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From Flashcardlet > Lab Final Zimmie09 > Flashcards

Lab Final Zimmie09 Flashcards

1
Q

What type of medium is phenol red fermentation

A

broth

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2
Q

what is in a phenol red fermentation medium

A

phenol red (ph indicator)
peptone- amino acid that can produce ammonia
carbohydrate- see if fermentation occurs in bacteria

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3
Q

what color is phenol red fermentation broth normally

A

red

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4
Q

what is also added to the phenol red fermentation test to detect gas

A

inverted durham tube

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5
Q

what causes the pH to increase in a phenol red fermentation test and what color

A

deamination of peptone amino acid produces ammonia

causes pink color

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6
Q

In phenol red fermentation test, how is fermentation determined

A

if acid is produced during fermentation which causes a yellow color change indicating a decrease in pH

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7
Q

In a phenol red fermentation test, how is gas present or determined

A

if there is a gas bubble in the inverted durham tube

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8
Q

What type of medium is a MR-VP test

A

broth medium

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9
Q

What ingredients are used in a MR-VP broth test

A

peptone, glucose and a phosphate buffer
peptone and glucose provide protein and fermentable carb
potassium phosphate resists pH changes in medium

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10
Q

What is the point of an MR test

A

see if organism is capable of performing mixed acid fermentation which is capable of overcoming potassium phosphate buffer and lower pH

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11
Q

What is added to MR test of MR-VP, and what are the end results

A

methyl red is added
red indicates ability to perform mixed acid fermentation (+)
yellow or orange shows not able to perform mixed acid fermentation (-)

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12
Q

what is the point of a VP test in MR-VP

A

see if organism can ferment glucose but also quickly convert acid products to acetoin and 2,3-butanediol

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13
Q

How is VP test performed of MR-VP, and what are the results

A

add KOH and alpha napthol which causes medium to oxidize acetoin to diacetyl which reacts with guanidine nuclei in peptone
red color produced due to fermentation of glucose and by products (+)
no change or copper color is no fermentation of glucose or by products (-)

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14
Q

The catalase test is part of what test

A

TSA slant

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15
Q

what is the point of catalase test in TSA slant

A

to see if organism produces enzyme catalase

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16
Q

in catalase test, what cycle does it use

A

ETC

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17
Q

a TSA test uses what medium

A

yellow slant

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18
Q

what shows a positive result in a catalase test (TSA)

A

H2O2 (hydrogen peroxide) is added
positive result would be immediate bubbles
this shows catalase enzyme is present

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19
Q

what shows a negative result in a catalase test (TSA)

A

no bubbles after hydrogen peroxide is added

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20
Q

The oxidase test is part of what test

A

TSA

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21
Q

what is the point of oxidase test in TSA slant

A

see if oxidase is present in organism can oxidize cytochrome C

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22
Q

How is oxidase test (TSA) performed and what are the results

A

take a q tip and take sample of bacteria grown on slant and add reagent to q tip
positive result- purple color change (+) oxidase present
negative result- no color change (-)

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23
Q

what type of medium is used in a nitrate reduction test

A

clear broth

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24
Q

what is the point of nitrate reduction test

A

to see if organism can convert nitrate to nitrite

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25
Q

what are the ingredients in the nitrate reduction test

A

beef extract, peptone, and potassium nitrate

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26
Q

what else is in the broth of a nitrate reduction test

A

inverted durham tube

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27
Q

what is the first step of the nitrate reduction test and what are the results

A

incubate broth and then after check to see if gas is present in durham tube
if gas present then (+) denitrification occurred, stop here
no gas present (-) then move to second step

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28
Q

what is the second step of the nitrate reduction test and what are the results

A

add A and B to broth
if red color change then (+) nitrite are present and react with reagent forming red color
no color change move to step 3

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29
Q

what is the third step of the nitrate reduction test and what are the results

A

add zinc powder (automatically causes nitrate to reduce to nitrite)
positive result would be no color change due to only nitrites being present
negative result is color change to red showing nitrates were present in the broth

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30
Q

what medium is used in a citrate test slant

A

green slant

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31
Q

what is the point of a citrate test

A

determine ability of organism to use citrate as its sole source of carbon also see if citrate-permease is present to convert citrate to pyruvate

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32
Q

what are the ingredients in a citrate test slant

A

sodium citrate (sole carbon source), ammonium phosphate (sole nitrogen source) and bromthymol blue dye (indicator)

