Mid Term Practical Flashcards

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1
Q

A species of pathogenic aerobic endospore forming bacterium is ____ and causes _______

A

Bacillus anthracis + anthrax

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2
Q

A species of anaerobic endospore forming bacterium is ____ + causes _____

A

Clostridium tetani + tetanus

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3
Q

Capsules may be made of

A

polysaccharides, glycoprotein, or polypeptides

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3
Q

Capsules may be made of

A

polysaccharides, glycoprotein, or polypeptides

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4
Q

Negative stains stain

A

everything except what you want to see

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5
Q

At the end of the endospore staining process

A

Endospores are green and vegetative cells are red/pink

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6
Q

At the end of the endospore staining process

A

Endospores are green and vegetative cells are red/pink

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7
Q

In acid fast stains

A

non-acid fast stains blue
acid-fast stains pink

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8
Q

What is used in acid-fast stain?

A

Carbol fuschin (2 min)
Acid alcohol
Methylene blue (1 min)

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9
Q

What is used in capsule stain?

A

Crystal violet (2 min)
Copper sulfate

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10
Q

What is used in endospore stain?

A

Malachite green (warmer) (5 min)
Malachite green (cooling) (5 min)
Safronin (2 min)

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11
Q

Who developed acid-fast stain?

A

Paul Ehrlich

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12
Q

Name of acid-fast used in the lab

A

Kinyoun

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13
Q

2 pathogenic species of mycobacterium are

A

Mycobacteria tuberculae + leprae
TB + leprosy

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14
Q

What parasite can be identified by acid fast?

A

Cryptosporidium and toxoplasma

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15
Q

Mycobacterium is filamentous (like a treebranch) and Nocardia is rod shaped

A

False

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16
Q

Where do used glass test tubes discard?

A

Discard rack

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17
Q

Where do used glass slides go?

A

Used slide basin

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18
Q

Where does broken glass go?

A

Broken glass container

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19
Q

Where do used latex gloves go?

A

Autoclave trash

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20
Q

Where do paper towels used in bench disinfection go?

A

Regular trash

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21
Q

Where does uncontaminated trash go?

A

Regular trash

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22
Q

Where do contaminated Petri plates go?

A

Autoclave trash

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23
Q

Where do qtip swab go?

A

Regular trash

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24
Q

How to calculate total magnification of an object?

A

Ocular (10x) x objective

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25
Q

Purpose of immersion oil

A

Decreases light refraction and loss of light

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26
Q

Define ubiquitous

A

Found everywhere

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27
Q

Define pure culture

A

Single genus + species

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28
Q

Define colony

A

Single bacteria divides by binary fission and grows large enough to be seen by naked eye

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29
Q

Define turbidity

A

Cloudiness which indicates growth

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30
Q

Define media

A

Nutrient material suitable for growth of microorganisms

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31
Q

Define innoculum

A

Sample of microbial culture

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32
Q

Define aseptic technique

A

Safely moving microbes without contaminating environment, oneself, or the microorganism

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33
Q

How much is in a pinpoint amt of bacteria?

A

1 mil

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34
Q

Why are agar plate labeled on bottom and inverted when incubated?

A

So condensation doesn’t disrupt colony formation

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35
Q

State the criteria used to describe a bacterial colony

A

Shape
Margin
Elevation
Colony

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36
Q

Order of gram stain

A
  1. CV (1 min) DIH2O
  2. Iodine (mordant) (1 min) DIH2O
  3. Acetone-alcohol (2-5 sec)
  4. Safronin (1 min) DIH2O
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37
Q

What color should Gram + and Gram -?

A

Gram + : purple (cocci)
Gram - : pink (bacilli)

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38
Q

What stains can show size, shape, and arrangment?

A

Simple and Differential

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39
Q

Two genera that can be seen in an acid-fast stain?

A

Mycobacterium + nocardia

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40
Q

What color should acid-fast and non-acid-fast bacteria?

A

Acid fast: Pink
Non acid fast: Blue

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41
Q

Two diseases caused by acid-fast bacteria?

A

TB + leprosy + nocardiosis + cyptosporidiosis

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42
Q

Why are acid fast bacteria acid fast and why do they stay pink?

A

The bacteria have a waxy wall made of mycolic acid that traps the carbol fuschin in. It resists decolorizing by acid-alcohol.

