Method of estimating microbial risk of development of bacterial intestinal infections transmitted by water way in case of direct release of causative agent of acute intestinal infections from water intended for different purposes
SUBSTANCE: bacteriological analysis of water by normalised indicators is performed with determination of pathogenic bacteria, additionally during water analysis determined are potentially pathogenic bacteria and their pathogenic and virulent properties, obtained data are used to estimate probability of infectious process development in people by formula: Hpatogen =(C•(100-Pt)•V•v/r)•T. After that, calculated is integral index of probability of development of bacterial intestinal infections transmitted by water way with direct release of causative agents isolated and identified during microbiological analysis of water by formula: After that, risk of development of bacterial intestinal infections transmitted by water way is carried out, it is considered to be acceptable if its value does not exceed 1x10-5, and microbial risk of development of bacterial intestinal infections transmitted by water way is considered to be low. If its value is from 1x10-5 and lower, microbial risk of development of bacterial intestinal infections transmitted by water way is considered to be high. If its value constitutes more than 1x10-5, risk of water contamination with pathogenic and potentially pathogenic microflora is estimated at population level by formula: Rp=Rv•100000.
EFFECT: application of claimed method makes it possible to reduce or prevent development of intestinal diseases and makes it possible to predict probability of BII development with direct release of bacterial causative agents from water for various purposes, taking into account their species composition and biological properties.
1 ex, 2 tbl
The present invention relates to medicine, more specifically to work on the epidemiological assessment of water for various purposes in order to prevent intestinal infections, waterborne, and allows on the basis of the determination and identification of microorganisms isolated from water for various purposes, conducting risk estimation and forecasting the incidence of intestinal infections of the population.
Morbidity of acute intestinal infections is beyond the scope of issues to be addressed by health care, and is directly related to the protection of the environment, improvement of environmental conditions and, in particular, supplied drinking water. One of the most important factors in the spread of infectious diseases is a water transmission, the probability of which in modern conditions is considerably expanding and represents a serious epidemiological problem (Rachmaninov Y.A., 2006; GP Craun and other, 2002 and others). According to UNESCO, more than 80% of human diseases are associated with water quality. The economic damage from the use of poor quality drinking water is hundreds of billions of dollars (Onishchenko GG, 2001).
Evaluation of epidemic risk associated with changes in species composition and biological properties of microbiocenosis under the influence of anthropogenous the CSO pollution, is one of the urgent problems. A number of studies revealed that in areas with higher levels of environmental pollution are often infectious disease with atypical signs of biochemical and genetic properties (Ginsburg A.L., 2006, Bondarenko V.M., 2004; Talaia YG 1973-2009; bayazitova LT, 2009 and others). This etiological significance can purchase a variety of "opportunistic" pathogens (Emelyanov I.G., 1994). The list of "legitimate" and potential pathogens of humans continuously replenished by yesterday's saprophytes (V.A. Pokrovsky, 2004; Prokopyeva M.V., 2004; Palkina S.V., 2004; Nilov LU, 2009), which often are pathogenic, virulent and antibiotic-resistant properties.
Development of a method of assessing the microbial risk associated with the spread of the causative pathogenic and potentially pathogenic taking into account the characteristics of the pathogens, as well as the infective dose and the time of development of infectious process, is one of the urgent problems in both scientific and practical terms.
The closest analogue of the prior art is not revealed.
The technical purpose of this invention is to reduce or prevent the incidence of intestinal infections associated with microbial contamination of water with p is izlagaemogo way through the many qualitative features with reliable probability forecast.
