Topic: Anaerobic organism

An anaerobic organism or anaerobe is any organism that does not require molecular oxygen
for growth. It may react negatively or even die if free oxygen is present. In contrast, an
aerobic organism (aerobe) is an organism that requires an oxygenated environment.
Anaerobes may be unicellular (e.g. protozoans, bacteria) or multicellular.

Most fungi are obligate aerobes, requiring oxygen to survive. However, some species, such as
the Chytridiomycota that reside in the rumen of cattle, are obligate anaerobes; for these
species, anaerobic respiration is used because oxygen will disrupt their metabolism or kill
them. Deep waters of the ocean are a common anoxic environment.

First observation
In his letter of 14 June 1680 to The Royal Society, Antonie van Leeuwenhoek described an
experiment he carried out by filling two identical glass tubes about halfway with crushed
pepper powder, to which some clean rain water was added. Van Leeuwenhoek sealed one of
the glass tubes using a flame and left the other glass tube open. Several days later, he
discovered in the open glass tube 'a great many very little animalcules, of divers sort having
its own particular motion.' Not expecting to see any life in the sealed glass tube, Van
Leeuwenhoek saw to his surprise 'a kind of living animalcules that were round and bigger
than the biggest sort that I have said were in the other water.' The conditions in the sealed
tube had become quite anaerobic due to consumption of oxygen by aerobic microorganisms.

In 1913 Martinus Beijerinck repeated Van Leeuwenhoek's experiment and identified

Clostridium butyricum as a prominent anaerobic bacterium in the sealed pepper infusion tube
liquid. Beijerinck commented: 'We thus come to the remarkable conclusion that, beyond
doubt, Van Leeuwenhoek in his experiment with the fully closed tube had cultivated and seen
genuine anaerobic bacteria, which would happen again only after 200 years, namely about
1862 by Pasteur. That Leeuwenhoek, one hundred years before the discovery of oxygen and
the composition of air, was not aware of the meaning of his observations is understandable.
But the fact that in the closed tube he observed an increased gas pressure caused by
fermentative bacteria and in addition saw the bacteria, prove in any case that he not only was
a good observer, but also was able to design an experiment from which a

conclusion could be drawn.'

Classification
For practical purposes, there are three categories of anaerobe:

Obligate anaerobes, which are harmed by the presence of oxygen. Two examples of obligate
anaerobes are Clostridium botulinum and the bacteria which live near hydrothermal vents on
the deep-sea ocean floor.

Aerotolerant organisms, which cannot use oxygen for growth, but tolerate its presence.
Facultative anaerobes, which can grow without oxygen but use oxygen if it is
present.However, this classification has been questioned after recent research showed that
human "obligate anaerobes" (such as Finegoldia magna or the methanogenic archaea
Methanobrevibacter smithii) can be grown in aerobic atmosphere if the culture medium is
supplemented with antioxidants such as ascorbic acid, glutathione and uric acid.

Energy metabolism
Some obligate anaerobes use fermentation, while others use anaerobic respiration.
Aerotolerant organisms are strictly fermentative. In the presence of oxygen, facultative
anaerobes use aerobic respiration; without oxygen, some of them ferment; some use
anaerobic respiration.

Fermentation

There are many anaerobic fermentative reactions.

Fermentative anaerobic organisms mostly use the lactic acid fermentation pathway:

C6H12O6 + 2 ADP + 2 phosphate → 2 lactic acid + 2 ATP + 2 H2OThe energy released in

this reaction (without ADP and phosphate) is approximately 150 kJ per mol, which is
conserved in generating two ATP from ADP per glucose. This is only 5% of the energy per
sugar molecule that the typical aerobic reaction generates taking advantage of the high energy
of O2.Plants and fungi (e.g., yeasts) in general use alcohol (ethanol) fermentation when
oxygen becomes limiting:

C6H12O6 (glucose) + 2 ADP + 2 phosphate → 2 C2H5OH + 2 CO2↑ + 2 ATP + 2 H2OThe

energy released is about 180 kJ per mol, which is conserved in generating two ATP from
ADP per glucose.

Anaerobic bacteria and archaea use these and many other fermentative pathways, e.g.,
propionic acid fermentation, butyric acid fermentation, solvent fermentation, mixed acid
fermentation, butanediol fermentation, Stickland fermentation, acetogenesis, or
methanogenesis.

Culturing anaerobes
Since normal microbial culturing occurs in atmospheric air, which is an aerobic environment,
the culturing of anaerobes poses a problem. Therefore, a number of techniques are employed
by microbiologists when culturing anaerobic organisms, for example, handling the bacteria in
a glovebox filled with nitrogen or the use of other specially sealed containers, or techniques
such as injection of the bacteria into a dicot plant, which is an environment with limited
oxygen. The GasPak System is an isolated container that achieves an anaerobic environment
by the reaction of water with sodium borohydride and sodium bicarbonate tablets to produce
hydrogen gas and carbon dioxide. Hydrogen then reacts with oxygen gas on a palladium
catalyst to produce more water, thereby removing oxygen gas. The issue with the GasPak
method is that an adverse reaction can take place where the bacteria may die, which is why a
thioglycollate medium should be used. The thioglycollate supplies a medium mimicking that
of a dicot, thus providing not only an anaerobic environment but all the nutrients needed for
the bacteria to thrive.Recently, a French team evidenced a link between redox and gut
anaerobes based on clinical studies of severe acute malnutrition. These findings led to the
development of aerobic culture of "anaerobes" by the addition of antioxidants in the culture
medium.

Multicellularity
Few multicellular life forms are anaerobic, since only O2 with its weak double bond can
provide enough energy for complex metabolism. Exceptions include three species of
Loricifera (< 1 mm in size) and the 10-cell Henneguya zschokkei.In 2010 three species of
anaerobic loricifera were discovered in the hypersaline anoxic L'Atalante basin at the bottom
of the Mediterranean Sea. They lack mitochondria which contain the oxidative
phosphorylation pathway, which in all other animals combines oxygen with glucose to
produce metabolic energy, and thus they consume no oxygen. Instead these loricifera derive
their energy from hydrogen using hydrogenosomes.

References