Lecture 4A: Culture Media, Biosafety and Aseptic Technique Flashcards

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1
Q

Culture _____ (sing.)

A

medium

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2
Q

Culture ____ (plu.)

A

media

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3
Q

_____ - is a nurtient-rich liquid or gel designed to support the growth of a target species or group of species.

A

Culture medium

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4
Q

is a ______________________ designed to support the growth of a target species or group of species.

A

nutrient-rich liquid or gel

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5
Q

What does culture medium/media typically contain?

A
  • water
  • energy source, usually a carbohydrate
  • nitrogen source (eg. amino acids, protein sequences such as pepton)
  • mineral salts (MgSO4, NaCl, KH2PO4)
  • special growth factors (if needed)
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6
Q

General composition of medium;

A
  • H-donors and acceptors
  • C-source
  • N-source
  • Inorganic nutrients
  • Trace elements
  • Growth factors
  • Solidifying agent
  • Solvent (usually distilled water)
  • Buffers
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7
Q

General composition of medium;
H-donors and acceptors

A

~1-15 g/L

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8
Q

General composition of medium;
C-source

A

~1-20 g/L

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9
Q

General composition of medium;
N-source

A

~0.2-2 g/L

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10
Q

General composition of medium;
Inorganic nutrients

A

e.g. S, P, (~50mg/L)

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11
Q

General composition of medium;
Trace elements

A

(0.1-1 μg/L

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12
Q

General composition of medium;
Growth factors

A
  • aminoacids, purines, pyrimidines, occasionally 50 mg/L,
  • vitamins occasionally 0.1-1 mg/L
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13
Q

General composition of medium;
Solidifying agent

A
  • eg. Agar (8-20 g/L) (if semi-solid or solid medium)
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14
Q

(3)Types of culture media based on physical state

A
  • Solid medium
  • semisolid medium
  • liquid (broth) medium
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15
Q
  • for isolation of colonies , which give distinct morphology on the surface of the medium
  • commonly employs agar as the solidifying agent, added at 1.5-2% (or 1.5-2.0 g/100ml of medium)
  • agar is a golden-yellow granular powder prepared from seaweeds
A

Solid medium

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16
Q

Solid medium melts at ___ and gels at ____

A

melts at 80°C and gels at <40°C

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17
Q
  • prepared with agar at concentrations of
    0.8% or less.
  • has a soft custard-like consistency and is useful for the cultivation of microaerophilic bacteria or for the determination of bacterial motility.
A

Semisolid medium

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18
Q

Semisolid medium has a soft custard-like consistency and is useful for the cultivation of _______ _______ or for the ____ _ ____ ____.

A

-microaerophilic bacteria
- determination of bacterial motility

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19
Q
  • contain specific amounts of nutrients but do not contain solidifying agent.
  • serves various purposes such as propagation of a large number of organisms, fermentation studies, and various other tests. e.g. sugar fermentation tests, MR-VR broth
A

liquid (broth) medium

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20
Q
  • serves various purposes such as propagation of a large number of organisms, fermentation studies, and various other tests. e.g. __ ____ __, __ __
A
  • sugar fermentation tests, MR-VR broth
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21
Q

Microoganism can be classified depending on ____

A

oxygen requirements

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22
Q

Classifications of microorganisms

A
  • Aerobic
  • Facultative
  • Anaerobic
  • Capnophilic
  • Microaerophilic
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23
Q

requires O2

A
  • aerobic
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24
Q

can grow in the presence or absence of O2

A
  • facultative
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25
Q

growth in the absence of O2

A
  • anaerobic
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26
Q

CO2 stimulates growth

A

capnophilic

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27
Q

require slightly decreased O2 tenstion

A

microaerophilic

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28
Q

(2) Types of culture media based on composition / ingredients

A
  • Complex
  • Defined /synthetic
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29
Q

with 1 or more unknown components, with complex ingredients
- eg. peptone, extracts, hydrolysates; rich medium

A

Complex

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30
Q

examples of complex media

A
  • potato dextrose agar/broth
  • nutrient agar/broth
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31
Q

proportion and identity of all components are known; pure chemical substances; may not be as rich as CM

A

Defined / Synthetic

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32
Q

Types of culture media based on purpose

A
  • General Purpose media
  • Enrichment media
  • Selective media
  • Differential Media
  • Transport media
  • Anaerobic media
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33
Q
  • support the growth of many in a microbial group
  • They do not contain any special additives.
A

General purpose media

34
Q

General Purpose media- ____ ___ ___ any special additives

A

do; not; contain

35
Q

support growth of most cultivatable bacteria

A
  • Nutrient agar (NA)
  • Tryptic soy agar (TSA)
36
Q

Nutrient agar (NA) and tryptic soy agar (TSA) support growth of most cultivatable _____________

A

bacteria

37
Q

supports growth of most cultivatable fungi

A

Potato dextrose agar (PDA)

38
Q

Potato dextrose agar (PDA) supports growth of most cultivatable _________.

A

fungi

39
Q

-provide nutrients and conditions that favor the growth of a particular microbe.

  • promote growth and dominance of a sensitive organism from a mixed culture.
A

Enrichment Media

40
Q

to target microbial group with particular metabolic capacity.

