Vol 92, No 2 (2015)

Aim. Evaluate antibacterial effects of millimole concentrations of copper and zinc cations used as sulfates or chlorides in S. aureus and P. aeruginosa cultures. Materials and methods. Suspension of S. aureus or P. aeruginosa containing 108 CFU/ml were lawn-seeded onto Petri dishes with nutrient agar. 30 minutes later salt solution of copper or zinc with concentrations by metal cation from 10-9 or 10-6 M to 5x10-1 M were applied to the surface of the lawn by 5 pl drops using a 36-channel stamp-replicator. Dishes with bacterial cultures were then incubated for 16 - 18 hours at 37°C, and diameter of growth inhibition zone was measured afterwards. For evaluation of the presence (absence) of viable bacteria in growth inhibition zones, seeding of the material from the center of the zone was carried out into tubes with nutrient broth that were thermostated up to 5 days at 37°C, clarity of the nutrient broth was then evaluated. Results. Inhibiting effects of zinc sulfate against S. aureus surpass effects of copper sulfate by 1.3 - 1.6 times (p<0.001 - 0.05) within metal concentrations from 50 to 500 MM. The effects of zinc chloride in S. aureus culture surpass effects of copper chloride by 1.2 - 1.6 times (p<0.02) for cation concentrations of 100 and 500 mM. In P. aeruginosa cultures, antibacterial effects of copper sulfate are comparable with effects of zinc sulfate. The effects of copper chloride on P. aeruginosa cells are 1.2 times more pronounced (p<0.05) than effects of zinc chloride for metal concentration of 500 mM. Material seeding from zones of culture growth suppression detects turbidity of nutrient broth in samples with specimens from wells treated with zinc salts and broth clarity in samples from wells treated with copper salts. Conclusion. In millimole concentrations, copper and zinc cations have pronounced antibacterial effects in cultures of S. aureus and P. aeruginosa. It is realized as bactericidal in the presence of copper cations and bacteriostatic - in the presence of zinc cations. S. aureus bacteria turn out to be more sensitive to the effects of zinc cations, evaluated by zones of growth inhibition, than P. aeruginosa. The latter show a higher, than S. aureus, tolerance to copper and zinc. Wherein, P. aeruginosa tolerance to copper cations is surmountable.

Aim. Detection of contemporary features of tularemia foci manifestations, determination of territories of high epidemic risk in various landscape zones and creation of a map of foci territories of Moscow Region for isolation of tularemia infectious agent cultures and registered human morbidity for justified planning of prophylaxis measures. Materials and methods. Report materials of epizootologic examinations of natural foci for 1965 - 2013, 156 maps of epidemiologic examination of cases of human infection with tularemia, results of studies of casting of predatory birds and dung of predatory mammals were used. Registered morbidity and isolation of tularemia infectious agent cultures from 1965 to date were applied to an electronic map of Moscow Region by sign method using modern GIS-technologies (MapInfo 10.5 program). Electronic maps Ingit at 1:200 000 scale, as well as Google Earth program were used to search for base points. Results. Analysis of morbidity has revealed structure change in human tularemia morbidity - an increase of the fraction of urban population and a decrease of the fraction of patients among rural inhabitants, unimmunized against this infection are mostly ill. The presence of DNA of tularemia causative agent in biological objects in the complex with serologic and bacteriological studies was shown to allow to detect flaccid epizootics even at low numbers of rodents. Conclusion. Cartographic reflection of registered morbidity and isolation of tularemia infectious agent cultures allowed to show territories with various degrees of epizootic activity and epidemic manifestation. Positive results of serologic and molecular-genetic studies of environmental objects gives evident on epizootic activity and constant risk of aggravation of epidemic situation for this infection.

