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General Microbiology > Isolation and Cultivation of Microorganism > Flashcards

Isolation and Cultivation of Microorganism Flashcards

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1
Q

a culture which contains a single
species of microorganism

A

pure culture

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2
Q

pure culture is also known as

A

axenic culture

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3
Q

a population of cells arising from a single cell

A

pure culture

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4
Q

increasing the population of
microorganisms by providing their
nutritional and physical
requirements

A

cultivation

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5
Q

extracellular substances which
provide the cell with materials for
building protoplasm and for
energy generation

A

nutrients

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6
Q

any nutrient material for growth
and cultivation of microorganisms
in the laboratory

A

culture medium

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7
Q

uses of culture media (3)

A

for growth and maintenance of
microbial cultures

  • to favor the production of
    particular compounds
  • to study microbial action on
    some constituents of the
    medium
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8
Q

types of culture media is categorized according to (3)

A

physical state
chemical composition
function, purpose, application

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9
Q

type of culture media according to physical state (3)

A

liquid
semi-solid
solid

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10
Q

media with no solidifying
agent

A

liquid (broth)

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11
Q

media wit h 0.1-0.5% solidifying agent

A

semi-solid

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12
Q

media with solidifying agent: agar or – 1.5 – 2.0% solidifying agent

A

solid media

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13
Q

type of culture media according to chemical composition

A

synthetic
complex

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14
Q

media where all components are
chemically
defined

A

synthetic

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15
Q

media not all components
are chemically defined

A

complex

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16
Q

example of complex medi

A

potato infusion
beef extract
yeast extract

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17
Q

types of culture media according to principal function, purpose, application

A

general
differential
selective
enrichment
assay

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18
Q

media that can support most or almost all types
of species

A

general purpose media

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19
Q

example of general purpose media

A

nutrient agar (NA)

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20
Q

media that distinguishes one type of bacteria from
another

A

differential media

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21
Q

media * with special reagents like pH indicators
or dyes

A

differential media

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22
Q

example of differential media

A

Eosin Methylene Blue Agar (EMBA)

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23
Q

media that allows the growth of a specific type of
microorganism only

A

selective media

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24
Q

media with selective agents

A

selective media

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25
Q

example of selective agents placed on selective media (3)

A

salts
dyes
antibiotics

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26
Q

example of selective media

A

Bacillus Cereus Agar (BCA)

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27
Q

media used to increase the number of
microorganisms with unusual
physiological characteristics

A

enrichment

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28
Q

media with special nutrients

A

enrichment media

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29
Q

example of special nutrients

A

blood

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30
Q

example of enrichment media

A

blood agar

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31
Q

is a variant of blood agar that contains red blood cells that have been lysed by heating to 80°C. The lysates provide growth factors like nicotinamide adenine dinucleotide (NAD) and hemin, which are needed for the growth of some bacteria. The heat also inactivates enzymes that could degrade NAD

A

chocolate agar

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32
Q

to grow fastidious organisms and to differentiate bacteria based on their hemolytic properties.

A

blood agar

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33
Q

is a medium used with supplements for the selective detection of Bacillus cereus in food

A

bacillus cereus agar

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34
Q

is a brand of chromogenic culture media used in microbiology to identify microorganisms. The color of the colonies formed indicates which substances in the media interacted with specific metabolic pathways in the microorganism.

A

chromagar

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35
Q

to identify organisms that are capable of producing the enzyme lipase

A

spirit blue agar

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36
Q

is the primary plating medium universally used for the selective isolation of vibrios causing cholera, diarrhea and food poisoning.

A

Thiosulfate Citrate Bile Sucrose (TCBS) agar

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37
Q

It is used in testing the quality of water, especially in determining if the water is contaminated by harmful microorganisms.

A

eosin methylene blue agar

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38
Q

media of prescribed composition used for assay
of vitamins, amino acids and antibiotics

A

assay

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39
Q

media used to determine qualitative/ quantitative
production of such a compound by an
organism

A

assay

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40
Q

example of assay media

A

fermentation media
triple sugar iron medium
antibiotic sensitivty testing

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41
Q

a tubed differential medium used in determining carbohydrate fermentation and H2S production

A

triple sugar iron medium

42
Q

isolation techniques (5)

A

plating
enrichment culture
serial dilution
single-cell isolation
membrane filter

43
Q

plating techniques

A

streak plating
spread plating
pour plating

44
Q

a macroscopically visible (surface
or subsurface) growth or cluster of
microorganisms on a solid medium

A

colony

45
Q

how much inoculum is used in pour plating

A

1 ml or 1000ul

46
Q

how much inoculum is used for spread plating

A

0.1 ml or 100 ul

47
Q

what is used for spread plating to spread the inoculum

A

glass spreader

48
Q

subsurface colonies are smaller/larger than surface colonies

A

smaller

49
Q

isolation of specific types of
microorganisms by a combination of
nutrient and physical conditions

A

enrichment culture

50
Q

used for the isolation of unusual
physiological types of microorganisms
which are present in small numbers and
which grow slowly

A

enrichment culture

51
Q

used if the desired microorganism is present at a
higher level than any other microorganism

A

serial dilution

52
Q

outcome is 10-fold reduction of cells/cfus in every
transfer.

