revision 2014

PRACTICAL REVISION

Scheme for slides

With background Without background

Brown Blue Red

Organism Gram +ve cocci Organism Gram -ve

Schemes for slides

With background Without background

Gram positive cocci Gram positive bacilli Gram negative

AND MIXTURES

Staphylococci in culture

Staphylococci in culture

Staphylococci in pus

Staphylococci in Pus

Streptococci in culture

Streptococci in culture

Streptococci in Pus

Streptococci in pus

S. pyogenes

Streptococci in pus

Pneumococci in tissue

Pneumococci in tissue

Pneumococci in tissue

Quellung reaction (Streptococcus pneumoniae)

Neisseria in culture

Pathogenic Neisseria in pus

Pathogenic Neisseria in pus

Pathogenic Neisseria in pus

Pathogenic Neisseria in pus

Corynebacterium diphtheriae (Gram stain)

Corynebacterium diphtheriae (M.B. stain)

Corynebacterium diphtheriae (M.B. stain)

Diphtheroids

Bacillus anthracis(Gram stain)

Bacillus anthracis(Gram stain)

B. anthracis(Spore stain)

B. anthracis(Spore stain)

B. anthracis McFadyean reaction(polychrome MB stain)

Clostridium tetani

Clostridium tetani

Mycobacterium tuberculosis (Acid Fast bacilli Z-N stain)

Mycobacterium tuberculosis (Acid Fast bacilli Z-N stain)

Mycobacterium tuberculosis (Acid Fast bacilli Z-N stain)

Mycobacterium leprae (Acid Fast bacilli with modified Z-N stain)

Mycobacterium leprae (Acid Fast bacilli with modified Z-N

stain)

Gram negative bacilli

Klebsiella in Tissues

Vibrio

Vibrio

Vibrio

Gram negative bacilli & Staphylococci

Mixture

Gram negative bacilli & Staphylococci

Mixture

Spirochaetes (Dark-field microscopy)

Spirochaetes (Fontana stain)

Spirochaetes(Fontana stain)

Spirochaetes (fontana stain)

Vincent’s angina

Candida

Candida

Mixture: Candida & Gram –ve bacilli

Mixture: Candida & Gram –ve bacilli

C. albicans(Positive Germ Tube test)

C. albicans

(Positive Germ Tube test)

Nutrient agar

Staphylococcus aureus on nutrient agar( showing golden yellow endopigment )

Staphylococcus aureus on nutrient agar( showing golden yellow endopigment )

Pseudomonas aeruginosa on nutrient agar(showing greenish exopigment)

Proteus on nutrient agar(showing swarming growth)

Proteus on nutrient agar(showing swarming growth)

Mannitol salt agar

Mannitol salt agar

Mannitol salt agar

Blood agar

Types of haemolysis on Blood agar

Alpha-haemolysis on blood agar

Beta-haemolysis on blood agar

No-haemolysis on blood agar

Bacitracin test

Streptococi other than group A

Bacitracin test

Streptococci other than group A (Resistant)

group A streptococci (Sensitive)

Optochin test

Optochin test

Sensitive (Pneumococci)

Resistant (Viridans streptococci)

Optochin test (sensitive) Pneumococci

Optochin test (resistant) Viridans streptococci

Corynebacterium diphtheriae on Blood Tellurite

Corynebacterium diphtheriae on Blood Tellurite

Loffler’s serumfor C. diphtheriae

Chocolate agarfor Neisseria and Haemophilus

Haemophilus or Neisseria on chocolate agar

Haemophilus influenzae on blood agar showing satellatism

(growth around staphylococcus culture)

Haemophilus influenzae (growth only around both factors X & V)

xv

Lowenstein Jensen mediumfor M. tuberculosis

Mycobacterium tuberculosis on Lowenstein Jensen medium

M. tuberculosis on L.J. medium

Mycobacterium Growth Indicator Tube (MGIT) (Positive tubes 1, 5 & 6)

