Lab 3 & 4.pdf - ~:luim'i L owls reacts ml Ferric 00d...

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Unformatted text preview: ~:léu’im'i L owl‘s reacts ml Ferric 00d Lab53&4 BIOiC EMICAL TE‘STS USED IN LAB THIS WEEK [ ‘ D ’r‘ nod-0. ' 83cm“): ammo + oefime ° Bile esculin hydrolysis Purpose: Diflereutiates microorganisms based on their ability to hydrolyze esculin to esculctin and dextrose. The esculetin reacts with ferric citrate in the medium to form a dark brown-black complex. Procedure: Using a loop, inoculate the media by streaking in a zig-zag pattern up the slant. Incubate for 24 hours at 37°C with loose cap. Interpretation: Positive: Blackening of the tube Negative: No color change or less than 1/2 of the slant is black . Catalase . Purpose: Differentiates microorganisms based on their ability to produce the catalase Enzyme. Catalase breaks hydrogen peroxide down to water and oxygen. (Caution: Too much hydrogen peroxide will “water down” the reaction and give a false negative result.) =‘ --- - " - Procedure: Using a loop, spread microorganism onto a slide. Add one drop of hydrogen peroxide. Interpretation: Positive: Bubbles Negative: No bubbles WW Purpose: Detection of coagulase that is excreted by Staphylococcus aurgys that causes lbl‘ifl clot formation: Procedure: Inoculate a loopfiil of organism into a coagulase (rabbit plasma) tube. Incubate for 24 hours at 37°C. Interpretation: Positive: Coagulation of the plasma (clot formation) (SW’b‘eb) Negative: No coagulation/clotformation(31\’\\\ \lQUiQ) 0 Hemolysis on SBA (Sheep Blood Agar) Purpose: Differentiates microorganisms based on their hemolytic activity on sheep blood agar. Procedure: Using a loop, streak the test organism onto a SBA plate (TSA with 5% sheep blood cells) using isolation technique. Incubate for ONLY 24 hours at 37°C. Observe for hemolytic activity. Interpretation: Alpha - incomplete hemolysis or greening of the media Beta - complete hemolysis or clearing of the media “ ,.=r-‘" . ""h'M-u. _E/M'MI'- -.' S ; . Q‘vae med‘ Q Labs 3 & 4 a MSA (Mannitol salt agar) Purpose: Differentiates microorganisms based on the' bility to grow at 7.5% NaCl and ferment mannitol. Procedure: Using a sterile loop, inoculate a MSA plate and streak for isolation. Incubate the plate at 37°C for 24 hours. Interpretation: Positive: Growth + / Acid +. Yellow colonies (acid produced by mannitol fermentation) L; CO“ fifm mom “0 \u'S‘l‘Oph Negative: Growth + / Acid — or Growth - 1’ Acid —. Colorless colonies (no yellow color i no acid production) or no growth. biasesteerht, on 8mm We. . ‘. mos “‘7" ”6333 RIP) ‘2 Phenol Red Carbohydrate Tubes (arabinose, lactose, trehalose, mannitol, sucrose, xylose, etc.) Purpose: Differentiates organisms based on carbohydrate fermentation. This media ccipntains specific carbohydrates and the pH indicator phenol red. When an organism is able to ‘ current the carbohydrate present, an acid or an acid and gas are produced and the indicator changes from red to yellow. If the carbohydrate is not fermented, the color will remain red. ' Procedure: Inoculate the tubes with a loopful of organism. Incubate at 37°C for 48 hdurs. Interpretation: _ {301 Positive (fermentation): Yellow color Negative: Red color —"- <1th .10 ton—eerie duo in SWIG“ 1 Starch hydrolysis Purpose: Difi‘erentiates microorganisms based on their ability to hydrolyze starch. Procedure: Using a sterile 100p, inoculate a starch agar plate by streaking one line across plate. Incubate the plate at 37°C for 24 hours. Add enough Gram’s iodine to just cover the surface of agar. Interpretation: Positive: Dark agar with clear zone around streak Negative: D- - - :. uni - the streak \ ... y _ . it «throws W OP Ugmfim ~ 39mm mocha Lab 3 and Lab 4: Gram Positive Cocci and Rods Gmmsmm Biochemical Test sm #1 Catalase - Bile Esculin Hydrolysis Hemolysis pattern on SBA plate Acid production from utilization ofvmannitol Unknown Or- " '- -- arm ._--..e_z:=-:: .'.'-E|?est=:r Catalase . Growth and Acid production on an MSA plate Coagulase Unknown Crush 7 End product! What is tested for Catalase ’65. 7‘ m I hf anism #2 - _ ism #3 - Hemolysis pattern on SBA plate m Growth and Acid production on an MSA plate Starch Hydrolysis - Identification of Unknown culture #1 Results End product I What is tested for End product i What is tested form-w - _'2 -a mm ldentification of Unknown culture #2 identification of Unknown culture #3 l 106 l M US C 3? ECU; n C\ dammw Ufimm 6(3er 5mm @KQHW" warms ® 3Co+ mo 3% 7943* *Cdflsudfl ‘13 Q VerrFurm )vw whn Fomw 52mg 7) Sewes \f ChorHo xderm‘fj W t mmTEEM , “Nb scaM’mn * 5-D qJesw-yun vxanow orgamsn ° on‘ 3 (mm W0?” P Wag/3:3“ W coagumse (4: ma PW?- ! \ \z’rmw , Mam \Cfifi? 3 *0 msam dthcpcm *9 . «Shame 0M. 0 S 0k P1 .969 Wefichqu/e {On ow magma ”mm mm, - vam‘mm 7i!" ...
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