Vol 89, No 2 (2012)

Aim. Determination of morpho-physiologi - cal changes in cells of a lactobacillus industry strain suppressed by antagonistically active clinical cultures of Staphylococcus aureus. Materials and methods. 5 clinical strains of Staphylococcus aureus detected among 42 strains isolated previously in atopic dermatitis were used. Bacteria of the industry strain Lactobacillus plantarum 8PA-3 that synthesize bacteriocinlike inhibitory substances (BLIS) were used as target-cells. Suppression of growth of lactobacilli cells by staphylococci was performed on modified agar nutrient medium MRS by delayed antagonism method. Ultrastructure changes in lactobacilli cells exposed to staphylococci BLIS were studied by various methods of transmission electron microscopy. Results. Destructive changes (exfoliation of small layered fragments of peptidoglycan layers) were detected in the cell walls of lactobacilli suppressed by staphylococci BLIS, and rearrangement of ultrathin structure of their cytoplasm with the destruction of protein-ribosomal complex was noticed. Conclusion. Clinical S. aureus strains that produce BLIS, suppress antagonistic activity of lactobacilli were revealed, and morpho-physiological changes in lactobacilli target-cells damaged by BLIS were detected. Local damage of cell components manifested by the destruction of cytoplasm and formation of «ell shadows »

Aim. Evaluate accumulation of capsule polysaccharide by Streptococcus pneumoniae 19A strain in semisynthetic nutrient medium including various amino acid sources. Materials and methods. Comparative evaluation of the production of capsule polysaccharide by the strain belonging to one of the most widespread S. pneumoniae serotype (19A) was performed by using rocket immunoelectrophoresis. The bacteria were cultivated in semisynthetic liquid nutrient media of varying composition. Results. Among 4 sources of nitrogen (aminopeptide, acid and pancreatic hydrolysate of casein, soy peptone) added to salt nutrient medium supplemented with glucose and vitamins, casein and soy peptone were shown to promote the maximum synthesis of capsule polysaccharide independently of the cultivation time. Supplementation of the medium with sulfates of iron, zinc and manganese, as well as pH decrease to acid values significantly reduced the level of capsule polysaccharide in the culture liquid. The maximum growth of bacteria was observed at 11 hours after the start of cultivation in a 10 L volume fermenter in semisynthetic nutrient medium with soy peptone. Accumulation of capsule polysaccharide in the culture liquid continued to the end of the observation period (24 hours) and by the end of the process reached 193 mcg/ml. Conclusion. Further study of influ ence of vitamins, carbohydrates, CO2 concentration on the synthesis of high molecular capsule polysaccharide by bacteria belonging to various pneumococcus serotypes is reasonable.

Aim. Study serologic diversity of Legionella pneumophila strains circulating in potentially dangerous water systems in Russian Federation by using an international panel of monoclonal antibodies. Materials and methods. Serotyping of 234 L. pneumophila strains isolated from coolers of industrial facilities and systems of hot water supply in Russian Federation in 2007 - 2011 was performed by enzyme immunoassay by using an international panel of monoclonal antibodies. Results. Membership of the isolated strains in 14 L. pneumophila serogroups and in 7 subgroups of serogroup 1 was established. Among the isolated cultures serogroup 1 and 6, and Oxford and Heysham subgroup strains were predominant. L. pneumophila serogroup 1 strains were predominant in cooler water, and serogroup 6 - in the hot water supply systems. 7 L. pneumophila strains of the serogroup 1 were typed by monoclone MAb 3/1 associated with LPS epitope that is characteristic for the most epidemically significant legionella strains. Conclusion. Typing by using international panel of monoclonal antibodies for characteristic and evaluation of epidemical significance of legionella strains being isolated form potentially dangerous water systems is the most informative and methodically accessible to a wide range of biological laboratories.