A

Enriching medium

41
Q

Enriching medium to target microbial group with particular ____________ ________________.

A

-metabolic capacity

42
Q

Enriched media are used to grow nutritionally exacting (____________) bacteria.

A

fastidious

43
Q

example of enriched medium

A
  • chocolate agar
  • blood agar
  • Loeffler’s serum slope
44
Q

also known as heated blood agar or lysed blood agar

A

chocolate agar

45
Q

Chocolate agar or ______

A

Blood agar

46
Q

from sheep (usually) created by adding 5-10% (by volume) blood to blood agar base

A

Blood agar

47
Q

first used enrichment to study Chemolithotrophy

A
  • Sergei Winogradsky
48
Q
  • promote the growth of organisms with the use of a favorable carbon
    source
  • and will reduce the growth of other organisms with the use of inhibitory substances
A

Selective media

49
Q

The ________________ is added to medium to eliminate unwanted microbes

A

inhibitory substances

50
Q

example of inhibitory substances

A
  • Antibiotics
  • Dyes
  • Chemicals
  • Alteration of pH
51
Q

Principle of selective media

A

Differential growth suspension

52
Q

state the 2 The principle of selective media’s Differential growth suspension

A
  • is designed to suppress growth of some microorganisms, while allowing the growth of others
  • is agar-based (solid) medium so that individual colonies may be isolated
53
Q

Examples of Selective media

A
  • Sabouraund’s Dextrose agar (SAB)
  • Phenylethyl Alcohol Agar (Phenylethanol agar)
54
Q
  • High concentrations of sugar
    promote yeast growth and inhibit bacterial growth.
  • In addition, low
    pH also inhibits bacteria
A

Sabouraund’s Dextrose agar (SAB)

55
Q
  • Promotes the growth of Gram positive cocci
  • Promotes the growth of Gram positive cocci
A

Phenylethyl Alcohol Agar (Phenylethanol agar)

56
Q

active against fungi, fungi cant grow it a medium with novobiocin

A

Novobiocin

57
Q
  • selective for Neisseria gonorrhoeae
A

Thayer Martin Medium

58
Q

Combination of antibiotics in Thayer Martin medium

A
  • Vancomycin
  • Colistin
  • Nystatin
  • Trimethoprim
59
Q

(antibiotic) which is able to kill most Gram-positive organisms

A

Vancomycin

60
Q

which is added to kill most Gram-negative organisms EXCEPT Neisseria.

A

Colistin

61
Q

(antibiotic) which can kill most FUNGI

A

Nystatin

62
Q

which inhibit gram-negative organisms, especially swarming ‘PROTEUS’

A

Trimethoprim

63
Q

selective for gram negative bacteria

A

Eosin methylene blue

64
Q

is used for isolation of Camplyobacter jejuni from fecal or rectal swab

A

Camplybacter agar

65
Q

Camplybacter agar is used for isolation of _____ ____ from fecal or rectal swab

A

Camplyobacter jejuni (scientific name)

66
Q

contains sucrose and lactose, utilized as fermentable carbohydrates substrates, which encourage the growth of some gram-negative bacteria, especially fecal and non-fecal coliforms.

A

EMB agar

67
Q

EMB agar contains __ and __, utilized as fermentable carbohydrates substrates, which encourage the growth of some gram-negative bacteria, especially fecal and non-fecal coliforms.

A

sucrose and lactose

68
Q

is possible due to the presence of the sugars lactose and sucrose and the ability of certain bacteria to _____________ the
lactose in the medium.

A

Differentiation of enteric bacteria

69
Q

green metallic sheen.

A

Lactose-fermenting gram-negative bacteria

70
Q

brown-pink colonies
- produces less acid.

A

Slow lactose-fermenters

71
Q

colorless or light pink colonies
- increase the pH of the medium by deamination of proteins.

A

Non-lactose fermenters

72
Q

Campylobacter Blood Agar ; supplies all the necessary growth factors

A

Peptone and extracts

73
Q

Campylobacter Blood Agar ; source of B vitamins

A

yeast extracts

74
Q

Campylobacter Blood Agar ; energy source

A

dextrose

75
Q

Campylobacter Blood Agar ; hemin and supplies other necessary
additional growth factors.

A

Sheep blood

76
Q

Campylobacter Blood Agar ; cephalothin, polymyxin B, trimethoprim, amphotericin B, and vancomycin, suppress the growth of the normal microbial flora in fecal specimens, thereby facilitating isolation of C. jejuni.

A

Antimicrobial agents

77
Q

Solidfying agent

A

Agar

78
Q

Solid medium used for Mycobacterium tubercolosis
- contain penicillin, nalidixic acid and malachite grene to inhibit growth of gram-positive and gram-negatuve bacteria, in order to limit growth to mycbacteria species only.

A

Lowenstein-Jenson medium

79
Q

Lowenstein-Jenson medium contains __, __ __ and __ __.

A
  • Pencillin
  • Nalidix acid
  • malachite green
80
Q

allow identification of microorganisms usually through the (visible) physiological reactions unique to those bacteria.

A

Differential Media

81
Q

The most practical media are those that both select for and differentiate common pathogens Usually contain an __________ that changes __________ depending on the metabolites produced by the culture on the medium during incubation

A
  • Indicator
  • color
82
Q
A