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43
Q

Steps in acid-fast stain?

A
  1. Carbol fuschin on filter paper (2 min on heat) DIH2O
  2. Acid alcohol (2-5 sec off heat) DIH2O
  3. Methylene blue (2 min off heat) DIH2O
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44
Q

What are 2 genera of bacteria that produce endospores?

A

Bacillus and clostridium

45
Q

Endospore stain process?

A
  1. Malachite green w/filter paper (5 min on warmer)
  2. Malchite green w/ filter paper (5 min off warmer) DIH2O
  3. Add safronin (2 min) DIH2O
46
Q

Difference between neg. stain and pos. stain:

A

Positive (+) Stain: stains everything you want to see
Negative(-) Stain: stains everything except for what you’re trying to see

47
Q

Why won’t capsules hold a stain?

A

Capsules have a neutral charge and stain have a positive charge

48
Q

Why don’t we heat fix negative or capsule stains?

A

Heat will denature the protein in the capsule and you won’t be able to see it

49
Q

Bacteria that grows in the candle jar is:

A

Microaerophile
Capnophile

50
Q

The two major types of microscopes that have been developed are the

A

Compound and electron microscope

51
Q

What are the 4 types of microscopes?

A

Phase contrast
Brightfield (we use these)
Darkfield
Fluorescent

52
Q

Two factors in determining how well you’ll see through the microscope is:

A

The magnification and
Resolving power

53
Q

Define resolving power

A

Resolving power is measured by how well your microscope can show two lines lying close together as two distinct lines

54
Q

Define refractive index

A

How well the lens is able to bend light

55
Q

State the function of the microscope condenser

A

concentrates the beam of light on the specimen

56
Q

State the function of the microscope iris diaphragm

A

Regulates the intensity of light entering the lens

57
Q

State three steps to achieve adequate lighting when the oil immersion objective is used to view a specimen

A

The condenser is raised to the highest position.
The iris and base diaphragm are open.
The rheostat is used to regulate light intensity

58
Q

The two components that influence the resolving power of a microscope are the

A

Wavelength of light
Numerical aperture

59
Q

As you go from low to high-dry objectives, the diameter of the iris diaphragm

A

Decreases

60
Q

The refractive index of glass and immersion oil are the same

A

True

61
Q

TSA(trypticase soy agar) is an example of nutrient media to grow bacteria

A

True

62
Q

What are the damaging by-products of oxygen during aerobic energy production (cellular respiration)?

A

Superoxide and peroxides

63
Q

Name three enzymes that neutralize the damaging by-products of oxygen during aerobic energy production (cellular respiration).

A

catalase, peroxidase, superoxide dismutase

64
Q

How does the Gas Pak jar achieve an anaerobic atmosphere?

A

The added packet of chemicals generates hydrogen. The hydrogen binds oxygen to produce water.

65
Q

How does the Candle Jar achieve an atmosphere to grow microaerophilic, capnophilic organisms?

A

The flame consumes oxygen and releases carbon dioxide

66
Q

chemicals produced by some bacteria and fungi that, in small quantities, can inhibit the growth of bacteria

A

Antibiotics

67
Q

the lowest concentration of an antibiotic that can inhibit the growth of a test organism

A

Minimal Inhibitory Concentration (MIC)

68
Q

E-test

A

Combination of Kirby-Bauer and Minimal Inhibitory Concentration tests

69
Q

Disadvantages of “broad-spectrum” antibiotics?

A

Broad spectrum antibiotics are probably contributing to the escalating drug resistance we are seeing in microorganisms. Broad spectrum antibiotics often wipe out a person’s normal microbiome as well as the pathogen they are intended to kill, resulting in superinfections from organisms

70
Q

What are some organisms that can cause super infections?

A

Clostridium difficile and Candida albicans

71
Q

List the factors that must be controlled to standardize the Kirby-Bauer test pt. 1

A

-stability of the antibiotic
-the rate of diffusion of the antibiotic
-the bacteria being tested
-the pH of the culture medium
-the depth of the culture medium

72
Q

List the factors that must be controlled to standardize the Kirby-Bauer test p. 2

A

-the inoculum density
-the incubation time
-the incubation temperature
-the concentration of the antibiotic

73
Q

What kind of agar plate do we use for Kirby-Bauer testing?

