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№ 9š(209),š12šoctoberš2017
áÎÄÒÅÊ óÅÒÇÅÅ×ÉÞ æáíéîãùî (1835–1918),
ÒÕÓÓËÉÊ ÂÏÔÁÎÉË É ÏÂÝÅÓÔ×ÅÎÎÙÊ ÄÅÑÔÅÌØ. áËÁÄÅÍÉË ðÅÔÅÒÂÕÒÇÓËÏÊ áî (1884; ÁÄßÀÎËÔ Ó 1878). ïÓÎÏ×ÁÔÅÌØ (1890) É ÄÉÒÅËÔÏÒ ÌÁÂÏÒÁÔÏÒÉÉ ÁÎÁÔÏÍÉÉ É ÆÉÚÉÏÌÏÇÉÉ ÒÁÓÔÅÎÉÊ áî (ÎÙÎÅ éÎÓÔÉÔÕÔ ÆÉÚÉÏÌÏÇÉÉ ÒÁÓÔÅÎÉÊ ÉÍ. ë.á. ôÉÍÉÒÑÚÅ×Á òáî). ïÓÎÏ×ÏÐÏÌÏÖÎÉË ÐÅÔÅÒÂÕÒÇÓËÏÊ ÛËÏÌÙ ÆÉÚÉÏÌÏÇÏ× ÒÁÓÔÅÎÉÊ É Á×ÔÏÒ ÐÅÒ×ÏÇÏ ÏÔÅÞÅÓÔ×ÅÎÎÏÇÏ ÕÞÅÂÎÉËÁ ÐÏ ÆÉÚÉÏÌÏÇÉÉ ÒÁÓÔÅÎÉÊ (1887). ïÓÎÏ×ÎÙÅ ÔÒÕÄÙ ÐÏ ÆÏÔÏÓÉÎÔÅÚÕ É ÏÂÍÅÎÕ ×ÅÝÅÓÔ× × ÒÁÓÔÅÎÉÑÈ. òÁÚ×É×ÁÌ ÔÅÏÒÉÀ ÓÉÍÂÉÏÇÅÎÅÚÁ.
Biology
Galaktionova L.V., Vasilchenko A.V., Anufrienko A.A., Terekhova N.A. RECOVERY OF THE HUMUS STATE OF THE STEPPE CHERNOZEMS UNDER THE CONDITIONS OF THE DEPOSITIn the middle of the last century, in order to provide the country’s population with food products, the country began implementing the plan for extensive development of agricultural production. Only in the Orenburg region from 1954 to 1962, about 2.2 million hectares of virgin and fallow lands were plowed up, and the total area of arable land reached 6.4 million hectares. Along with serious miscalculations in the technology of growing the main crops, excessive plowing of steppe chernozems caused the development of a number of degradation processes, such as dehumification, de-structuring, over consolidation, soil erosion, which was accompanied by a progressive decrease in their fertility. However, at the end of the twentieth century, significant political and economic changes took place in the country, affecting the agricultural production sector. One of his results was the transfer of more than two million hectares into the deposit. Thus, for the first time in the agricultural period, previously arable land was found in an unprecedented natural experiment on the natural restoration of chernozems. This article is devoted to the study of the humus state of steppe chernozems in the conditions of the deposit. The more than 20-year period of soil restoration led to a change in the humus content from 2.79 to 4,1š%, its reserves to 88.9š% of that of the virgin land, an increase in the proportion of humic acids to 39.6š% and a humification depth of 2.33. The studies made it possible to conclude that the system of indicators of humus condition of soils showed high sensitivity to cessation of arable use and can be recommended as a diagnostic indicator for assessing the degree of natural restoration of the soil cover of the steppe.Key words: the development trend, the deposit, the organic matter of soils, chernozems.
Elizareva E.N., Yanbaev Y.A., Redkina N.N., Kudashkina N.V., Baykov A.G. ASSESSMENT OF SOIL POLLUTION IN THE ZONE OF INFLUENCE OF METALLURGICAL ENTERPRISESThe total contents of heavy metals in soils selected in the zone of influence of Karabashmed CJSC, Satkinsky Iron-smelting Plant JSC and Uchalinsky Ore Mining and Processing Plant OJSC were analyzed. The excess of the standards for all metals except manganese has been established. The phytoremediation method — accumulation of heavy metals from soil by plants was tested on the investigated soils. It is shown that after growing plants in the soils, the content of iron, zinc and copper decreased. Assessment of soil contamination degree using the concentration coefficient showed that in the studied soils before planting the total amount of most metals is within the limits of natural fluctuation. A weak level of soil contamination with cadmium has been established (Uchaly), moderate contamination with copper (Uchaly) and zinc (Karabash), heavy pollution with cadmium and copper (Karabash). As a result of the use of phytoremediation technology, the concentration coefficients for iron, zinc and copper have significantly decreased. And according to the content of copper, Uchalv’s soil passed from the category of moderate pollution to the category of low pollution. To estimate the multielement influence the levels of heavy metal contamination were calculated using the Saet’s index and the index proposed by Yu.N.šVodyanitsky, taking into account the different toxicity of heavy elements and based on the average geometric parameters. It is shown that the use of phytoremediation technology decreases the level of total contamination of soils with heavy metals.Key words: heavy metals, soil, toxicity, total chemical contamination, phytoremediation.