The technical result is achieved in that the method of assessing the level of microbial risk of bacterial intestinal infections transmitted by water with the direct allocation of the pathogen, includes bacteriological analysis of water by standardized indicators with the definition of pathogenic bacteria and additionally for the analysis of water to determine potentially pathogenic bacteria and pathogenic and virulent properties, the data and assess the likelihood of human infection by the formula:
Hpatogen- the likelihood of human infection; V is the volume consumed by the individual fluid contaminated with foodborne bacterial gastrointestinal infections (conditionally on who is considered to be the 1 liter); Pt- purification of water from a specific pathogen to establish an acceptable risk; ν - generation bacterial growth; r - infective dose for a specific pathogen intestinal infections; T - time development of infectious process with regard to manifestations of bacteria pathogenic and virulent properties, then carry out the calculation of the integral indicator of the probability of occurrence of bacterial intestinal infections transmitted by water with the direct allocation of pathogens isolated and identifier the bathrooms when conducting microbiological analysis of water according to the formula:
Rν- the integral indicator of the probability of occurrence of bacterial intestinal infections transmitted by water with the direct allocation of pathogens isolated and identified when conducting microbiological analysis of water, M is the number of intestinal agents used in the assessment of sanitary conditions affecting the quality of drinking water; i - ordinal indicator; Xi- share index; Hpatogenis an indicator of the likelihood of bacterial intestinal infections in contamination of the water of pathogenic and potentially pathogenic microflora; W is the sum of the weights Xi, Rν- the integral indicator of the probability of occurrence of acute intestinal infections in contamination of water by pathogens of water-related infections, then assess the risk of water contamination by pathogenic and potentially pathogenic microflora at the population level by the formula:
Rνwith the direct selection pathogens were isolated and identified when conducting microbiological analysis of water; Rp- population risk, 100000 - calculation of population risk is 100,000 thousand people.
Method of assessment microbial risk including the AET such qualitative characteristics of microorganisms, as their pathogenic and virulent properties, i.e. the ability of microorganisms present in the water at a certain infective dose, initiate the infectious process in humans, especially with a weakened immune status.
The proposed method is implemented as follows.
Water quality assessment is performed on the normalized indicators under current law for the particular type of water taking into account not only pathogenic microorganisms (Salmonella, Shigella), but also potentially pathogenic micro-organisms (Klebsiella, and Pseudomonas), as well as identifying their pathogenic and virulent properties.
Currently, in accordance with the regulations determined by such factors as OKB (gram-negative, oxidatively that do not form spores, bacilli, able to grow on differentiated lactose environments, fermenting lactose to acid, aldehyde and gas at a temperature of 37°C)TKB (growth at 44°C) and coliphages. Use only ectoparasiticide representatives of the family Enterobacteriaceae exclude from consideration many species of bacteria, particularly Salmonella and Klebsiella, as shown by our research, most of which have the properties of pathogenicity and virulence, i.e. the ability to represent the risk of bacterial intestinal infections (BCI) in contact to identify Alannah doses in the human body. A false-positive result bacteriological analysis does not allow accurate prediction of the occurrence of the KJV. The hypothesis confirms that bacterial quality analysis of various types of water you need to consider and identify potentially pathogenic bacteria (Klebsiella, pseudomonad and other potentially pathogenic bacteria) as risk factors.
The results of experimental studies have paved the way to a fundamentally new solution to the question of the dependence between the level of bacterial contamination of water and the degree of risk of intestinal infections in the population, spreading through water.
Conducted research to identify the species composition of pathogenic and potentially pathogenic bacteria, as well as their biological activity has allowed us to develop a method for assessing the microbial risk of bacterial intestinal infections transmitted by water under the direct isolation of causative agent of acute intestinal infections of the water for various purposes, and to predict the likelihood of bacterial intestinal infections in the use of poor quality water for drinking, household and recreational purposes.
For the mathematical representation of the occurrence of infectious p is ocess, which initiate the microorganisms present in water sources and considered independently from each other, the most frequently used methods for the Poisson distribution (Meynell et all. 1968) or logarithmic normal distribution (Mechalas, 1972). The emergence of infectious process is characterized by a single parameter p, which plays the likelihood of BCI caused by each organism.
In the Handbook on drinking water (Geneva, volume 3, section 7) section to assess the biological risk presents a model for calculating the risk of infection on the content in the river water of some microorganisms (cryptosporidia, Campylobacter and rotavirus). In the present work on the basis of experimental data, the model who improved.
