loeffler's medium - Culture medium containing horse serum, beef extract,

dextrose, and proteose peptones that is used in
diagnostic bacteriology for the culture of
Corynebacterium diphtheriae.

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4. Selective media

- Selective medium is designed to suppress the growth of some microorganisms while

allowing the growth of others.
- Selective medium is agar-based (solid) medium so that individual colonies may be
isolated.

Examples Applications
1. Mannitol salt agar - Used as a selective medium to isolate
Staphylococcus spp.

2. MacConkey’s Agar - Used to isolate Gram-negative and enteric

(normally found in the intestinal tract)
bacilli.

3. Hektoen enteric agar - Used for the isolation of gram-negative

enteric pathogens such as Salmonella and
Shigella.

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4. Eosin Methylene Blue (EMB) Agar - Selective culture medium for Gram-
negative bacteria.
- Commonly used for the isolation and fecal
coliforms.

5. Xylose Lysine Deoxycholate agar - Used for the isolation of Salmonella and
(XLD agar) Shigella species from clinical samples and
from food.

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5. Differential media

- Differential media contain compounds that allow some groups of microorganisms to be

visually distinguished by the appearance of the colony or the surrounding media.
- Due to the presence of certain dyes or chemicals in the media, the organisms will produce
characteristic changes or growth patterns that are used for identification or
differentiation.
- This macroscopic differentiation of bacteria is based on the capability of the bacteria to
carry out specific biochemical processes.

Examples Applications
1. Blood agar (BA) - It is used to differentiate bacteria on the basis of
their hemolytic properties (β-hemolysis, α-
hemolysis, and γ-hemolysis (non-hemolytic)).
- Streptococcus pneumoniae is alpha-hemolytic.
- Group A beta-hemolytic streptococci is
Streptococcus pyogenes while Group B beta-
hemolytic streptococci is Streptococcus agalactiace.
- Staphylococcus saprophyticus and Staphylococcus
epidermidis display gamma hemolysis.

2. Mannitol salts agar (MSA) - Differential medium for mannitol-fermenting

staphylococci.
- It contain carbohydrate mannitol and the indicator
phenol red, a pH indicator for detecting acid
produced by mannitol-fermenting staphylococci.
- If an organism can ferment mannitol, an acidic
byproduct is formed that causes the phenol red in
the agar to turn yellow.
- Staphylococcus aureus produces yellow colonies
with yellow zones, whereas other coagulase-
negative staphylococci produce small pink or red
colonies with no colour change to the medium.

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3. MacConkey Agar - Used to differentiate lactose fermenting from non-
lactose-fermenting gram-negative bacteria.
- Lactose fermenting bacteria such as Escherichia
coli, Enterobacter and Klebsiella grow as red or pink
and may be surrounded by a zone of acid
precipitated bile. The red colour is due to
production of acid from lactose, absorption of
neutral red and a subsequent colour change of the
dye when the pH of medium falls below 6.8
- Non lactose-fermenting bacteria such as Shigella
and Salmonella are colourless and transparent and
typically do not alter appearance of the medium.

4. Eosin Methylene Blue (EMB) Agar - Used as a differential microbiological medium,

which distinguish gram-negative pathogenic
microbes.
- Will provides a color indicator distinguishing
between organisms that ferment lactose and non-
lactose fermenter.
- Organisms that ferment lactose are produce acid
and display "nucleated colonies"—colonies with
dark centers.
- E. coli colonies which is lactose fermenter have a
characteristic green sheen.
- Non-lactose fermenters such as Salmonella and
Shigella are thus either colorless or light lavender.

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6. Transport media

- Special media aims to preserve a specimen and minimize bacterial overgrowth from the
time of collection to the processing of the specimen.
- Depending on the type of organisms suspected in the sample, transport media may vary.
- Contains only buffers and salt.
- Does not contain any nutritional ingredients such as carbon, nitrogen, and organic growth
factors so as to prevent microbial multiplication.
- Addition of antibiotics and other substances like glycerol may be added for transporting
specimens for tissue culture.

Examples Applications
1. Cary and Blair Medium - Used to transport of clinical specimens (faeces)
suspected to contain enteric pathogens, including
Shigella, Salmonella, Vibrio cholerae, and
Escherichia coli O157:H7.
- Alkaline pH of the medium minimizes bacterial
destruction due to the formation of acid.

2. Amies transport medium - Used for the transportation of swab specimens

(sputum, pus, nasopharyngeal specimen and
urogenital swab) to the microbiology laboratory.
- Amies is a modification of Stuart’s medium in
which glycerophosphate is replaced by an inorganic
phosphate buffer and charcoal is added.
- Facilitates the growth of sensitive bacterial
pathogens such as Neisseria gonorrhoeae.

3. Stuarts transport medium - Used for transporting specimens suspected of

having gonococci.
- Also used for transporting throat, wound and skin
swabs that may contain fastidious organisms.
- Originally formulated for the conservation of
Neisseria gonorrhoeae and Trichomonas vaginalis
but it is also used for Haemophilus influenzae,
Streptococcus pneumoniae, Streptococcus
pyogenes and Corynebacterium diphtheria.

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 4. Glycerol Saline Transport medium - Used to transport faeces from patients suspected
to be suffering from bacillary dysentery that caused
by a group of Shigella bacteria

5. Anaerobic Transport Medium - Mineral salt base semi-solid media with reducing
(ATM) agents designed as a holding medium for
maintaining the viability of anaerobic bacteria such
as Actinomyces, Clostridium, Propionibacterium,
Bifidobacterium, Bacteroides, Fusobacterium and
Prevotella.
- It contains buffered mineral salts in a semi-solid
media with sodium thioglycolate and cysteine
added to provide a reduced environment.

References

Aryal, S., Khorshidian, G., Ramey, L., Shan, Sanjeeth, Semester, 4. Soles, E. (2018, October 26).
List of culture media used in microbiology with their uses. Retrieved June 25, 2020, from
https://microbiologyinfo.com/list-of-culture-media-used-in-microbiology-with-their-
uses/

Rijal, N., Says:, A., Says:, M., & Says:, S. (2020, March 23). Transport Medium used in Microbiology
Laboratory. Retrieved June 25, 2020, from https://microbeonline.com/transport-medium-
bacterial-viral-sample-transport-used-microbiology-laboratory/

Zunjarrao, K. (2013, December 28). Culture media & methods. Retrieved June 25, 2020, from
https://www.slideshare.net/KalpeshZunjarrao/cultural-media-methods

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