Microbiology Lab Practical 1 Flashcards

1
Q

What are the general nutritional requirements for growth?

A
  • water

- chemical element

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2
Q

The 4 physical factors that affect growth of an organism?

A
  • salt and sugar concentration
  • temperature
  • oxygen gas
  • pH
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3
Q

What ingredient is used as a solidifying agent in media?

A

agar

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4
Q

What type of organism makes up agar media?

A

algae

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5
Q

what percentage of organisms can be cultured in an artificial (laboratory) setting?

A

1 percent

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6
Q

By what process do most bacteria divide?

A
  • binary fision
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7
Q

The temperature (in celsius) for the the freezer is?

A

-20 degrees

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8
Q

The temperature (in celsius) for refrigerator is?

A

4-10 degrees

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9
Q

The temperature (in celsius) for room temperature is?

A

20-25 degrees

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10
Q

The temperature (in celsius) for body temperature is?

A

37 degrees

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11
Q

What percentage of organisms require oxygen for growth?

A

50%

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12
Q

Define negative control?

A
  • no growth
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13
Q

Define positive control?

A
  • expect growth
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14
Q

Why might an organism fail to grow?

A
  • none made it alive, dies before
  • culture is bad
  • physical conditions aren’t correct
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15
Q

Why might two different organisms show exactly the same growth features even if they aren’t supposed to?

A
  • cross contaimination
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16
Q

what is a meniscus?

A
  • the concave appearance artifact
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17
Q

What growth patterns are similar or different among the organisms tested?

A
  • bacillus only had a pellicle

- staph/e.coli were both turbid

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18
Q

Is it possible for an organism to produce more than one growth pattern in the same tube?

A

yes

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19
Q

A film growth on the surface is a what?

A

pellicle

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20
Q

Even cloudiness is called what?

A

uniform fine turbidity

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21
Q

Growth adhering to the lass at the liquid/air interface is called what?

A

a ring

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22
Q

Suspended flakes or particles are called?

A

flocculent

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23
Q

A layer of growth at the bottom of the broth is called?

A

sediment

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24
Q

How do you calculate total magnification?

A

objective x ocular = total magnification

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25
Q

If a student wants to learn morphology, arrangement and motility of an organism what type of preparations would they use?

A
  • wet mount preparation or the hanging drop procedure
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26
Q

Dyes that are positively charged are attracted to which components of cells?

A
  • negative components of living cells
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27
Q

What charged dye will cause the cell to pick up the color?

A
  • positively charged dye
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28
Q

Dyes that are negatively charged are attracted to what charge components?

A
  • positive components
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29
Q

What is a simple stain?

A
  • a procedure that uses one positively charged dye to give cells color
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30
Q

What are some examples of simple stains?

A
  • methylene blue, crystal violet, safranin, and manevals
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31
Q

What a negative stain?

A
  • a procedure that uses a negatively charged dye to stain the background while the cells remain virtually colorless
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32
Q

What are some examples of negatively charged acidic dyes?

A
  • eosin, india ink, migrosin and congo red
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33
Q

What is morphology?

A
  • shape
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34
Q

What are some advantages of making a heat fixed smear?

A
  • kills the organisms
  • organisms adhere to the slide
  • organisms take up the stain more readily
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35
Q

What are some disadvantages of making a heat fixed smear?

A
  • distorts the true size of the cell
  • may destroy heat- sensitive cell structures
  • may create artifacts
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36
Q

Why is simple staining not very useful for examining living organisms or to test an orgamism’s motility?

A
  • because many of the dyes re toxic to the cells and can paralyze flagella, cilia and so forth
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37
Q

Living cells have a net (negative or positive) charge and negative staining uses (negatively or positively) charged dye which is repelled by the cell’s charge.

A

Negative, Negative

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38
Q

Why is it important NOT to heat-fix a smear that will undergo negative staining?

A

bc with negative staining you’re usually trying to identify heat-sensitive structures and heat fixation can distort the morphology and size of a cell

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39
Q

Why do physicians rely on gram staining results for prescribing antibiotics?

A
  • some antibiotics will kill gram positive, some will kill/inhibit gram negative bacteria. Some will kill a wide array of bacteria including normal flora
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40
Q

Why do older cultures stain unreliably with gram stain protocol?