Zakirullin R.S., Odenbakh I.A. OPTIMIZATION OF SMART-WINDOW PARAMETERS WITH THE ACCOUNT OF GEOGRAPHICAL COORDINATES OF A BUILDINGThe results of calculation of the optimal angle of inclination of the filter gratings on the smart windows with angular selective light transmission at double glazing, a constant azimuth of the orientation of the windows to the cardinal, for different geographical latitudes with the same longitude of a building are presented. For comparison, the calculations are made for the middle of the hottest period of the year and the day with maximum solar radiation. Calculations have also been made for cities located in different hemispheres and having not only different latitudes but also longitudes. Angular selectivity of light transmission of a smart window is achieved by using in a window structure an optical filter with two inclined surface gratings consisting of absorbing, reflecting or scattering parallel strips. The possibility of angular selective filtering of direct solar radiation is shown taking into account the geographic coordinates of the building, the solar trajectory, the seasonal and daily distribution of solar radiation intensity. Smart windows with optical filters are designed to provide hygienic requirements for natural and artificial illumination and insolation, given in Sanitary and Epidemiological Regulations and Standards SanPiN 2.1.2.2645-10 “Sanitary and Epidemiological Requirements for Living Conditions in Residential Buildings and Premises”. Key words: solar radiation, grating optical filter, angular selective characteristic, directional light transmission, optimal inclination angle, latitude, longitude, hygienic norms.
Kozlenko E.A., Ukenov B.S. THE MAIN CHANGES IN THE PROPERTIES OF SOILS IN THE CHELYABINSK REGION AFTER PROLONGED ECONOMIC USELong-term economic use of soils becomes an important factor in changing their fertility and covers the entire process of soil formation. Depending on the duration of development and the level of crop cultivation, the soil undergoes significant changes. The study of the direction of changes in agrophysical, agrochemical, physico-chemical properties is one of the key tasks for the restoration and stabilization of soil fertility. By influencing the natural conditions of the environment, a person changes the basic properties and direction of development of soils. The processes of soil formation are associated with the natural productivity of soils, climatic conditions and anthropogenic factors. Abandoned soils — deposits are the object of research on the restoration of the basic properties of soils, since already 10 years later natural vegetation is restored and fertility is improved: the humus content is increased, the aggregate state is improved, and as a result, the basic agrophysical and agrochemical properties are improved. The article deals with the spatio-temporal changes in vegetation and fertility in the conditions of the deposit of the two most common types of soils in the Chelyabinsk region: leached chernozem and common black chernozem. Changes in the density, structure, and water permeability of soils are shown, and the regularity of the increase in the mass of vegetation in the fallow areas is established, which is related to the physiological characteristics of different communities of fallow plants. Also results of the carried out researches on definition of activity of a microflora of an arable land and a deposit are presented. Soil organisms are responsible for the decomposition of organic matter arriving with plant residues and supply plants with affordable resources. They also play an important role in the formation of stable soil aggregates, maintain the water-air balance, which is a necessary condition for the normal growth of plants. The data of the conducted field experiment gave an idea of the intensity of decomposition of plant material, which allows us to judge the state and activity of soil microflora.Key words: biological activity, density, structural state.
Kushnareva O.P., Kanygina O.N. ION-EXCHANGE PROPERTIES OF NATURAL MONTMORILLONITE CLAY CONTAINING ORENBURG REGIONClays are used in the production of paper, paints, drilling fluids, cleaning of petroleum products and in many other directions. A lot of research has been done on the hydrophilic, sorption and ion-exchange properties of clay materials. However, there is very little information on the properties of clays in the Orenburg region. Of interest is the study of ion exchange processes occurring on the surface of natural non-enriched montmorillonite containing clays of the Orenburg region. The influence of high-temperature treatment on the pH and ion-exchange properties of aqueous suspensions containing particles of natural clay of the Orenburg deposit in sizes from 160 to 630 microns was investigated. In order to separate the physical and chemical processes occurring at high temperatures in clay minerals, a series of successive stepwise firing was performed in the temperature range from 300 to 900œšC with a holding time of 2 hours at each temperature. The pH values of clay suspensions after this temperature action vary nonmonotonically from 8.90 in the initial sample to 12.46 after calcination at 900œšC. According to the results of potentiometric titration, the ion-exchange capacity is calculated as a function of pH. When the pH is changed from 3.5 to 7, a significant increase in the ion exchange capacity is observed. The transition to the alkaline zone sharply weakens the ion-exchange properties of the clay. In the pH range from 7 to 9, ion exchange capacity is reduced and becomes insignificant. Temperature treatment does not affect the preservation of ion-exchange properties of clay. The authors convincingly explain the results obtained by the loss of moisture during heating and the phase transformations that occur, as well as the chemical composition of the clay studied. In their opinion, the presence in the clay of a large number of iron, copper and cobalt ions in an alkaline medium leads to the formation of strong hydroxocomplexes of various composition in the montmorillonite structure, which hinders ion exchange.Key words: clay, calcination, acidity, ion-exchange properties.
