INFLUENCE OF THERAPEUTIC MUD ON THE VIABILITY AND PERSISTENCE

PROPERTIES OF BACTERIA
Abdrakhmanov A.R., Brudastov Yu.A., Abdrakhmanov R.A., Zhurlov O.S.
Research Institute of Cellular and Intracellular Symbiosis, Orenburg, Russia
The influence of therapeutic salt mud on the viability and some biological properties of
bacteria, responsible for their survival in the macroorganisms, was shown. Therapeutic mud had
low bactericidal properties, and enterobacteria were, on the whole, even less sensitive to these
properties than staphylococci. Therapeutic mud inhibited the capacity of bacteria for inactivating
complement, lysozyme, and the bactericidal component of the preparation of interferon and also
reduced the hydrophobic properties of bacterial cells. At the same time Escherichia were found
to be more susceptible to the modifying action of the mud than staphylococci. The greatest effect
on the hydrophobic properties and anticomplement activity of bacteria was observed after their
incubation in mud solution.
Zh. Mikrobiol. (Moscow), 1997, 4, P. 89-92.
Key words: bacteria, mud therapy, persistence
INTRODUCTION
The efficacy of mud therapy as a method of treatment of chronic infectious and inflammatory
processes [7, 10] puts on the agenda the question of regulating the properties of the components
of medical mud against the communicating parties of host-parasite. However, analysis of the
literature available to us today gives the impression of aloofness from problems peloterapii
systematic fundamental research with application of modern methodological approaches,
including microbiology. In this regard, the objectives of this study were: to study the sensitivity
of some representatives of the normal, opportunistic and pathogenic microflora to the effects of
human therapeutic mud; identify the range and severity of the effect of modifying therapeutic
mud on the biological properties of the bacteria responsible for their survival in the
microorganism.
When selecting the biological properties of indicator bacteria was preferred features relating
to the inactivation of natural resistance factors, because it is associated with these properties and
long persistence of resistant bacteria in humans [2, 11].

MATERIALS AND METHODS

We used 42 strains of Enterobacteriaceae (24 - Escherichia coli, 6 - Enterobacter cloacae, 8 Klebsiella pneumonia, 4 - Citrobacter freundii), isolated from urine with mucous genital and
bowel contents of patients with various urogenital infections; 36 strains of staphylococci (28 Staphylococcus aureus, 8 - S.epidermidis), isolated from the nasal mucosa bacteria carriers, from
the surface of healthy skin and wound discharge of patients with various purulent-inflammatory

diseases of soft tissues. Isolated strains of microorganisms identified using diagnostic test
systems MicroLaTest (Lachema, Czech Republic).
Isolation of strains in the course of an experimental study of bactericidal properties and
modifying therapeutic mud produced by direct sowing in the differential diagnosis Endo Agar
(for enterobacteria) and vitelline-salt agar (for staphylococci).
To evaluate the bactericidal properties of therapeutic mud used cleaned from sand and a large
particle of the sample does not detect the presence of enterobacteria and staphylococcus.
The reaction mixture consisted of 1g of dirt, 1 ml standard suspension test bacteria in 0.15M
NaCl solution, containing approximately 109 bacterial cells per 1 ml. All components are preheated to 37 C. Immediately after preparation the mixture was vigorously vortexed (1200 1500 rev / min), after which the sampled volume of 50 l to determine the initial concentration
of test bacteria. The mixture was incubated in vitro at 37 C under continuous shaking (shaker
400 - 500 rev / min), it periodically selecting sample volume of 50 l for determining the
concentration of bacteria tested, which were determined by serial dilutions of sowing in sterile
0.15M NaCl. Bactericidal effect was evaluated by the ratio of the number of surviving bacteria
(CFU) of their initial amount based breeding.
The desired ratio is expressed as the negative logarithm of which in turn subtracted the same
period, resulting in the control of a 0.15M NaCl solution. The resulting difference was defined as
the logarithm of the concentration of bacteria fall (Lg).
Determination of the hydrophobicity of bacteria were performed in the separation of a twophase system of aqueous solutions polyethylene glycol (PEG6000) and dextran (T500) with final
concentrations of respectively 6.2 and 4.5 of weight percent [8]. The degree of hydrophobicity
expressed as the hydrophilic-lipophilic balance (HLB) equal to the logarithm of the ratio of the
optical density of the top phase (PEG) to an absorbance lower (dextran).
Determination of anticomplementary activity of bacteria (ACA) was performed according to
the method described [1, 3]. Antilysozyme activity (ALA) has been evaluated using the method
of [4]. The ability to inactivate the bactericidal component of the preparation of human leukocyte
interferon was evaluated as described [6].
The treatment results produced using the methods of variation statistics and analysis of
variance [5].
RESULTS
Results of the study of bactericidal action of therapeutic mud against of enterobacterial and
staphylococcal in accordance with the species of bacteria are presented in Table. As can be seen,
incubation of bacteria in the mud 50% solution at 37 C resulted in a significant reduction in
viable microorganisms compared to control.

The logarithms of the bacterial concentration of maximum fall after 24 - 48 h of incubation.

