Physiological Tests PDF

Physiological Tests Test Name Carbohydrate Fermentation Tests Test Description Medium Used Provide energy Bact breakdown G,L,M to produce acid. pH decrease or no change Glucose PRB, Lactose PRB, Mannitol PRB IfPeptone is used,color changes to PINK! No Acid/Gas = -/-,Acid/Gas= A/G,Peptone=K Test Indi...

Physiological Tests Test Name Carbohydrate Fermentation Tests Test Description Medium Used Provide energy Bact breakdown G,L,M to produce acid. pH decrease or no change Glucose PRB, Lactose PRB, Mannitol PRB IfPeptone is used,color changes to PINK! No Acid/Gas = -/-,Acid/Gas= A/G,Peptone=K Test Indicator or pH indicator What Negative Test Looks Like Color Charge for results What Positive Test Looks Like no pH Change pH down Phenol Red (pH indicator) - acid: broth red  red/peach-orange; - gas: no bubbles Gelatin (test indicator) Solid  solid (no liquefaction) Solid  liquefaction + acid: broth red  yellow; + gas: bubbles EXOENZYME TESTS Use “2/3 ofneedle”1 time Gelatinase Test Add several drops ofGram’ s Iodine solution all over starch agar plate Amylase/Starch Hydrolysis Test Bact produces Gelatinase to breakdown gelatine Nutrient Gelatin Deep examine & record stab growth pattern in the medium (data entered under Cultural/Colony Characteristics -“Gelatin Deep” BACT PRODCES AMYLASE STARCH AGAR PLATE Breaks down Starch/Amylose POSITIVE NEGATIVE POSITIVE = Bact produced Amylase and used glucose producing CLEAR halo around the colony. Write results as "POSITIVE" TEST INDICATOR NEGATIVE = STARCH Present. Bact did not produce amylase and IODINE SOLUT. did not break down starch into glucose. Write results as "NEGATIVE" NO CLEAR HALO AROUND BACT keep screw cap loose! Urease Test 1 loopful ofculture with 1 fishtail streak ifbacteria can breakdown UREA by producing & secreting the enzyme URASE UREA AGAR SLANT The breakdown product ofUREA is AMMONIA Rapid diagnostic test for identification of Helicobacter pylori (urease producer) pH INDICATOR PHENOL RED Positive Test (tan to pink) Negative Test (tan to yellow) negative for ammonium compounds positive for ammonium compounds Increased pH (become alkaline) No change in pH (stays acidic) Write results as "NEGATIVE" Write results as "POSITIVE" No bubbles,looks like water Produces bubbles,looks white and bubbly For Results write "NEGATIVE" For Results write "POSITIVE" OXYDATIVE ENZYME TESTS add 1 drop of3% H2 O2 on well isolated colony Done on a glass "plate" Catalase Test rub applicator tip on well isolated colony;add 1 drop of oxidase reagent to applicator tip Oxidase Test ifbacteria produce the enzyme catalase (by adding hydrogen peroxide (H2O2) to the TSA plate) TEST INDICATOR TSA Plate *Catalase -detoxifies H2O2 by breaking it down into water and oxygen gas (H2O + O2) hydrogenperoxide (H2O2) Testing to see ifbacteria produce the enzyme cytochrome C oxidase by adding “oxidase reagent” TSA Plate (test indicator)to TSA plate or tip ofswab TEST INDICATOR OXIDASE REAGENT Swab doesnt change color For results write "NEGATIVE" TRYPTONE BROTH Indole Test DO NOT SHAKE Add 6 drops ofMethyl Red reagent rich in the amino acid TRYPTOPHAN *Testing to see which GLUCOSE FERMENTATION PATHWAY is used by bacteria Methyl Red Test Contains GLUCOSE SHAKE THE TUBE 2 loopfuls ofculture with 2 overlapping fishtail streaks Flame between loops Simons Citrate Test MR-VP BROTH TEST INDICATOR For results write "POSITIVE" When the electron donor is oxidized by cytochrome C oxidase it turns a dark purple ELECTRON TRANFER CHAIN Add 12 drops ofKovac’ s Reagent to tryptone broth.Wait *Testing to see ifbacteria produce enzyme TRYPTOPHANASE to breakdown tryptophan 5 min.Do it under fumehood into INDOLE Swab turns PURPLE YELLOW AT THE TOP CHERRY RED AT THE TOP For results write "POSITIVE" KOVAC' S REAGENT For results write "NEGATIVE" pH INDICATOR Ifcolor stays YELLOW Ifcolor changes to RED METHYL RED REAGENT For results write "NEGATIVE" For results write "POSITIVE" NO INDOLE YES INDOLE detects mixed acid fermentation ifbacteria can utilize citrate as a carbon source by producing enzyme citritase Citrate hydrolysis eventually produces an alkaline ammonium compounds Negative Test =GREEN,no change Positive Test = BLUE SIMMON' S pH INDICATOR negative for ammonium compounds positive for ammonium compounds CITRATE AGAR BROMOTHYMOLno change in pH (stays acidic) Increased pH (becomes alkaline) SLANT BLUE Write results as "POSITIVE" Write results as "NEGATIVE" contains sodium citrate as the sole source of carbon (energy source) Hydrogen Sulfide Test ifbacteria produce hydrogen sulfide (H2S)gas The amino acid cysteine contains sulfur cysteine desulfurase-producing bacteria will breakdown cysteine into H2S & other byproducts TEST INDICATOR KLIGLER' S IRON IRON DEEP OR TRIPPLE SUGAR IRON MEDIUM Negative Test (NO black) negative for H2S production Write results as "NEGATIVE" Positive Test (black precipitate) positive for H2S production Write results as "POSITIVE" 2/3 ofneedle TSA PLATE -Non-selective growth media;Used for isolation,observing colony morphology,developing a pure culture,and achieving enough growth for further biochemical testing and culture storage IMViC tests -a group of4 biochemical reaction tests used to determine the Genus name offacultative anaerobic,Gram-Negative bacilli