VISUALIZATION OF BIOCHEMICAL ACTIVITY OF MICROORGANISMS GENUS AEROCOCCUS SPECIES AEROCOCCUS VIRIDANS

Introduction. Aerococci are catalase-negative Gram-positive
microorganisms, widespread in the environment, in
the cold-blooded microbiocenoses [1, 2] and warmblooded
organisms [3]. Currently, there are seven species
of the Aerococcus genus [4]. Type A. viridans got its
name as a result of greening blood agar around the growing
colonies. It was found that one of the products of A.
viridans growth is hydrogen peroxide, has an antagonistic
effect on various kinds of bacteria "in vitro" and "in vivo"
[5]. It was subsequently found that hydrogen peroxide and
superoxide radical are produced as a result of NAD-independent
lactatoxidase [6,7] and piruvatoxidase functioning[
8]. The oxidative ability of A. viridans is a distinctive
feature and allows to visualize these properties using a
specific indicator medium [9]. We have developed an indicator
that enables to visualize oxidation and reduction
properties of aerococci.
Material & methods. Auto-symbiotic cultures of A. viridans
was used for studying of oxidase - reductase activity,
inoculated from human body, an industrial strain of A.
viridans 167, used for the preparation of "A-bacterin",
culture of Aureobasidium pullulans B5, isolated from the
soil and having glucose oxidase activity [10]. Designed
indicating medium were also tested in the inoculation of
aerococci crops, having lactatoxidase activity of bio
materials from birds and mammals. As a basis for the indicating
media (IM) IM1 media was selected with following
composition (g per 1 liter of water) Iodide / potassium
26.0 Soluble starch 10.0 Nutrient agar 30.0 For visualization
of oxidase-reductase activity of aerococci acellular
components IM4 was developed based on IM3, in which
distilled water after double purification was used instead
of tap water and highly purified agar-agar was used instead
of standard nutrient agar. Results & discussion. IM1 is used to test the oxidative properties of aerococci crops. IM manifests the appearance
of a dark purple color, after processing the surface of
cups with 10% sulfuric acid. Dark purple areas with lactatoxidase
activity in the growth zone of the culture are
clearly visible. IM2 with high content of KJ can be used
for isolation of aerococci colonies from contaminated biological
material. IM3 with sodium selenite at a concentration
of 1 g / l was used to visualize the reductase activity
of microorganisms. Selenium recovered with the advent
of the red color in the areas of crop growth and the colonies
while growth of aerococcuses. Nutrient indicator medium
IM3, containing an increased concentration of KJ,
can be used for isolation and studying the oxidizing ability
of different microorganisms which produce hydrogen
peroxide in the oxidation of biological substrates. IM4 allows
us to study the biochemical activity of the enzyme
complex (EC) obtained by ultrasound (US) from washed
thoroughly biomass of aerococcuses [11] and biological
substrates. Conclusion. 1. Indicator nutrient mediums
IM1, IM2, containing the KJ and soluble starch allows to
visualize the oxidizing power of aerococci and isolate
them together with another microorganisms from contaminated
biological products (water, air, soil, biological fluids).
2. The indicator nutrient media IM3 allows (simultaneously
with oxidase activity of microorganism) to visualize
reductase activity in the reduction of selenium with
classification on biological types. 3. Minimal (hungry)
IM4 allows us to observe the interaction between aerococci
enzyme complex and substrates of oxidation and reduction,
with further extrapolation of the results to the
role of aerococci in microbiotope of host organism. The
proposed indicator media can be used for microbiological
and genetic studies, microbiological determination of
amino acids, vitamins and nitrogenous bases.