A
  • waste products accumulate and break down cell walls as cultures age. Cells that usually stain gram positive will look gram negative
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41
Q

What genera of bacteria may not gram stain reliably?

A
  • the mycobacterium species wont stain. They’ll look barely visible
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42
Q

How does iodine function?

A

iodine acts as a mordant. It forms a molecular bond with the crystal-violet

43
Q

Gram negative cells have ________ cell walls surrounded by _________. They will _______ the crystal-violet-iodine complex during decolorization.

A

very thin, outter lipid membrane called lipopolysaccharide, not retain

44
Q

Gram positive cells have ______________ cell walls. They will _______ the crystal-violet-iodine complex during decolorization.

A

very thick, retain

45
Q

if a gram positive cell is overdecolorized, it will be the color _____.

A

red

46
Q

if a gram negative cell is underdecolorized it will be the color ______.

A

purple

47
Q

Skipping the iodine step means that gram positive cells will appear _________ at the end of the protocol.

A

red

48
Q

What cell wall structure causes the stringing in the KOH test?

A

the DNA

49
Q

Why is the gram stain result important for treating physicians?

A
  • for prescribing an appropriate antibiotic and to learn the morphology and arrangement of the organism
50
Q

Which test(s) yields the quickest result ? KOH, Vancomycin or gram stain

A
KOH
#2 gram
#3 vancomycian
51
Q

What is the advantage of performing all 3 test (KOH Vancomycian and gram stain) on an unknown specimen?

A
  • the three tests together provide a more accurate result
52
Q

What are the two genera of endospore-forming bacteria mentioned in exercise 9?

A

bacillus spp.

clostridium spp.

53
Q

Endospores are (metabolically active or inactive) while vegetative cells are (metabolically active or inactive) ?

A

inactive, active

54
Q

What is the purpose of steaming the slide while staiing the Malachite Green?

A

it increases the porosity of spore coat so stain can penetrate

55
Q

Define sporogenesis

A

the creation of an endospore

56
Q

Define germination

A

the conversion of an endospore to a vegetative cell

57
Q

What color will endospores and free spores be following the staining protocol?

A

green

58
Q

What color will vegetative cells be following the staining protocol?

A

red

59
Q

The role of water in the endospore staining protocol is (2)..

A
  • to rinse the slide of excess stain, and to decolorize malachite green from vegetative cells
60
Q

Why might a young (18 hour) culture of endospore-forming bacteria fail to have endospores? explain

A

because to produce an endospore the cell must be in harsh conditions. A young cell is not yet nutritionally stresses.

61
Q

What are three functions that capsules serve?

A
  • protect cell from desiccation
  • protect from phagocytosis
  • allows organisms to stick to each other
62
Q

Water functions as a _____ in the capsule stain.

A

decolorizer

63
Q

Is congo red a negative or positive stain? does it stain the organism or the background?

A

negative stain, stains the background

64
Q

Is maneval’s a negative or positive stain? does it stain the background or the organism?

A
  • positive stain, stains the organism
65
Q

How does TTC function in motility media?

A
  • as a tracking dye
66
Q

What might cause false positive results in the test using motility medium?

A

you could have contaminated probe…it could have moved a bit inside the medium, gas bubbles could’ve exploded and shot organisms outward

67
Q

Why is motility medium semi-solid?

A

it ensure that the motile organism will be able to swim away from the stab line

68
Q

when performing hanging drop, how is it possible to distinguish true motility from brownian motion?

A

make sure they are moving in a purposeful direction not not just vibrating in place. make sure you’re not mistaking dehydration for movement

69
Q

Describe proteus vulgaris flagellar arrangement.

A
  • had flagella all around the edges, peritrichous
70
Q

Describe spirillum voluntans flagellar arrangement.

A
  • only had flagella on the opposite ends (amphirichous)
71
Q

What was the purpose of flame sterilizing the inoculation loop in between quadrants during the streak plate technique?

A
  • to ensure you don’t pick up too much bacteria into the next quadrant. flaming it ensures you start with a clean loop into the next quadrant.
72
Q

If you have growth in quadrant one but none in 2-4 what may have caused this? streak plate technique

A
  • not waiting for the inoculation loop to cool can kill the organisms not allowing it to grow
73
Q

What sample from your own body could you have used as a mixed culture during the streak plate technique?