Miroshnikova E.P., Kvan O.V., Sergeeva V.A., Miroshnikova M.S. THE INFLUENCE OF PROBIOTIC PREPARATIONS AND NANOCOPPER ON THE HEMATOLOGICAL PARAMETERS OF BLOOD OF CHICKENSThe provision of microbiota in the chemical elements is solved through the introduction in the diet of mineral salts of the individual trace elements and their complexes. However, with the development of nanotechnology was more suitable to combine giving probiotics in conjunction with ultrafine preparations of trace elements. Studied the combined use of the probiotic Vetom 1.1. (Bacillus subtilis strain 7092) and ultrafine particles of copper (Cu) (dš=š55šnmš±š15 nm) on chickens-broilers cross “Smenaš8”. The control group received the basic diet, I experienced in addition to the basic diet received ODC Cu at a dosage of 1.7šmg/kg diet, a II — nano Cu (1.7šmg/kg of feed) and Vetomš1.1. (Bacillus subtilis strain 7092), in the dose of 1.5šmg/kg of feed. As follows from the data of the joint feeding of the two drugs was accompanied by a significant increase in the content of total protein in blood serum, the growth parameters of glucose. Moreover, the combined effect of the drugs was accompanied by a significant rise in the blood levels of lymphocytes and leucocytes. The combined effect of the two drugs in contrast to the group receiving only probiotic, characterized by increase in ALT. While feeding, Vetom was associated with a decrease in this indicator. The level of creatine kinase at the age of 28 days was significantly increased in the II experimental group. The carried out researches allow to assume the perspective sharing nano Cu and probiotic preparation.Key words: nanoparticles, probiotic, blood morphology and biochemistry.
Notova S.V., Kiyaeva E.V., Ermakova N.V., Kazakova T.V., Marshinskaya O.V. CORRELATION OF PSYCHOLOGICAL CHARACTERISTICS WITH PARAMETERS OF THE METABOLISM OF DISABLED ATHLETESóurrently the Paralympic games are the largest event of world sports, which increases the level of competition between participants. In the preparation of athletes to the games should pay increased attention to their psychological state and a number of indicators of metabolism of the body. Using this data and taking into account their relationship can increase the level of sporting achievements and preserve the health of Paralympic athletes. Conducted psychological research in General had satisfactory results. It was established the absence of significant negative emotions in connection with the peculiarities of physical and psycho-emotional stresses sledge hockey team. This may indicate a sufficient level of adaptation of athletes to training activities. The study of the elemental composition of hair showed that for the majority of the surveyed athletes was characterized by an excess of P, Na, K, Zn, and deficiency of Co, Cr, Ca and Se. In biochemical indicators of blood revealed elevated values of glucose, ALT, direct bilirubin and alkaline phosphatase from 40, 33, 47 and 47š% of patients, respectively. In the course of correlation analysis the highest correlation was observed between the studied psychological indicators and the level of Be, Mg, Se, Si, Al and CA in hair. Most of the indicators of the mental state was significantly correlated with the level of pancreatic amylase, γ-GT, AST, total protein and blood glucose. The study shows the link of the elemental composition of hair and biochemical indicators of psychological characteristics. Despite the fact that the psychological status of the athletes was normal, the indicators of the elemental balance and biochemical profile, in contrast, had deviations, which subsequently can lead to deficient growth and manifestations of neuropsychiatric diseases. Therefore, a questionnaire analysis of psychological status cannot give a full picture of the condition of the athletes, indicating the need for a parallel analysis of the indices of protein, carbohydrate, mineral and lipid metabolism and their timely correction.Key words: athletes, sledge hockey, biochemical indexes of blood, elemental status, psychological characteristics, adaptation.
Ponomareva P.A. DETERMINATION OF EQUILIBRIUM PARAMETERS OF EXTRACTION OF IODINE BY MIXTURES OF EXTRACENTS OF VARIOUS NATUREComplex processing of poor mineral and hydromineral raw materials is a rather urgent task for today due to various reasons. The use of new sources of valuable components can be associated with the depletion of existing deposits, with the inability to process traditional raw materials due to geopolitical, economic, logistical or other reasons. Due to the fact that iodine does not form any significant deposits in the earth’s crust in the form of minerals, since it is a very mobile migrant, but tends to accumulate in groundwater near oil and gas condensate deposits, the use of the latter for iodine mining can be promising. Extraction extraction of iodine from reservoir and drilling waters of natural and technogenic origin is quite effective and economically advantageous. To study the extraction process, model aqueous solutions were used, which in composition are as close as possible to the composition of natural waters. The main equilibrium parameters of the extraction process were determined depending on the composition of the extraction composition. Mixtures of extractants of the aliphatic series — isooctane and cyclohexane and aromatic series — benzene and toluene were investigated. The mineralization of solutions was set by adding sodium chloride in the concentration range from 0 to 4šmol/l. The iodine content corresponded to the minimum industrial concentrations. The method of “isomolar series” and the method of “variable volumes” were used as the basic methods of investigation. In the course of the study, it was determined that, with an optimal ratio of organic and aqueous phases of 1 to 10, the greatest synergistic effect is achieved when using a mixture of extractants of aliphatic and aromatic series in a ratio of 7 to 3. The use of mixtures of this composition allows to achieve the greatest distribution coefficient and extraction ratio.Key words: iodine, extraction, synergetic effect, equilibrium, degree of extraction, distribution coefficient.
