Oxygen Requirements Biol G210L PDF

Oxygen Requirements Biol G210L Hernandez Oxygen Requirements Biol G210L Hernandez baceria can be placed in 1 od 2 groups based on oxygen  Aerobes - Utilize oxygen (O2) (if a bacterium utilizes oxygen just mean that it uses...

Oxygen Requirements Biol G210L Hernandez Oxygen Requirements Biol G210L Hernandez baceria can be placed in 1 od 2 groups based on oxygen  Aerobes - Utilize oxygen (O2) (if a bacterium utilizes oxygen just mean that it uses it for its metaboliism and will use it asn electron transport chain) - 2 types: 1. Obligate aerobes: require O2 2. Microaerophiles: prefer low O2 and high CO2 -- low amount  Anaerobes - Don’t utilize O2 dont use oxygen in electron trasport chjain - 3 types: 1. Obligate anaerobe: cannot tolerate O2 will kill anareobic 2. 3. Loading… Aerotolerant: cannot utilize O2, but can tolerate it fairly well Facultative anaerobe: can live with or without O2, but prefers it still arent utilizing oxygen Culturing Anaerobes - Thioglycollate Broth - Candle Jar - Anaerobic Jar Thioglycollate Broth - Sodium thioglycollate: reducing agent combine with any free oxygen and converting and reducing it to water - Small amount of agar: reduces diffusion of O2 purpose is to reduce the amount of diffusion with oxygen - Resazurin: O2 indicator - Pink: O2 present Loading… - Colorless: O2 absent Thioglycollate Broth: Aerobes * Obligate Aerobe Microaerophile  Thioglycollate Broth: Anaerobes higher concentration at the top evenly throughout, can * tolerate oxyegn Obligate anaerobe Aerotolerant Facultative anaerobe Candle Jar - Put petri plates in jar with a candle - Light candle, close lid - Candle flame will consume most of the O2 in the jar and generate CO2 - microaerphiles require less amount of oxyegn and high amount of cO2 Anaerobic Jar - Creates anaerobic environment - Add petri plates to Anaerobic Jar - Add sachet - Absorbs O2 - Generates CO2 - Add O2 indicator (pink pill) - Active ingredient: methylene blue - Purple: aerobic - Pink: anaerobic  Oxygen Requirements Procedure: Thioglycollate Broth 1. Label broth 2. Aseptically inoculate one loopful of culture into Thioglycollate Broth 3. Incubate at 37oC, 24 hours  C-Jar Oxygen Requirements Procedure: Candle Jar, Anaerobic Jar, 37oC 1. Plates Divide a TSA plate into 4 sections. Label the plate C- Jar. Write names of orgs in each section. A-Jar 2. Label another TSA plate as described in Step 1. Label this plate A-Jar. Loading… 3. Label another TSA plate as described in Step 1. Label this plate 37oC. 4. Aseptically streak cultures into appropriately labeled 37oC sections on each plate. 5. Place C-Jar plate in Candle Jar. Incubate at 37oC, 24 hours. 6. Place A-Jar plate in Anaerobic Jar. Incubate at 37oC,  24 hours. Thioglycollate Results (pg. 67 Lab Manual) Alcaligenes faecalis Staphylococcus aureus Escherichia coli Clostridium sporogenes Results: 37oC, Candle Jar, Anaerobic Jar Lab Evaluation #5, pgs 23 – 24