1 2 3 4 5 6

Mycobacterium Growth Indicator Tube (MGIT)

Positive fluorescenceNegative

fluorescence

MacConkey’s medium

Lactose & Non-lactose fermenter on MacConkey’s medium

Lactose fermenter on MacConkey’s medium

Lactose fermenters on MacConkey’s medium

Klebsiella (mucoid & LF) on MacConkey’s medium

Lactose non-fermenter on MacConkey’s medium

Lactose non-fermenter on MacConkey’s medium

LNF on MacConkey’s medium

Salmonella on Salmonella-Shigella (S-S) agar

T.C.B.S. medium

Vibrio cholerae on T.C.B.S.

Blood culture technique

1

2

35 – 10 ml blood

Added to 50 – 100 ml fluid medium

Subculture on Solid medium

Candle Jar ( 5 – 10% CO2)

Candle Jar ( 5 – 10% CO2)

Cooked meat medium for anaerobic cultivation

Anaerobic Jar

Anaerobic Jar

Triple sugar iron( TSI )

TSILactose Fermenter + gase.g. E. coli, Klebsiella

Glucose Fermentere.g.Shigella, Salmonella

Glucose Fermenter + H2Se.g.Salmonella

Non-fermentere.g. Pseudomonas

TSI

Lf +gase.g.E.coli,Kleb

NLe.g.Ps

TSI gg Gl fere.g.Sal,Shig

gl-ferm+H2Se.g.Salmonella

TSI

Non fermenter e.g. Pseudomonas

Lactose ferm + gase.g. E. coli, Klebsiella

Glucose fermentere.g.Shigella, Salmonella

Glucose fermenter + H2Se.g.Salmonella

Glucose Fermenter +H2S e.g.Salmonella

Urease test

-ve +vee.g. Proteus

Catalase Test

e.g Staphylococci

e.g.Streptococci

Catalase Test

e.g Staphylococci

e.g.Streptococci

Coagulase Test

Staphylococcus aureus

Coagulase negative staphylococci, e.g. S. epidermidis

Coagulase test

Staphylococcus aureus Coagulase negative staphylococci, e.g. S. epidermidis

Oxidase test

Oxidase positivee.g. Neisseria, Pseudomonas

Oxidase test

Oxidase positivee.g. Neisseria, Pseudomonas

Oxidase test

+ ve - ve

Oxidase positivee.g. Neisseria, Pseudomonas

Indole test

- vee.g. Klebsiella

+ vee.g. E.coli

MR & VP tests

MR test VP test

-ve +ve

Citrate test

B.R. of E. coli

I M V C + + - - TSI

MR VP Indole Citrate

B.R. of Klebsiella

I M V C - - + + TSI

VP

Nagler’s Reaction(Clostridium perfringens)

Antitoxin No antitoxin

Nagler’s Reaction (Clostridium perfringens)

Analytical Profile Index (API)Biochemical identification system

Analytical Profile Index (API) Biochemical identification system

Analytical Profile Index (API) Biochemical identification system

String Test (Positive in Vibrio cholerae)

String Test (Positive in Vibrio cholerae)

Tube agglutination test(Titre:1/320)

TITRE

1/20 1/40 1/80 1/160 1/320 1/640 1/1280 - ve control

Brucella tube agglutination test showing prozone phenomenon

(Titre:1/320)

TITRE

1/20 1/40 1/80 1/160 1/320 1/640 1/1280 - ve control

Widal Test (Recent infection with S. typhi) (Titre:1/320)

O

HCT

CPA

CPB

C

1/20 1/40 1/80 1/160 1/320 1/640 1/1280

H

H

Widal Test (Old infection with S. typhi)

O

HCT

CPA

CPB

C

1/20 1/40 1/80 1/160 1/320 1/640 1/1280

H

H

Widal Test (Post vaccination)

O

HCT

CPA

CPB

C

1/20 1/40 1/80 1/160 1/320 1/640 1/1280

H

H

Latex Agglutination Test (A: Positive)