Romanenko N.A., Sviridova ô.G., Salnikova å.V. ADHESION OF CELLS OF ESCHERICHIA COLI K-12 ON A SURFACE OF THE ORGANOMODIFIED SORBENTS ON THE BASIS OF SILICA GELAdhesion of cells of microorganisms on the surface organomodification sorbents for microextraction when working with the solutions containing chemical and biological pollyutant is an obligatory link in a chain of in the chain of physico-chemical interactions in the quantitative analysis. At the same time questions of efficiency of definition of chemical analytes in such systems are often connected with deterioration in accuracy and reproducibility of an analytical method. It is defined first of all by features of a structure of sorbents — porosity, high surface area, existence and the structure of the imparted functional groups. So, by preparation of tests of water by method of solid-phase microextraction for gas chromatography definition of organochlorine pesticides (the alpha and scale of isomers of hexachlorocyclohexane (α–HCH and γ-HCH) and a 4.4-dichlorodiphenyltrichloroethane (4.4-DDT)), is possible adhesion cells of Escherichia coli K-12 which are contained in natural sources as on silica gels with the surface modified by C18-hydrocarbonic groups, and on the classical granulated silica gel without modification. Besides, the question of efficiency of microextraction of pesticides from the model solutions containing a suspension of E.šcoli cells demands studying. As a result of a research the high connecting ability of silica gels in relation to microorganisms has been shown. Timed establishment of sorption balance didn’t exceed 30 minutes, and the coefficient of distribution of microorganisms Vs when using unmodified silica gel was higher, in comparison with that for silica gel with a surface, containing the imparted functional C18 groups that demonstrates smaller adhesive ability of the last. At joint sorption of microorganisms and organochlorine pesticides the distribution coefficient significantly didn’t differ from that, calculated earlier. At the same time indicators of efficiency of extraction 4.4-DDT decreased against the background of minor change of extent of extraction of HCH isomers. Thus, the situation of joint sorption of the studied pesticides and cages of E.šcoli without loss for quality of the analysis is represented possible.Key words: organomodification silica gel, adhesion, Escherichia coli K-12, organochlorine pesticides, extraction.
Balkin A.S., Cherkasov S.V. DEVELOPMENT OF COCULTIVATION MODEL OF FREE-LIVING AMOEBA ACANTHAMOEBA CASTELLANII WITH BACTERIA SALMONELLA ENTERICA SEROVAR TYPHIMURIUM 14028SAbility of salmonella to persist and replicate within free-living amoebae allows bacteria to survive adverse environmental conditions. However, many aspects related to the survival of salmonella in the environment and the interactions with eukaryotes remain poorly understood.
In this study, the interaction of Salmonella enterica serovar Typhimurium 14028S with cells of free-living amoebas of Acanthamoeba castellanii was studied. Established, that bacteria of this strain are able to penetrate and survive inside A. castellanii phagosome. Most of salmonella is absorbed by amoebas during the first hour of cocultivation. Cytotoxic effect of salmonella is observed with respect to acanthamoeba after 8 hours of co-cultivation and reaches a maximum after 16 hours.
Developed model of interaction between Salmonella and Acanthamoeba is promising for studying the survival strategy of pathogenic microorganisms in environmental conditions. This model can be used to evaluate expression of salmonella genes during phagocytosis by protozoa.Key words: Salmonella, Acanthamoeba, virulence, phagocytosis.
Bekpergenova A.V., Khlopko Y.A., Ivanova E.V., Perunova N.B. FORMATION OF ASSOCIATIONS OF OBLIGATE-ANAEROBIC BACTERIA OF THE GUT HUMANExamination of infection as a form of symbiotic relationship between microorganisms and humans allowed to substantiate the idea of infection as a model system of associative symbiosis. Microsymbiocenosis is one of the vectors of associative symbiosis, where the formation of associations of microorganisms with different types of bonds takes place. A significant contribution to understanding infectious symbiology has been made by new methodological approaches using the persistent potential of microbes in the evaluation of microsymbiocenosis. The study of the mechanisms of intermicrobial interactions has shown that the basis of the formation of microsymbiocenosis is the relationship that determines the nature of the relationship — synergistic and antagonistic.
Since the results of the in vitro experiments revealed that under the action of supernatants, both the increase and decrease in biological properties occur to varying degrees, and there is no change in the indices studied (indifferent effect), to obtain synergistic and antagonistic relationships, the resulting factual material was processed with discriminant analysis, which integrated the investigated properties: growth properties, biofilm formation, antilisozime activity. This made it possible to single out associations with a “synergistic” type of bond: B.šbifidumš/ B.šlongum; B.šlongumš/ E.šlimosum; E.šlimosumš/ B.šfragillis; C.šperfringensš/ B.šfragilis and “antagonistic” — E.šlimosumš/ C.šperfringens.
In natural conditions, the vast majority of microorganisms exist in the form of associations, the formation of which gives a certain advantage to members of the microbial community when surviving in various biotopes of the human body. The carried out researches made it possible to establish that in addition to the growth properties in the associations of obligate anaerobic bacteria, there is a change in biological properties (biofilm formation, antilisozime activity) that make up the system-forming factor of microsymbiocenosis, which contributes to understanding the mechanisms of the formation of associative symbiosis of man and can make a difference in the development of microbial compositions of new synbiotics.Key words: obligate-anaerobic bacteria, intermicrobial interactions, antilysozyme activity, the biofilm formation, growth properties, associative symbiosis.
