Microbiological culture
Microbiological culture
A microbiological
culture, or microbial culture, is a method of multiplying microbial
organisms by letting them reproduce in predetermined culture
media under controlled laboratory conditions. Microbial cultures are used
to determine the type of organism, its abundance in the sample being tested, or
both. It is one of the primary diagnostic methods of microbiology and
used as a tool to determine the cause of infectious disease by
letting the agent multiply in a predetermined medium. For example, a throat
culture is taken by scraping the lining of tissue in the back of the
throat and blotting the sample into a medium to be able to screen for harmful
microorganisms, such as Streptococcus pyogenes, the causative agent of
strep throat. Furthermore, the term culture is more generally used
informally to refer to "selectively growing" a specific kind of
microorganism in the lab.
Microbial cultures
are foundational and basic diagnostic methods used extensively as a research
tool in molecular biology. It is often essential to isolate a pure culture
of microorganisms. A pure (or axenic) culture is a population of cells or multicellular
organisms growing in the absence of other species or types. A
pure culture may originate from a single cell or single organism, in which case
the cells are genetic clones of one another.
For the purpose of
gelling the microbial culture, the medium of agarose gel (agar) is used. Agar
is a gelatinous substance derived from seaweed. A cheap substitute for
agar is guar gum, which can be used for the isolation and maintenance of thermophiles.
Bacterial culture
Microbiological
cultures can be grown in petri dishes of differing sizes that have a thin
layer of agar-based growth medium. Once the growth medium in the petri dish is
inoculated with the desired bacteria, the plates are incubated at the best
temperature for the growing of the selected bacteria (for example, usually at
37 degrees Celsius for cultures from humans or animals, or lower for
environmental cultures).
Another method of
bacterial culture is liquid culture, in which the desired bacteria are
suspended in liquid broth, a nutrient medium. These are ideal for preparation
of an antimicrobial assay. The experimenter would inoculate liquid broth with
bacteria and let it grow overnight (they may use a shaker for uniform growth).
Then they would take aliquots of the sample to test for the antimicrobial
activity of a specific drug or protein (antimicrobial peptides).
As an alternative, the
microbiologist may decide to use static liquid cultures. These cultures are not
shaken and they provide the microbes with an oxygen gradient.
Culture collections
Microbial culture
collections focus on the acquisition, authentication, production, preservation,
catalogueing and distribution of viable cultures of standard reference microorganisms, cell lines and other materials for research
in microbial systematics Culture collection are also repositories of type strains.
Major national culture
collections
ATCC (American Type Culture Collection)- Manassas, Virginia
NCTC-(National Collection of Type Cultures)-
Health Protection Agency,
London, United Kingdom
BCCM (Belgium
Coordinated Collection of Microorganism)- Ghent, Belgium
CIP (Collection d'Institut
Pasteur)- Paris, France
DSMZ (Deutsche
Sammlung von Mikroorganismen und Zellkulturen)-
Braunschweig, Germany
JCM (Japan Collection of Microorganisms)-
Tsukuba, Ibaraki, Japan
NCCB (Netherlands Culture Collection of
Bacteria)- Utrecht, Netherlands
NCIMB (National Collection of Industrial, Food
and Marine Bacteria)-
Aberdeen, Scotland
STCC (Spanish type culture collection, Valencia University)- Valencia, Spain
Virus and
phage culture
Virus or phage cultures require host
cells in which the virus or phage multiply. For bacteriophages, cultures are grown by
infecting bacterial cells. The phage can then be isolated from the resulting
plaques in a lawn of bacteria on a plate. Virus cultures are obtained from their
appropriate eukaryotic host cells.
Isolation of pure cultures
For single-celled eukaryotes, such as
yeast, the isolation of pure cultures uses the same techniques as for bacterial
cultures. Pure cultures of multicellular organisms are often more easily
isolated by simply picking out a single individual to initiate a culture. This
is a useful technique for pure culture of fungi,
multicellular algae,
and small metazoa, for example. m/M;
Developing pure culture techniques is
crucial to the observation of the specimen in question. The most common method
to isolate individual cells and produce a pure culture is to prepare a streak
plate. The streak plate method is a way to physically separate the microbial
population, and is done by spreading the inoculate back and forth with an
inoculating loop over the solid agar plate. Upon incubation, colonies will
arise and single cells will have been isolated from the biomass.
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