Latex agglutination Test (2 & 4: Positive )

Single radial immunodiffusion

Single radial immunodiffusion

Elek’s Test

Toxigenic C. diphtheriae Non-toxigenic C. diphtheriae

Elek’s Test

Immunofluorescence

Immunofluorescence

ELISA plate

ELISA plate

Tuberculin Skin Test (TST)

Positive

Antibiotic sensitivity (Disc diffusion)

12

3

6

54

The organism is sensitive to 4, 5 & 6

Resistant to 1, 2 & 3

Antibiotic sensitivity (Disc diffusion)

Sensitive to………… Resistant to………..

E-Test (MIC determination)

E-Test (MIC determination)

Phage-typing

1 2

3 4 5 6

7 8 9 10

11 12 13 14

15 16 17 18

19 20

The organism is sensitive to phages 4, 5, 6,……..etc.

Gel Electrophoresis DNA Marker 1 2 3 4 5 6

Gel Electrophoresis

Gel Electrophoresis

Western Blot For diagnosis of HIV

Sterile Cotton Swab

Sterile cup

for collection of urine & sputum specimens

Sterile Gloves For surgery & invasive procedures

Non- Sterile Gloves

For skin protection against blood & body fluids

Heavy-Duty Gloves

For environmental cleaning

Standard mask

Used for protection against droplet infections

N95 mask

Used for protection against airborne infections e.g, M.tuberculosis

For protection against splashes of blood or body fluids

Aprons and Gowns

Eye Goggles

For protection against splashes of blood or body fluids

Rota virus (wheel-like appearance)

Rabies virus (bullet-shaped)

Coronavirus (crown-like surface projection)

Tissue culture bottlefor cultivation of viruses

Plaques in a cell-monolayer

Examples of data show Exams

Choose a test

Choose and name the test used to detect microbial antigen.

ab

c d

a: Slide agglutination test

Choose a medium

Choose & name the selective medium used for isolation of Mycobacterium tuberculosis.

(A)

(B)

(C)

(D)

D: Lowenstein-Jensen medium

Infection Control

Infection control The most important PPE when dealing with patients having diarrhoea is:

173

a.

c.

b.

d.

d: Non-sterile gloves

Final Practical

Examination

Final Practical Examination

NO Mobile Phones are allowed in the department (waiting rooms & exam. halls)

Practical exam. : Total 43 marks

Composed of 2 parts:

• Part 1: 13 marks• Part 2: 30 marks

Part 1: Demonstration: Data show (total 13 Marks)

Formed of: 3 different photos (3 marks/ 1 mark each) ANY photo included in the modified revision CD (version 2014) 2 Choose a test 2 Choose a medium 1 Infection control 5 Photos

Part 2: Demonstrations (total 30 Marks)Formed of 15 different stations (2 marks each) In case of a) microscopical slides: mention organism + stain b) plain culture media: mention name + use c) inoculated culture media: mention name + the organism/effect d) biochemical reactions: mention result + organism e) serological test: mention name + result/titer f) others: mention name + reading (e.g. phage-typing)

Name: Date: No: Microbiology Practical Exam

• 1)………………………………………………………………………………………………………………………..• 2)………………………………………………………………………………………………………………………..• 3)………………………………………………………………………………………………………………………..• 4)………………………………………………………………………………………………………………………..• 5)………………………………………………………………………………………………………………………..• 6)………………………………………………………………………………………………………………………..• 7)………………………………………………………………………………………………………………………..• 8)………………………………………………………………………………………………………………………..• 9)………………………………………………………………………………………………………………………..• 10)………………………………………………………………………………………………………………………• 11)………………………………………………………………………………………………………………………• 12)………………………………………………………………………………………………………………………• 13)………………………………………………………………………………………………………………………• 14)………………………………………………………………………………………………………………………• 15)……………………………………………………………………………………………………………………… ………………………………………………………………………………………………………………………….. 30 Marks (2 marks each)