Sterilization refers to any process that eliminates In order to grow microorganisms in the laboratory, Bacto Skim Milk - for
to Skim Milk - for propagation of organisms
(removes) or kills all forms of life, including
transmissible agents (such as fungi, bacteria, viruses,
spore forms, etc.) present on a surface, contained in a
fluid, in medication, or in a compound such as
biological culture media. Sterilization can be achieved
by applying the proper combinations of heat,
chemicals, irradiation, high pressure, and filtration.
The term has evolved to include the disabling or
destruction of infectious proteins such as prions related
to Transmissible Spongiform Encephalopathies (TES).
Methods of Sterilization of Culture Media
Steam under pressure (instructor may demonstrate).
Autoclaving is by far the best method of sterilizing
most media. All hospitals are equipped to carry out
this procedure; the home pressure cooker can also be
used. The usual process is to subject the media to 15
lbs. pressure (121ºC.) for 20 minutes. Certain media,
for example those containing blood and special
proteins or sugars, cannot withstand the high
temperature and should not be autoclaved.
Fractional Sterilization. Media may also be sterilized
by using flowing steam. The temperature in this
procedure never rises above 100ºC., but must be
repeated for 3 or 4 consecutive days. An Arnold
sterilizer or an ordinary double boiler may be used for
this method. The usual practice is to expose the
material for one hour for 3 consecutive days.
Filtration. Bacteriologic filters are used whenever heat
cannot be used for sterilization. Examples of materials
which require filtration are: blood serums, ascetic
fluid, various sugar and amino acid solutions, enzymes
and exotoxins. Filters used are of three main types: (1)
diatomaceous earth filters, (2) unglazed porcelain
filters, and (3) asbestos filters.
Culture Media
various materials must be provided for the occurring in milk to demonstrate coagulation and
nourishment of the cell. These materials must be of proteolysis of casein
such a nature as to enhance the growth of specific
micro-organisms by supplying readily assimilated Sabouraud’s Agar - for cultivation and identification of
food. Some organisms require special substances to fungi
maintain growth. On the whole, however, the basic Media for Cultivation of Pathogens:
materials found in culture media are water and the
soluble constituents of meat. To this base many other Brain heart Infusion - liquid media for streptococci,
chemicals may be added to encourage the growth of pneumococci and meningococci
specific micro-organisms: for example, salt, agar,
Brain Veal Agar - principally for gonococci but good
peptone, sugar, gelatin, blood and serum. The pH of
for other pathogenic
the media is important, since most organisms require a
neutral base. The pH must be adjusted to bacteria considered difficult to cultivate
approximately 7.0 before tubing and sterilization.
Bordet-Gengou - used for cough plates in determining
For practical purposes and to ensure uniformity, most Hemophilus pertussis
hospitals and laboratories use commercially prepared
media. This simplifies the preparation considerably, Agar Base
since the addition of water is all that is usually needed.
Löffler Blood Serum - used in cultural diagnosis of
Following is a list of the more common prepared
diphtheria
media * and their uses. Complete directions for the
preparation of the media are given on the label. Liver Infusion Agar - used for cultivation of
Endameba histolytica
Classification of Media
* Supplied by Difco Laboratories, Inc., Detroit, Mich.
General Culture Media:
Differential Media:
Nutrient Agar - general culture media for less
fastidious micro-organisms Bismuth Sulfate Agar - isolation of Salmonella
typhosa from excreta
Nutrient Broth - general liquid media
S.S. Agar - isolation of Shigella and Salmonella
Tryptone Glucose - for determining number of bacteria
in Milk Extract Agar Crystal Violet Agar - used to inhibit staphylococci
Lactose Broth - presumptive test for water analysis
Endo Agar - confirmation for coliform group in water
and milk analysis
Nutrient Gelatin - for determining gelatin liquefaction