MICROBIOLOGY PRACTICAL

BY; NURHAFIZAH BT HATTA C 111 09 865

1. Lab coat 2. Name tag 3. Practical book with coloured photo 4. Mask 5. Handscoen (1 pair) 6. Ruler 1/group 7. Hand sanitizer 1/group 8. Colour pencil 3/group

ASEP METRIKA (801 - 809) MOH. DIMAS ISMAIL (810 - 819) MUH. AYYUB PRIMADI (820 - 828) MUH. YOGI PRATAMA (829 - 838) ANA ZAHARINA HASYIM (839 - 847) FATIMAH YUNIKARTIKA AKBAR (848 - 857) NURHAFIZAH BT. HATTA (858 - 867) WINDA AGUSTIN (868 - 876) SUHAILAH BINTI MOHD JAMIL(877- 885)

Mycobacterium tuberculosis
Observing demonstration of Gram staining of Sputum Slide Klebsiella pneumoniae

Characteristics;

Cannot be classified as Gram positive or Gram negative Thin straight rods Categorized as Acid-fast bacilli Obligate aerobes Grows well in dark and warm condition Replicating time is about 18hours Non-motile, non-capsulated, non-spore Stain using Ziehl-Nielson / Tanzil method Causative agent for Tuberculosis disease

Grow on egg-based media such as; LOWENSTEIN-JENSEN, KUDOH, OGAWA, BACTEC

Components of Lowenstein-jensen media - White-egg (ducks/ chickens) - Malachite-green ( to inhibit other bacteria growth) - Asparagine, KH2PO4, Magnesium citrate, MgSO4, glycerine - selective antibiotics ( to inhibit other bacteria growth)

Growth on Lowenstein jensen media Dry Rough Yellow Shaped as cauliflower

1. 2. 3. 4.

To confirm the diagnosis To evaluate the treatment response To determine the potential for transmission of the disease
Sputum taken : SPS - S : Sewaktu (arrived) - P : Pagi (the next morning) - S : Sewaktu (arrive on the next day) Upper tract specimens (Throat swab, blood) Lower tract specimens (Sputum, brush bronchus)

NON-CONCENTRATED SPUTUM
Leukocyte present ( + ) Other bacteria present ( + ) Acid Fast bacilli ( Few ; 4-5 cells)

CONCENTRATED SPUTUM
Leukocyte absent ( - ) Other bacteria absent ( - ) Acid Fast bacilli ( Many ; >10 cells)

Ziehl-Nielson method
Carbolfuchsin

Tanzil method
Kinyoun Solution

Acid-alcohol
(3% HCl)

Acid-alcohol (3% HCl)

Methylene blue

Methylene blue

Characteristics;

Gram negative bacteria, Coccobacilli shape Facultative anaerobes Grows well on MacConkey Agar + ferment lactose Indole ( negative) distinguish with E.coli Capsulated, non-motile Can be stained using Gram staining

Has 2 antigens (K:capsular polysaccharides; O:Lipopolysaccharides)

One of the causative agent for Pneumonia disease

Triple sugar iron (TSI) reaction - Observe slant, butt, H2S, Gas production - Yellow (acid), Orange (neutral), Red (alkali) - Black precipitate formed (H2S positive)
Sulphur Indole Motility (SIM) reaction - Sulphur positive (black precipitate formed) - Indole positive ( Red ring formed on the surface); bacteria used trypthophanase enzyme to degrade tryptophan to indole - Motility positive ( Middle line production/cloudy)

Methyl Red test ( Red = positive reaction)

Voges-proskauer test ( Red = positive reaction)

Citrate test ( Blue = positive reaction)

Urea test ( Pink = positive reaction) ; bacteria used urease enzyme to degrade urea to NH4 Sugar test ; Glucose, Lactose, Sucrose, Maltose ( Yellow = positive reaction)

 Aims;

- To compare the bacterial sensitivity towards the types of antibiotic with KirbyBauer method. ~ Do some measurements of clear zone

around antibiotic disc (inhibition zone) then determine sensitivity characteristics from each antibiotic.

Types of bacteria may be found in respiratory tract infections ;

1. 2. 3. 4. 5.

Mycobacterium tuberculosis Streptococcus pneumoniae Klebsiella pneumoniae Escherichia coli Bordetella pertussis

Hospital-acquired infections - infections that gained by staying in hospital, also known as NOSOCOMIAL INFECTIONS
Community-acquired infections - infections gained by exposure to the environment, normally as a pathway of the air-borne transmitted disease

Practical Work Guidelines, Respiratory System 2011 Medical Microbiology 24th Edition, McGraw Hill (LANGE)

QUIZ!!!!
(CODE A)

Give at least 5 characteristics of Mycobacterium tuberculosis

Name 4 types of egg based medium that being used to culture Mycobacterium tuberculosis.

What are the reagents that used in the Tanzil method & write down their functions.

Write down the differences between concentrated & non-concentrated sputum.

Write down at least 5 characteristics of Klebsiella pneumoniae.

Using biochemical tests, how are we going to differentiate between Klebsiella pneumonia & Escherichia coli?

Which colour indicates the positive results of acid fast staining?

What is the name of this test & how to indicate the results.

What are the common pathogenistic microorganisms that can infect the respiratory system?

How the colonies of Mycobacterium tuberculosis look like on the LowensteinJensen medium?

QUIZ!!!!!
(CODE B)

Give at least 5 characteristics of Mycobacterium tuberculosis

Name 4 types of egg based medium that being used to culture Mycobacterium tuberculosis.

a) Write down the characteristics of this colony.

b) What medium is this?

Give the examples of specimens that can be taken from upper & lower respiratory tracts.

What are the reagents that used in the ZiehlNielson method & write down their functions.

State 2 differences of using Ziehl-Nielson & Tanzil method.

When will be the right time to take the specimen (sputum) for the sputum examination?

What are the characteristics of nonconcentrated sputum?

List down at least 5 tests in the biochemical tests & their positive indications.

What are the components of LowensteinJensen medium?