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Cameron Lecture study guides > Lab exam FINAL > Flashcards

Lab exam FINAL Flashcards

1
Q

what are the four steps of differential staining?

A
  1. primary stain - colors the target cell
  2. mordant - helps color stick in target cells
  3. decolorizer - removes color from background and non-target cells
  4. counterstain - stains non-target cells
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2
Q

what are the four chemicals used for gram stain?

A
  1. primary stain: crystal violet
  2. mordant: iodine
  3. decolorizer: acetone/alcohol
  4. counterstain: safranin
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3
Q

what is the primary stain in gram staining?

A

crystal violet

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4
Q

what is the mordant in gram staining?

A

iodine

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5
Q

what is the decolorizer used in gram staining?

A

alcohol and acetone

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6
Q

what is the counterstain used in gram staining?

A

safranin

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7
Q

what were the target and non-target cells used in gram stain? what color should they have been?

A

target: Staphylococcus aureus - purple

non-target: Escherichia coli - pink

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8
Q

what are the four chemicals used for acid fast stain?

A
  1. primary stain: basic fuchsin
  2. mordant: phenol + dimethyl sulfoxide
  3. decolorizer: acid-alcohol
  4. counterstain: methylene blue
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9
Q

what were the target and non-target cells used in acid fast stain? what color should they have been?

A

target: Mycobacterium smegmatis - pink

non-target: Staphylococcus aureus - blue

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10
Q

what are the four chemicals used for endospore stain?

A
  1. primary stain: malachite green
  2. mordant: steam
  3. decolorizer: distilled water
  4. counterstain: safranin
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11
Q

what were the target and non-target cells used in endospore stain? what color should they have been?

A

target: endospores - green

non-target: vegetative cells - pink

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12
Q

what is the primary stain in capsule staining?

A

congo red

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13
Q

what is the purpose of the acid-alcohol for the capsule stain? how is this different than how it was used for acid-fast?

A

In capsule staining, acid-alcohol chemically fixes the cells to the slide

In acid-fast, acid-alcohol is used as the decolorizer

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14
Q

what is the secondary stain used in capsule staining?

A

carbol fuchsin

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15
Q

How do capsules appear at the end of capsule staining?

A

background will be dark blue/purple and magenta
capsules will look like a clear halo around the pinkish color of the bacillus shaped cell

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16
Q

what was the target bacteria in capsule stain? what color?

A

Klebsiella pneumoniae - mostly colorless

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17
Q

what is the purpose of streak plating?

A

to produce pure, isolated colonies from a mixed sample

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18
Q

what antibiotic was most and least effective?

A

Most: Chloramphenicol
Least: Bacitracin

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19
Q

Which bacterial type is the most and least susceptible to antibiotics?

A

most susceptible: Mycobacterium smegmatis

least susceptible: Escherichia coli

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20
Q

what does the death zone of a susceptible organism look like?

A

large death zone

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21
Q

what does the death zone of a resistant organism look like?

A

small/no death zone

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22
Q

what antiseptic was most and least effective?

A

Most effective – 1% formaldehyde

Least effective – 70% isopropyl alcohol

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23
Q

which bacterial type is the most and least susceptible to antiseptics?

A

most susceptible: Mycobacterium smegmatis

least susceptible: Escherichia coli

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24
Q

what is the equation to convert Celsius to Fahrenheit

A

°F = (°C x 1.8) + 32

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25
Q

what is refrigerator temp in Celsius?

A

4 °C

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26
Q

what is room temp in Celsius?

A

25 °C

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27
Q

what is body temp in Celsius?

A

37 °C

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28
Q

what is the definition of selective media?

A

inhibits growth of unwanted microbes (only the selected microbes live)

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29
Q

what is the definition of differential media?

A

allows organisms to grow but the added chemicals make them look different (they change different colors and/or precipitate is visible)

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30
Q

What did MSA stand for? Whats the original color?

A

MSA: Mannitol salt agar

red

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31
Q

How is MSA selective?