Zdvizhkova I.A., Andryuschenko S.V. SCREENING OF THE GENETIC DETERMINANTS OF PATHOGENIC POTENTIAL OF ENTEROBACTERIACurrently, to determine the microbiological safety of environmental objects requires expertise, which takes several days in the case of a bacteriological method. As a result of research on existing techniques, only species identification of microorganisms or, at best, determination of virulence factors, unconditionally pathogenic microorganisms, is carried out. Therefore, at the present time, not only the detection of pathogenic and conditionally pathogenic species of microorganisms becomes an urgent task, but also a one-stage determination of their pathogenic potential; first of all, the main factors of the persistence of microorganisms in relation to the host organism. Screening of the investigated genetic determinants among 30 strains of Escherichia coli and Klebsiella pneumoniae revealed 1 strain of Escherichia coli, carrying both determinants of hemolytic activity, as well as enterotoxin C gene, 3 strains of Escherichia coli carrying the markers of aerobactin production and 1 carrier strain detected in our work marker genes of colibactin. It has been established that three strains of Klebsiella pneumoniae, previously described as carriers of the megaplasmid of virulence, demonstrate simultaneous presence of all detected genes of three determinants of persistence: aerobactin, colibactin and periplasmic inhibitor of lysozyme.
The carried out researches allowed to establish that the prevalence of known carrier strains of genetic markers of known determinants of persistence and pathogenicity of enterobacteria in intestinal microbiota of clinically healthy subjects does not exceed 10š%. In this case, the adhesion of several determinants of different nature is observed in the detected carrier strains, including the marker of hemolytic activity The latter fact complements the data obtained with respect to the virulence factors of diarrheic Escherichia, allowing to divide the list of markers of highly virulent and conditionally pathogenic genetic profiles of enterobacteria.
In addition the developed test system will simultaneously reveal genetic markers of five known determinants of the persistence of opportunistic enterobacteria using a single amplification algorithm, which greatly simplifies the procedure of analysis.Key words: enterobacteria, PCR analysis, pathogenic potential.
Kartashova O.L., Pashkova T.M., Pashinina O.A. INFLUENCE OF MEDICINAL PREPARATIONS OF PLANT ORIGIN ON ANTILIZOZYMIC ACTIVITY OF PRIORITY TYPES OF UROFLORA ALLOCATED FROM UROLITHIASIS PATIENTSOne of the most common urological diseases is urolithiasis, the therapy of which uses herbal medicines, effective in the presence of a disease associated with the inflammation in the urinary tract. A pathogenetically determined relationship between the presence of urinary stones and the occurrence of a chronic urinary infection is established in which the causative agent has a set of persistent properties that allow it to persist for a long time in the host organism. Phytopreparations have a different effect on the body, which is due to the biologically active substances that make up their composition. At the same time, the effect of herbal medicines on the persistent potential of microorganisms, in particular, antilysozyme activity, which is one of its significant factors, has not been studied.
In this connection, the effect of a number of phytopreparations, differing in composition and used for the treatment and prevention of urinary tract infections, on the antilysozyme activity of priority uroflora species isolated from patients with urolithiasis (P.šaeruginosa and E.šcoli) was studied in vitro. The study of the possibility of phytopreparation regulation of the antilysozyme sign of bacteria in an in vitro experiment revealed their unidirectional inhibitory effect and found that the maximum reduction in antilysozyme activity in P.šaeruginosa occurred under the action of the drug Gentos, and in E.šcoli — Kanefron.
Thus, the revealed ability of phytopreparations to unilaterally reduce the antilysozyme activity of Pseudomonas sp. and Escherichia sp. under in vitro conditions can be considered as one of the possible mechanisms of their biological activity in vivo.Key words: Medicinal preparations of vegetable origin, Pseudomonas aeruginosa, Escherichia coli, antilysozyme activity, urolithiasis disease.
Klimova T.A., Barysheva E.S., Morozova N.V., Korobova I.V. APPLICATION OF PROBIOTIC PREPARATIONS BASED ON BACILLUS OF THE GENUS BACILLUS IN THE IRON AND ZINC SYSTEM OF DELIVERY IN THE ORGANISM OF LABORATORY ANIMALSStudies in the field of iron and zinc deficiency are caused by the high prevalence of iron deficiency and deficient zinc states according to official statistics of the Ministry of Health of the Russian Federation. It is known that metals in comparison with other elements have the ability to bioaccumulate. Microorganisms are able to extract and concentrate metals, in particular, this ability is possessed by microorganisms that are part of probiotic preparations based on bacteria of the genus Bacillus.
The results of the study of the possibility of delivering iron and zinc to the organism of laboratory animals using probiotic preparations based on bacteria of the genus Bacillus are presented. The probiotic strains were the living and inactivated bacteria of the genus Bacillus: B.šsubtilis 534, B.šcereus 5832, B.šlicheniformis 7038 and B.šsubtilis 7048. The data obtained indicate that in the iron delivery system, the most effective probiotic strains studied are inactivated with B.šlicheniformis 7038 and B.šsubtilis 7048 (increase in iron in the skin of laboratory animals by 31.7š%, in bone tissue — 80.8š%, in muscle tissue — 75.9š%). In the zinc delivery system, the B.šsubtilis strain 534 proved to be the most effective, while the zinc content in the skin of laboratory animals increased by 60š%, in bone tissue by 78.5š%, and in muscle tissue by 43.6š%.
In the iron delivery system, the most effective of the drugs being tested is “Vetom-2”, and the least “Sporobacterin”. In the zinc delivery system, the most effective is “Sporobacterin”, and the least effective is “Bactisubtil”.Key words: iron, zinc, probiotic strains, delivery.
Kochkina E.E., Pashkova T.M., Sycheva M.V., Kartashova O.L. CHARACTERISTICOF BACTERIA OF THE GENUS ENTEROCOCCUS BIOPROFILES ISOLATED FROM ANIMALSThe increasing clinical significance of enterococci highlights the issue of the safety of their use as anti-infective biologics probiotic and requires the search algorithm for the effective differentiation of clinically significant strains from representatives metallisticheskoy microbiota.