Example: the Proposed method is tested on the material collected data (table 1).
When conducting microbiological analysis of water was isolated and identified 2 colonies of Klebsiella related to potentially pathogenic bacteria Hpatogen=(2·(100-99,999)·1·0,15):100·24=7,7×10-5where 100-99,999=10-3(water purification for the establishment of acceptable risk); ν generation of bacterial growth 0.15; 100 - infectious dose by Klebsiella, resulted from studies in volunteers, the 24 h time once the development of infectious process with regard to manifestations of bacteria pathogenic and virulent properties and one colony found H patogen=(1·(100-99,999)·1·0,62):100·24=1,5×10-4where 100-99,999=10-3(water purification for the establishment of acceptable risk); ν generation of bacterial growth is equal to 0.62; 100 - infectious dose for Pseudomonas established by calculating the Poisson, 24 h - time development of infectious process with regard to manifestations of bacteria pathogenic and virulent properties. Rν- the integral indicator of the probability of occurrence of acute intestinal infections in the water: 2 colonies of Klebsiella and 1 colony pseudology equal to Rp=Rν·100000=6,1 per 100 thousand population (see table 1).
In the experimental studies were calculated constants generation of bacterial growth of some microorganisms (table 2). The use of the constant generation of bacterial growth gave an opportunity with greater reliability to calculate the likelihood of human infection with consideration of the properties of the pathogen (BCI).
In accordance with the guidelines for drinking-water quality [Recommendations, who, Geneva, 2004] recommended value acceptable level of microbial risk of bacterial intestinal infections that spread through water, is 1×10-5all risk values below this level is a low level of microbial risk value is of above this level is a high level of microbial risk of bacterial intestinal infections, propagating through water.
The data obtained show prospects for the use of the developed method to assess the level of microbial risk, to predict the likelihood of bacterial intestinal infections with the direct allocation of bacterial pathogens from water of different types of water use, taking into account changes in their species composition (pathogenic and potentially pathogenic bacteria) and biological properties.
|The calculation of the risk assessment of microbial contamination of water with the direct selection potential pathogen from water of different water|
|no water sample||Microbial contamination by Klebsiella||The likelihood of BCI||Microbial contamination by Pseudomonas||The likelihood of BCI||The total risk of the KJV||Population risk per 100 thousand population (1/year)|
|1. Moscow region, Bolshevo (drinking well)||2||1||of 1.5×10-4||of 6.1×10-5||6,1|
|2. The river Pakhra, Moscow region, Podolsk district, the village of Krasnaya Pakhra river (below the town, 300 m from the discharge of wastewater)||4||of 1.5×10-4||2||3×10-4||of 2.6×10-4||26|
|3. Pond water (the convent)||3||of 1.1×10-4||4||6×10-4||4,8×10-4||48|
|4. Waste water after disinfection (Kurianovskaya WWTP)||19||7,3×10-4||11||of 1.64×10-3||of 1.4×10-3||141,25|
|5. Voskresensk, Moscow region (a well on the outskirts of the village)||5||of 1.9×10-4||0||0||the 5.7×10-5||the 5.7|
|Constant generation of bacterial growth of some microorganisms, obtained experimentally and their weights|
|Pathogens OKA||Escherichia||Klebsiella||Found||Staphylococcus||Salmonella, Shigella|
|Constant generation of bacterial growth||0,18||0,15||0,62||0,19||0,28|
|The weighting factor Xi||0,1||0,5||1||1,5||2|
Method of assessment microbial risk of bacterial intestinal infections transmitted by water with the direct allocation of the pathogen, including bacteriological analysis of water by standardized indicators with the definition of pathogenic bacteria and additionally for the analysis of water to determine potenziale pathogenic bacteria and pathogenic and virulent properties,
according to the data, assess the likelihood of human infection by the formula:
where Hpatogen- the likelihood of human infection; V is the volume consumed by the individual fluid contaminated with foodborne bacterial gastrointestinal infections (conditionally on who is considered 1 l); Pt- purification of water from a specific pathogen to establish an acceptable risk; ν - generation bacterial growth; r - infective dose for a specific pathogen intestinal infections; T - time development of infectious process with regard to manifestations of bacteria pathogenic and virulent properties, then carry out the calculation of the integral indicator of the probability of occurrence of bacterial intestinal infections transmitted by water with the direct allocation of pathogens isolated and identified when conducting microbiological analysis of water according to the formula:
where Rν- the integral indicator of the probability of occurrence of bacterial intestinal infections transmitted by water with the direct allocation of pathogens isolated and identified when conducting microbiological analysis of water; M - the number of intestinal agents used in the assessment with the nitarno hygienic conditions, affecting the quality of drinking water; i - ordinal indicator; Xi- share index; Hpatogenis an indicator of the likelihood of bacterial intestinal infections in contamination of the water of pathogenic and potentially pathogenic microflora; W is the sum of the weights Xi, Rνwith the direct selection agents, selected and identifitsirovannykh when conducting microbiological analysis of water, then assess the risk of bacterial intestinal infections that spread through water, the microbial risk of bacterial intestinal infections that spread through water, is considered acceptable if its value does not exceed 1·10-5the microbial risk of bacterial intestinal infections that spread through water, is considered low, if its value is 1·10-5and below, the microbial risk of bacterial intestinal infections that spread through water, is considered high if its value is greater than 1·10-5then assess the risk of water contamination by pathogenic and potentially pathogenic microflora at the population level by the formula:
where Rν- the integral indicator of the probability of occurrence of acute intestinal infections in Contamines is the water pathogens of water-related infections; Rp- population risk, 100000 - calculation of population risk is 100,000 thousand population.
SUBSTANCE: method involves collecting samples directly in the region of functioning of the pond and in the control zone, and then determining content of chemical compounds in the samples and determining main quantitative indicators of components of the aquatic ecosystem based on said content. Quantitative indicators of the biota - number and biomass of phytoplankton, zooplankton and benthos - are also determined in the samples from the control zone and the region of functioning of the pond. The ratio of the quantitative indicators of the biota in the pond region to those in the control zone is then calculated. A reference table of ranges of the calculated chemical indicators of the ratios of components of the aquatic ecosystem is then created while adding rows with ratios of quantitative indicators of the biota into the table. By comparing the calculated ratios with ranges of identical ratios in the reference table, the effect of the pond on the state of the aquatic ecosystem is then determined.
EFFECT: high reliability of results.
1 ex, 4 tbl
SUBSTANCE: during realisation of the method the test objects are soaked in tested solutions, parameters of test objects survivability are registered, and using them, threshold concentrations of tested pesticide toxicity are calculated, besides, pathomorphological modifications are registered in test objects, the average percentage of malformations is calculated, and the threshold concentration of teratogenic impact is established as the pesticide concentration with minimum teratogenic impact at test objects, and extent of pesticides toxicity is estimated on the basis of the threshold concentration coefficient, which is calculated using the formula Kn emb - coefficient of threshold concentrations of pesticides toxicity taking into account their teratogenicity, LC16 - limit concentration of pesticides toxicity causing death of 16% embryos, EterC16 - threshold concentration of teratogenic effect of pesticides. At the same time if values of the coefficient Kn emb >10, the class of pesticides hazard is established as I, i.e. extremely hazardous, 5-10 - the class of pesticides hazard is set as II, i.e. highly hazardous, 1-5 - the class of pesticides hazard is established as III, i.e. hazardous, <1 - the class of pesticides hazard is IV, i.e. moderately hazardous. Test objects are embryos of sturgeons.
EFFECT: increased accuracy and validity of assessment.
2 cl, 1 ex, 6 tbl
SUBSTANCE: enteroviruses are concentrated by introduction into an analysed water sample of a magnetic sorbent microparticles coated with polymeric silicon dioxide with amino poropyl groups in proportions 1:1000-3000 of water sample volume. It is incubated at constant stirring for 1-2 hours. The sorbent is collected with a magnet, a supernatant is removed, and a sorbent-enterovirus complex is produced. Enteroviruses are eluted by 0.5M NaCl and 0.05M Tris (pH-10.5) solution. Enteroviruses are identified by immunochemical, cultural and molecular methods.
