Manipal spotters

1. Streptococcus
this
medium.
4.
Gi
v
e
t
w
o
examples
of
fastidious
bacteria
growing
on
3.
Mention
t
w
o
uses
of
this
medium.
b)
Demonstration
of
hemolytic
property
a)
Enhances
growth
of
fastidious
bacteria
3.Two
uses:
this?
2.
Based
on
nutritive
property,
what
ki
n
d
of
medium
is
2.
Enriched
medium
1.
Blood
agar
1.Identify
t
h
e
culture
medium.
1.
BLOOD
AGAR-
ANSWERS
1.
BLOOD
AGAR
QUESTIONS
pneumoniae
4.
StreptococcUS
PyOgenes

2. 4.
Blood
agar,
Loeffler's
serum
slope.
type.
4.
Give
examples
of
t
w
o
other
culture
media
of
similar
providing
3.
Mention
t
w
o
uses
of
this
medium.
Neisseria
spp.
a)
Enhances
growth
of
fastidious
bacteria
like
3.
Two
uses:
this?
2.
2.
Enriched
medium.
identification
property.
1.Identify
t
h
e
culture
Dark-brOWn
colored,
1.
Chocolate
agar.
2.
CHOCOLATE
AGAR
QUESTIONS
2.
CHCOLATE
AGAR
ANSWERS
X
and
V
factors.
b)
Promotes
growth
of
Haemophilus
spp.
by
Based
on
nutritive
property,
what
kind
of
medium
is
medium.
Mention
t
h
e
opaque,
solid
medium

3. 3. MACCONKEY'S AGAR-QUESTIONS
1. Identify the culture medium.
2. Based on function, what kind ofmedium is this?
3. Mention two uses of this medium.
4.Give two examples for lactose fermenting and non
lactose fermenting Gram negative bacilli of clinical
importance.
3.
MACCONKEY'SAGAR ANSWERS
1.MacConkey's agar
2.Differential medium/Indicator medium
3.Two uses:
a)
Differentiation of lactose and non-lactose
fermenting bacteria
b)Inhibition of swarming growth of Proteus spp.
4.Lactose fermenting: Escherichia coli, Klebsiella
pneumoniae
Non-lactose fermenting: Salmonella Typhi,
Pseudomonas aeruginosa

4. 4. TCBS AGAR-QUESTIONS
1. Identify the culture medium with identification
properties in support.
2. What type ofculture medium is this?
3. What is the use of this medium?
4. Give examples oftwo other mediaused for the
same purpose.
4. TCBS AGAR ANSWERS
1. Thiosulphate citrate bile salt sucrose agar (TCBS
agar).
It isa translucent, green coloured, solid medium.
2. Selective medium
3. Isolation of Vibrio cholerae from stool
specimen.
4.Alkaline bile salt agar
Monsur's gelatin taurocholate trypticase tellurite
agar (GTTA)

5. 5. ROBERTSON'S COOKED MEAT BROTH
-QUESTIONS
1. Identify the medium.
2.What kind ofmedium is this?
3.What is the use ofthis medium?
4.Give examples oftwo bacteria growing in this
medium.
5.ROBERTSON'S COOKED MEAT ROTH
ANSWERS
1.Robertson's cooked meat broth
2. It is a liquid anaerobic culture medium.
3. It is used to grow anaerobic bacteria.
4. Clostridium perfringens, Bacteroidesfragilis

6. 6.THIOGLYCOLLATE BROTH-QUESTIONS
1. ldentify the medium.
2. Name one use ofthis medium.
6. THIOGLYCOLLATE BROTH- ANSWERS
1. Thioglycollate broth.
2. It is used to grow anaerobes.

7. 7. GASPAK JAR-QUESTIONS
1. Identify the instrument.
2. Name two bacterial pathogens that can be
cultivated using this.
3. Name two culture media uscd for similar
purpose.
7. GASPAK JAR-ANSWERS
1.Gaspak jar
2. Clostridium tetani, Bacteroides fragilis
3. Robertson's cooked meat broth, Neomycin
blood agar

8. 8.INOCULATION
LOOP-
ANSWERS
8.INOCULATION
LOOP-
QUESTIONS
ldentify
the
item
1.
1.
2.
Mention
2.
It
is
used
to
inoculate
clinical
specimens
on
to
solid
culture
media.
3.
How
is
it
sterilized?
3.
Flame
sterilization
(till
the
loop
becomes
red
hot)
Inoculation
loop.
the
uses.