Studied the biological properties of bacteria of the genus Enterococcus isolated from the feces of clinically healthy productive animals and from animals with infectious and inflammatory diseases. Determined bioprofile characterizing pathogenic and non-pathogenic isolates. In addition to the genetic determinants of the virulence factors of the strains isolated in the pathology detected hemolytic, gelatinase activity and the factors of persistence (ALA, Acre), with higher characteristic values than fecal cultures. Strains isolated from the intestines of healthy animals was characterized by a high activity of proteases and not had the ability to form biofilm.
The obtained results can be used for differentiation of pathogenic variants Enterococcus spp. from symbiotic cultures included in the composition of the intestinal normality.Key words: Enterococcus spp., differentiation, factors of persistence, factors of virulence, polymerase chain reaction.
Lavina A.M., Khakimova L.R., Matniyazov R.T., Vershinina Z.R., Baymiev A.Kh. THE OBTAINING OF THE RECOMBINANT STRAINS OF RHIZOBIA CARRYING AN EXTRA COPY OF GENES PSSA AND ROSR LABELED WITH FLUORESCENT PROTEIN GFPThe establishment of specific symbiotic relationships between rhizobia and leguminous plants is a complex process that requires the exchange of signaling intermediary molecules between both partners. The flavonoids, secreted by the roots of leguminous plants, induce rhizobium expression of nodule-forming genes — Nod-factors. Synthesis of Nod factors is necessary, but not sufficient for the morphogenesis of effective nitrogen fixing nodules. Since no less important factor in the formation of nodules is the synthesis of rhizobacteria exopolysaccharides (EPS), which play a decisive role in the symbiotic interactions of nodule bacteria with leguminous plants. In addition, in free-living cells, rhizobia EPSs are responsible for attachment to abiotic and biotic surfaces and for the formation of biofilms that ensure the adaptation of bacteria to changing environmental conditions. Currently, the formation of biofilms by rhizobia and the relationship of this process with effective nodule formation is being intensively studied. Special attention should be paid to the rosR gene, which encodes the transcriptional regulator involved in EPS biosynthesis, as well as the pssA gene responsible for initiating the process of EPS biosynthesis in rhizobia.
To study the genes responsible for the synthesis of EPS in the context of their impact on the formation of biofilms by rhizobia, in particular in artificial symbiotic systems, where colonization of plant roots plays an important role, it is necessary to visualize the interaction of nitrogen fixing bacteria with plant roots. In view of this, the aim of this work was to obtain recombinant rhizobial strains labeled with fluorescent protein GFP by the pssA and rosR genes. To do this, a genetically engineered construct containing the fluorescent protein gene GFP was used, which serves to visualize the interaction of rhizobia with plant roots. Cloning of pssA and rosR genes into plasmid pJB658GFP, transformation of rhizobium with this construct, microscopic examination of the obtained recombinant strains was carried out.
It has been shown that recombinant rhizobial strains with an additional copy of the pssA or rosR gene are characterized by a large mismatch of the cell walls, compared to wild strains, which indicates their high production of exopolysaccharides and, as a consequence, indicates an increase in the competitiveness of the strains.Key words: R. leguminosarum, rhizobia, exopolysaccharides (EPS), pssA, rosR, biofilms.
Pashkova T.M. CHARACTERISTICS OF ANTI-CYTOKINE ACTIVITY OF ENTEROCOCCUS SPP., ISOLATED FROM ANIMALSEnterococcus spp. are typical synanthropic microorganisms that, during evolution, have adapted well to survival in a macroorganism, including through the ability to inactivate the factors of innate immunity. Possessing the factors of persistence, the bacteria of the genus Enterococcus can act as the etiological factor of endogenous infectious and inflammatory diseases of animals that arise against the background of a decrease in the natural resistance of the macroorganism. A number of studies have been devoted to the characterization of the persistent properties of Enterococcus spp. of various origin, but certain properties of Enterococcus spp., in particular, anticytokine activity, have not yet been characterized.
In this regard, the anti-cytokine activity of microorganisms of the genus Enterococcus isolated from healthy and infectious-inflammatory diseases (acute and chronic endometritis, mastitis, soft tissue abscesses) in agricultural animals has been studied and the high prevalence of the trait is shown. A higher level of anti-cytokine activity against IL-4 and IL-8 was detected in Enterococcus spp. isolates isolated from infectious inflammatory diseases in comparison with strains isolated from the intestines of healthy animals. Expression of anti-cytokine activity in clinical enterococci was 1.5–1.7 times higher than in intestinal isolates (pš<š0.05). Data on the high level of anti-cytokine activity in clinical isolates Enterococcus spp. in comparison with intestinal ones testify to the possible influence of their exometabolites on the local cytokine balance, which supports the inflammatory process.
Thus, the results obtained on the modification of Enterococcus spp. of different origin of cytokine dynamics can be used to differentiate pathogenic strains of Enterococcus spp.Key words: Enterococcus spp., anti-cytokine activity, agricultural animals, intestinal microflora, infectious-inflammatory diseases.