Enterobacteriaceae generally showed less sensitivity to dirt bactericidal components compared
with staphylococci (Fig. 1). Logarithms fall concentrations determined after 2, 4, 6, 12, 24 and
48 hours of incubation, averaged 0.110.14, respectively; 0.320.16; 1.120.26; 1.920.17;
2,890,47 and 7,920,82. In other words, after 4 hours of incubation the concentration of
enterobacteria fell an average of about 2-fold after 6 hours - 10 times lower, in 12 hours - 100
times lower in 24 hours - 1,000 times lower, after 48 hours of incubation in 85.7% cases, sowing
stopped. At the same time no significant differences in species and pathovariant clearance
parameters studied enterobacteria. Species differences in sensitivity to the bactericidal action of
therapeutic mud consisted only slightly greater resilience to the representatives of species
E.cloacae and K.pneumoniae in the first days of incubation.
Several different trends were observed in the study of bactericidal therapeutic mud against
staphylococci.
The logarithms of the fall of their concentration determined for the above incubation periods,
were respectively 0.960.34; 2.080.53; 2.610.74; 4.840.72, and after 24 h and 48 h
staphylococci not inoculated. The differences in the parameters of the "purification" of
therapeutic mud from bacteria between S.aureus and Staphylococcus epidermidis were
significant only in the first 6 - 12 h of incubation. And more sensitive to the bactericidal action of
mud components proved strains of Staphylococcus aureus.
Study modifying effect mud culturing medium for bacteria and hydrophobicity properties,
responsible for the inactivation of natural resistance factors was conducted for 6 uropathogenic
E.coli strains, 6 strains of S.aureus and 6 strains S.epidermidis, isolated from bacteria carriers.
These properties of the bacteria were detected in 10 - 16 clones each of the test strains.
6-hour incubation of the test bacteria in the mud solution greatly affected the properties of
the hydrophobic surface as well as anti-ALA and inactivate the bactericidal component of the
preparation of human leukocyte interferon (anti-BCI) of E.coli and S.aureus, AKA all 3 test
species of bacteria. An analysis of inter- and intra-distribution of sensitivity to therapeutic mud
showed that E. coli (from the point of view of the studied properties) showed the greatest
susceptibility to modifications. E.coli characterized with a greater proportion of strains
significantly reduced the expressiveness of test features, the maximum relative shift. The mean
absolute level shifts ALA, anti-BCI, AKA and HLB amounted to 0.48 ug / ml (p = 0,002); MBC
-0.39 (p = 0.002); Anti-CH50 -2.67 (p = 0.000); -0.179 (P = 0.003).
Among the staphylococci, showed great inertia with respect to the modifying effect of the
mud, the most sensitive were generally members of the species S.aureus: Appropriate absolute
shifts were equal to 0.32 g / ml (p = 0.069); MIC -0.36 (p = 0,033); Anti-CH50 -2.12 (p = 0.005);

-0.373 (p= 0.029). The least affected by incubation in a muddy solution turned Staphylococcus
epidermidis for which similar changes ability to inactivate lysozyme and anti-BCI identified as
invalid (respectively -0.49 mg / ml, p = 0,1 and 12 -0.29 MIC, p = 0.097). Shifts the ACA and
HLB S.epidermidis were lowest in the sample strains (-1.16, p = 0.001 and -0.120, p = 0.045,
respectively).
Moreover, significant differences between the analyzed properties of the bacteria (Fig. 2). In
particular, the most unstable were hydrophobic properties of the surface of bacterial cells. The
relative magnitude of the shift of the grounds of -22.3%; -37.6% and -64.2% for S.epidermidis,
S.aureus and E.coli, respectively. Most inert properties of bacteria were studied and their ALA
anti-BCI (relative values drop: -15.5% -13.7% -16.7% -11.6% and, -19.9%; - 16.0%,
respectively).
DISCUSSION
These results allow us to characterize the therapeutic mud lake Tuzluchnoe as a factor that
has a relatively low potential microbicides. The spectrum of bactericidal action of therapeutic
mud covers both gram-positive and gram-negative bacteria, including the most typical
representatives of normal and conditionally pathogenic microorganisms. These findings are
broadly consistent with the results obtained in the microbiological analysis of therapeutic mud
used [11, 12]. According to these results, samples of spent mud staphylococci significantly rare
than enterobacteria pollution which, at the same time, usually insignificant and not recorded 48
hours after the initial detection.
The bactericidal action of therapeutic mud contribute, perhaps, a low redox potential of mud
solution, and high ionic strength.
At the same time, especially the Enterobacteriaceae and Staphylococci survival in mud
solution suggests the presence in the therapeutic mud essentially components with antibacterial
properties. The search for such substances could be the basis of a very promising area of future
research. In addition, with appropriate refinement data on the dynamics of survival sanitaryindicative microorganisms in mud medium can be used to create a recreational microbiological
models.
The most important results of this study were obtained in the study of the modifying effects
of therapeutic mud on the biological properties of the bacteria responsible for their survival in
the microorganism. It was found that 6-hour (relatively short) incubation of the bacterial
suspension in the mud solution significantly reduces the persistent potential E. coli and
staphylococcus. Thus there is a marked reduction HLB on the surface of bacterial cells (decrease
of the surface energy), which is known to lead to a decrease in the adhesiveness to the cells
corresponding to bacterial microorganism [9].

These facts, firstly, broaden and complement our understanding of the possible mechanisms
for the implementation of the therapeutic action peloterapii secondly, offer the promise of indepth study of the interaction of bacterial cells with host cells in order to identify the key
conditions for such cooperation and the development of criteria for the rational use of peloterapii
practice treatment chronic infectious-inflammatory processes.
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