Bacterial and Fungal Culture Characteristics

Presentation
on
BACTERIAL AND FUNGAL
COLONY CHARACTERISTICS IN
DIFFERENT CULTURE MEDIA
Welcome
Basics of different bacteriological culture media
 Culture media contains nutrients and physical

growth parameters necessary for microbial
growth.
Classification of different bacteriological culture media

a) On the basis of consistency
1. Solid medium
-contains agar at a concentration of 1.5-2.0% , mostly inert
solidifying agent.
-allows bacteria to grow in as colonies or in streaks.
-useful for isolating bacteria or for determining the colony
characteristics of the isolate.
2. Solid medium
-contains agar at a concentration of 1.5-2.0% , mostly inert solidifying
agent.
-allows bacteria to grow in as colonies or in streaks.
-useful for isolating bacteria or for determining the colony characteristics
of the isolate.
3. Semisolid media
-prepared with agar at concentrations of 0.5% or less.
-useful for the cultivation of micro-aerophilic bacteria or for determination
of bacterial motility.
4. Liquid (Broth) medium

-contains specific amounts of nutrients but don’t have trace of gelling
agents such as gelatin or agar.
b) On the basis of composition
1. Synthetic or chemically defined medium:

-prepared from purified ingredients and therefore whose exact
composition is known.
2. Non synthetic or chemically undefined medium

-contains at least one component that is neither purified nor
completely characterized nor even completely consistent
from batch to batch. Nutrient broth, for example, is derived
from cultures of yeasts.
c) On the basis of purpose/ functional use/ application
1. General purpose media/ Basic media

-are basically simple media that supports most bacteria.
-peptone water, nutrient broth and nutrient agar are considered as basal
medium.
2. Enriched medium (Added growth factors):

-addition of extra nutrients in the form of blood, serum, egg yolk etc, to
basal medium makes them enriched media.
-Enriched media are used to grow nutritionally exacting (fastidious)
bacteria.
-blood agar is prepared by adding 5-10% (by volume) blood to a blood
agar base.
-chocolate agar is also known as heated blood agar or lysed blood agar.
3. Selective and enrichment media

- help to recover pathogen from a mixture of bacteria.
-while selective media are agar based, enrichment media are liquid
in consistency.
4. Differential/ indicator medium
-designed in such a way that different bacteria can be recognized on
the basis of their colony colour.
-differential media allow the growth of more than one
microorganism of interest but with morphologically distinguishable
colonies.
5. Transport media
-transport media used to transport the sample to laboratory
-such media prevent drying (desiccation) and inhibit overgrowth of
unwanted bacteria.
Examples of selective media include
Mannitol Salt Agar and Salt Milk Agar used to recover Staphylococcus aureus contains
10% NaCl.
MacConkey’s Agar used for Enterobacteriaceae members contains bile salt that inhibits
most gram positive bacteria.
Pseudosel Agar (Cetrimide Agar) used to recover Pseudomonas aeruginosa contains

cetrimide (antiseptic agent).
Examples of differential media include
Mannitol salts agar (mannitol fermentation = yellow)
Blood agar (various kinds of hemolysis i.e. α, β and γ hemolysis)
Mac Conkey agar (lactose fermenters, pink colonies whereas non- lactose fermenter
produces pale or colorless colonies.
6. Anaerobic media
-anaerobic bacteria need special media for growth because they need
low oxygen content, reduced oxidation –reduction potential and
extra nutrients.
-Robertson Cooked Meat (RCM) medium that is commonly used to
grow Clostridium spp.
-Thioglycollate broth contains sodium thioglycollate, glucose,
cystine, yeast extract and casein hydrolysate.
Bacterial growth on Nutrient agar
Fig: Different bacterial growth (Left: E. Coli, Right: Others) on Nutrient agar
Bacterial growth on Nutrient agar
Fig: Different bacterial growth (Left: E. Coli, Right: Staphylococcus aureus) on Nutrient
agar
Bacterial growth on MacConkey’s agar (MCA)
Expected Colony characteristics in MacConkey Agar
Lactose-fermenting organisms grow-

-as pink to brick red colonies
-with or without a zone of precipitated bile.
-Citrobacter sp., Escherichia sp., Enterobacter sp., Klebsiella sp.
Non-lactose fermenting organisms grow-

-as colorless or clear colonies
-Shigella sp., Yersinia sp., Proteus sp.,Salmonella sp.
Lactose fomenters
Non lactose fermenter

Fig: Klebsiella (pink) and E. coli (brick Fig: E. coli (brick red) and Salmonella
red) colonies (Colorless) colonies
Bacterial growth on Mannitol Salt Agar (MSA)
Fig- (Left) Yellow colonies; may have yellow halo around colonies.
(Right) Colorless to pink colonies
Fig-:Yellow colonies; may have yellow halo around colonies.

Bacterial growth on Cetrimide Agar
The presence of growth is indicative of a

positive reaction.
Visual examination may also reveal the typical

yellow-green to blue color which indicates the
production of pyocyanin. Both pyocyanin and
fluorescein are typically produced by strains
of Pseudomonas aeruginosa .
A negative reaction is denoted by no growth.

Bacterial growth on Cetrimide Agar
Fig: Typical yellow-green to blue color colony in both images.

