1.

A gram-negative bacillus was recovered from the urine of a child with a history of recurrent urinary
tract infections. The organism was oxidase negative, lactose negative, urease positive, and motile and
phenylalanine positive. The most likely identification of this agent would be:

A. Escherichia coli

B. Klebsiella pneumoniae

C. Proteus mirabilis (it was the phenylalanine that I saw that gave me clue: PPM)

D. Pseudomonas aeruginosa

E. Klebsiella oxytoca

F. None of the above

2. State 2 components of the MacConkey agar that inhibit gram positive bacteria: Crystal violet and bile
salt

2. The bottom layer from the indole test using tryptone broth produces a red color. This tells us that the
microorganism contains what enzyme? _____none_(it is on the upper layer, specifically the xylene
floating on the inoculum, that will have a color change) _____

3. Describe the color and or transparency of the E. coli: O157:H7 colony on Sorbitol MacCnlckey Agar:
Yellow transluscent colonies, because O157:H7 does not utilize sorbitol and so no acid will be produced
that will react with the neutral red to accumulate on the colony to give a pink coloration of the colony.

4. Phenylalanine deamination occurs on the deep or the slant part of an agar? What’s the positive
result? Provide 3 genera positive for Phenylalanine deamination: PPM

- Phenylalanine deamination happens on the slant part of LIA while Phenylalanine

decarboxylation happens in the butt part of LIA.
- Purple coloration
- Proteus, Providencia, and Morganella
5. Shigella sonnei is differentiated from other Shigella sp. By?

A. A positive ONPG

- Shigella sonnet is a group D Shigella and is characterized by its ability to ferment lactose. S. sonnei
is ONPG positive but is a delayed lactose fermenter. The genus Shigella is characterized biochemically by
being negative for citrate, urease, motility, and lysine decarboxylation.

B. Its positive phenylalanine deaminase reaction

C. Its negative oxidase reaction

D. Its ability to demonstrate motility at 22°C

6. Which of the following reactions is typical for Escherichia coli?

A. Beta-hemolytic on sheep blood agar (especially EHEC and O157:H7)

B. Colorless colonies on MacConkey agar (nope kasi lactose fermenter siya so pink ang colony)

C. Colorless colonies on xylose-lysinedesoxycholate agar (lactose fermenter so will produce a yellow to

yellow-red colonies)

D. Green colonies with black centers on Hektoen enteric agar (E. coli does not produce an enzyme that
could lead to formation of Hydrogen Sulfide (H2S)

7. Which one of the following organisms would produce a yellow slant and a yellow butt on TSI agar
after incubating 18 hours?

A. Escherichia coli ( remember, E. coli in TSIA or KIA is A/A g [don’t forget gas production])

B. Proteus mirabilis

C. Salmonella Typhimurium

D. Shigella sonnei

8. A positive indole reaction is characteristic of

A. Escherichia coli

B. Proteus mirabilis
C. Salmonella Choleraesuis

D. Serratia marcescens

9. A positive DNase would be seen with

A. Escherichia coli

B. Klebsiella oxytoca

C. Proteus mirabilis

D. Serratia marcescens (remember this… Serratia marcescens and S. aureus is positive for DNAse test)

10. A 6-year-old female patient was admitted to the hospital following 2 days of severe diarrhea.

Cultures from three consecutive stool samples contained blood and mucus. Patient history revealed a
hamburger lunch at a fast-food restaurant 3 days earlier. Which pathogen is most likely responsible for
the following results?

Growth on:

XLD agar = yellow colonies (lactose fermenter)

HE agar = yellow colonies (lactose fermenter)

Mac agar = light pink and dark pink colonies (lactose fermenter)

Mac with sorbitol agar – few dark pink and many colorless colonies ( O157:H7 does not utilize sorbitol)