Aim. Detection of nucleotide sequences of genes controlling synthesis of pathogenicity factors in clinical strains of opportunistic enterobacteria isolated from children with acute intestinal infections (AII), as well as their association with resistance to antibiotics and the course of the infectious process. Materials and methods. 175 clinical strains obtained from children with AII undergoing treatment in Irkutsk state infectious diseases hospital (2007-2010) were studied. Primers to a number of genes detected in Escherichia coli pathogenicity islands, cont rolling type S and type 1 adhesion; formation of hemolysins; iron-regulatory protein synthesis; capsule formation were used in the study. PCR products analysis was performed by agar gel electrophoresis. Results. Genetic determinants of pathogenicity were detected in bacteria genera Klebsiella, Citrobacter, Enterobacter, Proteus, Kluyvera, Morganella, Pantoea, Serra tia. Fragments of hlyA and hlyB genes (hemolysin production) were detected more frequently; less frequently - sfaA, sfaG, fimA (adhesion), as well as irp-2 gene (synthesis of iron-regulatory protein). The largest set of genetic determinants of pathogenicity was noted in clinical strains of Klebsiella spp. Cultures with DNA fragments specific to genes of E. coli pathogenicity clusters were obtained predominately from children aged up to 3 years, had multiple antibiotic resistance and were isolated significantly more frequently in severe forms of AII when compared with strains in which these determinants were not detected. Conclusion. The studies performed showed that clinical strains of opportunistic bacteria isolated from patients with AII have a certain pathogenic potential, as evidenced by the presence of genetic pathogenicity markers in them.

Aim. Perform pharmacoeconomic analysis of effectiveness of implementation of program of pre-exposition vaccination against HPVassociated diseases in the Russian Federation. Materials and methods. Pharmacoeconomic analysis consisted of comparison of vaccination costs and assumed benefits of prevented losses from HPV-associated diseases from the society perspective as a whole under the condition of cohort vaccination of girls aged 12 years. A population model executed in MS Excel that allows to perform calculations and obtain results when initial data are changed was adapted for the analysis. The number of prevented HPVassociated diseases was determined by multiplying the number of disease cases observed per year and the effectiveness of the vaccine against each type of the disease. The resulting number of prevented cases was then multiplied by the cost of every HPV-associated condition, and as a result the benefit of vaccine use against human papilloma virus was determined. Results. The cervical cancer (CC) morbidity parameter in the Russian Federation is 13 807. By method of simple proportional extrapolation based on the European data, a calculation of morbidity in the Russian Federation was performed: atypical squamous cells of undetermined significance (ASCUS) - 1 454 015; low grade cervical intraepithelial neoplasia (CIN 1) - 660 207; high grade CIN2, CIN3 - 142 153. Vaccine effectiveness against CIN1, CIN2, CIN3 and CC would presumably be 45.9%, 65.1%, 85.7%, respectively. Vaccine effectiveness against these diseases caused by HPV type 16 and 18 would be 98%, and HPV type 31, 33, 45, 52, 58, 35, 39, 51, 56, 59 cumulatively - 47.7%. The cost of HPV-associated diseases (per case) was 8200 RUR (Russian ruble) for ASCUS, 25 200 RUR for CIN1, 60 020 RUR for CIN2 and CIN3, average weighted cost of CC - 188 772 RUR. The number of prevented cases of ASCUS, CIN1, CIN2, CIN3 and CC when vaccine was used was 322 791, 303 050, 92 496 and 11 827, respectively. The cost of vaccination of 1 cohort was 3.4 milliards RUR, and the prevented losses in Russian Federation were 19.4 milliards RUR. Conclusion. The results of pharmacoeconomic analysis allow to assume that the cost of implementation of a program of vaccination against CC and other HPV-associated diseases in the Russian Federation are 5.4 times less compared with yearly preventable losses, that has a significant social and economical effect.

