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Bacte Lab (Midterm) > 1.1.2 Culture media commonly used in Mirobiology Lab > Flashcards

1.1.2 Culture media commonly used in Mirobiology Lab Flashcards

1
Q

A general purpose, nutrient medium used for the cultivation of microbes supporting growth of a wide range of non-fastidious organisms

A

Nutrient Agar/Broth

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2
Q

What are the ingredients of Nutrient agar?

A

15g/L Pancreatic digest of casein
5 g/L Papaic digest of soyabean meal
5 g/L Sodium chloride
15 g/L Agar
Final pH at 25 C (7.4 +- 0.2)

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3
Q

What is the purpose of pancreatic digest of casein and papaic digest of soyabean meal?

A

Provides amino acid and long chains peptides for the growth of microorganisms

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4
Q

What is the purpose of sodium chloride

A

Maintains the osmotic balance

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5
Q

What is the positive control for nutrient agar?

A

E. coli
Pseudomonas aeruginosa

(Expected growth at 35)

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6
Q

A general purpose, nonselective growth medium that supports the growth of most non-fastidious gram-negative and gram positive bacteria as well as many yeast and molds

Can be use for longer storage, Maintenance purposes, Cultivation and transport

Can be use as base if you want to add blood or antibiotics

A

Trypticase soy agar/Broth

Tryptone soy agar

Tryptic soy agar

Soybean casein digest agar medium

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7
Q

What are the ingredients of TSA

A

300 g/L Beef infusion
17.5 g/L Casein acid hydrolysate
1.5 g/L Starch
17 g/L Agar
Final pH 7.3 +- 0.2

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8
Q

What is the purpose of Beef infusion and Casein acid

A

Main provider of nitrogenous compounds, sulfur, carbon and etc.

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9
Q

One of the ingredients that provides a Protective colloid against toxic substance present in the medium

A

Starch

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10
Q

What happen if there is hydrolysis on starch?

A

Yields a dextrose

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11
Q

What is the purpose of dextrose?

A

Serves as the source of energy

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12
Q

Observation for the growth of FASTIDIOUS organisms and study of hemolytic reaction is done after addition of defibrinated sheep blood

A

BAP

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13
Q

What is the procedure in creating a BAP?

A
  1. Provide blood agar base (TSA)
  2. Autoclave
  3. Cool down in 45 - 50 C
  4. Add defibrinated sheep blood (5%)
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14
Q

How to prepare defibrinated sheep blood?

A

100ml of TSA molten agar = 5 mL blood

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15
Q

Blood agar base is has similar ingredient to what agar?

A

TSA

Beef infusion
Casein acid hydrolysate
starch
agar

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16
Q

3 type of hemolytic pattern

A

Beta hemolysis (Complete)
Alpha hemolysis (Partial)
Gamma hemolysis (None)

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17
Q

What is the positive control for BAP?

A

Streptococcus pneumoniae (Alpha)
Streptococcus pyogenes (Beta)
Staphylococcus epidermidis (Gamma)

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18
Q

FASTWhat is the positive control for BAP?

A

Streptococcus pneumoniae (Alpha)
Streptococcus pyogenes (Beta)
Staphylococcus epidermidis (Gamma)

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19
Q

Observation for the growth of FASTIDIOUS organisms is done after addition of defibrinated sheep blood and heated at 80-100 C.

A

Chocolate agar plate

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20
Q

Procedure of CAP is similar to?

A

BAP but heated at the end (80-100 C)

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21
Q

What is the rationale in heating at 80 - 100 C?

A

To release growth factors NAD (Factor V) and Hemin (Factor X) for nutrients

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22
Q

What bacteria uses CAP?

A

Neisseria and Haemophilus

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23
Q

What is the Positive Controls for CAP?

A

Neisseria gonorrhoeae (5-10% CO2)
Haemophilus influenzae (5 -10% CO2)

In candle jar (Carbon dioxide enrichment environment)

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24
Q

Used as selective media for isolating gram-negative bacteria and differential media for the isolation and differentiation of lactose fermenting and non-lactose fermenting enteric bacilli

A

MacConkey agar

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25
Q

2 function of MacConkey agar

A

Differential (Differentiate Lactose to non Lactose) and Selective (Isolating Gram negative)

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26
Q

What are the ingredients of MacConkey agar?