Polyudova T.V., Yurkina N.O., Eroshenko D.V., Korobov V.P. INFLUENCE OF BLOOD COMPONENTS ON ADHESION OF NON-TUBERCULOUS MICROBACTERIACurrently, diseases caused by non-tuberculosis mycobacteria (NTMB) are increasingly diagnosed in people with reduced immune competence. The most common form of diseases associated with NTMB is infection caused by the biofilm formation on the surface of long-term medical devices. However, the mechanisms of adhesion of NTMB on abiotic surfaces upon contact with internal media of the organism are still unclear.
The effect of human plasma and hemoglobin, fetal bovine serum (FBS), bovine serum albumin (BSA), fibronectin, DNA, lysozyme and lactoferrin on the adhesion of Mycobacterium smegmatis mc2 155, M.šsmegmatis GISK 607, M.šavium GISK 168 to polystyrene has been studied. The presence of plasma (10š%), serum (10š%), albumin (0.5šmg/ml), lysozyme (0.1šmg/ml), and lactoferrin (0.1šmg/ml) significantly reduced the amount of adherent cells of all strains studied. The greatest suppressing effect on mycobacteria was detected in the presence of BSA and FBS. The presence of cattle DNA (0.5šmg/ml) into the incubation medium also resulted in a significant decrease in the number of adherent cells. However, the introduction of fibronectin (2.5šmg/ml) and hemoglobin into the medium revealed heterogeneity among NTMBs for the ability to bind to the polystyrene. Thus, the adhesion of M.šsmegmatis GISK 607 and M.šavium HISK 168 was enhanced in the presence of fibronectin and did not change for M. smegmatis mc2 155. M.šHemoglobin has both inhibitory (M.šsmegmatis mc2 155 and M.šavium GISK 168) and stimulating (M.šsmegmatis GISK 607) effect on the adhesion of NTMB.
The obtained data about the influence of blood components on the attachment of NTMB to the abiotic surface allow us to indirectly judge the biofilm formation in vivo.Key words: adhesion, albumin, DNA, non-tuberculous microbacteria, serum blood.
Sidorov R.Yu., Tkachenko A.G. INFLUENCE OF EXPRESSION OF REL GENE FROM MYCOBACTERIUM SMEGMATIS ON RPOS-LACZ-FUSION ACTIVITY IN ESCHERICHIA COLIGuanosine tetraphosphate (ppGpp) is an important regulatory bacterial molecule involved in stringent response, regulation of persistence, stress adaptation, pathogenicity. The Rel protein from Mycobacterium smegmatis is responsible for synthesis and hydrolysis of ppGpp. The study is devoted to intracellular level detection of guanosine tetraphosphate via gene fusion of ppGpp-dependent rpoS gene and reporter gene lacZ in strain harbouring expression plasmid pMind::rel. The method of detection allows to quantify low concentrations of guanosine tetraphosphate, which is difficult to achieve when using analytical chemistry methods. At the moment gene fusion method is poorly developed for mycobacteria, so there is utility for finding approaches on E. coli. The mycobacterial expression plasmid pMind with a rel gene insert from Mycobacterium smegmatis have been constructed in following study. The resulting plasmid and precursory vector have been transformed into rpoS-lacZ-fusion strain. The cultures have been compared by measuring β-galactosidase activity.
An increase in the expression of rpoS was observed in E. coli strain carrying the plasmid pMind with rel insert in comparison with the control strain containing plasmid without the insert. An addition of tetracycline, inducer of pMind expression, did not affect the expression of rpoS.
Based on the data obtained, we can conclude that there is expression from tetA promoter in the pMind expression plasmid and keeping of mycobacterial Rel enzyme activity in Escherichia coli. The absence of tetracycline induction may result from the inability of the TetR repressor from pMind bind to regulatory sites in E. coli.Key words: guanosine tetraphosphate, persistence, expression plasmid pMind.
Khakimova L.R., Serbaeva E.R., Lavina A.M., Vershinina Z.R., Baymiev A.Kh. GROWTH-STIMULATING ACTIVITY OF BACTERIA RHIZOBIUM LEGUMINOSARUM, SELECTED FROM LEGUMINOUS PLANTS OF THE SOUTH URALOne of the promising methods for the developmrntally-oriented farming is the use of PGPR microorganisms. They can act as an effective growth promoter and, as a result, plant productivity by improving mineral nutrition and protecting against phytopathogens. One of the most important and promising groups of PGPR are the nodule bacteria of the genus Rhizobium, which are widely used in agriculture as associative microsymbionts for nonleguminous economically significant cultures.
The strains R.šleguminosarum LSy10 (D18), LVe13 (Inz 2008), PVu5 isolated from legumes of the South Urals were tested on the growth-stimulating activityon seeds of cucumber and physalis plants. It has been shown that the strains R.šleguminosarum LSy10 (D18), LVe13 (Inz 2008), PVu5 have growth-stimulating activity and can be used as biofertilizers for physalis and cucumber in the concentration of 107cfu / ml and 105cfu / ml. In addition, it was found that all strains have antagonistic activity against the phytopathogenic fungus F.šsolani and have a positive effect on plant growth.
Thus, these strains can be used as biofertilizers to protect useful crops from phytopathogenic fungi, as well as to improve plant growth.Key words: rhizobiÁ, PGPR, biofertilizers, F.solani.
Khaova E.A., Kashevarova N.M., Tkachenko A.G REGULATION OF BACTERIAL PERSISTENCE BY POLYAMINES THROUGH THE STIMULATION OF EXPRESSION OF RPOS, YQJD AND RMF GENES Persisters are rare dormant variants of regular cells and are highly tolerant to antibiotics. Persisters are able to resume growth. These observations suggest that persister cells may be the main culprit responsible for the recalcitrance of chronic infectious disease to antimicrobial therapy. General stress responses and polyamines are described to responsible for the formation of persister cells.