9. 9.STERILECOTTON SWAB-QUESTIONS
1. Identify the spotter.
2. Namne any two other types ofswabs used for
specimen collection.
3. How are the cotton swabs sterilized?
9. STERILE COTTON SWAB - ANSWERS
1.
2.
3.
Sterile cotton Swab.
Dacron swab, Rayon swab.
InHot air oven - 160°C for 2 hrs.

10. 10. EMBRYONATED EGG-QUESTIONS
1. ldentify the spotter.
2.What is it's application in diagnostic
microbiology?
3. Enumerate various routes of inoculation
giving example for each.
141 220101104
10. EMBRYONATED EGG- ANSWERS
1.Embryonated egg
2. a) Cultivation of rickettsiae and viruses
b) Preparation of viral vaccines
3. Various routes of inoculation.
a) Chorioallantoic membrane: Vaccinia
virus, Variola virus, Herpes simplex
virus
b) Allantoic cavity: Influenza virus,
Parainfluenza virus
c) Amnioticcavity: Influenza virus
d) Yolk sac: Rabies virus, Rickettsiae,
Chlamydia

11. 11.SYNCYTIUM FORMATION IN CELL
CULTURE -QUESTIONS
1.What are the different kinds of cell cultures?
2.Mention four methods of detecting viral
growth in cell culture.
3.Mention two important uses of cell cultures.
4.Mention two methods for removal of bacteria
from contaminated clinical samples before
inoculating into cell cultures.
1Q. What are the different kinds of cell cultures?
1A. Different kinds of cellculture are
a. Primary cellculture- Monkey kidney cell line
b.Secondary or diploid cell culture- Human
fibroblast cell line
c. Continuous cell lines- Hep-2 cell lines
20. Mention four methods of detecting viral
growth in cell culture?
2A. Four methods are
a. Demonstration ofcytopathic effect in cell lines
b.Viral interference
Detection of viral antigens by direct
immunofluorescence assay
d. Viralgene detection by PCR
30.Mention two important uses ofcell
cultures.
3A. Two important uses ofcell culture
a. Isolation of viruses
b.Growth of viruses for vaccine
production
40. Mention two methods ofremoval of
bacteria from contaminated clinical samples
before inoculating into cellcultures.

12. 4A.Two methods of removal of bacteria fronm
contaminated clinical samples before
inoculating into cell culture
a. Filtration
Addition of antibiotics
Spotter No. 11
Syncytium format:on in cellculkuve
12/3/2023

13. 12. SAB0URAUD'S DEXTROSE AGAR
WITHMOULD GROWTH-QUESTIONS
1.Identify the culture medium.
2.Name two organisms which can be grown
on this medium.
12.
SAB0URAUD'SDEXTROSE AGAR
WITH MOULD GROWTH- ANSWERS
1.
Sabouraud's dextrose agar
2. a) Rhizopus species
b) Aspergillus species

14. 13. AUTOCLAVE -QUESTIONS
1.Identify the spotter.
2.What is the clinical significance ofthis
equipment in hospital setting?
3. What are the quality control methods to be
followedtoconfim the proper working of
the equipment?
13. AUTOCLAVE - ANSWERS
1. Autoclave
2. Clinical significance
" Sterilization of surgical instruments,
implanted medical devices, surgical drapes
and linen.
" Decontamination of biomedical waste
before disposal.
3.Quality control methods
"Physical control - digital displays on the
equipment displaying temperature, time
and pressure
" Biological indicators
Geobacillus stearothermophilus
spores of
" Chemical indicators - eg: Autoclave tapes

15. 14. MEMBRANE FILTER-QUESTIONS
1. Identify the spotter.
2. What are the two important advantages?
3. Give examples for two other types of filters.
14. MEMBRANE FILTER ANSWERS
1.Membrane Filter
2. Advantages
" Water purification and analysis
Preparation ofsolutions for parenteral use
3. Examples for filters
Candle filters
EPA flters