A

3 g/L Peptones (Meat and casein)
17 g/L Pancreatic digest of gelatin
10 g/L Lactose monohydrate
1.5 g/L Bile salts
5.0 g/L Sodium chloride
0.001 g/L Crystal Violet
0.03 g/L Neutral red
13.5 g/L Agar
pH 25 = 7.1 +- 0.2

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27
Q

What is the purpose of Pancreatic digest of gelatin and peptones?

A

Provides the nutrient such as nitrogenous factors and vitamins

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28
Q

What is the purpose of Lactose monohydrate

A

Fermentable source of carbohydrate for the bacteria

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29
Q

What is the purpose of Bile salts and crystal violet

A

Use to inhibit most gram positive bacteria

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30
Q

What is the purpose of Sodium chloride

A

Maintains the osmotic balance

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31
Q

What is the purpose of Neutral red

A

pH indicator

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32
Q

What is the positive indicator for MacConkey agar?

A

Pink or Red = Acid (<6.8)

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33
Q

What is negative indicator for MacConkey agar?

A

Clear colonies

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34
Q

What is the positive control for MacConkey?

A

E. coli (Pink-Red with bile precipitate)

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35
Q

What is the control for Non-Lactose fermenter

A

Pseudomonas aeruginosa (Can grow in MacConkey)

36
Q

What is the negative control for MacConkey?

A

Gram positive cocci

37
Q

Example of Bacteria grows on MacConkey agar

A

EEK

38
Q

Isolation and differentiation of lactose fermenting and non lactose fermenting enteric bacilli

A

Ethylene methylene blue agar (EMB)

39
Q

2 functions of EMB?

A

Selective and Differential

40
Q

What are the ingredients for EMB?

A

10 g/L Peptic digest of animal tissue
2 g/L Dipotassium phosphate
5 g/L Lactose
5 g/L Sucrose
0.4 g/L Eosin -Y
0.065 g/L Methylene blue
13.5 g/L Agar
pH 7.2 +- 0.2

41
Q

What is the purpose of peptic digest of animal tissue

A

Source of nutrients such as carbon, vitamins and minerals

42
Q

What is the purpose of dipotassium phosphate

A

Use as a buffer

43
Q

What is the purpose of Lactose and Sucrose

A

Source of energy by being the fermentable carbohydrates

44
Q

What is the purpose of Eosin-Y and Methylene blue

A

Use to inhibit the growth of gram positive bacteria

45
Q

What is the positive control for EMB?

A

E.coli (Purple colonies with green metallic sheen)
Enterobacter aerogenes (Purple growth)

46
Q

What is the reason of purple colonies

A

Lactose fermentation

47
Q

What is the negative indicator for EMB?

A

Colorless or pink colonies

48
Q

Used for selective differentiation of staphylococci

A

Mannitol salt agar

49
Q

Mannitol salt agar is what type of agar?

A

Selective and differential agar

50
Q

What are the ingredients of MSA?

A

5.0 g/L Enzymatic Digest of Casein
5.0 g/L Enzymatic Digest of animal Tissue
1.0 g/L Beef extract
10.0 g/L D-Mannitol
75.00 g/L Sodium chloride
0.025 g/L Phenol Red
15 g/L Agar
pH 7.4 +- 0.2

51
Q

What is the purpose of Enzymatic digest of Casein and animal tissue, and Beef extract?

A

Main provider of nutrients such as carbons, nitrogen

52
Q

What is the purpose of D-mannitol

A

Carbohydrate source (Use to differentiate mannitol fermenters)

53
Q

What is the purpose of Sodium chloride

A

Inhibits most bacteria except Staphylococci (Reason for being Selective)

54
Q

What is the purpose of Phenol red

A

pH indicator

55
Q

What are the positive control for MSA?

A

S. aureus (Yellow colonies with yellow halo - Mannitol fermenter)

S epidermidis (pink Colonies with pink medium - Non-mannitol fermenter)

56
Q

What is the negative control for MSA?

A

E. coli due to High concentration of MSA

57
Q

Used as differential and selective medium for the isolation and differentiation of Salmonella and Shigella spp. from other gram-negative enteric bacilli?

A

Hektoen Enteric agar

58
Q

What are the ingredients for HEA?

