BHT. 2022. No 1 Biota. Human. Technology
33
UDC 631.461.1
Nataliia Tkachuk, Liubov Zelena, Yevheniy Olkhovik
ISOLATION OF ACTINOBACTERIES FROM THE SOIL
FERROSPHERE AND THEIR IDENTIFICATION
ВИДІЛЕННЯ АКТИНОБАКТЕРІЙ З ФЕРОСФЕРИ ҐРУНТУ
ТА ЇХ ІДЕНТИФІКАЦІЯ
DOI: 10.5281/zenodo.7110904
This is an Open Access article distributed under the terms of the Creative Commons Attribution License, which permits
unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
© Tkachuk N., Zelena L., Olkhovik Ye., 2022
ABSTRACT
Actinobacteria is one of the bacterial groups important in terms of biofilm formation. Among
them, representatives of Streptomyces sporoactinobacteria, some of the most active soil ammonifying
bacteria, deserve attention. It is known that ammonifying bacteria are involved in microbially induced
corrosion in the first stages of biofilm formation and form ammonia – a corrosion-dangerous metabolite.
However, their diversity in soil ferrosphere remains poorly understood. The purpose of the work is the
isolation of an actinobacterium strain from the soil ferrosphere and its identification.
The work was carried out by conventional methods: general biological (preparation of mounts
"crushed drop", stained smears, microscopy), microbiological (preparation of medium for the cultivation
of bacteria, the method of ten-fold dilutions, Koch’s method, seeding microorganisms in a liquid and agar
medium, methods of staining of cells of bacteria and their structures (staining with fuchsin, methylene
blue, Gram staining method in Kalina’s modification), methods of determining physiological and
biochemical properties (tests for catalase, oxidase, utilization of citrate, casein, fats, starch, urea,
formation of indole, ammonia, hydrogen sulfide, MRVP-test, relation to oxygen and temperature),
molecular-genetic (isolation of DNA from bacterial cells, polymerase chain reaction with primers for 16S
rRNA gene, 16S rRNA gene sequencing, horizontal agarose gel electrophoresis, phylogenetic analysis
using GenBank database and MEGA 6.0 computer program).
Scientific novelty – from the soil ferrosphere is isolated a strain of actinobacteria, which by the
complex of microbiological, physiological and biochemical properties and based on the sequence of the
16S rRNA gene fragment (according to the results of phylogenetic analysis) is classified as Streptomyces
canus. It develops and expands the understanding of the diversity of corrosive bacteria and a set of test
cultures for the study of microbial-induced corrosion processes.
Conclusions – according to a number of microbiological, physiological, biochemical and genetic
characteristics, the NUChC F2 strain is classified as Streptomyces and identified as Streptomyces canus. The
nucleotide sequences of the 16S rRNA gene were registered in the GenBank database as Streptomyces сanus
MG924748 and MG924855. The isolated strain is ammonifying, thus potentially corrosion-active, and
can be used in the study of microbial-induced corrosion processes. According to the level of biosafety,
bacteria of the S. canus species belong to the 1st risk group (according to the German Technical Rules for
biological agents) and are safe for human health.
Key words: ferrosphere, actinobacteria, phenotypic characteristics, 16S pRNA gene.
АНОТАЦІЯ
Однією з бактеріальних груп, важливих з точки зору формування біоплівки, є
актинобактерії. Серед них на увагу заслуговують представники спороактинобактерій роду
Streptomyces – одні з найбільш активних амоніфікувальних бактерій ґрунту. Відомо, що