16. 15. BIOLOGICAL INDICATOR
QUESTIONS
1.Identify the spotter.
2.What is principle of this test?
3.Name two bacteria used as indicators in this
test.
20101029NDA V
PRASAD
149 2201013
KRISHNA VENI A
15.BIOLOGICAL INDICATOR-
ANSWERS
1. Biological indicator
2. Principle
" Biological indicator tubes containing
thermophilicbacterial spores are kept in
autoclaveduringsterilizationcycle.
"After the completion ofsterilization, the
tubesareincubatedat37°C.
" No color change indicates adequate
sterilizationwhichhaskilled thespores.
to yellow indicates
change
" Color
inadequatesterilizationwhich has failed to
killthespores.
3.Two bacteria usedasindicators in this test.
Geobacillusstearothermophilus
"Bacillus atrophaeus

17. 3.The
purpose
of
this
test
is
to
demonstrate
t
h
e
susceptibility
of
bacteria
to
different
antibiotics.
3.
What
is
t
h
e
purpose
of
doing
this
test?
2.
Muller
Hinton
agar.
2.
What
is
t
h
e
medium
used
for
this
test?
1.
Identify
this
test.
disk
diffusion
method.
1.
Antibiotic
sensitivity
t8
Bauer
SHOWING
DIFFUSION
TEST:
QUESTIONS
KIRBY
16.
ANTIBIOTIC
DIFFUSION
TEST:
ANSWERS
SHOWING
KIRBY
16.
ANTIBIOTIC
SENSITIVITY
PLATE
BAUER
DISK
SENSITIVITY
PLATE
BAUER
DISK

18. 16. ANTIBIOTIC SENSITIVITY PLATE
SHOWING KIRBY BAUER
DIFFUSION TEST: ANSWERS
DISK
1. Antibiotic sensitivity testing by Kirby Bauer
disk diffusion method.
2. Muller Hinton agar.
3. The purpose of this test is to demonstrate the
susceptibility ofbacteria to different antibiotics.

19. 17. TUBE
AGGLUTINATION TEST
(WIDAL TEST)-QUESTIONS
A male patient aged 30 years, suffering from fever of
14 days duration was admitted to the hospital. On
examination he had step ladder type of fever, rose
spots and
splenomegaly. Appropriate test was done
and results are provided herewith:
Result:S. Typhi"0" antibodytitre -640
S. Typhi "H" antibody titre - 640
S.ParatyphiH" antibody titre- 20
1. Identify theserologicaltest.
2. In which clínical condition would you like to do this
test?
3. Interpretthetest resultandgive youropinion.
4.Give an exampleofanother serological test based on
thesame principleandclinical
limplicationofthetest.
17. WIDAL TEST- ANSWERS
1. Widal test-done by tube agglutination method.
2. Enteric fever.
3.S. Typhi"0" antibodytitre-640and S. Typhi "H"
antibody titre- 640.
So the test is positive for S. Typhi "0" antibody
and S. Typhi "H" antibody and the above titre is
suggestive of Typhoidfever.
4. Standard agglutination test which is used for
serological diagnosis of Brucellosis.

20. 18. ELISA- QUESTIONS
1.Identifythe serologicaltest.
2. Whatis theprinciple ofthistest?
3.Mentiontheclinicalapplications ofthis test.
18. ELISA- ANSWERS
1. The test is Enzyme linked Immunosorbentassay.
2. It is an immunoassay that detects eitherantigen
or antibodies in the specimen, by using enzyme
substratechromogen systemfor detection.
3. Detection of antibodiesto HIV.
Detection of HepatitisBsurface antigen.

21. 19. IMMUNODOT (HIV TRIDOT)
TEST - QUESTIONS
1. Identifytheserological test.
2. In which clinicalcondition would you like to do
this test?
3. Interpret the test result and give your opinion.
19. IMMUNODOT (HIV TRID0T)
TEST - ANSWWERS
1. Immunodot assay.
2. This test is done to detect antibodies to HIV type
1and HIV type 2.
3. The observed test is reactive for HIV Iantibody.
The test result needs to be confirmed by other two
serological tests ofdifferentprinciple like ELISA
and immunochromatography.

22. 20. IMMUNOCHROMATOGRAPHY
TEST (NSI
ANTIGEN TEST):
QUESTIONS
1. What type ofAg-Abreaction is this testbased
on?
2. Interpret the test result.
3. Mention two applicationsoftheabove test format.
DENGU
NS1 Aetonn
ibigen
CONTROL LINE
TEST LINE
SAMPLE WINDOW
20. IMMUNOCHROMATOGRAPHY
TEST: ANSWERS
1. This test is based on Immunochromatography.
2. The test result is positive for NS1 antigen.
3. Detection of antibodies to HIV
Detection ofHepatitis B surface antigen.