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33
Q

what are the results of a citrate test

A

positive result shows use of citrate and ammonium phosphate converted to ammonia and ammonia hydroxide causing alkalinity increasing pH changing color to blue
no color change indicates negative result

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34
Q

what medium is used in a malonate test

A

green broth

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35
Q

what is the purpose of a malonate test

A

see if organism can use malonate as sole carbon source if it contains enzyme succinate dehydrogenase

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36
Q

what are the ingredients in a malonate test broth

A

sodium malonate, yeast extract, glucose (promote gowth) and bromthymol blue dye (indicator)

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37
Q

what is a positive result on a malonate test

A

if organism utilizes malonate then it alkanizes medium causing increase in pH and change to blue color

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38
Q

what is a negative result on a malonate test

A

if organism cant use malonate but ferment glucose then broth turns yellow/ green color or there is no color change

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39
Q

what type of medium is a phenylalanine deaminase test

A

clear slant

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40
Q

what ingredients does a phenylalanine deaminase test slant have

A

phenylalalnine

41
Q

what enzyme needs to be present for Ferric chloride to turn green

A

phenylalalnine deaminase

42
Q

what is added to a phenylalanine deaminase test slant to see what result you would get

A

ferric chloride FeCl3

43
Q

what re the results of a phenylalanine deaminase test

A

positive- FeCl3 turns green due to enzyme

negative- FeCl3 remains yellow, enzyme not present

44
Q

what type of medium is used in a starch hydrolysis test

A

clear agar

45
Q

what is starch made of

A

alpha-d-glucose subunits attaced by acetal linkages

has to be broken down because to large to pass through cell membrane

46
Q

what is the point of a starch hydrolysis test

A

to see if organism hydrolyzes starch

47
Q

what enzymes are present in order to hydrolyze starch

A

alpha-amylase and oligo-1,6-glucosidase

help break glycosidic linkages between sugar subunits

48
Q

what is in a starch hydrolysis test agar

A

beef extracr, soluble starch and agar

49
Q

what is added to starch hydrolysis test to see if enzymes are present

A

iodine

50
Q

what are the results of a starch hydrolysis test after iodine is added

A

positive- hydrolysis will prevent iodine from attaching leaving halo around innoculated area
negative- iodine will cover innoculated area

51
Q

what type of medium is used in a urea hydrolysis test

A

orange broth

52
Q

what are the ingredients in a urea hydrolysis test broth

A

urea, phenol red, peptone, glucose and potassium phosphate
peptone and glucoseprovide nutrients
potassium phosphate used as buffer to keep broth frm alkanizing due to peptone metabolism
phenol red used as pH indicator

53
Q

what enzyme is present to convert urea into ammonia and carbon dioxide

A

urease

54
Q

what are the results of a urea hydrolysis test

A

positive- pink indicates urease is used

negative- yellow or orange, urease is not present

55
Q

what type of medium is used in a gelatin hydrolysis test

A

clear gelatin in a tube

56
Q

what gelatin derived from

A

collagen

57
Q

what are the ingredients in a gelatin hydrolysis test

A

gelatin, peptone and beef extract

58
Q

how do you innoculate a gelatin hydrolysis test

A

stab the butt

59
Q

what is the point of a gelatin hydrolysis test

A

see if gelatinase is produced by organism

60
Q

how do the process for a gelatin hydrolysis test occur

A

stab the butt, incubate, comes out liquid, refrigerate and then see results

61
Q

what are the results for a gelatin hydrolysis test

A

after refrigeration,
positive- gelatin is liquified gelatinase is present
negative- gelatin is sold, gelatinase is not present

62
Q

what type of medium is used for a SIM test

A

clear/ yellow gelatin

63
Q

what does sim stand for

A

sulfur, indole, motility

64
Q

what ingredients are in a SIM medium

A

casein, animal tissue as a source of amino acids, iron containing compound, sulfur in the form of sodium thiosulfate

65
Q

what two enzymes are present in order for sulfur to be reduce to H2S (hydrogen sulfide gas)