A

12 Protease peptone
3 Yeast extract
12 Lactose
2 Sucrose
9 Saliclin
9 Bile salt mixture
5 Sodium chloride
5 Sodium thiosulfate
1.5 Ferric ammonium citrate
0.1 Acid - fuchsin
0.065 Bromothymol blue
14 Agar
pH 7.5 +-0.2

59
Q

provide Nutrients such as carbon, nitrogen, and amino acids for growth in HEA

A

Protease peptone

60
Q

What is the Vitamin source for HEA?

A

Yeast extract

61
Q

What is the purpose of Lactose, sucrose, Salicin

A

Cab source or being fermented

62
Q

Inhibits gram positive and HEA

A

Bile salts
acid fuchsin

63
Q

What is the purpose of Sodium chloride

A

Maintain osmotic balance

64
Q

What is the purpose of Ferric ammonium citrate

A

Source of iron, production of hydrogen sulfide (Blackening)

65
Q

What is the purpose of Bromothymol Blue

A

pH indicator

66
Q

What is the Positive indicator for HEA?

A

E. coli (Partial to complete inhibition, Yellow to salmon orange colonies)

Shigella flexneri (Green colonies)

Salmonella typhimurium (Green colonies with black centers)

67
Q

What is the Negative control of HEA?

A

Enterococcus faecalis

68
Q

Selective and differential medium for the isolation of pathogenic enteric bacilli particularly Salmonella and shigella from stool, Food and Clinical Material

A

Salmonella shigella agar

69
Q

What are the ingredients of Salmonella Shigella agar?

A

5.0 Beef extract
2.5 Enzymatic Digest of casein
2.5 Enzymatic digest of animal tissue
10 Lactose
8.5 Bile salts
8.5 Sodium Citrate
8.5 Sodium Thiosulfate
1.0 Ferric Citrate
0.00033 Brilliant Green
0.025 Neutral red
13.5 Agar
pH 7.0 +- 0.2

70
Q

What is the source of nutrients for SSA?

A

Beef extract
Enzymatic digest of Casein
Enzymatic digest of animal tissue

71
Q

What is the carbohydrate source of SSA?

A

Lactose

72
Q

Inhibits gram positive and coliform bacteria, and Swarming motility

A

Bile salts
Sodium citrate
Brilliant green

73
Q

Ingredients for H2S producers of SSA

A

Sodium Citrate
Sodium Thiosulfate

74
Q

pH indicator of SSA

A

Neutral red

75
Q

Unique to SSA from other media?

A

Does not autoclave

76
Q

What is the Positive control for SSA?

A

Salmonella typhimurium (Colorless colonies with Black centers)

Shigella flexneri (Colorless colonies)

77
Q

What is the negative control for SSA

A

Enterococcus faecalis

78
Q

Selective differential medium for isolating and cultivating Vibrio cholerae and other Vibrio species from clinical specimens and other materials

A

TCBS agar

79
Q

Ingredients of TCBS agar

A

5 Yeast extract
5 Enzymatic digest of casein
5 Enzymatic digest of animal tissue
10 Sodium Citrate
10 Sodium Thiosulfate
5 Oxbile
3 Sodium cholate
20 Sucrose
10 Sodium Chloride
1 Ferric Citrate
0.04 Bromothymol blue
0.04 Thymol blue
14 Agar
pH 8.6 +- 0.2

80
Q

TCBS requires autoclave?

A

No

81
Q

Nutrient provider for TCBS agar

A

5 Yeast extract
5 Enzymatic digest of casein
5 Enzymatic digest of animal tissue

82
Q

Selective agents that creates alkaline pH, suppresses coliform (E.coli)

for TCBS agar

A

Sodium citrate
Sodium Thiosulfate
Oxbile
Sodium Cholate

83
Q

Carbohydrate in TCBS

A

Sucrose

84
Q

Act as the indicator for H2S producers in TCBS agar

A

Sodium Thiosulfate
ferric Citrate

85
Q

pH indicators for TCBS

A

Bromothymol blue
Thymol blue

86
Q

What is the positive Control for TCBS?

A

Vibrio alginolytious (Yellow, Sucrose fermenter)
Vibrio parahaemolyticus (Green, non sucrose fermenter)

Bacte Lab (Midterm) (9 decks)

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