A

selects for gram +

selects against gram -

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32
Q

How is MSA differential? (fermentation?)

A

differential for mannitol fermentation

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33
Q

what are the possible bacteria and colors produced by inoculating MSA plate?

A

fermenter: growth turns yellow (Staphylococcus aureus)

nonfermenter: growth remains red (Staphylococcus epidermidis)

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34
Q

What did EMB stand for? Whats the original color?

A

EMB: Eosin methylene blue

dark purple

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35
Q

how is EMB selective?

A

selects for gram -

selects against gram +

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36
Q

how is EMB differential? (fermentation?)

A

differential for lactose fermentation

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37
Q

what are the possible bacteria and colors produced by inoculating EMB plate?

A

strong fermenter: growth turns shiny metallic green (Escherichia coli)

weak fermenter: growth turns pink around the edges of colonies (Enterobacter aerogenes)

nonfermenter: growth remains dark purple (Proteus vulgaris)

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38
Q

What did MAC stand for? Whats the original color?

A

MAC: MacConkey agar

raspberry

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39
Q

how is MAC selective?

A

selects for gram -

selects against gram +

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40
Q

how is MAC differential? (fermentation?)

A

differential for lactose fermentation

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41
Q

what are the possible bacteria and colors produced by inoculating MAC plate?

A

strong fermenter: entire plate turns bright pink (Escherichia coli)

weak fermenter: growth turns light pink with precipitate and background turns light brown (Enterobacter aerogenes)

nonfermenter: entire plate turns brown (Proteus vulgaris)

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42
Q

which plate used in lab this semester was not selective or differential?

A

TSA, Tryptic soy agar

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43
Q

which organisms used in the S&D exercise are Gram + ?

A

Staphylococcus aureus
Staphylococcus epidermidis

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44
Q

which organisms used in the S&D exercise are Gram - ?

A
  • Escherichia coli
  • Enterobacter aerogenes
  • Proteus vulgaris
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45
Q

what genus is used for the gram + sphere project?

A

Staphylococcus

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46
Q

Gram + Sphere Unknown (Staphylococcal)

what are the potential species used within the gram + sphere project?

A

Staphylococcus aureus
Staphylococcus capitis
Staphylococcus epidermidis
Staphylococcus haemolyticus
Staphylococcus lentus
Staphylococcus saprophyticus

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47
Q

Gram + Sphere Unknown (Staphylococcal)

What type of media was used to test for growth at elevated temperature?

A

TSB

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48
Q

Gram + Sphere Unknown (Staphylococcal)

what was a positive/negative result for the test for growth at elevated temperature?

A

positive: turbid = indicated that growth was present

negative: clear = growth not present

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49
Q

Gram + Sphere Unknown (Staphylococcal)

Which elevated temperature was used to differentiate between species of Staphylococcus?

A

45C or 113F

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50
Q

what tool did we use to inoculate the 45 degree experiment?

A

cotton swab

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51
Q

Gram + Sphere Unknown (Staphylococcal)

Is urea a product or reactant for this test?

A

Urea is the reactant

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52
Q

what tool was used to inoculate the urea agar slant?

A

loop

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53
Q

Gram + Sphere Unknown (Staphylococcal)

What are the two products of the enzyme urease? Know how to write out the molecular formula of each of these products

A

ammonia + carbon dioxide

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54
Q

what is the original media color of urea agar slant?

A

peach color

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55
Q

Gram + Sphere Unknown (Staphylococcal)

Which product of urease changes the pH of the sample tube?

A

ammonia

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56
Q

Gram + Sphere Unknown (Staphylococcal)

Will a tube that is positive for urease activity become more acidic or more alkaline?

A

Alkaline

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57
Q

Gram + Sphere Unknown (Staphylococcal)

What is the weak acid/weak base pair that is produced by urease activity?

A

carbon dioxide = weak acid

ammonia = weak base

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58
Q

what was the reaction for the substrate and products of the urea agar slant?