A

cysteine deslfurase and thiosulfate reductase

66
Q

what is formed when H2S combines with iron

A

creates Ferric Sulfide which is a black precipitate

67
Q

what are the reactions for sulfur in the SIM medium

A

positive if black precipitate is formed due to enzymes producing H2S

68
Q

what enzyme needs to be present to produce indole

A

tryptophanase

69
Q

when tryptophan reacts with tryptophanase what are its products

A

indole, ammonia, and pyruvate

70
Q

what reagent do you add to a SIM test to determine if indole is present

A

kovacs reagent

71
Q

what are the possible reactions for the indole part of the SIM test

A

positive- kovac reagent reacts with indole to produce red top layer
negative- no color change to kavoc reagent due to no indole present

72
Q

what are the possible reactions for motility in a SIM test

A

positive- if organism moves away from stabbed area

negative- if organism stays where it was initially stabbed

73
Q

what does TSI test stand for

A

triple sugar iron

74
Q

what is the purpose of a TSI test

A

see if glucose, lactose and sucrose fermentation occur as well as sulfur reduction

75
Q

what type of medium is used for a TSI test

A

red slant

76
Q

what ingredients does the TSI test contain

A

animal proteins for carbon and nitrogen sources
contains ferrous sulfate and sodium thiosulfate sources of oxidized sulfur
contains phenol red as pH indicator
iron in ferrous sulfate which is used as indicator for sulfide

77
Q

why do you innoculate a TSI slant by stabbing and running it up the slant

A

show anaerobic fermentation and aerobic fermentation

78
Q

if glucose is fermented in the butt of a TSI slant what happens

A

butt of slant will turn yellow due to pH decreasing

if cant ferment butt stays red

79
Q

if organism can ferment sucrose and lactose in TSI slant what happens

A

slant becomes yellow due to decrease in pH

80
Q

what else can occur in TSI slant due to fermentation of carbohydrates

A

can cause gas production which will appear as fissures and lift the agar from bottom of tube

81
Q

if H2S is produced by organism in TSI slant what happens

A

ferrous sulfate reacts with H2S causing black precipitate

showing sulfur reduction and fermentation

82
Q

if organsim does not ferment carbohydrates but utilizes peptone and amino acids in TSI slant what happens

A

slant turns pink

83
Q

what are possible reactions in TSI slant test

A

yellow butt/ pink slant= glucose fermentation and acid production in butt but alkaline products on slant
yellow butt and slant= glucose, lactose and/or sucrose fermentation with acid accumulation
Pink butt and slant= no fermentation but peptone catabolized aerobically and anerobically with alkaline products
black precipitate in agar= sulfur reduction
agar breakage= gas production

84
Q

what are the ingredients in a litmus milk test

A

skim milk and azolitmin (ph indicator)
skim milk provides nutrients for gowth, lactose for fermentation and casein
azolitmin with is litmus

85
Q

what are the four reactions that can occur in a litmus test

A

lactose fermentation, reduction of litmus, casein coagulation and casein hydrolysis

86
Q

how does lactose fermentation occur

A

beta-galactosidase is present causing lactose to split into glucose and galactose
turns litmus pink and decrease pH

87
Q

how does casein coagulation occur in a litmus test

A

accumulation of acid, which causes acid clot

appears white at bottom and pink band at top

88
Q

how is reduction of litmus present in a litmus test

A

turns white

89
Q

how does digestion occur in a litmus test

A

occurs when enzymes are present and can digest acid clots and curds leaving a clear/brownish fluid

90
Q

what happens to partially digest casein in a litmus test

A

ammonia is produced causing the pH to increase and turning litmus blue
alkaline reaction

91
Q

what ingredients are in a mannitol salts agar

A

mannitol (carb), sodium chloride and phenol red (indicator)

92
Q

what type of medium is used for a mannitol salts agar

A

red agar

93
Q

if organism turns yellow in mannitol salts agar what happened

A

mannitol was used for fermentation which produced an acid causing decrease in ph and chage color to yellow

94
Q

what is the point of sodium chloride in a mannitol salts agar

A

makes it selective because salt concentration is so high can kill bacteria

95
Q

what happens if a pink color is produced in a mannitol salts agar

A

there is no fermentation of mannitol but organism can grow in sodium chloride

96
Q

results of gram stain

A

g- red

g+ purple/blue

97
Q

results of acid fast stain

A

acid fast + red

acid fast - blue/gree

98
Q

endospore stain results

A

spore producer + red with green dot in center

non spore producer- red

99
Q

capsule stain results

A

capsule + halo

capsule- no halo

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