Aim. Determination of epidemiologic features of genital HPV infection in women living in the Republic of Belarus, and detection of phylogenetic features of HPV-16. Materials and methods. 1035 samples of epithelial cervix smears from patients with benign pathology, pre-cancer and cervix cancer were included into the study. 35 samples containing HPV-16 type DNA were selected for sequencing. HPV L1 gene sequencing (300 n.p.) was performed by using 3100-Avant (Applied Biosystems). Sequencing results analysis was performed by using ClustalX (1.8) and BioEdit. MEGA 4.1 was used for phylogenetic analysis of the fragments obtained and tree construction. Results. HPV-16 type is a dominant genotype that is responsible for the development of cervix cancer, its frequency of occurrence is 56.7% among patients with the specified pathology living in the Republic of Belarus. Predominance of East-Asian and European HPV-16 subtypes that can be attributed to a single evolutionary branch are the territorial features. At the same time detection of HPV-16 subtypes that have 0.003 - 0.007 p-distance between probes may indicate the presence of the several progenitors circulating in the country. A unique HPV-16 subtype was sequenced in the Gomel Region that could be one of the progenitors of HPV-16 circulating on the European continent. Conclusion. Further studies performed in the field of molecular epidemiology will allow to give a qualitative characteristic of the epidemic process, determine the direction and time of introduction of the virus into the country, control the circulation of the virus in the population.

Aim. Study microecologic features of intestine microbiocenosis of frequently ill children. Materials and methods. Composition of intestine microbiocenosis of 86 children aged 3 - 7 years living in the city of Karaganda (Kazakhstan) by bacteriological method. A number of microecologic coefficients was calculated: species saturation index, consistency index, relative occurrence coefficient, modified Sanders dominance index, ƒ2 criteria, Jaccard similarity coefficient. Results. The constants members of the biocenosis were Escheri chia coli with high enzymatic activity, enterococci, lactobacilli and bifidobacteria. Additional species were opportunistic bacteria and fungi of the Candida genus. A tendency to collective existence between members of the normal microflora (E. coli, lactobacilli and bifidobacteria) and opportunistic bacteria and Candida genus fungi was detected. Conclusion. A change of characteristics of intermicrobial interactions in intestine microbiocenosis with a formation of pathologic connections between members of normal and opportunistic microflora and Candida genus fungi was noted.

Literature data on the evaluation of clinicalepidemiologic and social effectiveness of implementation of programs of vaccination against pneumococcal infection in various countries are presented in the review. A prognosis of pharmacoeconomic effectiveness of a universal vaccination of children at the first years of life during realization of program in Russia, where more than 3000 children yearly suffer from pneumococcal bacteremia, around 39 000 have pneumococcal pneumonia and more than 700 000 - pneumococcal otitis is given. The frequency of lethal outcomes from pneumococcal meningitis in Russia is 20-40% and exceeds similar parameters of meningitis of other etiology. Cost for the state from the therapy of pneumococcal infections is no less than 30 646 milliards of RUR (Russian ruble) based on 2009 calculations. In Russia 5 years after an implementation of yearly mass immunization in children aged from 2 months against pneumococcal infection an overall economic effect of the program of mass vaccination would be 39.19 milliards of RUR. With direct costs reducing by 19.69 milliards of RUR, and indirect costs (temporarily disability allowance payment, reduction of losses of gross domestic product) - by 37.4 milliards of RUR.

General problems of classification and nomenclature of human diseases caused by agents of biological nature are reviewed. From biological and epidemiological position, prion diseases that belong to non-infectious human pathology are analyzed. Directions of improvement of ecologic-etiologic and ecologic-epidemiologic classifications and nomenclature of human diseases caused by infectious, invasive and prion agents are determined.

Data on staphylococci species producing staphylococcus enterotoxins (SE) are presented in the review. Genetics of toxin formation, SE biosynthesis, factors influencing SE formation (pH, temperature, effect of inductors and repressors), physical-chemical properties of SE, influence of temperature on enterotoxin stability, enterotoxin structure, immunologic properties, super antigen properties, SE mechanism of action, role of SE in nosocomial infections, intestine dysbacteriosis, atopic dermatitis, enterotoxin toxicity, clinical manifestations are examined.