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Lab Two, Part 2: Blood and MSA, pg 429 Lab Two, Part 1: Coagulase Test, pg 429

New This Week: Gram (+) Bacteria ID Review Gram + and - tests. Last week:. Exp 62: ID of Human Staph. and Strep. Pathogens. Lab Two, Part 2: Blood and MSA, pg 429 Lab Two, Part 1: Coagulase Test, pg 429. Physical Factors Exp 15: Temperature Exp 16: pH Exp 17/18: O 2 Requirements.

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Lab Two, Part 2: Blood and MSA, pg 429 Lab Two, Part 1: Coagulase Test, pg 429

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  1. New This Week: • Gram (+) Bacteria ID • Review Gram + and - tests Last week: Exp 62: ID of Human Staph. and Strep. Pathogens • Lab Two, Part 2: Blood and MSA, pg 429 • Lab Two, Part 1: Coagulase Test, pg 429 Physical Factors Exp 15: Temperature Exp 16: pH Exp 17/18: O2 Requirements Exp 29: Catalase Test • Lab Two, pg 196 • add H2O2 • Gram staining practice Week 10W

  2. Syllabus Corrections • Pg 3 • Week 12: Assign. #7 due • Week 13: Assign. #6 due (not # 8) • Pg 8 • Week 11: Prepare Assign. #7 (not #6) • Week 12: Quiz 9 covers definitions from Assign. #7 (not #6) : Prepare Assign. #6 (not #8)

  3. Exp 62: ID of Human Staph. And Strep. Pathogens: Blood and MSA Review Red blood cells on an agar plate are used to diagnose infection. On the left is a positive Staphylococcus infection, on the right a positive Streptococcuspyogenesculture. An MSA plate with Micrococcus sp. (1), Staphylococcus epidermis (2) and S. aureus (3) colonies.

  4. Exp 62: ID of Human Staph. And Strep. Pathogens: Coagulase Test Coagulase reacts with the fibrinogen in plasma, and precipitates into fibrin cells clump together , look “lumpy”. Test useful for differentiating Staphylococcus aureus from other Gram positive, catalase + cocci. Any degree of clotting, from a loose clot suspended in the plasma to a solid, immovable clot is a positive result. Plasma which liquid remained is a negative result.

  5. Exp 62: ID of Human Staph. And Strep. Pathogens: Coagulase Test fibrin

  6. Exp. 29: Catalase Test • Hydrogen peroxide (H2O2) • accumulation is toxic to cells. • Presence of bubbling or foaming • when adding H2O2 to a culture, • indicates that the hydrogen • peroxide is being broken down • by catalase, with the release of • oxygen gas. • Bubbling = pos. catalase • Absence of foaming/bubbling= • neg. catalase • (catalase is not present in cell)

  7. Grand Prismatic Spring, Yellowstone National Park (thermophiles) Bacteria Around Nuclear Reactors PHYSICAL FACTORS Below Freezing Salt Water Pockets in Antarctica Seafloor bacteria on ocean-bottom rocks are more abundant and diverse than previously thought. Biofilm build-up on tank walls—guaranteed tocontaminate any fuel

  8. Exp 15: Physical Factors: Temperature

  9. Exp 16: Physical Factors: pH of the Extracellular Environment

  10. Exp 17/18: Physical Factors: Atmospheric Oxygen Requirements How is distribution of growth affected by atmospheric oxygen ……..?

  11. TABLES 1 & 2 • Exp 15: Temperature • Media: 12 NUT slant • Culture: SA , ML, BST • Procedure: (refer to pg. 111) • Label each tube w/ org. and temp. • Inoculate with loop 4 slants w/each org. • Incubate 1 slant from each org. at either 4oC, 25oC, 37oC, and 55oC for 24 hrs. • TABLES 3 & 4 • Exp 16: pH of the Extracellular Environ. • Media: • 2 NUT broth @ pH 2 • 2 NUT broth @ pH 5 • 2 NUT broth @ pH 7 • 2 NUT broth @ pH 9 • Culture: EC & SA • Procedure: (refer to pg. 116) • Label each tube w/ • org. and pH. • Inoculate with ea. org in various pH tubes. • Incubate at 37oC for 24 hrs. BST- Bacillus stearothermophilus CS- Clostridium sporogenes • TABLES 5 & 6 • Exp. 17/18: Atmospheric • Oxygen Requirements • Media: • 3Thioglycollate broth • 3 NUT broth • Culture: EC, ML & CS • Procedure: (refer to pg. 127) • Label ea. tube w/org & media. • Inoculate each org. in • thioglycollate and NUT broth. • Place thioglycollate tubes in • anaerobic GasPack chamber. • Incubate all at 37oC for 24 hrs. SUC GLU LAC Week 10W • Gram staining practice

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