A
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59
Q

Gram + Sphere Unknown (Staphylococcal)

what gives a pos/neg result in the urea agar slant?

A

pos: hot pink color change = ^ pH

neg: yellow color change = v pH

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60
Q

Gram + Sphere Unknown (Staphylococcal)

What kind of molecule is trehalose?

A

plant carbohydrate

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61
Q

Gram + Sphere Unknown (Staphylococcal)

Is trehalose a product or a reactant for this test?

A

Trehalose is the reactant

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62
Q

Gram + Sphere Unknown (Staphylococcal)

  • Which pH indicator is used in a trehalose agar stab?
A

Phenol red

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63
Q

Gram + Sphere Unknown (Staphylococcal)

  • Which tool is used to inoculate an agar stab?
A

needle

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64
Q

Gram + Sphere Unknown (Staphylococcal)

Trehalose stab:

  • What color will this pH indicator be with a positive test for fermentation?
A

Yellow color change (pH goes down)

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65
Q

Gram + Sphere Unknown (Staphylococcal)

What is the name of the toxin that destroys red blood cells?

A

hemolysin

66
Q

Gram + Sphere Unknown (Staphylococcal)

What would a positive blood agar test result look like for the BLood agar test?

What does this indicate?

A

Red color disappears under colonies

Indicates that hemolysin production occurred

67
Q

What tools did we use to inoculate the blood agar plate? what technique?

A

Cotton swab
Zag plating technique

68
Q

Gram + Sphere Unknown (Staphylococcal)

Which Greek letter is used to describe a positive test result?

A

Beta patter (clearance zone)

69
Q

Gram + Sphere Unknown (Staphylococcal)

Which Greek letter is used to describe a negative test result?

A

Gamma pattern (no clearance zone)

70
Q

Gram + Sphere Unknown (Staphylococcal)

What type of agar is used to screen for antibiotic sensitivity of our unknown?

what is the test called?

what plating technique was used?

A

TSA
Novobiocin resistance test
Lawn plate technique

71
Q

Gram + Sphere Unknown (Staphylococcal)

How does novobiocin inhibit bacterial growth?

A

Novobiocin is an antibiotic that acts as DNA synthesis inhibitor to bacteria

72
Q

Gram + Sphere Unknown (Staphylococcal)

What would a pos/neg test result for novobiocin resistance look like?

A

pos: no death zone = resistant

neg: large death zone >4 mm = susceptible

73
Q

Gram + Sphere Unknown (Staphylococcal) (nitrate broth)

Which form of metabolism utilizes nitrate salt as a terminal electron acceptor?

A

anaerobic respirator

74
Q

what did the nitrate broth test for?

A

tests for nitrate respiration

75
Q

Gram + Sphere Unknown (Staphylococcal)

Is nitrate an oxidized or a reduced salt? what does this mean?

A

oxidized salt

it can accept electrons

76
Q

Gram + Sphere Unknown (Staphylococcal)

what is the equation for nitrate being successfully oxidized in the nitrate test?

A

Nitrate –> Nitrite –> Nitrous Oxide —> Nitrogen gas –> Amonia
No3- –> No2- –> N2O —————-> N2(g) ————> NH3
(color change) ( trapped in Durham tube )

77
Q

Gram + Sphere Unknown (Staphylococcal)

Which of these reduced products are gases?

A

nitrous oxide (N2O)

nitrogen gas (N2)

78
Q

Gram + Sphere Unknown (Staphylococcal)

Which color is produced by a positive nitrite test?

A

Color changes to Red and maybe gas bubbles

79
Q

Gram + Sphere Unknown (Staphylococcal)

  • Is a positive nitrite test a positive or negative result for nitrate reduction?
A

positive nitrite test (red color change) indicates that your organism reduced nitrate to nitrite. (positive for nitrate reduction)

80
Q

Gram + Sphere Unknown (Staphylococcal)

What is the function of a Durham tube?

A

to trap any potential gasses

81
Q

Gram + Sphere Unknown (Staphylococcal)

Is the presence of bubbles in the Durham tube positive or negative for nitrate reduction?

