MICROBIOLOGY LAB 11 LECTURTER

THIRD STAGE DR. ASMAA QASSIM

ENTEROBACTERIACEAE

Enterobacteriaceae are a large family of Gram-negative bacteria present in the

human intestinal tract and are a normal part of the gut flora that includes a number
of opportunistic pathogens Klebsiella, Enterobacter, Citrobacter, Salmonella,
Escherichia coli, Shigella, Proteus and Serratia. All species are end-toxigenic
since they have lipopolysaccharide outer cell wall.

According to the lactose fermentation, they divided into two groups; Lactose
ferment appear as pink to red colonies and Lactose non-ferment appear as pale
colonies.

General characteristics

1. Citrobacter spp., are gram-negative, facultative anaerobic bacteria that

appear as rods or coccobacilli, (Catalase + and Oxidase -)
2. Salmonella Gram negative, facultative anaerobes, motile, non-spore rode.
All except S. typhi are non- capsulated.(Motility +)
3. Shigella Gram negative, non-motile rods and non-capsulate. (Motility –)
4. Escherichia coli gram negative flagellated-rode motile, sometimes
capsulate, facultative anaerobic and bile-tolerant.(Ureas – and Indol +)
5. Proteus is gram-negative, rod-shaped, and facultative anaerobic,
characteristic swarming motility that will become evident on blood agar
plates. (Ureas + and Motility +)
6. Klebsiella is a gram-negative, lactose-fermenting, non-motile, aerobic rod-
shaped bacterium. (Motility – )
7. Enterobacter is a gram-negative, lactose-fermenting, motile, aerobic rod-
shaped bacterium. (Motility + )

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 MICROBIOLOGY LAB 11 LECTURTER
THIRD STAGE DR. ASMAA QASSIM

8. Serratia is a motile, short rod-shaped, Gram-negative, facultative anaerobe

bacterium (Motility + and Urease -)
9. Yersinia Gram-negative, coccobacilli bacteria, and are facultative anaerobes
and they are actively motile at 22–25 °C but not motile at 37 °C. (Motility –
and Catalase +)

Lab Diagnostic Tests

Colonial Morphology

1- MacConkey agar (MAC)

Selective: Bile salts and crystal violet inhibit most gram-positive organisms and
permit growth of gram-negative rods.

Differential: Lactose serves as the sole carbohydrate. Lactose fermenters produce

pink or red colonies, may be precipitated bile salts may surround colonies. Non–
lactose fermenters appear colorless or transparent. This color differentiation is due
to the presence of neutral red indicator (yellow in alkaline pH and pink in acid pH).

Klebsiella: It appears as a mucoid lactose fermenter on MacConkey agar.

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 MICROBIOLOGY LAB 11 LECTURTER
THIRD STAGE DR. ASMAA QASSIM

Enterobacter: lactose-positive colonies are colored pink to red

Escherichia coli Colonies on MacConkey agar palte are pink to dark pink, dry and
donut-shaped, surrounded by a dark pink area of precipitated bile salts.

Citrobacter: small, circular, convex dark pink colonies on MacConkey agar.

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 MICROBIOLOGY LAB 11 LECTURTER
THIRD STAGE DR. ASMAA QASSIM

Serratia: The organism grew as dark red colonies against the light pink
background of the media.

Proteus: pale or colorless due to lack of lactose fermentation

Yersinia: translucent, small colonies, pale non-lactose fermenting colonies are seen

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 MICROBIOLOGY LAB 11 LECTURTER
THIRD STAGE DR. ASMAA QASSIM

Salmonella-Shigella agar or SS agar

 On the use of bile salt, sodium citrate and brilliant green for the inhibition
of gram-positive bacteria.
 On lactose fermentation and hydrogen sulphide (H2S) production for the
differentiation of organisms

Salmonella: Salmonella will not ferment lactose, but produce hydrogen sulfide
(H2S) gas. The resulting bacterial colonies will appear colorless with black
centers.

Shigella do not ferment lactose or produce hydrogen sulfide gas, so the resulting
colonies will be colorless.

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 MICROBIOLOGY LAB 11 LECTURTER
THIRD STAGE DR. ASMAA QASSIM

2- Gram Stain Reaction

Generally, the enterobacterase group appease under microscopic examination as

Gram negative rod (bacillus to cocoobacillus)