EFFECT: high degree of virus concentration in the eluate, decreased amount of the eluating solution.
SUBSTANCE: fixed and mobile monitoring sites equipped with measuring instrumentations are located. Various environmental parameters are registered and subjected to analysis. More specifically, hydrophysical field signals are being registered, chemiluminescence, chromatographic, ion-selective, spectral and radiometric analysis is performed. Besides, bed acoustic impendance is registered, molecular spin interactions of seawater protons are detected, artifacts resulting from the magnetohydrodynamic, bioelectric and concentration effect are detected, synthetic surfactant content in the aquatic environment, chlorophyll concentrations, microorgasnisms, phytoplankton, zooplankton is determined. The collected data is further transferred to the archivers and modeling is performed. In the course of modeling the industrial facility environment and infrastructure is divided into a number of areas and a material balance model and a forecast model are created for each of them. For the purposes of the method implementation a system comprising a water withdrawal line equipped with hydrophysical field sensors, a filtering plant for chlorophyll concentration, a filtering plant with a Seitz funnel for microorganisms sampling, a Nageotte chamber for counting the phytoplankton content, a Bogorov Counting Chamber for enumerating zooplankton, a centrifugal apparatus to determine chlorophyll content, a geophone, spectral sensor of proton spin echo is proposed. Besides, the proposed system comprises devices for chemiluminescence, chromatographic, ion-selective, spectral and radiometric analysis, a radiation spectrometer, an atomic absorption spectrophotometer, an X-ray fluorometric analyser, TV sensors, infrared sensors, heat sensors, a metrological module, a sidescan sonar, multiple-beam echo sounder, water quality evaluator by TropoSample parameters and bed deposits characteristics, a lidar (a light radar), a penetrometer, methane and hydrogen detection sensors.
EFFECT: enhanced functional capabilities.
2 cl, 11 dwg
SUBSTANCE: invention relates to a method of concentrating salicylic acid from aqueous solution, involving extraction with trioctylamine oxide solution in hexane, deposited on foamed polyurethane tablets in amount of 75-80% of the weight of foamed polyurethane.
EFFECT: invention increases concentration coefficient of salicylic acid.
2 tbl, 4 ex
SUBSTANCE: system for rapid biological monitoring and indication consists of measurement-detection, analytical and signal units. The measurement-detection unit is n apparatus for measuring reactions of aquatic indicator organisms, where n=2, 3, 4, for two or more aquarium in which there are indicator organisms, into which water enters from a distributing aquarium, said water being pumped by a pump from the tested underwater horizon of the water body or from water pipe. Parameters of functional characteristics of the indicator organisms are calculated from signals of measuring apparatus coming into the analytical unit which comprises a computer with software, containing a data base of parameters of the state of functional characteristics of different indicator organisms under normal conditions, configured for constant population and editing. Values of the measured parameters are continuously processed by the computer in real time, separately for each individual indicator organism. Upon deviation of average values from standard values, the signal unit is automatically switched off and a three-step alarm signal is generated - upon deviation from the standard on one parameter, on three parameters and on all parameters for all indicator organisms.
EFFECT: high accuracy and reliability of continuous indication of the quality of water.
3 cl, 7 dwg
FIELD: processing procedures.
SUBSTANCE: invention can be used in analytic chemistry for sorption concentration and successive determination of heavy metals in water solutions. The procedure for production of sorption material consists in impregnation of surface of a cellulose filter with an analytic reagent wherein thio-semi-carbazone of picoline aldehyde is used as such. Impregnation is carried out with conditioning cellulose material in solution of the reagent in ethanol containing 2.5 % of cetyl alcohol with successive extraction and drying in air. Produced cellulose material is applied for sorption-roentgen-fluorescent analytic determination of heavy metals in water solutions. Metals are extracted with cellulose material for roentgen-fluorescent determination at pH 7.5-10.5, preferably, at pH 10.0.