A

Mueller-Hinton Agar Plate

74
Q

E. coli and Pseudomonas are gram ____ bacteria?

A

Gram negative

75
Q

Staphylococcus epidermidis is gram _____ bacteria?

A

Positive

76
Q

Negative Beta-Lactamase Result

A

-No pH change=organism did not produce beta-lactamase.
-Organism is sensitive to penicillin
-Turns PINK

77
Q

Positive Beta-Lactamase Result

A

-pH change
-Organism produce beta-lactamase
-Organism is resistant to penicillin

78
Q

Which gene allows jellyfish to glow in the dark?

A

GFP

79
Q

able to take up the DNA (e.g. plasmid) from the environment

A

Competent Cells

80
Q

Define “competent” cells

A

Able to pick up DNA from the environment

81
Q

What laboratory procedure is done to encourage cells to become competent?

A

growth in the presence of calcium chloride

82
Q

What is the relationship between a gene and a protein?

A

A gene is a piece of DNA which provides the instructions for making a protein

83
Q

Compare “chromosomal DNA” and “plasmid”

A

Chromosomal DNA contains essential information for bacterial growth and reproduction. Plasmid DNA are small, circular DNA separate from its chromosome which contains non-essential infornation

84
Q

List the genes contained in the pGLO plasmid

A

GFP, BLA, Ara-C

85
Q

How is the expression of the Green Fluorescent Protein gene regulated?

A

The BLA gene and Ara-C gene are continually transcribed and translated into protein. The gene for GFP, on the other hand, is regulated by the Repressor protein and will not be transcribed without the addition the sugar arabinose to the cells’ nutrient medium. The arabinose will inactivate the repressor protein thus allowing the GFP gene to be transcribed.

86
Q

Genes can be transferred from one organism to another by

A

Transformation

87
Q

Aa plasmid that contains the gene for green fluorescent protein and resists ampicillin

A

pGLO

88
Q

Able to take up a plasmid

A

Competent bacteria

89
Q

Explain why certain organisms are “acid-fast”

A

Waxy wall enables the bacteria to resist decolorizing with acid alcohol

90
Q

What is the difference between the Ziehl-Neelsen stain and the Kinyoun stain?

A

Ziehl-Neelsen stain uses steam Kinyoun uses a concentrated dye, heat, and longer stain time

91
Q

List four acid fast organisms and their associated disease names

A

Mycobacterium tuberculosis-tuberculosis

Mycobacterium leprae-leprosy or Hansen’s disease

Nocardia asteroides-nocardiosis

Cryptosporidium-cryptosporidiosis

92
Q

Name the dyes used in the Schaeffer-Fulton endospore stain

A

Malachite green and Safranin

93
Q

Define vegetative cell

A

Metabolically active, replicating cell

94
Q

Define vegetative cell

A

Metabolically active, replicating cell

95
Q

Media used for antibiotic sensitivity test

A

Mueller-Hinton

95
Q

Media used for antibiotic sensitivity test

A

Mueller-Hinton

96
Q

Genes located on the pGLO plasmid

A

BLA, GFP, ARA-C

97
Q

Ara-C protein is a

A

Repressor protein that regulates transcription of GFP gene

98
Q

Beta-lactamase does what?

A

Breaks apart ampicillin (targets beta-lactam ring)

99
Q

GFP is

A

Gene that translate to become fluorescent protein that glows under UV light

100
Q

Ara-c block what?

A

Transcription of GFP

101
Q

What does arabinose do to Ara-c?

A

Inactivates it and allows RNA polymerase to transcribe and translate the GFP protein

102
Q

Difference between slime layer and capsule

A

Slime layer: loosely bound
Capsule: tightly bound, easier to visualize

103
Q

Capsule in virulence

A

Decreases phsagocytosis, aids in attachment, can be a source of nutrition, helps prevent drying

104
Q

State two differences between the smear preparation for a Gram stain and a capsule stain

A

Do not heat fix
Do not rinse with water

105
Q

Anthonys capsule stain method

A

Stains the background blue

106
Q

Most common stains in capsule/negative stain

A

Nigrosin and india ink

107
Q

How does the thioglycollate broth grow anaerobic bacteria as well as aerobic bacteria?

A

Agar prevents diffusion of oxygen into the medium. Thioglycollate binds oxygen. Together they produce a decreasing level of oxygen in the media.

108
Q

Methylene blue is blue in color in the presence of oxygen

A

True