The Ames Test for mammalian
environmental mutagenicity
The AmesTest
combines a bacterial revertant mutation assay
with a simulation of mammalian metabolism to produce a highly sensitive
test for mutagenic chemicals in the environment.
A rat liver homogenate
is prepared to produce a metabolically active extract (S9). [Above] The extract is combined
with a strain of his-
Salmonella bacteria: in the absence of
histidine, the bacteria are
unable to grow on minimal
medium (control result). [Below] The homogenate and bacterial strain
are combined with a suspected mutagenic substance (X). The
induction of revertant colonies
indicates that some his-
bacteria have mutated (reverted) to his+
, and therefore that substance X is a mutagen.
Different bacterial strains are sensitive to different types of mutation.
Initial experiments used the reversion assay without
a liver homogenate. However, It is important to realize that
mutagenicity, unlike toxicity, is not the result of ingestion of a
suspect substance, but rather the accumulation of the substance and its
breakdown products in the body. Use of a liver homogenate
simulates the metabolic breakdown of the suspected mutagen in a
mammalian system, and more accurately predicts mutagenicity of
substances ingested by humans. For example, sodium
nitrate (NaNO3), which occurs naturally in smoked
meat such as bacon, hot dogs, ham, etc., is not itself mutagenic.
However, when acted upon by HCl in
the
stomach,it
is converted to nitrous acid (HNO2),
which
has
been demonstrated to be a powerful mutagen by the Ames Test.
Bruce Ames (1928 - ) and his undergraduate
students tested large numbers of commercial products in student labs at
UC Berkeley when the test was first introduced in the 1970s. Many
common items such as hairspray and
food colours were discovered to be
mutagenic and were withdrawn from the market. Ames also established
that many mutagenic compounds are also carcinogenic.
Illustration
after Griffiths et al. © 1996; All text
material © 2010 by Steven M.
Carr