Aerotolerant anaerobes use fermentation to produce ATP. They do not use oxygen, but they can protect themselves from reactive oxygen molecules. In contrast, obligate anaerobes can be harmed by reactive oxygen molecules.
There are three categories of anaerobes. Where obligate aerobes require oxygen to grow, obligate anaerobes are damaged by oxygen, aerotolerant organisms cannot use oxygen but tolerate its presence, and facultative anaerobes use oxygen if it is present but can grow without it.
An obligate aerobe is an organism that requires oxygen to grow. Through cellular respiration, these organisms use oxygen to metabolise substances, like sugars or fats, to obtain energy. In this type of respiration, oxygen serves as the terminal electron acceptor for the electron transport chain. Aerobic respiration has the advantage of yielding more energy (adenosine triphosphate or ATP) than fermentation or anaerobic respiration, but obligate aerobes are subject to high levels of oxidative stress.
Thioglycolate broth is a multipurpose, enrichment, 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 medium along its depth. The oxygen concentration at a given level is indicated by a redox-sensitive dye such as resazurin that turns pink in the presence of oxygen.
This allows the differentiation of obligate aerobes, obligate anaerobes, facultative anaerobes, microaerophiles, and aerotolerant organisms. For example, obligately anaerobic Clostridium species will be seen growing only in the bottom of the test tube.
Obligate anaerobes are microorganisms killed by normal atmospheric concentrations of oxygen (20.95% O2). Oxygen tolerance varies between species, with some species capable of surviving in up to 8% oxygen, while others lose viability in environments with an oxygen concentration greater than 0.5%.
Obligate anaerobes, which die when normal amounts of oxygen are present, are contrasted with obligate aerobes, which die without oxygen. Bacteria that fall in between these two extremes may be classified as either facultative anaerobes, which can use oxygen but also survive without it, or microaerophiles, which need lower levels of oxygen. Aerotolerant organisms are indifferent to the presence or absence of oxygen.
A microaerophile is a microorganism that requires environments containing lower levels of dioxygen than those present in the atmosphere (i.e. < 21% O2; typically 2–10% O2) for optimal growth. A more restrictive interpretation requires the microorganism to be obligate in this requirement. Many microaerophiles are also capnophiles, requiring an elevated concentration of carbon dioxide (e.g. 10% CO2 in the case of Campylobacter species).
The original definition of a microaerophile has been criticized for being too restrictive and not accurate enough compared to similar categories. The broader term microaerobe has been coined to describe microbes able to respire oxygen "within microoxic environments by using high-affinity terminal oxidase".