A

positive

82
Q

Gram + Sphere Unknown (Staphylococcal)

What would the results look like for an organism that did not use the nitrate?

A

Neg: color remains clear and no bubbles

83
Q

Gram + Sphere Unknown (Staphylococcal)

Gelatin is what type of biological molecule?

A

protein

84
Q

Gram + Sphere Unknown (Staphylococcal)

  • What is the name of the enzyme that hydrolyzes (breaks down) gelatin?
A

protease

85
Q

Gram + Sphere Unknown (Staphylococcal)

How did we inoculate the gelatin agar so that it could be incubated?

A

needle

86
Q

Gram + Sphere Unknown (Staphylococcal)

If we were to analyze the tubes for gelatin hydrolysis directly after removing the cultures from the incubator our results would be incorrect. What do we need to do before analyzing the results of our hydrolysis test?

A

allow them to chill

87
Q

Gram + Sphere Unknown (Staphylococcal)

Briefly describe the appearance of a tube that is positive for gelatin hydrolysis.

Whats negative look like?

A

pos: liquid after chilled = break down into amino acids occurred

neg: solid after chilled = break down did not occur

88
Q

Gram + Sphere Unknown (Staphylococcal)

what does the Vogel Johnson Agar test for?

A

mannitol fermentation

coagulase production

89
Q

what chemicals are in Vogel Johnson Agar? what do they serve as?

A

tellurite and lithium chloride
selective chemicals

90
Q

what tool and what technique is used within the Vogel Johnson Agar experiment?

A

cotton swab
zag plating technique

91
Q

what causes the broth to coagulate?

A

Coagulase

92
Q

Gram + Sphere Unknown (Staphylococcal)

In the Vogel Johnson Agar test, how are black colonies formed?

A

tellurite gets reduced to tellurium when coagulase is present

93
Q

Gram + Sphere Unknown (Staphylococcal)

What concentration of NaCl is used in our hypertonic broth?

A

6.5%

94
Q

Gram + Sphere Unknown (Staphylococcal)

what was the pH indicator of the sodium chloride broth?

A

bromocresol purple

95
Q

What tools did we use for the sodium chloride broth?

A

loop

96
Q

Gram + Sphere Unknown (Staphylococcal)

What was the color of the sodium chloride broth before incoluation?

A

purple

97
Q

Gram + Sphere Unknown (Staphylococcal)

What color is expected with a positive test result in the NaCl broth test?

A

Yellow color change (pH goes down)

Bacteria survived and grew

98
Q

Gram - Rod Unknown

what are a few of the potential genera used in the gram - rod unknown experiment?

A

Citrobacter freundii
Enterobacter aerogenes
Escherichia coli
Klebsiella pneumoniae
Proteus vulgaris
Pseudomonas aeruginosa

99
Q

Gram - Rod Unknown

what does a pos/neg test look like for the catalase test?

A

pos: bubble formation after H2O2 is added

neg: no bubbles formed after H2O2 is added

100
Q

What type of media was used for the catalase test?

A

TSA
(In a slant)

101
Q

Gram - Rod Unknown

write out the complete catalase reaction, including the names and molecular formulas of all substrates and products.

A

H2O2 –catalase–> H2O + O2 (forms bubbles)

102
Q

Gram - Rod Unknown

Which product of catalase activity is responsible for the visible reaction?

A

oxygen gas

103
Q

Gram - Rod Unknown

Which substrate is broken down by catalase?

A

Hydrogen peroxide (H2O2)

104
Q

Gram - Rod Unknown

what is a pos/neg result for the simmons’ slant?

A

pos: blue color change

Neg: color remains green

105
Q

What is the pH indicator of the simmons slant?

A

Bromothymol blue

106
Q

Gram - Rod Unknown

What color is a Simmons’ slant that is positive for nutrient utilization?

A

Blue

107
Q

Gram - Rod Unknown

What color is the Simmons slant tube before it is incubated?

