Axenic
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In
biology Biology is the scientific study of life. It is a natural science with a broad scope but has several unifying themes that tie it together as a single, coherent field. For instance, all organisms are made up of cells that process hereditary i ...
, axenic (, ) describes the state of a
culture Culture () is an umbrella term which encompasses the social behavior, institutions, and norms found in human societies, as well as the knowledge, beliefs, arts, laws, customs, capabilities, and habits of the individuals in these groups.Tyl ...
in which only a single species, variety, or strain of
organism In biology, an organism () is any living system that functions as an individual entity. All organisms are composed of cells (cell theory). Organisms are classified by taxonomy into groups such as multicellular animals, plants, and ...
is present and entirely free of all other contaminating organisms. The earliest axenic cultures were of
bacteria Bacteria (; singular: bacterium) are ubiquitous, mostly free-living organisms often consisting of one biological cell. They constitute a large domain of prokaryotic microorganisms. Typically a few micrometres in length, bacteria were among ...
or unicellular
eukaryote Eukaryotes () are organisms whose cells have a nucleus. All animals, plants, fungi, and many unicellular organisms, are Eukaryotes. They belong to the group of organisms Eukaryota or Eukarya, which is one of the three domains of life. Bacte ...
s, but axenic cultures of many multicellular organisms are also possible. Axenic culture is an important tool for the study of
symbiotic Symbiosis (from Greek , , "living together", from , , "together", and , bíōsis, "living") is any type of a close and long-term biological interaction between two different biological organisms, be it mutualistic, commensalistic, or parasit ...
and
parasitic Parasitism is a close relationship between species, where one organism, the parasite, lives on or inside another organism, the host, causing it some harm, and is adapted structurally to this way of life. The entomologist E. O. Wilson has c ...
organisms in a controlled environment.


Preparation

Axenic cultures of
microorganism A microorganism, or microbe,, ''mikros'', "small") and ''organism'' from the el, ὀργανισμός, ''organismós'', "organism"). It is usually written as a single word but is sometimes hyphenated (''micro-organism''), especially in olde ...
s are typically prepared by
subculture A subculture is a group of people within a culture that differentiates itself from the parent culture to which it belongs, often maintaining some of its founding principles. Subcultures develop their own norms and values regarding cultural, poli ...
of an existing mixed culture. This may involve use of a
dilution series Dilution may refer to: * Reducing the concentration of a chemical * Serial dilution, a common way of going about this reduction of concentration * Homeopathic dilutions, Homeopathic dilution * Dilution (equation), an equation to calculate the rate ...
, in which a culture is successively diluted to the point where subsamples of it contain only a few individual organisms, ideally only a single individual (in the case of an asexual
species In biology, a species is the basic unit of classification and a taxonomic rank of an organism, as well as a unit of biodiversity. A species is often defined as the largest group of organisms in which any two individuals of the appropriate s ...
). These subcultures are allowed to grow until the identity of their constituent organisms can be ascertained. Selection of those cultures consisting solely of the desired organism produces the axenic culture. Subculture selection may also involve manually sampling the target organism from an uncontaminated growth front in an otherwise mixed culture, and using this as an
inoculum In biology, inoculum refers to the source material used for inoculation. ''Inoculum'' may refer to: * In medicine, material that is the source of the inoculation in a vaccine * In microbiology, propagules: cells, tissue, or viruses that are used t ...
source for the subculture. Axenic cultures are usually checked routinely to ensure that they remain axenic. One standard approach with microorganisms is to spread a sample of the culture onto an
agar plate An agar plate is a Petri dish that contains a growth medium solidified with agar, used to culture microorganisms. Sometimes selective compounds are added to influence growth, such as antibiotics. Individual microorganisms placed on the plate wil ...
, and to incubate this for a fixed period of time. The agar should be an enriched medium that will support the growth of common "contaminating" organisms. Such "contaminating" organisms will grow on the plate during this period, identifying cultures that are no longer axenic.


