2. Oxygen Requirements 1. Aerobes 2. Anaerobes 3. Facultative 4. Microaerophilic

3. Agar Stabs & Agar Shakes 1. Liquify 6 NA Deeps in boiling water 2. Boil for 10 minutes to drive out dissolved oxygen 3. Allow 3 NA Deeps to cool and solidify Agar Stabs 4. Temper 3 NA Deeps in water bath at 55 degrees C Agar Shakes

4. Agar Stabs 1. Single stab with Inoculating needle – straight down, straight back up 1. Micrococcus luteus 2. Staphylococcus epidermidis 3. Clostridium sporogenes Clostridium rubrum Clostridium butrycum

5. Agar Shakes 1. Inoculate with Inoculating Loop - “shake” test tube so bacteria are equally distributed 1. Micrococcus luteus 2. Staphylococcus epidermidis 3. Clostridium sporogenes Clostridium rubrum Clostridium butrycum

6. Agar Stab Aerobic

7. Agar Stab Anaerobic

8. Agar Stab Facultative

9. Agar Stab Microaerophilic

10. Fluid Thioglycollate Broths (FTB) Sodium thioglycollate Binds to and removes free oxygen Resazurin Anaerobic indicator O2 - PINK No O2 - Clear

11. Oxygen Reqirements

12. Aerobic Microaerophilic Anaerobic