Culture & Culture media

Question 1

Acc. to Consistency
1. do not contain any amount of agar
- example (3)
- indication of growth
2. semi solid medium contains ___ agar (clot like consistency)
- agar is made of (3)
- example (1)
- indole positive (red ring)
- sulfide positive (black)
3. solid contains ___ agar
- example (3)
- melting temp
- solidifying temp
- cooling temp (temp distributed to Petri dishes)
- volume on plate

Answer 1

1. Liquid Medium (Broth)
   - brain heart infusion, trypticase soy broth, thioglycollate
   - turbidity
2. 0.5% - 1.0% agar
   - D-galactose, D-glucoronic acid, 3,6-anhydro-L-galactose
   - Sulfide Indole Motility (SIM) Media
   - e.coli
   - p.mirabilis
3. 2-3% agar
   - Triple Sugar Iron Agar, MacConkey Agar, Blood Agar
   - >95C
   - 40-50C
   - 55-60C
   - 20-25mL

Question 2

Acc. to Composition
1. all substances are known to the uses
- aka
- used for ______
- isolation of _____
2. some unknown substances
- isolation of ______
- example (3)
3. isolation of obligate intracellular bacteria (Ricketssia, Chaplamydia)
- fibroblast cell for vaccine production
- fibroblast cell from mouse
to culture Riccketsia
- (immortal cell=cervical cell

Answer 2

1. Synthetic
   - Defined Medium
   - research purposes
   - cyanobacteria & chemoorganotrophs
2. Non-synthetic
   - medically significant bacteria (fastidious)
   - Nutrient broth, TSB, MacConkey agar
3. Tissue Culture Medium
   - W138
   - McCoy cells
   - HELA 229 cells

Question 3

Acc to. Dispense/ Distribution
1. dispensed in Petri dish
2. tube media
- butt = ____
- slant = ____

Answer 3

1. Plated media
   2.
   - anaerobic
   - aerobic

Question 4

Acc. To Use

1. support the growth of most nonfastidious bacteria (with meat and soybean extracts)
   - examples (3)
2. contains specific nutrients w/o additional supplements
   - alkaline peptone water (w/ thiosulfate salt)
   - selenite F (w/ selenite)
   - tetrathionate
   - GN Broth (w/ sodium citrate & sodium desoxylate)
   - LIM broth

Answer 4

1. Simple/ Supportive/ General Isolation/ General Purpose/ Basal
   Culture
   - Nutrient Agar, Trypticase Soy Agar, Nutrient Broth
2. Enrichment Medium (Liquid)
   - Vibrio
   - Salmonella
   - Salmonella & Proteus
   - Salmonella & Shigella
   - Group B streptococcus

Question 5

Acc. To Use

_____: w/ additional supplements

1. allows differentiation of hemolysis/hemolytic pattern
   - for most ______
   - ___ w/ ___
2. enriched medium for Haemophilus & Neisseria
   - supplies ___ & ___ factors
   - incubate in ____

Answer 5

Enriched Medium

1. Blood Agar
   - fastidious
   - tryptic soy agar w/ 5% sheep blood
2. Chocolate agar
   - X & V factors
   - increased CO2

Question 6

Acc. To Use

_____: distinguishes group of organisms based on cultural characteristics

1. allows visualization of _______
2. lactose fermenters from non lactose fermenters
   - lactose fermenter color
   - non lactose fermenter color
3. hemolytic patterns
   - _____ vs _____
   - very lysis color (beta)
   - partial lysis color (alpha)
   - no hemolysis color (gamma)
4. inhibits gram positive (lactose vs non lactose
   - inhibitors (2)
5. differentiate enteric gram neg pathogens
   - differentiates ____ & ____
   - EMB w/ clear colonies
   - EMB w/ black colonies

Answer 6

Differential Medium

1. metabolic differences
2. MacConkey
   - pink
   - colorless
3. BAP
   - Streptococcus vs Staphylococcus
   - clear
   - greenish
   - none
4. EMB
   - EOSIN Y & Methylene blue
5. Hektoen Enteric Agar
   - Salmonella & Shigella
   - Shigella flexeri
   - Salmonella typhi

Question 7

_______: selects the growth of a particular organism

1. incorporated w/ _____
2. Gentamycin blood agar
3. Bacitracin Chocolate agar
4. Blood agar plate w/ ampicillin
5. Phenyl ethyl alcohol
   - this inhibits the growth of gram neg
6. GN Broth
7. Thayer Martin Agar
8. HEA lactose
   - positive
   - negative color (organisms)
9. CNA
   - disrupts cell membrane of gram neg
   - blocks DNA replication

Answer 7

Selective Medium

1. antibiotics
2. Streptococci
3. Hemophilus
4. Aeromonas
5. gram positive
   - 5% sheep blood
6. Salmonella & Shigella
7. Neisseria gonorrhea
   8.
   - yellow
   - green/blue
   9.
   - colistin
   - nalidixic

Question 8

_____: isolate bacteria w/ specific growth requirement

1. for mycobacteria
   - contains ____,_____,____
   - which one out of 3 inhibits most of bacteria
2. for vibrio

Answer 8

Special media

1. LJ medium
   - arginine, glycerine, malachite green
   - malachite green
2. TCSB

Question 9

INHIBITORY SUBSTANCES

1. inhibitory to gram positive
2. inhibitory to gram negative
3. inhibitory to swarming
4. preferred blood for BAP order
5. human blood is not preferred because it contains nonspecific inhibitors:
   - inhibit grown of beta hemolytic strep
   - alter type of hemolysis
6. blood is added at what temp

Answer 9

1. Crystal/gentian violet, Basic Fuchsin, Bile salt (sodium desoxycholate)
2. Potassium tellurite, Sodium azide
3. alcohol, chloral hydrate
4. Sheep > Horse > Rabbit
   5.
   - citrate
   - dextrose
5. 40-50C