Showed that polyamines putrescine, spermidine and cadaverine upregulate the formation of Escherichia coli persister cells in a dynamic fashion through the stimulation of expression of rpoS, yqjD and rmf genes, relating to general stress responses. Results showed that rpoS gene is the most upregulated by spermidine. The expression of yqjD gene is slightly dependent on polyamines. We obtained that the expression of yqjD is dependent on rpoS. These results are in agreement with previously published data. So, although yqjD is not referred to as a “polyamine modulon”, polyamines still can upregulate its expression indirectly through positive effects on rpoS expression. The persister cells frequency of knockout strains decreased versus their parent during the time periods of stationary phase that were specific for each gene knockout mutant. The persister cell frequency of ΔrpoS strain exhibited maximal decrease versus parental strain at early stationary phase. Therefore, the persister formation at early stationary phase is primarily dependent on rpoS gene. The persister cell frequencies of rmf and yqjD mutants decreased nearer to the lowest values versus parent at late stationary phase. These results show that rmf and yqjD genes can significantly contribute to the persister formation at late stationary phase. These data are confirmed by results, obtained with strains harboring double and triple knockout mutations.
Therefore, rpoS gene is the most upregulated by spermidine and is involved in the persister cell formation predominating at early stationary phase. rmf gene, also relating to polyamine modulon, and yqjD gene, whose expression is slightly dependent on polyamines, are involved in the persister cell formation predominating at late stationary phase. Key words: persisters, polyamines, general stress response.
Hassan G.O.O., Yagudina I.R., Karamova N.S. ANTIMICROBIAL POTENTIAL OF ENDOPHYTIC ACTINOBACTERIA FROM MEDICINAL PLANTSThe progressive increase of resistant microorganisms causing different infectious diseases forces the search for new natural sources of compounds with antimicrobial activity.
In this work we isolated endophytic actinobacteria from twelve medicinal plants growing in Republic of Tatarstan, Russia and studied their antimicrobial activity against five Gram-positive bacteria associated with respiratory tract infections (RTI). 68 isolates from different parts (root, leaf, and stem) of plants were obtained. The isolates were found to be prevalent in roots (38.24š%) followed by stem (32.35š%) and leaves (29.41š%). It was shown that 14 isolates exhibit antimicrobial activity against Staphylococcus aureus, Staphylococcus epidermidis, Enterococcus faecalis, Corynebacterium striatum, Bacillus subtilis isolated from respiratory tract of patients with RTI symptoms in Kasr El-Aini, El-Rahma Masr El Gedida hospitals, Egypt. The highest number of actinobacteria with antimicrobial activity was isolated from the roots of Achillea millefolium and isolate 8R7 inhibited the growth of all bacteria tested. Antimicrobial isolates 8R7, 5R1 (from the roots of Mentha piperita) and 6R1 (from the roots of Hypericum perforatum) demonstrated greatest growth inhibiting activity against Bacillus subtilis, Enterococcus faecalis and Corynebacterium striatum (inhibition zones are 18.3, 18.0 and 17.5 mm respectively).
Thus, data obtained indicate that secondary metabolites of endophytic actinobacteria isolated from the roots of Mentha piperita, Hypericum perforatum and Achillea millefolium have the potential to be considered for development as new agents for treatment of respiratory tract diseases.Key words: endophytic actinobacteria, antimicrobial activity, respiratory infections, medicinal plants.
Shchuplova E.A. THE ROLE OF MICROBIAL ASSOCIENTS IN THE PROCESSES OF INTERACTIONS BACTERIA WITH ERYTHROCYTESRecently, more and more often there are works describing the presence of bacteria inside the red blood cells. Different microorganisms developed different types of strategies for interaction with the host cell membrane and various mechanisms of penetration into the cell. The presence of microorganisms in the blood of humans and animals, as well as inside erythrocytes, leads to the development of bacteremia and sepsis with high mortality. As a hemoculture in bacteremia, gram-positive microorganisms, coagulase-negative staphylococci, are mainly isolated, second place is occupied by enterobacteria and other conditionally pathogenic microorganisms, and the isolated hemocultures are both a monoinfection and an association of two, three or even four pathogens. It should be noted that the study of biological properties in microorganisms is especially important when they are found in associations and even more so when bacteria interact with the cells of the human body, in particular with erythrocytes.
Using the modern method of confocal laser scanning microscopy, it was possible to study the features of the interaction of erythrocytes with Staphylococcus epidermidis cells, pretreated with exometabolites of microbial associates. The results of the study showed that the exometabolites of the pathogenic S.šaureus strain, as well as supernatants of Candida albicans and Klebsiella pneumoniae, strengthened the adhesion processes to the erythrocytes of S.šepidermidis cells and their intra-erythrocyte localization. Under the influence of exometabolites conditional-pathogenic microorganisms such as S.šsaprophyticus, Escherichia coli and Corinebacterium spp. observed a decrease in the studied processes.
Thus, under the influence of exometabolites of bacterial associates, a multidirectional effect on the processes of adhesion and penetration into the erythrocytes of Staphylococcus epidermidis cells was observed.Key words: Íicrobes-associates, exometabolites, erythrocytes, interaction.
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Editor-in-chief |
Sergey Aleksandrovich
MIROSHNIKOV |
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