rods, mostly single or double, held
together by a polysaccharide called a
glia. Some of the cells resemble
Oenococcus or Pediococcus, but
Acetobacter is noticeably larger when
compared to reference cultures of the
other bacteria (for example, to freezedried
Oenococcus used for cellar additions).
In wine, Acetobacter is not always
visually distinct from Lactobacillus.
However, Acetobacter cells are often
wider than Lactobacillus, and taper at
the ends. Acetobacter chains also tend
to be composed of shorter cells than
Lactobacillus though not always. In culture,
though not in wine, Acetobacter
produces strange involution forms
that resemble crudely formed, lumpy
snakes made out of clay. These forms
distinguish Acetobacter in culture from
other rod-shaped microbes.
In culture, surface film yeasts grow
readily on yeast media, often producing
wrinkled colonies, though a few
species produce smooth colonies.
While species can be distinguished by
genetic sequencing, this not commonly
useful to winemakers because
the remedies (eliminate headspace,
increase molecular SO2) do not differ.
The common practice of pushing the
film out the top by adding wine to
make the container overflow contaminates
the cellar and is not recommended.
Acetobacter grows on bacterial
media and on most yeast media.
Acetobacter is a normal part of red
wine flora in the cellar. A large population
does not necessarily mean the
wine is spoiling; it means it has the
potential to spoil if air is, or becomes,
available.
Lactic acid bacteria
Wines not intended for malolactic
fermentation (MLF) should be kept
below pH 3.4, with a molecular SO2 of
at least 0.6 to 0.8 ppm. Test malic acid
enzymatically before MLF to obtain a
starting level; if any CO2 or haziness
occurs during cellaring, check microscopically
for bacteria and test for a
decrease in malic acid.
If residual sugar is present, certain
Lactobacillus species make acetic acid
from sugar, raising the VA in stuck
wines, but there is no smell because
they do not produce ethyl acetate. In
stuck or sweet
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wines, monitor VA very
closely and check microscopically for
Lactobacillus every day or two. Many
stuck wines are attacked by “ferocious”
Lactobacillus, which produce
acetic acid from sugar. Also monitor
VAduring MLF and afterwards, and if
possible, test D-lactic acid, produced
from sugar. Check population microscopically;
lactobacilli do not grow
well in culture, and PCR-based tests
do not presently detect all species.
After malic acid is gone, citric acid
can be metabolized stoichiometrically
to produce 0.1 to 0.15 g/100 ml acetic
acid. Biogenic amines, regulated by
some European countries, may be produced
by various lactic acid bacteria
even after MLF is finished.
Many heterofermentative lactic
acid bacteria can produce the N-heterocycle
compounds of “mousiness,” a
rare taint characterized by a very
unpleasant aftertaste of mouse urine
or rancid nuts. The compounds are
only slightly volatile at wine pH so the
wines exhibit a “popcorn” or “basmati
rice” aroma, even if the taint is intense
once it is put into the mouth and the
higher pH allows the compounds to
volatilize. Studies have been proposed
to distinguish a "transient mousiness"
following MLF and cured by SO2
addition from a permanent taint
which resists curative measures.
Saccharomyces and
Zygosaccharomyces
These fermentation yeasts require
residual fermentable sugars. The classic
minimum level for activity is 0.5
g/L measured by enzymatic tests of
glucose and fructose, though this is a
arbitrary concentration. Prediction of
refermen-
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tation also depends on glucose/
fructose ratio, so these sugars
should be measured separately, not
together. Zygosaccharomyces bailii is
resistant to SO2, sorbate, and benzoate,
giving it a formidable advantage.
Both yeasts grow on unselective
yeast media. Zygosaccharomyces cells
tend to be slightly more oval than
Saccharomyces, but its identity is confirmed
microscopically when the sexual
stage of conjugation begins in culture
in one to three weeks.
When preparing to conjugate, cells
grow a projection called a “shmoo
tip,” distinguished from a bud by the
lack of a membrane between the cell
and the projection. Conjugating cells
join their cytoplasm and exchange
genetic material, making odd shapes
often resembling dumbbells or more
bizarre structures.
Most Zygosaccharomyces strains
grow on acetic acid agar while Saccharomyces
do not. PCR-based genetic
techniques can identify Zygosaccharomyces
cultures before conjugation is
observed. However, in some identification
systems (such as API), certain
Candida yeasts cross-react with
Zygosaccharomyces,
so all methods, including
PCR primers, should be rigorously
checked against a large number of
wine-related Candida species to ensure
specificity. Cultures identified as Zygosaccharomyces
by any technique including
PCR should be kept until conjugation
is observed to confirm their
identity.
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