A
  • nose, mouth, feces, sputum, saliva, sweat
74
Q

Why is it so important to isolate organisms from a mixed culture into a pure culture?

A

for species identification, antibiotic susceptibility testing and other necessary tests
- can’t carry out copulates with a mixed culture

75
Q

Define viable

A

only counts the LIVING cells

76
Q

what is dilution?

A
  • dilution is reducting the concentration of a substance.

- it’s a fraction and in scientific notation is a negative number

77
Q

what is dilution factor?

A
  • the FACTOR by which the substance has been diluted.

- the reciprocal of the dilution

78
Q

How is MacConkey Agar both selective and differential?

A
  • it is used in selective for detecting enteric bacteria (gram negative). It is used as a differential media when differentiating between lactose fermenters (pinkish colonies)and non-lactose fermenters (beige color)
79
Q

How would you characterize a dilution series plate containing 419 CFUs?

A

too numerous to count (TNTC)

80
Q

How would you characterize a dilution series plate containing 28 CFUs?

A

too few to count (TFTC)

81
Q

How many CFUs/ml are generally present in the urine of young women?

A

10^2 - 10^3 mixed bacterial colonies

82
Q

For the urine test, why might an uncircumcised male yield higher colony counts than an circumcised male?

A

the difference in anatomy

- mucous membranes cover urethra and bacteria is in this

83
Q

How are quantitative and semi-quantitative counts different?

A

quantitative count are used to count the NUMBER of living bacteria in a unit volume
- semi-quantitative is a rough estimate in an unknown volume

84
Q

What color are the colonies produced by E.coli on BBL chrome agar orientation medium

A

dark rose-pink

85
Q

What color are the colonies produced by KES on BBL chrome agar orientation medium

A

medium blue to dark blue

86
Q

What color are the colonies produced by PMP on BBL chrome agar orientation medium

A

pale-beige colonies surrounded by dark halos

87
Q

Steps of gram stain

A
  • Crystal violet
  • Rinse with water
  • Iodine (mordant)
  • Rinse with water
  • Decolorize with ethanol/acetone
  • Rinse with water
  • Counterstain with safranin
88
Q

Steps of capsule stain

A
  • Congo red to emulsify cells (negative stain that gives background color
  • Maneval’s stain (positive stain to give cells color)
  • Decolorize with water (to decolorize the capsule)

*smears are not heat-fixed

89
Q

Steps of endospore staining?

A
  • Steam smear and stain with Malachite Green (steam drives MG into spore coat)
  • Decolorize vegetative cells with water
  • Counterstain vegetative cells with Safranin
90
Q

Antibiotics are ___ that word to inhibit or ____ microorganisms.

A

chemicals, kill

91
Q

Antimicrobials that target bacteria are usually called _____

A
  • antibiotics
92
Q

An antimicrobial that kills bacteria

A

bactericidial

93
Q

an antibiotic that inhibits bacteria

A

bacteriostatic

94
Q

The antibiotic targets in bacterial cells

A

peptidoglycan, 70’s ribosome, bacterial nucleic acid synthesis enzymes, antimetabolites, cell membrane

95
Q

Penicillin is ___ and inhibits ________

A

bactericidal, peptidoglycan synthsis

96
Q

Synthetic sulfonamides are ___ and are considered antimetabolites

A

bacteriostatic

97
Q

Quinolones interfere with _____replication

A

DNA

98
Q

Tetracycline interferes with bacterial ______ by inhibiting the _______________ ribosome.

A

protein synthesis, 70s

99
Q

Define broad spectrum

A

antibiotics that are effective against a wide array of bactria

100
Q

Define narrow spectrum

A

antibiotics that are effective aainst a narrow array of bacteria

101
Q

Define antibiotic

A

substance produced by one microorganism (typically a fungal or bacterial organism) that inhibits or kills another microorganism.

102
Q

Define semisynthetic antibiotic

A

chemically altered version of a naturally-produced antibiotic.

103
Q

Define zone of inhibition

A

the clear ring around an antimicrobial disk. It represents an area of growth inhibition.

104
Q

Define Mueller-Hinton agar

A

Medium used; contains added buffers to prevent wide fluctuations in pH, that might otherwise alter the effectiveness of the antimicrobials being tested.