4th test - MYCOBACTERIUM, Nocardia, Actinomyces, Trueperella, Actinobaculum, Dermatophilus, Corynebacterium, Rhodococcus, Reinibacterium

Question 1

Impact of Mycobacteria worldwide

Answer 1

• Human Tuberculosis
• Human Leoprosy

Question 2

Impact of Mycobacteria worldwide
- Human Tuberculosis

Answer 2

• 1/4 of the human population is infected with M.TUBERCULOSIS
• *- 1,6 million people die in TB each year**
• 10 million new cases (2017)

Question 3

Human leprosy

Answer 3

• 12 million infected patients
• 230-300.000 (2012) new cases
• *- Africa, Asia, America**

Question 4

Who discovered M.Tuberculosis

Answer 4

Robert KOCH

Question 5

Mycobacterium
1. HABITAT

Answer 5

Infected host
Environment

Question 6

Mycobacterium
2. Morphology

Answer 6

Rods, Coccoid
Cellwall structure
Species differences

Question 7

Mycobacterium
3. Staining

Answer 7

GRAM + (theoretically)
Fluorescence dyes (AURAMIN staining)
Ziehl-Neelsen positive (RED)

Question 9

Ziehl-Neelsen staining

Steps

Answer 9

1.

Strong carbol fuchsin - about 10 minutes (heat treatment 3x till steaming)

(phenol)

2.

wash

3.

Sulphuricacid (5%) and alcohol (96%) treatment - several drops

4.

wash

5.

counter staining with methylene blue - 1-2 minutes

6.

wash

Question 10

Ziehl-Neelsen staining

Ziehl-neelsen POSITIVE

Colour

Answer 10

RED

Question 11

Ziehl-Neelsen NEGATIVE

with sulphuricacid (5%) and alcohol (96%) treatment

Colour

Answer 11

Colourless

Question 12

Zeel-Neelsen staining

counter staining with methylene blue

Color

Answer 12

Zeel-Neelsen POSITIVE = RED

Zeel-Neelsen NEGATIVE = BLUE

Question 13

MYCOBACTERIUM

4. Culture

Answer 13

1• obligate aerobic

2• broth:

• Henley, Sauton (cord)

3• solid:

• egg-enriched media (Kertai, Dorset, Petragnani, Löwenstein-Jensen)
• glycerol potato

4• selective:

• Petragnani
• Acid (alkaline) treatment (5% oxalic acid or 2-4% NaOH)

5• incubation time, growth characteristics (species differences)

Question 14

Cultural differences

M. tuberculosis

Incubation time

Answer 14

3-6 weeks

Question 15

Cultural differences

M. tuberculosis

Shape

Answer 15

slender rods

Question 16

M. tuberculosis

Cultural differences

Solid medium

Answer 16

dry colonies

Question 17

Cultural differences

M. tuberculosis

Broth

Answer 17

thin film

Question 18

Cultural differences

M. bovis

Incubation time

Answer 18

4-8 weeks

Question 19

Cultural differences

M. bovis

Shape

Answer 19

Short rods

Question 20

Cultural differences

M. bovis

Solid medium

Answer 20

wet colonies

Question 21

Cultural differences

M. bovis

Broth

Answer 21

Islands

Question 22

Cultural differences

M. avium subsp. avium

Incubation time

Answer 22

2-3 weeks

Question 23

Cultural differences

M. avium subsp. avium

Shape

Answer 23

Polymorph

Question 24

Cultural differences

Solid Medium

Answer 24

Cream like colonies

Question 25

Cultural differences

M. avium subsp. avium

Broth

Answer 25

Thick-film, sediment

Question 26

Cultural differences

M. avium
subsp. paratuberculosis

Incubation time

Answer 26

6-12 weeks

Question 27

Cultural differences

M. avium
subsp. paratuberculosis

Shape

Answer 27

Short rods

Question 28

Cultural differences

M. avium
subsp. paratuberculosis

Solid Medium

Answer 28

Wet Colonies

Question 29

Cultural differences

M. avium
subsp. paratuberculosis

Broth

Answer 29

Thin film

Question 30

M. tuberculosis - COLONIES

Answer 30

(cauliflower like colonies)

Question 31

Biochemistry of Mycobacterium

• catalase +, oxidase –

Question 32

Mycobacterium Antigens

Answer 32

• glycolipids, lipoproteins
• shared antigens, immuno-electrophoresis
• Intradermal tuberculin test – PPD (purified protein derivate)