A

serial dilution

53
Q

how many ml of broth in serial dilution for each tube

A

9 ml

54
Q

uses a micropipette or a
microprobe to physically pick a
single cell and transfer it on an
agar medium

A

single-cell isolation technique

55
Q

for samples with low population

A

membrane filter technique

56
Q

uses a sterile membrane filter
having a pore size that retains
microorganism

A

membrane filter technique

57
Q

steps in preparing pure cultures

A

isolation
transfer
verify the purity
make stock cultures

58
Q
  • to retain the viability of the
    stock culture for a long period of time
    while maintaining its purity and trait
    of being “true-to-type”
A

culture preservation

59
Q

culture preservation methods (5)

A

periodic transfer to fresh media
overlaying cultures with mineral oil
freeze-drying (lyophilization)
freezing with liquid nitrogen
drying

60
Q

considerations for the tranfer of fresh media

A

time interval of transfers
proper medium
proper storage temperature

61
Q

aim to limit the availability of O2

A

overlaying of mineral oil to cultures

62
Q

advantages of putting mineral oil over cultures

A

culture

enables one to remove some growth
under the oil and inoculate it in a
fresh medium and still preserve the
initial culture

63
Q

disadvantage of overlaying cultures with mineral oil

A

viability of microorganisms varies
with species

64
Q

temperature of lyophilization freeze-drying

A

-70C

65
Q

is a water removal process typically used to preserve perishable materials, to extend shelf life or make the material more convenient for transport.

A

lyophilization

66
Q

Employs
* rapid drying in frozen state
* dry ice in alcohol

A

lyophilization

67
Q

advantages of lyophilization

A

long-term survival
* less opportunity for changes in the
characteristics of culture
* smallness of storage containers

68
Q

temperature of freeze-drying with liquid nitrogen

A

-196C

69
Q

considerations for freezing with liquid nitrogen

A

cryoprotective agent (glycerol)
* liquid-nitrogen refs

70
Q

drying temperature

A

45C

71
Q

limitation for drying preservation method

A

for spore and cyst formers

72
Q

organizations which maintain authentic
pure cultures of microorganisms

A

culture collections

73
Q

provide ‘type’ strains to microbiologists
throughout the world

A

culture collections

74
Q

example of culture collection

A

American Type Culture Collection (ATCC)
National Collection of Type Cultures
Japanese Type Culture Collection
Philippine National Collection of Microorganism

75
Q

ATCC is located in

A

Maryland

76
Q

NCTC is located in

A

London

77
Q

JTCC is located in

A

Japan

78
Q

PNCM is located in

A

Biotech-UPLB Philippines

79
Q

Enumeration of microorganisms is especially important in what industries

A

dairy microbiology
food microbiology
pharmaceutical microbiology
water microbiology

80
Q

done to evaluate the effects of antimicrobial agents or the decontamination processes

A

enumeration

81
Q

methods for counting microorganisms (3)

A

viable
total
rapd

82
Q

viable counts include (4)

A

pour plate
surface spread/spread plate
membrane filter method
MPN

83
Q

total count method (4)

A

direct microscopic counting
turbidity methods
dry weight determination
nitrogen, protein, nucleic acid determination

84
Q

rapid method include (4)

A

epifluorescence with image analysis

ATP testing
impedance method
manometric method

85
Q

is a counting procedure enumerating both living and dead cells.

A

total count

86
Q

are possible using special slides known as counting chambers, consisting of a ruled slide and a coverslip.

A

direct microscopic count

87
Q

Bacteria can be counted efficiently and accurately with the (direct count)

A

Petroff-Hausser counting chamber

88
Q

This is a special slide accurately ruled into squares that are 1/400 mm² in the area; a glass coverslip rests 1/50 mm above the slide so that the volume over a square is 1/20,000 mm² i.e. 1/20, 000, 000 cm².

A

Petroff Hausser counting chamber

89
Q

A suspension of unstained bacteria can be counted in the Petroff-Hausser counting chamber using what kind of microscope

A

phase-contrast

90
Q

advantages of direct microscopic count

A

Rapid, simple, and easy method requiring minimum equipment.

Morphology of the bacteria can be observed as they are counted.

Very dense suspensions can be counted if they are diluted appropriately.

91
Q

limitation of direct microscopic count

A

Dead cells are not distinguished from living cells.

Small cells are difficult to see under the microscope, and some cells are probably missed.

Precision is difficult to achieve
A phase-contrast microscope is required when the sample is not stained.

The method is not usually suitable for cell suspensions of low density i.e., < 107 Cells per ml, but samples can be concentrated by centrifugation or filtration to increase sensitivity.

92
Q

are the most common means of estimating the total number of bacteria present in a sample

A

turbidity measurements

93
Q

Measuring the turbidity using a ____ or colorimeter and reading the concentration from a calibration plot is a simple means of standardizing cell suspensions for inocula in antibiotic assays or other tests of antimicrobial chemicals.

A

spectrophotometer

94
Q

records living cells alone.

A

viable count

95
Q

is defined as one that can divide and form offspring

A

viable cell

96
Q

or this reason, the viable count is often called the

A

plate count
colony count

97
Q

It uses fluorescent dyes that either exhibit different colors in living and dead cells (e.g., acridine orange) or appear colorless outside the cell but become fluorescent when absorbed and subjected to cellular metabolism (e.g., fluorescein diacetate).

A

epiflourescent technique

98
Q

Living cells generate adenosine triphosphate (ATP) that can readily be detected by enzyme assays, e.g., luciferin emits light when exposed to firefly luciferase in the presence of ATP; light emission can be measured and related to bacterial concentration.

A

aTP testing

99
Q

The resistance, capacitance, or impedance of a culture medium changes due to bacterial or yeast growth and metabolism, and these electrical properties vary in proportion to cell concentration.

A

impedance technique

100
Q

are appropriate for monitoring the growth of organisms that consume or produce significant quantities of gas during their metabolism, e.g., yeasts or molds producing carbon dioxide from fermentation.

A

manometric

General Microbiology (20 decks)

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