ISSN 1022-7954, Russian Journal of Genetics, 2008, Vol. 44, No. 1, pp. 27–34. © Pleiades Publishing, Inc., 2008.
Original Russian Text © S.Z. Mindlin, V.S. Soina, M.A. Petrova, Zh.M. Gorlenko, 2008, published in Genetika, 2008, Vol. 44, No. 1, pp. 36–44.
On the basis of conventional notions, many of anti-
biotic resistance genes of contemporary bacterial clini-
cal strains were originated from antibiotic producers,
mainly, actinomycetes. Later, resistance determinants
were disseminated via sequential rounds of horizontal
transfer among Gram-positive and Gram-negative bac-
teria, which occupied the same ecological niches as act-
inomycetes, and were eventually introduced into bacte-
ria inhabiting human and animal organisms .
Most simple method for verifying this hypothesis
seems to be study of environmental strains of soil and
aqueous bacteria resistant to antibiotics. Indeed, bacte-
ria resistant to many identiﬁed antibiotics were recently
detected among strains of soil bacteria, and it should be
noted that some of them were resistant to more than two
antibiotics [2–4]. Based on the results obtained, it was
concluded that environmental bacteria are a natural res-
ervoir of resistance genes in clinical bacterial strains
inhabiting human and animal organisms .
Given the validity of this hypothesis, it cannot be
ruled out, however, that antibiotic resistance determi-
nants detected in present-day environmental strains of
bacteria in fact appeared in these bacteria as a result of
environmental pollution due to foreign bacteria, such as
commensals or pathogenic bacteria.
A unique possibility for studying the origin of resis-
tance determinants and the mechanism underlying their
transfer in environmental strains is the use of bacterial
communities of permafrost sediments, which were not
exposed to anthropogenic factors and appeared long
before the mass application of antibiotics for the thera-
peutic treatment of diseases. As follows from the previ-
ously published data [5–7], microbial communities can
preserve viability under stable conditions of subzero
temperatures over thousand and million years.
To date, only few works have been reported in which
the presence of strains resistant to antibiotics was
revealed among bacteria isolated from permafrost sed-
iments [8–10]. However, no systematic research, in par-
ticular, molecular–genetic studies, was undertaken in
this direction for comparative examination of antibiotic
resistance determinants in ancient bacterial strains and
As shown in previous works, we isolated from per-
mafrost soils of Kolyma lowland bacterial strains resis-
tant to mercury compounds and showed that they contain
resistance determinants (
-operons) highly homolo-
gous to those of contemporary bacteria [11–13].
The purpose of this work is to isolate bacterial
strains resistant to different antibiotics from arctic per-
mafrost subsoil sediments of various genesis and age,
determine their afﬁliation to systematic groups, and, in
the case of successful isolation of streptomycin-resis-
tance strains, to determine the molecular structure of
the corresponding determinants.
MATERIALS AND METHODS
Isolation of antibiotic resistance strains from sam-
ples of permafrost sediments.
Sampling sites and the
age of the corresponding sediments are listed in Table 1.
The procedure of aseptical removal of samples from
permafrost sediments, their transportation, storage con-
Isolation of Antibiotic Resistance Bacterial Strains
from Eastern Siberia Permafrost Sediments
S. Z. Mindlin
, V. S. Soina
, M. A. Petrova
, and Zh. M. Gorlenko
Institute of Molecular Genetics, Russian Academy of Sciences, Moscow, 123182 Russia;
Soil Science Faculty, Lomonosov Moscow State University, Moscow, 119899 Russia
Received July 5, 2007
—A collection of bacterial antibiotic resistance strains isolated from arctic permafrost subsoil sedi-
ments of various age and genesis was created. The collection included approximately 100 strains of Gram-pos-
) and Gram-negative bacteria (
) resistant to aminoglycoside antibiotics (gentamicin, kanamycin, and streptomycin), chloramphenicol and
tetracycline. Antibiotic resistance spectra were shown to differ in Gram-positive and Gram-negative bacteria.
Multidrug resistance strains were found for the ﬁrst time in ancient bacteria. In studies of the molecular nature
of determinants for streptomycin resistance, determinants of the two types were detected:
ing for aminoglycoside phosphotransferases and genes
encoding aminoglycoside adenylyltransferases.
These genes proved to be highly homologous to those of contemporary bacteria.