ACTIVITY 12

METABOLIC ACTIVITIES OF MICROORGANISMS

MEB32 Group 4
Abille, Jaydhelle Mae
Bechayda, Andrew
Gayeta, Nicole
Jove, Mary Jane D.

May 22, 2017

 Tests

Phenylalanine
Methyl Red

Production

Production

Production
Utilization
Proskauer
BMlasuc

Catalase
BMman

Oxidase
BMmal

Sulfide
BMglu

Lysine

Citrate
Microorganisms

Voges

Indole

Urea
Pseudomona
aeruginosa
Micrococcus
-
luteus
Bacillus
stearothermophil
us
Escherichia
+ - + + - + - + - + - - +
coli
Staphylococcus
+ + + + -
aureus
Proteus
+ - + + - + +/- +
vulgaris
 FERMENTATION OF CARBOHYDRATES

I. Fermentation of Glucose, Maltose, Lactose, Sucrose and Mannitol

Medium Basal medium

Fermentation of (Glu, Mal, Suc , Man, Lac,)
Test for
(Glucose, Maltose, Lactose, Sucrose and Mannitol)
Reagent Bromcresol Green

Active component of reagent Bromcresol Green

Positive indication/
Yellow/ Acidic, Fermentor of given Carbohydrate
conclusion
Negative indication Blue/ Basic, Non-Fermentor
II. Methyl Red Test

Medium MRVP Broth (Methyl Red Voges Proskauer Broth)

Test For Mixed Acid Fermentation Pathway

Reagent 5 Drops Methyl Red

Active Component of Reagent Methyl Red

Positive Indication/ Conclusion Stays Red/ Acidic, Mixed Acid Fermentation

Negative indication Yellow/ Basic, Non-Mixed Acid Fermentor

III. Voges Proskauer Test

Medium Mrvp Broth (Methyl Red Voges Proskauer Broth)

Test For Butelyne Glycol/ Butane Glycol Pathway

Reagent 10 Drops 40% KOH, 15 Drops Alpha Napthol Reagent

Active Component of Reagent Alpha Napthol

Dark Brown/ Acetoin & 2,3-Butanediol Products Of
Positive Indication/ Conclusion
Butanediol Fermentation Are Reduced To Guanidium Group
Negative indication Copper or Light Brown/ No Acetoin
 RESPIRATION OF CARBOHYDRATES

I. Oxidase Test

Medium NA (Nutrient Agar Plate)

Test For Cytochrome C Oxidase

Reagent Oxidase Reagent

Active Component of Reagent Tetramethyl-p-Phenylenediamine (TMPD)

Formation of Black Spots/ Cytochrome C Oxidase present,
Positive Indication/ Conclusion
TMPD reduced to TMDP+
Negative indication No Color Formation/ Cytochrome C Oxidase Absent
II. Catalase Test

Medium NA (Nutrient Agar Plate)

Test For SOD, Catalase

Reagent Catalase Reagent

Active Component of Reagent Hydrogen Peroxide

Positive Indication/ Conclusion Formation of Bubbles/ O2

Negative indication No Bubbles

 UTILIZATION OF AMINO ACIDS

I. Indole Production

Medium SIM butt (Sulfide Indole Motility)

Test For Tryptophanase/ Utilization of Tryptophan

Reagent 10 Drops Kovac's Reagent

Active Component of Reagent (P)-Dimethylamino Benzaldehyde

Red/ Indole was Removed from Tryptophan, Reacted W/
Positive Indication/ Conclusion
PDMB
Negative indication Yellow
II. Hydrogen Sulfide Production

Medium SIM butt (Sulfide Indole Motility)

Test For Sulfur Reduction

Reagent Sodium Thiosulfate in Medium

Active Component of Reagent Sodium Thiosulfate

Positive Indication/ Conclusion Black Precipitate/ Sodium Thiosulfate is Reduced to H2S

Negative indication Yellow

III. Lysine Decarboxylase

Medium LDC broth (Lysine Decarboxylase)

Test For Lysine Decarboxylase Enzyme/ Utilization of Lysine

Reagent Bromcresol Purple

Active Component of Reagent Bromcresol Purple

Purple/ Basic, CO2 Removed (Decarboxylized) from
Positive Indication/ Conclusion
Lysine, Lysine Broken Down to Cadaverine
Negative indication Orange/Acidic
IV. Phenylalanine Deamination

Medium PA slant (Phenylalanine Agar)

Test For Presence of Phenylalanine Deaminase

Reagent 10 Drops Ferric Chloride

Active Component of Reagent Ferric Chloride

Black Or Green Precipitate/ Ammonia Was Removed
Positive Indication/ Conclusion (Deamination) Fr Phenyalalanine, Resulting Phenylpyruvic
Acid Reacts with Iron to Produce Black Precipitate
Negative indication No Precipitate
 CITRATE UTILIZATION
Medium SCA slant (Simmons Citrate Agar)

Test For Citrate Utilization as Carbon Source

Reagent Bromthymol Blue

Active Component of Reagent Bromthymol Blue

Blue/ Basic, Citrate Broken Down to Oxaloacetate And
Acetate, Oxaloacetate is Broken Down to Acetate + Lactate
Positive Indication/ Conclusion
and CO2, CO2 Reacts with Sodium in the Medium to
Produce Sodium Carbonate W/C is Basic
Negative indication Green (No Color Change)
 UREA HYDROLYSIS
Medium CUA slant (Christensens Urea Agar)

Test For Urea Hydrolysis

Reagent Phenol Red

Active Component of Reagent Phenol Red

Positive Indication/ Conclusion Pink/ Urea is Broken Down into Ammonia, Basic

Negative indication Yellow/ Acidic, No Ammonia Produced