EFFECT: simple and safe procedure for production of sorption cellulose material used for efficient concentration of heavy metals with successive determination of each of them separately and in whole.
4 cl, 2 tbl, 2 ex
SUBSTANCE: in order to realise the method, aromatic amines are extracted from waste water with an emulsion of aqueous solution of an inorganic acid with ionisation constant higher than 10 in an organic solvent.
EFFECT: high degree of extraction of aromatic amines from waste water with minimum consumption of reagents and combining extraction and re-extraction processes at one step.
SUBSTANCE: method of determining crystallisation of heavy isotope types of water during volumetric, uniform cooling of natural water and formation of ice of heavy water involves determining and recording changes in optical properties of water using a laser beam and two photocells. The photocells are placed at different heights and the laser beam and its scattered radiation are picked up. The laser beam is pulsed with pulse duration of up to 1 second and period between pulses of 30-200 seconds. Measurements are taken after lowering temperature of the processed water to +4°C. Before each measurement, the water aeration process is stopped completely or only on the area under the beam for the period of time when bubbles surface.
EFFECT: invention increases quality of water and preserves its salt composition.
SUBSTANCE: nanobacteria are counted in a human nephrolith. A fixed mass is separated from the latter, mechanically powdered and divided into j=5 weight fractions pj. The powder is poured into j=5 sterile cells, water infiltrate at pore size not exceeding 0.05 mcm is added. The concentrations of nanobacteria is set between 102 to 106 cells in 1 ml by varying the water volume Vj or weight fractions pj of a powder mineral mass in each cell with using the formula. It is followed with mixing poured into j measuring cells. A nutrient medium - calves' fetal serum is added in the ratio 1:9. Two electrodes are inserted in each cell, then the measuring cells with the mixture is placed in an autoclave wherein constant temperature within 30°C≤T≤40°C is maintained. A mixture impedance (R) is periodically measured, and a point of measurement time (t) is determined until a mixture impedance slump is observed. A calibration diagram of an impedance variation time (timpj) to the concentration of nanobacteria in an initial sample (timpj) is presented. Thereafter, the above-stated stages of the method are conducted for analysed water as well. The derived impedance time (timpj) values are projected on the calibration diagram on the axis (timpj), then on the axis (lgnj).
EFFECT: invention allows evaluating the water concentration of nanobacteria.
3 dwg, 1 ex
SUBSTANCE: invention relates to field of medicine. Claimed are immunologically active fragments of polypeptide coded by nucleic acid TOM 34, which possess ability to induce cytotoxic T-cells, containing amino acid sequence SEQ ID NO:7, in which second amino acid from N-end is substituted and represents phenylalanine, tyrosine, methionine or tryptophan, and/or where 1 or 2 amino acids are added on N-end and/or C-end, and/or where C-end amino acid is substituted and represents phenylalanine, leucine, isoleucine, tryptophan or methionine.
EFFECT: invention provides novel epitope for efficient immunotherapy of large intestine cancer.
3 cl, 10 dwg, 3 tbl
SUBSTANCE: invention relates to microbiology. Biological or artificial fluid medium containing the selected microorganism is centrifuged. Supernatant fluid is filtered. A series of diluted samples corresponding to increase in filtrate dilution of up to 10-15 is obtained. The samples are exposed to an electric, magnetic and/or electromagnetic excitation field. Electric signals detected by a solenoid and the digital record of said electric signal after passing through an analogue-to-digital converter are analysed. Diluted samples for which characteristic electric signals, whose amplitude is 1.5 times higher than that of the background noise signals emitted by water, are obtained are selected. Test tubes with equal volume of diluted samples are placed in protective jackets to protect the diluted solutions from external electromagnetic fields. The solution contained in test tube T1 is used as the standard solution. Test tube T2 is placed in the immediate proximity of the sample or is brought into contact with sample X which is presumed to contain the selected microorganism (e.g., E.coli). The obtained electromagnetic signals are compared. Suppression of the signal indicates presence of the microorganism in sample X.