Bacterial growth on Xylose-Lysine Deoxycolate (XLD) Agar
Colony characteristics in XLD Agar
Enterobacter aerogenes : Yellow

Escherichia coli : Yellow
Proteus vulgaris : Grey with black centers
Salmonella paratyphi : Red
Salmonella sp.: Red colonies with black centres (strong
H2s producer)
Shigella dysenteriae , Shigella flexneri & Shigella
sonnei : Red
Staphylococcus aureus : No growth
E. coli on XLD
Salmonella on XLD
Fig: Salmonella on XLD (Black), E. coli on XLD (White)

Proteus on XLD
Salmonella on XLD
Fig: Proteus on on XLD (Grey), Salmonella on XLD (Dark

black)
Shigella sp. on XLD Enterobacter sp. On XLD
Proteus sp. on XLD
Fig: Proteus on on XLD (Grey), Shigella on XLD (Red to

colorless) and Enterobacter on XLD (Yellow)
Bacterial growth on Salmonella-Shigella (SS)Agar
Typical colonial morphology on SS Agar is as follows-
E.coli ……….………………..Slight growth, pink or red

Enterobacter/Klebsiella …..………..Slight growth, pink
Proteus …………..Colorless, usually with black center
Salmonella……….Colorless, usually with black center
Shigella .…………………………………..Colorless
Pseudomonas .……………….Irregular, slight growth
Salmonella sp.
on SS
Shigella sp. on SS
Fig: Salmonella sp. on SS (Black centre), Shigella

sp. on SS (Colorless)
Proteus sp. on SS
Fig: Proteus sp. on SS (Black centre)

E. coli on SS
Fig: E. coli on SS (Pink or red)

Bacterial growth on Eosin Methylene Blue (EMB) Agar
Expected colony characteristics of organisms in EMB agar
Escherichia coli : Blue-black bulls eye; may

have green metallic sheen.
Pseudomonas aeruginosa : Colorless.

E. coli on EMB
Fig: E. coli on EMB Agar (Green metallic sheen)

Pseudomonas sp. on EMB
Fig: Pseudomonas aeruginosa on EMB agar (Colorless).

Bacterial growth on Modified Semi-solid Rappaport Vassiliadis
(MSRV) medium
MSRV interpretations-
Positive:
-Growth of migrated cells is visible as a grey-white, turbid
zone extending out from the inoculated drop.
-Test sample is considered presumptively positive for motile
Salmonella spp.
Negative:
-Medium remains blue-green around inoculation drops, with
no grey-white, turbid zone extending out from the drop.
-Test sample is considered negative for motile Salmonella spp.
(MSRV) medium
(MSRV) medium
Fig: Suspected Salmonella spp. on MSRV medium

(MSRV) medium
Fig: Suspected Salmonella spp. on MSRV medium

Bacterial growth on Triple Sugar Iron (TSI) Agar
Slunt
Fungal growth on Potato Dextrose Agar (PDA)/ Sabouraud
Dextrose Agar (SDA)
Fig: Aspergillus flavus

Dextrose Agar (SDA)
Fig: Penicillium chrysogenum

Dextrose Agar (SDA)
Fig: Candida albicans

Bacterial and Fungal Culture Characteristics

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Bacterial and Fungal Culture Characteristics

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Presentation

 Culture media contains nutrients and physical

Classification of different bacteriological culture media

4. Liquid (Broth) medium

b) On the basis of composition

1. Synthetic or chemically defined medium:

2. Non synthetic or chemically undefined medium

1. General purpose media/ Basic media

2. Enriched medium (Added growth factors):

3. Selective and enrichment media

Pseudosel Agar (Cetrimide Agar) used to recover Pseudomonas aeruginosa contains

Examples of differential media include

Mannitol salts agar (mannitol fermentation = yellow)

Blood agar (various kinds of hemolysis i.e. α, β and γ hemolysis)

Expected Colony characteristics in MacConkey Agar

Lactose-fermenting organisms grow-

Non-lactose fermenting organisms grow-

Non lactose fermenter

Fig-:Yellow colonies; may have yellow halo around colonies.

The presence of growth is indicative of a

Visual examination may also reveal the typical

A negative reaction is denoted by no growth.

Fig: Typical yellow-green to blue color colony in both images.

Enterobacter aerogenes : Yellow

Fig: Salmonella on XLD (Black), E. coli on XLD (White)

Fig: Proteus on on XLD (Grey), Salmonella on XLD (Dark

Shigella sp. on XLD Enterobacter sp. On XLD

Proteus sp. on XLD

Fig: Proteus on on XLD (Grey), Shigella on XLD (Red to

Typical colonial morphology on SS Agar is as follows-

E.coli ……….………………..Slight growth, pink or red

Fig: Salmonella sp. on SS (Black centre), Shigella

Fig: Proteus sp. on SS (Black centre)

Fig: E. coli on SS (Pink or red)

Expected colony characteristics of organisms in EMB agar

Escherichia coli : Blue-black bulls eye; may

Pseudomonas aeruginosa : Colorless.

Fig: E. coli on EMB Agar (Green metallic sheen)

Pseudomonas sp. on EMB

Fig: Pseudomonas aeruginosa on EMB agar (Colorless).

Fig: Suspected Salmonella spp. on MSRV medium

Fig: Suspected Salmonella spp. on MSRV medium

Fig: Aspergillus flavus

Fig: Penicillium chrysogenum

Fig: Candida albicans

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