A

Green

108
Q

Gram - Rod Unknown

What is the carbon source in the Simmons’ slant?

A

citrate

109
Q

Gram - Rod Unknown

What is the nitrogen source in the Simmons’ slant?

A

ammonium

110
Q

Gram - Rod Unknown

  • How does nutrient utilization affect the pH of the Simmons’ slant?

what happens to the ammonium as well?

A

citrate utilization removes citrate (acidic) from media which increases pH

ammonium (NH4+) gets reduced to ammonia (NH3)

111
Q

Gram - Rod Unknown

Litmus milk broth contains two key ingredients, which are..

A

skim milk and litmus powder

112
Q

Gram - Rod Unknown

What are the two functions of the litmus powder in the litmus milk broth?

A
  1. pH indicator –> change color based on fermentation of lactose
  2. Used as electron acceptor (can be reduced)
113
Q

Gram - Rod Unknown

What are the three reactions that can occur with the litmus powder?

A

Alkaline result: No lactose fermentation / purple color

Acidic result: Positive fpr lactose fermantation / pink color change

Reduction of litmus: White band that sinks to the bottom of tube

114
Q

WHat does an alkaline result look like with the litmus powder?

A

Alkaline result: No lactose fermentation / purple color

115
Q

WHat does an acidic result look like with the litmus powder?

A

Acidic result: Positive fpr lactose fermantation / pink color change

116
Q

WHat does a reduction of litmus result look like with the litmus powder?

A

Reduction of litmus: White band that sinks to the bottom of tube

117
Q

Gram - Rod Unknown

What are the two possible reactions that can occur with the milk protein in the litmus milk broth?

A

Protein results (peptonization) media is more clear

Coagulation (solid formation due to casin in the milk coagulating w/acid)

118
Q

Gram - Rod Unknown

WHat is peptonization?

A

Protein is utilized which removes the cloudiness

119
Q

Gram - Rod Unknown

What causes the broth to coagulate in the litmus test?

A

casein not being used, and instead clumping with the acids

120
Q

Gram - Rod Unknown

  • What does the abbreviation ‘SIM’ stand for?
A

Sulfide Indole Motility

121
Q

Gram - Rod Unknown

Which of the three tests did we not record data for? why?

A

motility
because some results would be black in color and would be too hard to see microbes being motile

122
Q

Gram - Rod Unknown

  • Which amino acid is degraded to produce ‘S’?
A

cysteine

123
Q

Gram - Rod Unknown

  • What is the enzyme that is responsible for ‘S’?
A

cysteine desulferase

124
Q

Gram - Rod Unknown

  • What are the three products that are produced as part of ‘S’?
A
125
Q

Gram - Rod Unknown

Which specific product of the three is detected by ‘S’?

A

hydrogen sulfoxide

126
Q

Gram - Rod Unknown

What color is a positive/neg ‘S’ test?

A

pos: black color change

neg: no color change (remains plain broth)

127
Q

Gram - Rod Unknown

Which amino acid is degraded to produce ‘I’?

A

tryptophan

128
Q

Gram - Rod Unknown

  • What is the enzyme that is responsible for ‘I’?
A

tryptophanase

129
Q

Gram - Rod Unknown

  • What are the three products that are produced as part of ‘I’?
A

Tryptophan – tryptophanase –> Nh3 + Indole + Pyruvate

130
Q

Gram - Rod Unknown

Which specific product of the three is detected by ‘I’?

A

indole

131
Q

Gram - Rod Unknown

What reagent is used to test for ‘I’?

A

Kovac’s reagent

132
Q

Gram - Rod Unknown

what does a pos/neg look like for ‘I’ test?

A

pos: red ring on top of stab

neg: no color change but drops of reagent remain yellow

133
Q

Gram - Rod Unknown

What does the abbreviation ‘MRVP’ stand for?

A

Methly Red Vogues Proskaver

134
Q

what does the MR in MRVP test for? what reagent is testing for this?