EFFECT: group of inventions enables to obtain reagents and a system for detecting microorganisms in a sample, and detect infection in humans or animals.
3 ex, 8 dwg
SUBSTANCE: invention relates to biotechnology and virology. Compositions and methods used to induce immune response against the influenza virus in canines using novel strains, polynucleotides or polypeptides thereof are described. The invention can be used in veterinary.
EFFECT: versions of an influenza virus, which can infect canines and cause a respiratory disease in canines are disclosed.
38 cl, 14 dwg, 25 tbl, 16 ex
SUBSTANCE: what is offered is a molecular genetic diagnostic technique for nonsyndromic deafness (NSD) that implies detecting 17 disease-related GJB2 and GJB6 gene mutations by PCR amplification of appropriate regions conducted in 8 reaction mixtures with the use of specific primer pairs that is followed by analysing the produced amplicons either without preliminary endonuclease cleavage (in identifying the mutations c.312dell4, c.333-334delAA, ΔGJB6-D13S1830, ΔGJB6-D13S1854), or following the hydrolysis with appropriate specific restrictases (in identifying the mutations IVS1+1G>A, c.35delG, c.71G>A, c.79G>A, c.167delT, c.235delC, c.224G>A, c.299-300delAT, c.360delGAG, c.341A>G, c.269T>C, c.101T>C, c.109G>A).
EFFECT: use of the invention enables more accurate, objective diagnosis of inherited autosomal-recessive hearing loss.
2 tbl, 8 dwg, 3 ex
SUBSTANCE: mice are infected by the oral introduction of a suspension 0.1 ml of a two-day swine Rhodococcus equi, pathogenic of culture containing 5×107 CFU in normal saline 1.0 ml. The infection is followed the oral five-fold introduction of a probiotic preparation 1-5×105-6 CFU in 1.0 ml every 24 hours; in 5 days after the infection, the mice are killed, and dense egg medium are inoculated of the internals suspensions. In 1-2 cultivation days at optimal temperature, but not later than in 5 days, the grown colonies are controlled by acid fast stain. Antagonist activity is evaluated by a growth block index that is a relation of Rhodococcus colony count grown at inoculation of the internals suspensions of the animals prescribed with no probiotic preparations to Rhodococcus colony count grown at inoculation of the internals suspensions of the animals prescribed with the probiotic preparations. The efficacy of the probiotic preparation is concluded if the value exceeds 3.
EFFECT: invention provides fast and nocardioform actinomycete adequate screening of the probiotic preparations in vivo.
15 tbl, 14 ex
SUBSTANCE: microbial cultures are grown on LB-broth that is followed by McFarland standartisation to 0.5. The grown microbial cultures are introduced in wells of a polystyrene plate and added with LB-broth to form a monofilm or added with a broth culture to form a mixed biofilm. The plates are kept in a thermostat at 37°C for 48 hours to form biofilms. The formed biofilms are washed with distilled water and coloured with 0.1% aqueous gentian violet 200 mcl for 45 minutes in the dark, and the biofilms are thoroughly washed for three times with distilled water that is followed by drying the plate and colour intensity biofilm testing. For this purpose, the colourant is eluted in 96% alcohol; the eluate is analysed for an optical density in a spectrophotometer; this value matches to a film formation level; the average optical densities of the eluate of the mixed biofilms and the total optical densities of the eluate of the monoculture biofilms are compared, and if observing no reliable differences, the neutral pattern of interactions are stated; if the value for the mixed biofilm is less than the total value for the monofilms, the antagonistic pattern of interactions is concluded, while the value for the mixed biofilm exceeding the total values for the monofilms enables stating a synergistic pattern of the intermicrobial interaction. The total optical densities of the eluate of the monoculture biofilms shall not exceed 4.
EFFECT: invention enables evaluating the pattern of intermicrobial interaction and predicting a developing mixed infection.