A. Salmonella spp.

B. Shigella spp.

C. E. coli O157:H7

D. Yersinia enterocolitica
ENTEROBACTERIAL ID POSITIVE RESULT Reagent added/ test Ph indicator
TESTS indicator
(Write the expected
result) (choose from the (choose from the
choices provided) choices provided)
Citrate Utilisation 1. Prussian blue color/ 2. Bromothymol blue/
heavy growth on slant ***** Bromothymol blue
(X/A1)
MUG test 3. Blue fluorescence of 4. 4- *******
the solution methylumbelliferyl-B-
D-glucuronide (J)
ONPG test 5. Yellow color of the 6. O-nitrophenyl-beta- 6. if you got the other
solution D-galactopyranoside number 6, then you got
(B) this, too. Pampakaba
lang.
Indole test with SIM 7. Pink color on the 8. KOVAC’S/Ehrlich’s
reagent dropped reagent (C/A) *******
Phenylalanine 9. green-colored 10. 10% ferric chloride
Deamination test complex (D1) ********
Lysine deamination 11. red slant (because 12. Bromcresol purple
test using Lysine Iron Lysine deamination ****** (Z)
Agar happens on the slant,
while Decarboxylation
happens in the butt
portion of the LIA)
Urease test 13. magenta/pink color 14. Phenol red (Y)
of the Christensen’s ********
urea agar
Methyl red test 15. Red color of the 16. Methyl red 17. Methyl red (U)
solution
Voges-Proskauer test 18. Red color after the 19. 40% KOH and alpha
addition of potassium naphtol or otherly *******
hydroxide known as Barrit’s (A)
reagent (G)

Catalase test (result for 20. Bubbling/ 21. 10% h202 (H) 21. None (D1)
most Enterbactia) effervescence
Indole test with 22. Pink/red on the 23. Kovac’s / erlich’s 24. None (D1)
tryptone broth interface of the rgt. and xylene (A &/ K or
and inoculum or in the C)
Xylene
H2S production with 25. Black precipitate 26. (indicator present
SIM in the medium) ******

Ferrous salt (M)

Ferric ammonium
citrate
Etc…
Glucose utilization 27. Yellow color of 28. Phenol red (Y)
using TSIA medium *******
Sucrose utilization 29. Can’t be 30. Phenol red (Y)
using KIA determined (no ******
 sucrose in KIA)
MUG test 31. Blue fluorescence 32. Peptone 32. None (D1)
4-methylumbelliferyl-
B-D-glucuronide (J)

Nitrate reduction 33. Red color of the 34. alpha-

solution naphthylamine in ******
(no addition of zinc acetic acid (referred to
dust yet) as reagent A), and
sulfanilic acid in acetic
acid (referred to as
reagent B)
(F)
Oxidase test (result for 35. Blue-purple color 36. tetramethyl-para-
most Enterbactia) phenylenediamine ******
dihydrochloride (the
oxidase reagent) in
dimethyl sulfoxide
(DMSO) (N)
DNAse test 37. 38. BONUS. Will just
Without indicator: ******** give this as bonus than
clearing/fading of the answer D1 since
Green color around the Methyl green is not a
colony after the Ph indicator but cation
addition of HCl binder that binds to
the negatively-charged
With indicator DNA.
(Toluidine Blue O):
development of a pink If the organism that
or red halo around the grows in the medium
colony or the well in produces
the agar. Deoxyribonuclease, it
breaks down DNA into
smaller fragments.
When the DNA is
broken down, it no
longer binds to the
methyl green, and
green color fades and
the colony is
surrounded by a
colorless zone.
 Agar Inhibitory Sulfur Lactose Lactose Carbohydrate Ph
medium components source positive negative component indicato
MacConckey Bile salt and none Pink colony Translucen 1. Lactose 1.
agar crystal violet surrounde t colonies Neutral
d with red
darker
shade of
pink on the
pedge
Salmonella- 2. Brilliant 3. Sodium 4. Red/ 4. Yellow 4. Lactose Neutral
Shigella agar green and citrate/ pink translucent red
Bile salt sodium colonies colonies
thiosulfate
Xylose lysine 5. Bile salts 5. sodium 6. yellow 6. 7. Lactose 8.
deoxycholate thiosulfate Translucen Sucrose Phenol
t colonies Xylose red
on red
medium
Hektoen 9. Bile salts 9. sodium 10. Bright 10. Green 11. Lactose 11.
enteric agar thiosulfate orange/ to blue Sucrose Thymol
salmon blue
pink color
EMB 12. Aniline 12. none 13. Dark Yellow/ Lactose and Aniline
dyes ( eosine pink, translucent sucrose dyes
and violet/ colonies
methylene greenish
blue) sheen
Brilliant 14. Brilliant 14. none 15. pink 15. yellow- Lactose and 16.
green agar green green sucrose Phenol
colony red