A

methyl red reagent (pH indicator) - tests for mixed acids (lactic, formic, etc.)

135
Q

Gram - Rod Unknown

  • Which reagent tests for the production of mixed acids by fermentation?
A

Methly red, after incubation

136
Q

Gram - Rod Unknown

How does the color of the reagent that tests for mixed acid fermentation change with decreasing pH? Mvrp

A

Red color change (acidic result)

137
Q

How does the color of the reagent that tests for mixed acid fermentation look like with negative change?

A

Yellow color change (neutral)

138
Q

What goes the Vogues Proskaver test for?

A

Testing for alcohol fermentation

139
Q

Gram - Rod Unknown

  • How many days does the 2,3-butanediol test using an MRVP tube incubate for?
A

5 days

140
Q

Gram - Rod Unknown

  • Why doesn’t 2,3-butanediol react with a pH indicator, like phenol red?
A

Because alcohol has a ph of 7

141
Q

Gram - Rod Unknown

  • What is the exact procedure for performing the VP test in lab?
A
142
Q

Gram - Rod Unknown

  • What is the purpose of shaking the small tube?
A

To oxidize the acetoin

143
Q

Gram - Rod Unknown

-Which final chemical product will be created in the vogues proskaver test?

A

diacetyl (wat we detect)

144
Q

Gram - Rod Unknown

  • How long does the VP test sit before the color change is complete?
A

30mins

145
Q

Gram - Rod Unknown

  • What color is a positive VP test? What color is a negative test result?
A

Positive: Deep red

Neg: Brown

146
Q

Gram - Rod Unknown

  • What kills Gram + & some Gram – in the Brilliant Green Agar?
A

brilliant green dye

147
Q

Gram - Rod Unknown

-What sugars are found in the Brilliant Green?

A

Lactose and Sucrose

148
Q

Gram - Rod Unknown

-What is the pH indicator for the BG agar?

A

Low pH = ferments sugar

High pH = Prefers peptides

149
Q

Gram - Rod Unknown

  • What color is brilliant green agar that is positive for fermentation (Acidic)? Is there growth?
A

Lime green color change with no grown

150
Q

Gram - Rod Unknown

  • What color is brilliant green agar that is negative for fermentation (Basic)?
A

Bright pink color change

151
Q

Gram - Rod Unknown

  • What color is brilliant green agar when the bacteria is dead (Also negative)?
A

Plate stays orange (original color)

152
Q

What does the Oxidase slant test for?

A

Testing for cytochrome oxidase

153
Q

Gram - Rod Unknown

What is the protein cytochrome oxidase normally used forin the oxidase slant?

A

transfers electrons from cytochrome C (protein) into oxygen

154
Q

Gram - Rod Unknown

Where specifically is cytochrome oxidase located?

A

Cytochrome oxidase is used in the electron transport chain (ETC)

155
Q

Gram - Rod Unknown

what is the reaction associated with oxidase test?

A

2e- + 2H+ + 1/2O2 —> H2O

156
Q

Gram - Rod Unknown

  • What color is a pos/neg cytochrome oxidase test?
A

pos: dark purple color change w/in 20 seconds

neg: no color change w/in 20 seconds (remains plain agar)

157
Q

Gram - Rod Unknown

  • What type of media was used for the oxidase test?
A

TSA slant

158
Q

Gram - Rod Unknown

-What is in the Hektoen agar that makes it selective?

A

bile

159
Q

Gram - Rod Unknown

  • What three carbohydrates does it contain?
A

Lactose, sucrose, salicin

160
Q

Gram - Rod Unknown

-What are the pH indicators in the Hektoen agar?

A
161
Q

Gram - Rod Unknown

  • What is the fermentation color change in the Hektoen agar? is there growth? Is it alkaline or acidic?
A

Orange color change (growth and acidic)

162
Q

Gram - Rod Unknown

-What is the non-fermenter color change for Hektoen agar?

A

Blue/Green color change

Cameron Lecture study guides (23 decks)

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