1 tbl, 3 ex
SUBSTANCE: early diagnostic technique for sexually-transmitted infections involves sampling from a cow's cervical canal in 50-60 postpartum days within the preparation for insemination; inoculation of meat infusion agar with added 5% sheep erythrocytes, of yolk-saline agar, Endo medium, Ploskirev's medium, Enterococcus agar, Saburaud medium, and Zeisler agar with the latter to be incubated in an anaerobic culture apparatus. It is combined with inoculation of liquid high-column Blikfeldt medium and Bifidus medium, sugar broth. The inoculations are grown at temperature 37°C within a period optimal for each microorganism to be detected. The smears are Gram stained to identify the detected cultures by conventional techniques.
EFFECT: technique enables early diagnosis of the sexually-transmitted infections caused by opportunistic microflora, prevention of aborts, reproductive disorders, and well-timed prevention and treatment.
SUBSTANCE: adhesive activity index (A), invasive activity index (I) and toxic activity index (T) of pathogenic bacteria is determined on continuous cells of a tissue culture of buffalo green monkey (BGM) kidney cells. A dose containing 107 colonies of bacterial cells is added to the culture monolayer of BGM cells and held at 37°C for 24 hours. The monolayer cells are washed 8 times with 10 ml of a physiological solution at pH 7.2. Index A is determined using a formula after first visually counting in Gorjaev's chamber living and dead cells stuck with bacterial growth and coloured by 0.1% trypan blue solution. Index (I) is determined using a formula after first replacing the culture medium in vials with a medium containing an antibiotic with concentration 50 mcg/ml and incubated in a temperature-controlled cabinet for 1 hour at 37°C. BGM cells are washed from bacteria 8 times with 10 ml of a physiological solution and lysed with 0.1% Triton X-100. Seeding of 0.1 ml is made from the obtained suspension based on the required cultivations per cup with Endo medium. Incubation is carried for 24 hours in a temperature-controlled cabinet at 37°C and the number of grown bacterial cells is counted. Index (T) is determined using a formula after first incubating bacteria in the BGM cell monolayer for 1 day. The suspension is centrifuged and filtered through membrane filters with pore diameter 0.22 mcm. The obtained substrate is put into a pure monolayer of continuous cells and the number of living and dead BGM cells is counted after one day.
EFFECT: invention enables to determine the degree of epidemic hazard of pathogenic bacteria isolated from water.
3 tbl, 4 ex
SUBSTANCE: potentially affected tissue and adjoining histologically normal tissue are examined for a level of RHOV gene transcription. A higher level of RHOV gene transcription in potentially affected tissue in comparison with the same in adjoining histologically normal tissue serves as a diagnostic sign of non-small-cell lung cancer. A polymerase chain reaction for the purpose of RHOV gene transcription is conducted by using a set of primers having sequences SEQ ID NO: 1 and 2.
EFFECT: invention provides high-reliable diagnosis of non-small-cell lung cancer, including squamous cell carcinoma and adenocarcinoma, including at the early stage of progressing cancer transformation.
12 cl, 1 tbl, 7 ex, 2 dwg
SUBSTANCE: invention may be used for human genotype determination by 4343 gene BRCA1 (rsl799950) position polymorphism The method consists in using allele-specific primers with real-time PCR values recording with using ﬂuorescence marked probes. A reaction mixture contains the primers different for each allele, the primer and the probe common for two alleles, and also the common fluorescent probe.
EFFECT: invention provides cutting expenses for genetic researches and making them more available due to the use of standard equipment and only one marked probe.
FIELD: medicine, psychiatry.
SUBSTANCE: one should isolate DNA out of lymphocytes of peripheral venous blood, then due to the method of polymerase chain reaction of DNA synthesis one should amplify the fragments of hSERT locus of serotonin carrier gene and at detecting genotype 12/10 one should predict the risk for the development of hallucino-delirious forms of psychoses of cerebro-atherosclerotic genesis.
EFFECT: more objective prediction of disease development.