Aerobic and anaerobic bacteria can be identified by growing them in test tubes of thioglycollate broth:
1: Obligate aerobes need oxygen because they cannot ferment or respire anaerobically. They gather at the top of the tube where the oxygen concentration is highest.
2: Obligate anaerobes are poisoned by oxygen, so they gather at the bottom of the tube where the oxygen concentration is lowest.
3: Facultative anaerobes can grow with or without oxygen because they can metabolise energy aerobically or anaerobically. They gather mostly at the top because aerobic respiration generates more ATP than either fermentation or anaerobic respiration.
4: Microaerophiles need oxygen because they cannot ferment or respire anaerobically. However, they are poisoned by high concentrations of oxygen. They gather in the upper part of the test tube but not the very top.
5: Aerotolerant organisms do not require oxygen as they metabolise energy anaerobically. Unlike obligate anaerobes however, they are not poisoned by oxygen. They can be found evenly spread throughout the test tube.
Thioglycollate broth is a multi-purpose, enriched, differential medium used primarily to determine the oxygen requirements of microorganisms. Sodium thioglycolate in the medium consumes oxygen and permits the growth of obligate anaerobes. This, combined with the diffusion of oxygen from the top of the broth produces a range of oxygen concentrations in the media along its depth. The oxygen concentration at a given level is indicated by a redox sensitive dye like resazurine that turns pink in the presence of oxygen.
Thioglycollate broth medium is recommended to isolate strict anaerobes should an anaerobic infection be suspected.
^ abLeijh PC; van Zwet TL; ter Kuile MN; van Furth R (November 1984). "Effect of thioglycolate on phagocytic and microbicidal activities of peritoneal macrophages". Infection and Immunity46 (2): 448–452. PMC261553. PMID6500699.