Experimental use

As axenic cultures are derived from very few organisms, or even a single individual, they are useful because the organisms present within them share a relatively narrow
gene pool The gene pool is the set of all genes, or genetic information, in any population, usually of a particular species. Description A large gene pool indicates extensive genetic diversity, which is associated with robust populations that can surv ...
. In the case of an asexual species derived from a single individual, the resulting culture should consist of
identical Two things are identical if they are the same, see Identity (philosophy). Identical may also refer to: * ''Identical'' (Hopkins novel), a 2008 young adult novel by Ellen Hopkins * ''Identical'' (Turow novel), a 2013 legal drama novel by Scott T ...
organisms (though processes such as
mutation In biology, a mutation is an alteration in the nucleic acid sequence of the genome of an organism, virus, or extrachromosomal DNA. Viral genomes contain either DNA or RNA. Mutations result from errors during DNA or viral replication, mi ...
and
horizontal gene transfer Horizontal gene transfer (HGT) or lateral gene transfer (LGT) is the movement of genetic material between Unicellular organism, unicellular and/or multicellular organisms other than by the ("vertical") transmission of DNA from parent to offsprin ...
may introduce a degree of variability). Consequently, they will generally respond in a more uniform and
reproducible Reproducibility, also known as replicability and repeatability, is a major principle underpinning the scientific method. For the findings of a study to be reproducible means that results obtained by an experiment or an observational study or in a ...
fashion, simplifying the interpretation of
experiment An experiment is a procedure carried out to support or refute a hypothesis, or determine the efficacy or likelihood of something previously untried. Experiments provide insight into Causality, cause-and-effect by demonstrating what outcome oc ...
s.


Problems

The axenic culture of some pathogens is complicated because they normally thrive within host tissues which exhibit properties that are difficult to replicate ''in vitro''. This is especially true in the case of intracellular pathogens. However, careful replication of key features of the host environment can resolve these difficulties (e.g. host
metabolite In biochemistry, a metabolite is an intermediate or end product of metabolism. The term is usually used for small molecules. Metabolites have various functions, including fuel, structure, signaling, stimulatory and inhibitory effects on enzymes, c ...
s,
dissolved oxygen Oxygen saturation (symbol SO2) is a relative measure of the concentration of oxygen that is dissolved or carried in a given medium as a proportion of the maximal concentration that can be dissolved in that medium at the given temperature. It ca ...
), such as with the
Q fever Q fever or query fever is a disease caused by infection with ''Coxiella burnetii'', a bacterium that affects humans and other animals. This organism is uncommon, but may be found in cattle, sheep, goats, and other domestic mammals, including ...
pathogen, ''
Coxiella burnetii ''Coxiella burnetii'' is an obligate intracellular bacterial pathogen, and is the causative agent of Q fever. The genus ''Coxiella'' is morphologically similar to ''Rickettsia'', but with a variety of genetic and physiological differences. ''C. ...
''.


See also

*
Asepsis Asepsis is the state of being free from disease-causing micro-organisms (such as pathogenic bacteria, viruses, pathogenic fungi, and parasites). There are two categories of asepsis: medical and surgical. The modern day notion of asepsis is deriv ...
*
Gnotobiotic animal Gnotobiosis (from Greek roots ''gnostos'' "known" and ''bios'' "life") refers to an engineered state of an organism in which all forms of life (i.e., microorganisms) in or on it, including its microbiota, have been identified. The term gnotobioti ...
**
Germ-free animal Germ-free organisms are multi-cellular organisms that have no microorganisms living in or on them. Such organisms are raised using various methods to control their exposure to viral, bacterial or parasitic agents. When known microbiota are intr ...
*
Sterilization (microbiology) Sterilization refers to any process that removes, kills, or deactivates all forms of life (particularly microorganisms such as fungi, bacteria, spores, and unicellular eukaryotic organisms) and other biological agents such as prions present in or ...


References

{{Bacteria Bacteria Bacteriology Biotechnology Cell biology Cell culture Microbiology techniques Microbiology terms