DIPLOMA IN MEDICAL LABORATORY TECHNOLOGY

PROGRAM
DEPARTMENT OF MEDICAL SCIENCES & TECHNOLOGY
FACULTY HEALTH SCIENCES
COURSE CODE PDML 4233

COURSE NAME Clinical Microbiology II

LECTURER Miss Nurul Aini binti Ahmad

Case study 2:
Gram Negative Bacilli

STUDENT NAME STUDENT ID.

NUR HAZIRAH AMANI BINTI HALIM MLT07190003
 Bacteria belonging to the family Enterobacteriaceae are the most commonly
encountered organisms isolated from clinical specimens. Medically important
members of this family are commonly referred to as fermentative, gram-negative,
enteric bacilli, because they are gram-negative rods that can ferment sugars. Some
of the more common clinically important genera of the family Enterobacteriaceae
include Escherichia, Klebsiella, Salmonella, Shigella and Proteus. Many are normal
flora of the intestinal tract of humans and animals while others infect the intestinal
tract.

Biochemical tests that have been mentioned in the journal in the below table.

Test E.Coli Shigella Salmonella

Gram positive cocci, in purple
Gram negative E. Coli, in red
1. Gram stain
Anaerobe bacteria
Gram positive cocci, purple
Gram negative, pink
Anaerobe bacteria
Gram positive: stain purple.
Thick cell wall without an
outer membrane
Gram negative: stain red.
Thin cell wall with an outer
membrane. Salmonella

Anaerobe bacteria

Shows green metallic sheen.

2. Eosin Rapid fermentation of lactose
Luxurian growth: colorless Luxurian growth: colorless
Methylene Blue and production of strong acids,
colonies colonies
agar-culture reduction in pH.

3. Mac conkey- Appear pinkish colour of
culture colonies
TSI recommended for use in
the differentiation of
Enterobacteriaceae by their
ability to ferment glucose,
lactose, and sucrose, and
ability to produce hydrogen
sulfide.Growth under 35oC for
4. TSI 18-24hr
Yellow slant, yellow butt:
lactose and glucose
fermentation.
Gas positive, No H2S produced.
Indicator: phenol red
Appear pinkish colour of
colonies
Shigella: shows alkaline
slant and acidic butt which
red/yellow. Glucose
fermentation only no bubble
no production of H2S.
Indicator: phenol red
Colourless, non lactose
fermenting colonies.
Salmonella: shows slant
alkaline, butt acid which is
red/yellow with bubble and
black precipitate. Glucose
fermentation. Gas
produced and H2S produced.
Indicator: phenol red.

5. Indole Positive: Formation of a pink to positive: pink colored pink Positive: formation of a pink

red color in the reagent layer
on top of the medium within
seconds of adding the reagent
Negative: No color change even
after the addition of
appropriate reagent.
Positive: E. coli shows red
coloration as a result of high
acid production and decrease
6. Methyl Red in the pH of the culture
Negative: has a yellow color
indicating a less acidic medium.
Positive: color change to pink
due the bacteria has butenodial
fermentation and cam split
glucose to acetoin.
7. Voges- Negative: E. coli result shows
Proskauer Test no color change
Indicator: Potassium hydroxide
(KOH) added react with alpha-
naphtol to forms a pink
complex.
Positive: growth with color
change from green to intense
blue along the slant.
8. Citrate
Negative: no growth and no
color change slants remain
green.
Motile: E. coli shows turn as
red color as the result of the
bacteria metabolizing with
added colored dye
9. Motility
tetrazolium.
Non-motile: the result will
see the straight stab line.
ring after the addition of an
appropriate reagent.
Negative: no color changes
even after the addition of an
appropriate reagent.
Positive: Shigella shows red
color as result of high acid
production and decreased in
ph culture.
Negative: yellow color
indicating less acidic
medium.
Indicator: methyl red.
Positive: Color change to
forms pink (red) color
Negative: no color change
Positive: growth with color
change from green to
intense blue along the slant
Negative: no growth and no
color change, slants remains
green
Motile: No straight stab line
Motile: Salmonella shows red
result, spread the inoculum
Non-motile: Shows straight
from the inoculation line.
stab line. Shigella do not has
flagella or flagella damaged
Non-motile: straight stab line.
due to heating, shaking.
to ring red color. Able to
metabolize the amino acid
tryptophan into indole add
pyruvic acid
Negative: stay in base
condition
Indicator: Kovac’s indole
reagent.
Positive: Salmonella shows
red color as a result of high
acid production and decrease
in the pH culture.
Indicator: methyl red
Positive: detect the presence
of acetoin. It will turn to pink
color.
Negative: salmonella show
negative result which no color
change because methyl-red
added has been inoculated
with entrobacteria.
Positive: growth with color
change from green to intense
blue along the slant.
Negative: no growth and no
color change slants remain
green.
References
1. Clinical Microbiology notes.
2. Slide presentation
3. https://microbeonline.com/triple-sugar-iron-agar-tsi-principle-procedure-
and-interpretation/
4. https://microbiologyinfo.com/biochemical-test-and-identification-of-
salmonella-typhi/
5. https://www.austincc.edu/microbugz/eosin_methylene_blue_agar.php
6. https://microbenotes.com/biochemical-test-of-escherichia-coli-e-coli/
7. https://www.youtube.com/watch?app=desktop&v=PzCTn3g4K-Q
8. https://microbiologyinfo.com/biochemical-test-and-identification-of-
salmonella-typhi/