BIOD171 Final Lab Notebook

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Santa Fe College *

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171

Subject

Biology

Date

Dec 6, 2023

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pdf

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3

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Lab 9 Notebook Back to Home Page Title: KD09 Unknown Pathogen Analysis Objective: To identify an unknown pathogen(s) based on the given assays developed during this course. The goal is to correctly identify the 2 pathogens. Procedure: · Test 1: Gram staining – gram positive (purple); gram negative (pink) · Test 2: MacConkey plate – only gram negative grows; grows lactose fermenters · Test 3: Blood Agar Plate (BAP) – non-selective differential media contains RBCs, important nutrient source, tryptic soy agar yeast + RBCs · Test 4: EMB plate – (grows for gram negative, dark red color, lactose fermentation) selective and differential media; colonies turn green, partial lactose fermentation; colonies turn pink, no color change in the bacteria, doesn’t produce any enzymes that can break down lactose · Test 5: Catalase test (negative control had no bubbles) – determines the presence or absence of catalase (an enzyme used to breakdown hydrogen peroxide) · Test 6: Oxidase Test – (if oxidase negative it will be clear; if positive it will turn purple) determine the presence or absence of cytochrome c activity in bacteria · Test 7: Coagulase Test – (if it forms clumps it is positive; if it stays liquid then negative) determines the presence of absence of coagulase which are enzymes that play a role in the formation of blood clots Notes:
Two samples from the hospital was brought to your lab for diagnosis. After culturing the samples on LB agar plates another co-worker began the analysis. However, due to an emergency your co-worker has left the remaining work to you. Before leaving your co-worker informs you the 2 liquid cultures were accidently combined before the Gram strain was started. However, the 2 cultures remain separated on LB plates for future tests. Results: · Test 1: Gram staining o The cultures were mixed and viewed together. There is gram positive bacteria (purple in color/round – coccus clusters) and gram negative bacteria (pink in color/ bacillus rod-shaped) · Test 2: MacConkey plate – only gram negative grows; grows lactose fermenters o Culture A: no growth o Culture B: did grow; the plate is mostly red with light clearing around the streaking · Test 3: Blood Agar Plate – non-selective differential media o Culture A: growth; clearing around the streak – beta hemolysis o Culture B: growth; no clearing bacteria, white in color – gamma hemolysis · Test 4: EMB plate (grows for gram negative, dark red color, lactose fermentation) o Culture A: no growth (not able to ferment lactose) o Culture B: culture has a green metallic sheen (able to ferment) · Test 5: Catalase Test – (negative control had no bubbles) determines the presence or absence of catalase o Culture A: catalase positive (bubbles) o Culture B: catalase positive (bubbles)
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