Hydrateam Test D

Front 1

Name the FIVE test tube tests that use broths and what they are used for.

Back 1

>Carbohydrate broths - use or fermentation of sugars
(lactose, glucose, sucrose, mannitol)
>Urea broth - breakdown of urea by urease to ammonia and CO2
>MR/VP broth - MR/VP glucose broth w/2 tests for glucose utilization (MR) & production of acetoin (VP)
> Nitrate Reduction - breakdown of nitrate to nitrite or N2
>MPN - lactose broth testing for facultative anaerobic Gram neg rods that can ferment lactose at 44.5C

Front 2

Name the SIX test tube tests that use agar (including slants) and what they are used for.

Back 2

>Simmons Citrate - use of citrate or not
>Deaminase - deamination of amino acid: phenylanaline
>Indole Test - production of indole from tryptophan using tryptophanase (in either SIM or tryptone broth)
>Sulfate Reduction - reduction of sulfate by cysteine desulfurase to hydrogen sufide then to iron sulfide in SIM.
> SIM or TTC motility tests (In TTC growth = red)

Front 3

Describe this test tube test, what it tests for, any indicators it uses and anything unusual about it:
Carbohydrate Broths

Back 3

Test for use or fermentation of sugars including lactose, glucose, sucrose and mannitol.
Primary indicator is phenyl red which turns yellow due to the acid of sugar fermentation.
Secondary indicator is gas formation (CO2) trapped by inverted Durham Tube.
Test can be unreliable (false color) if left too long and organism starts to process proteins and/or other nutrients

Front 4

Describe this test tube test, what it tests for, any indicators it uses and anything unusual about it:
Urea Broth

Back 4

Test for differentiation based on production of urease to break down urea to ammonia (basic) and CO2.
Indicator is phenol red which changes from light orange to hot pink which is positive for urea hydrolysis.

Front 5

Describe this test tube test, what it tests for, any indicators it uses and anything unusual about it:
MR of MR/VP broth

Back 5

One MR/VP glucose broth for two tests. After innoculation in the broth, it is split into two tubes for the two tests. For MR test, Methyl Red reagent is added. Red color is positive for mixed acid fermentation of glucose. Yellow or orange is negative.

Front 6

Describe this test tube test, what it tests for, any indicators it uses and anything unusual about it:
VP of MR/VP broth

Back 6

One MR/VP glucose broth for two tests. After innoculation in the broth, it is split into two tubes for the two tests. For the VP test, Barritt's A & B are added and left for 1 hour. Test is for production of acetoin during the neutral metabolism of pyruvate to butylene glycol. A red ring at the top of the tube is positive.

Front 7

Describe this test tube test, what it tests for, any indicators it uses and anything unusual about it:
Nitrate Reduction broth

Back 7

Tests for ability to reduce nitrate (NO3) (or not) to one of three states (need to know if fermenter or not):
Nitrate to Nitrogen gas (N2) (yellow & N2 gas if non-F)
Nitrate to Nitrite (NO2) (add Nitrate A&B) red = positive
Nitrate to ammonia (NH3) (add Zn) yellow = positive
Red after adding zinc = true negative for reduction of NO3

Front 8

Describe this test tube test, what it tests for, any indicators it uses and anything unusual about it:
Simmons Citrate Slant.

Back 8

Tests for use of citrate or not as a sole carbon source. The conversion of citrate to pyruvate produces CO2 which combines with sodium to make NaCO2 which raises the pH changing the bromthymol blue indicator from neutral green to basic royal blue. Separates fecal coliforms (neg) from non-fecal coliforms (pos).

Front 9

Describe this test tube test, what it tests for, any indicators it uses and anything unusual about it:
Demainase Slant

Back 9

Tests for ability to deaminate the amino acid phenylalanine to form phenylpyruvic acid (PPA) and ammonia. After growth on the slant, 10-15 drops of ferric chloride (FeCl3) is added. Color change to avocado green is positive.

Front 10

Describe this test tube test, what it tests for, any indicators it uses and anything unusual about it:
Indole of SIM agar (or in tryptone broth)

Back 10

Tests for indole production from tryptophan using the enzyme tryptophanase in either SIM agar or tryptone broth. Done in conjunction with Sulfate reduction test. Ten drops of KOVAC's reagent is added. If indole is present a bright red ring or layer will be at the top of the tube. Yellow = negative.

Front 11

Describe this test tube test, what it tests for, any indicators it uses and anything unusual about it:
Sulfate Reduction of SIM agar

Back 11

Tests for reduction of sulfate. Done in conjunction with Indole test. Sulfate is reduced by the enzyme cysteine desulfurase to hydrogen sulfide which then combines with the iron in the media to become iron sulfide which is a black precipitate. A red ring at the top and black below is positive for both indole and hydrogen sulfide (H2S).

Front 12

Describe this test tube test, what it tests for, any indicators it uses and anything unusual about it:
Motility Test of SIM agar

Back 12

SIM agar's third test is for motility. Non-motile organisms will only grow along a stab line. Motile will spread throughout.

Front 13

Describe this test tube test, what it tests for, any indicators it uses and anything unusual about it:
TTC

Back 13

TTC agar contains a tetrazolium salt that changes color to red where bacterial growth is located. Non-motile organisms will only grow along a stab line. Motile will spread throughout.

Front 14

Describe this test tube test, what it tests for, any indicators it uses and anything unusual about it:
MPN

Back 14

MPN=Most Probable Number, which is the means of estimating the presence and/or quantity of fecal coliforms in water analysis. Lactose broth in both single and double strength is the medium in tubes with inverted Durham tubes to capture fermentation gas of facultative anaerobic Gram neg rods that can ferment lactose at 44.5C.