Culture Media Preparation Flashcards

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1
Q

– can also be prepared for fungi and bacteria

A

culture media

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2
Q

– virus can be grown in this

A

living cells

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3
Q
  • the survival of microorganisms depend on
A

 available nutrients
 favorable growth environment

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4
Q

nutrient preparations that are used for growing microorganisms

A

media or culture media

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5
Q

microbes that grow and multiply or on a culture medium
* three physical forms of media

A

culture

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6
Q
  • three physical forms of media
A

liquid or broth media
semisolid media
solid media

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7
Q

ex. Nutrient broth, tryptic soy broth, brain-heart infusion broth

A

liquid or broth media

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8
Q

– have agar (ex. Motility medium)

A

semisolid media

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9
Q

have agar (ex. Nutrient agar, blood agar)

A

solid media

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10
Q

complex carbohydrate (polysaccharide) extracted from seaweed

A

agar

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11
Q

 a solidifying agent

A

agar

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12
Q

 solid agar melts about

A

100 degrees c

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13
Q

 liquid agar solidifies at about

A

42 degrees c

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14
Q

 while in the liquefied state, semi solid and solid media can be poured into either a test tube or petri dish/plate

A

agar

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15
Q

type of medium (5)

A

broth
agar slant
agar deep
agar stab
agar plate

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16
Q

autoclaving container for

broth
agar slant
agar deep
agar stab
agar plate

A

culture tube (broth - agar stab)
~200ml culture bottle or EM flask (agar plate)

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17
Q
  • types of Culture Media (2)
A

 chemically defined or synthetic
 Chemically complex or non-synthetic

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18
Q

composed of known amounts of pure chemicals

A

chemically defined or synthetic

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19
Q

 used in culturing autotrophic microorganisms such as algae or non-fastidious heterotrophs

A

chemically defined or synthetic

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20
Q

composed of complex materials that are rich in vitamins and nutrients
 beef extract, yeast extract, and peptones

A

chemically complex or non synthetic

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21
Q
  • culture media preparation
A
  1. calculation and weighing
  2. mixing with water and heating
  3. sterilization
  4. fine-tuning (supplements and pH)
  5. dispensing
  6. packing and storage
  7. quantity control
  8. use of dispensed culture media
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22
Q

temperature higher than 50oC poured in media can cause this

A

condensates

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23
Q

used to prevent buildup of condensation

A

inverted media

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24
Q
  • Sterilization of media and materials for microbial culture
A

 Culture dishes
 Test tubes
 Flasks
 Pipettes
 Transfer loops
 media

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25
Q

using an oven, this kind of sterilization is carried out at 150oC to 180oC for 1 to 4 hours

A

dry heat sterilization

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26
Q

 Can be used for glassware, fixed oils, thermostable powders

A

dry heat sterilization

27
Q

 sterilization using this requires higher temperatures and longer

A

dry heat sterilization

28
Q

 heat transfer is slow, small volumes of oil and thin layers should be used

A

dry heat sterilization

29
Q

in the presence of moisture, microorganisms are destroyed at a lower temperature than in dry heat

A

moist heat sterilizaton

30
Q

 autoclave is using heat + pressure

A

moist heat sterilization

31
Q

 180 degrees at 150 psi
 121.0 °C / 249.8 °F at 15 lbs / square inch (psi) for 15 minutes
 Method of choice when product can withstand treatment (heat stable)

A

dry heat sterilizaton

32
Q
  • Steam sterilization applications
A

 Solutions sealed in containers ampules, vials
 Media
 Bulk Solutions
 Glass wares (ex. tubes with media)
 Surgical Dressing
 Instruments

33
Q
  • Advantages of steam sterilization:
A

 Rapid, Inexpensive, Effective, Large volumes

34
Q
  • Disadvantages of steam sterilization:
A

 Cannot be used for oily preparation (oil base ointment)
 Cannot be used for moisture sensitive preparations

35
Q

where culture medium is sterilized in

A

autoclave

36
Q

varies from original value when subjected to heat sterilization (can increase/decrease_

A

pH of media

37
Q

most bacteria grow best at this pH

A

pH 6.5-7.5

38
Q

– pH should be adjusted after this

A

sterilization

39
Q

very little grow at this pH

A

pH 4

40
Q

may grow in pH as low as 1

A

acidophiles

41
Q
  • must be adjusted after sterilization if the pH of medium strays too far from neutral.
A

pH

42
Q
  • Preparation of media from commercial dehydrated products
A

 Each bottle of dehydrated medium has instructions for preparation on its label
 Nutrient agar 23g/1000ml

43
Q

formula to find the g needed

A

=(how many g (how many mL))/1000mL

44
Q

n order to make 400 mL of medium with a concentration of 15 g/L, how much medium powder would you weigh in grams

A

6g

45
Q
  • In order to make 650 mL of medium with a concentration of 20 g/L, how much medium powder would you weigh in grams?
A

13g

46
Q
  • To fill up 10 agar plates that contains 20 mL of medium with a concentration of 36 g/L, how much medium powder would you weigh in grams?
A

7.2g

47
Q
  • Preparation of Nutrient Broth (NB), a chemically complex medium
A

 Prepare 300 ml of NB
 Weigh appropriate NB powder and dissolve in 300 ml dH2O
 Check initial pH of medium
 Divide contents into 3 flasks
 = 100 ml in each

48
Q
  • Sterilization of media:
A

 1st flask – in autoclave (121⁰C, for 15 mins.)
 2nd flask – in oven (170 ⁰C, for 1 hr.)
 3rd flask – no sterilization

49
Q
  • Preparation of Glucose Minimal Salth Broth (GMSB)
A

 Weigh media powder appropriate for 300 ml.
 Combine media and dH2O in flask.
 Check initial pH of medium
 Divide contents into 3 flasks
 = 100 ml in each
 Sterilize. - autoclave

50
Q

retain coloration and structure of bacteria during staining

A

mordant (gram’s iodine)

51
Q

helps in the removal of primary stains from bacteria

A

decolorizer

52
Q

escherichia coli

shape
gram positive/negative

A

Gram-negative, rod-shaped

53
Q

Bacillus subtilis

gram positive/negative

A

gram positive, rod-shaped

54
Q

Proteus vulgaris

gram/positive negative

A

gram-negative, rod-shaped

55
Q

Serratia marceschens

gram/positive negative

A

rod-shaped, Gram-negative

56
Q

Pseudomonas aeruginosa

gram/positive negative

A

rod-shaped Gram-negative rod

57
Q

Staphylococcus aureus

gram positive/negative

A

Gram-positive, spherical or cocci-shaped

58
Q

dye for positive staining

A

methylene blue

59
Q

is a simple tool used primarily by microbiologists to take and transfer a small sample (inoculum) of a microorganism culture

A

inoculation loop

60
Q

view experiment results on gc

A

+1

61
Q

20 petri dishes will be used that will have 20mL NA/plate

28.0g/1000mL

compute for the grams

A

(28g)(20x20)/1000mL

11.2g

62
Q

Normal Saline Solution abbreviation

A

SNSS

63
Q

20 test tubes will be filled with 10 mL per test tubes

How much NaCl will be used to make 0.9% SNSS soln?

A

wt% = g solutes/ml of soln *100

0.009(250) = g
g= 2.25g