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Lab Final Zimmie09 Flashcards

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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
what are the ingredients in the nitrate reduction test
beef extract, peptone, and potassium nitrate
26
what else is in the broth of a nitrate reduction test
inverted durham tube
27
what is the first step of the nitrate reduction test and what are the results
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
28
what is the second step of the nitrate reduction test and what are the results
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
29
what is the third step of the nitrate reduction test and what are the results
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
30
what medium is used in a citrate test slant
green slant
31
what is the point of a citrate test
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
32
what are the ingredients in a citrate test slant
sodium citrate (sole carbon source), ammonium phosphate (sole nitrogen source) and bromthymol blue dye (indicator)
33
what are the results of a citrate test
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
34
what medium is used in a malonate test
green broth
35
what is the purpose of a malonate test
see if organism can use malonate as sole carbon source if it contains enzyme succinate dehydrogenase
36
what are the ingredients in a malonate test broth
sodium malonate, yeast extract, glucose (promote gowth) and bromthymol blue dye (indicator)
37
what is a positive result on a malonate test
if organism utilizes malonate then it alkanizes medium causing increase in pH and change to blue color
38
what is a negative result on a malonate test
if organism cant use malonate but ferment glucose then broth turns yellow/ green color or there is no color change
39
what type of medium is a phenylalanine deaminase test
clear slant
40
what ingredients does a phenylalanine deaminase test slant have
phenylalalnine
41
what enzyme needs to be present for Ferric chloride to turn green
phenylalalnine deaminase
42
what is added to a phenylalanine deaminase test slant to see what result you would get
ferric chloride FeCl3
43
what re the results of a phenylalanine deaminase test
positive- FeCl3 turns green due to enzyme | negative- FeCl3 remains yellow, enzyme not present
44
what type of medium is used in a starch hydrolysis test
clear agar
45
what is starch made of
alpha-d-glucose subunits attaced by acetal linkages | has to be broken down because to large to pass through cell membrane
46
what is the point of a starch hydrolysis test
to see if organism hydrolyzes starch
47
what enzymes are present in order to hydrolyze starch
alpha-amylase and oligo-1,6-glucosidase | help break glycosidic linkages between sugar subunits
48
what is in a starch hydrolysis test agar
beef extracr, soluble starch and agar
49
what is added to starch hydrolysis test to see if enzymes are present
iodine
50
what are the results of a starch hydrolysis test after iodine is added
positive- hydrolysis will prevent iodine from attaching leaving halo around innoculated area negative- iodine will cover innoculated area
51
what type of medium is used in a urea hydrolysis test
orange broth
52
what are the ingredients in a urea hydrolysis test broth
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
what enzyme is present to convert urea into ammonia and carbon dioxide
urease
54
what are the results of a urea hydrolysis test
positive- pink indicates urease is used | negative- yellow or orange, urease is not present
55
what type of medium is used in a gelatin hydrolysis test
clear gelatin in a tube
56
what gelatin derived from
collagen
57
what are the ingredients in a gelatin hydrolysis test
gelatin, peptone and beef extract
58
how do you innoculate a gelatin hydrolysis test
stab the butt
59
what is the point of a gelatin hydrolysis test
see if gelatinase is produced by organism
60
how do the process for a gelatin hydrolysis test occur
stab the butt, incubate, comes out liquid, refrigerate and then see results
61
what are the results for a gelatin hydrolysis test
after refrigeration, positive- gelatin is liquified gelatinase is present negative- gelatin is sold, gelatinase is not present
62
what type of medium is used for a SIM test
clear/ yellow gelatin
63
what does sim stand for
sulfur, indole, motility
64
what ingredients are in a SIM medium
casein, animal tissue as a source of amino acids, iron containing compound, sulfur in the form of sodium thiosulfate
65
what two enzymes are present in order for sulfur to be reduce to H2S (hydrogen sulfide gas)
cysteine deslfurase and thiosulfate reductase
66
what is formed when H2S combines with iron
creates Ferric Sulfide which is a black precipitate
67
what are the reactions for sulfur in the SIM medium
positive if black precipitate is formed due to enzymes producing H2S
68
what enzyme needs to be present to produce indole
tryptophanase
69
when tryptophan reacts with tryptophanase what are its products
indole, ammonia, and pyruvate
70
what reagent do you add to a SIM test to determine if indole is present
kovacs reagent
71
what are the possible reactions for the indole part of the SIM test
positive- kovac reagent reacts with indole to produce red top layer negative- no color change to kavoc reagent due to no indole present
72
what are the possible reactions for motility in a SIM test
positive- if organism moves away from stabbed area | negative- if organism stays where it was initially stabbed
73
what does TSI test stand for
triple sugar iron
74
what is the purpose of a TSI test
see if glucose, lactose and sucrose fermentation occur as well as sulfur reduction
75
what type of medium is used for a TSI test
red slant
76
what ingredients does the TSI test contain
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
why do you innoculate a TSI slant by stabbing and running it up the slant
show anaerobic fermentation and aerobic fermentation
78
if glucose is fermented in the butt of a TSI slant what happens
butt of slant will turn yellow due to pH decreasing | if cant ferment butt stays red
79
if organism can ferment sucrose and lactose in TSI slant what happens
slant becomes yellow due to decrease in pH
80
what else can occur in TSI slant due to fermentation of carbohydrates
can cause gas production which will appear as fissures and lift the agar from bottom of tube
81
if H2S is produced by organism in TSI slant what happens
ferrous sulfate reacts with H2S causing black precipitate | showing sulfur reduction and fermentation
82
if organsim does not ferment carbohydrates but utilizes peptone and amino acids in TSI slant what happens
slant turns pink
83
what are possible reactions in TSI slant test
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
what are the ingredients in a litmus milk test
skim milk and azolitmin (ph indicator) skim milk provides nutrients for gowth, lactose for fermentation and casein azolitmin with is litmus
85
what are the four reactions that can occur in a litmus test
lactose fermentation, reduction of litmus, casein coagulation and casein hydrolysis
86
how does lactose fermentation occur
beta-galactosidase is present causing lactose to split into glucose and galactose turns litmus pink and decrease pH
87
how does casein coagulation occur in a litmus test
accumulation of acid, which causes acid clot | appears white at bottom and pink band at top
88
how is reduction of litmus present in a litmus test
turns white
89
how does digestion occur in a litmus test
occurs when enzymes are present and can digest acid clots and curds leaving a clear/brownish fluid
90
what happens to partially digest casein in a litmus test
ammonia is produced causing the pH to increase and turning litmus blue alkaline reaction
91
what ingredients are in a mannitol salts agar
mannitol (carb), sodium chloride and phenol red (indicator)
92
what type of medium is used for a mannitol salts agar
red agar
93
if organism turns yellow in mannitol salts agar what happened
mannitol was used for fermentation which produced an acid causing decrease in ph and chage color to yellow
94
what is the point of sodium chloride in a mannitol salts agar
makes it selective because salt concentration is so high can kill bacteria
95
what happens if a pink color is produced in a mannitol salts agar
there is no fermentation of mannitol but organism can grow in sodium chloride
96
results of gram stain
g- red | g+ purple/blue
97
results of acid fast stain
acid fast + red | acid fast - blue/gree
98
endospore stain results
spore producer + red with green dot in center | non spore producer- red
99
capsule stain results
capsule + halo | capsule- no halo

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