Aim. Frequency of detection determination for past and current hepatitis E virus (HEV) infection markers in children with immune suppression, as well as children with normal immune status. Materials and methods. The presence of HEV markers (anti-HEV IgG and IgM, HEV RNA) was studied in 609 sera samples of children with neurologic pathologies, 87 samples - from children with immune deficiencies, as well as 3122 samples from conditionally healthy children of 6 regions of Russia. The children were divided into 5 age groups. Anti-HEV IgG and IgM determination was carried out in EIA, HEV RNA - by RT-PCR. Results. The frequency of detection of anamnestic anti-HEV IgG turned out to be significantly higher among immune-compromised children compared with healthy children (5.7% against 1.4%, p<0.05). Anti-HEV IgM, that testify to current or recent infection, were also detected significantly more frequently among children with immune-suppression (1.1 - 1.6%) compared with healthy children (0.25%, p<0.05). HEV RNA was detected in 1 child with the absence of anti-HEV IgM and IgG. Nucleotide sequence analysis of HEV confirmed membership of this isolate in genotype 3, that is prevalent in non-endemic territories. Conclusion. The data obtained have demonstrated, that HEV-infection is prevalent among children in Russia and its course is, probably, asymptomatic in most cases. Immune suppression is a factor of increased risk of infection of children with HEV.

Aim. Evaluation of the ability to form biofilms by various M, T and MT-types of group A streptococci (GAS), as well as study of the effect of various antibiotics on biofilm formation. Materials and methods. 43 strains of various M and T type GAS were studied. The cultures were grown in Todd-Hewitt broth with the addition of 0.5% yeast extract. Comparative evaluation of the ability to form biofilm was carried out using photometry. Benzylpenicillin, oxacillin, cephalosporin, cefuroxime and ceftriaxone antibiotics were used at various concentrations. Results. GAS differ significantly by their ability to form biofilms. The highest ability was noted in 8 strains - 2М, 9М, 12М, 13М, 19М, 30М, 36М-types and 6МТ type. Simultaneous introduction of GAS cultures and antibiotics into the culture well, except for ceftriaxone, resulted in growth inhibition of both plankton cells and biofilms. Conclusion. The ability of GAS to form biofilm depends on streptococci serotype. During simultaneous introduction of GAS with antibiotics into the well, the biofilm does not form.

Aim. Study the circulation of respiratory viruses in children with community-acquired pneumonia (CAP) during the period from October 2012 to May 2013. Materials and methods. 136 children with CAP aged from 3 months to 16 years with ARI symptoms at the disease debut were studied. RNA/DNA of influenza A, B, parainfluenza (PI), adeno-, rhino-, RS-viruses, corona-, metapneumo- (MPV) and bocaviruses were detected in nasopharynx smears by PCR with hybridization-fluorescent detection in real time. Antibodies against influenza viruses A/H1N1/pdm09 California/07/09, epidemic reference strains of influenza viruses A/H1N1/Brisbane/59/07, A/ H3N2/Victoria/361/2011, B/Wisconsin/1/10, against PI viruses type 1, 2, 3 were determined in paired sera by HAI. Results. In February-March 2013 the number of children protected by antibodies against influenza decreased, and circulation of influenza viruses A/H3N2 and A/H1N1/ pdm09 was detected. Rhinoviruses and PI viruses were determined throughout the epidemic season, bocavirus and adenoviruses - during the autumn-winter period, RS-virus and MPV - during winter-spring. Coronaviruses were not detected. The peak of virus detection was established in February, when the threshold of influenza and ARI morbidity was exceeded. The main pathogens of children of the first 3 years of life are rhinoviruses, RS-virus, PI viruses and bocavirus. RS-virus infection at the debut of CAP in children younger than 3 years in 55.5% of cases is associated with the development of broncho-obstructive syndrome. Bocavirus infection in 50% of cases progresses with laryngo-tracheitis and bronchiolitis. Conclusion. The fraction of viruses in etiologic structure ofARI in children varies depending on immune layer, season and age ofchildren. Etiology of viral infection at the debut of CAP could only be proven using specialized laboratory studies.

Aim. Study of the role of P. aeruginosa in the development of osteomyelitis of sternum and ribs in cardio-surgery patients, and analysis of the main biological properties of the isolated bacterial strains. Materials and methods. 132 bacterial cultures were isolated from 83 hospital patients as a result of bacteriological examination during 2007 - 2013. Wound discharge was the study material. Sampling, seeding and identification of the isolated cultures was carried out by using the respective test-systems; antibiotic sensitivity was studied by disc-diffusion method. Results. The proportion of P. aeruginosa was 10.6% (n=14) that is comparable with data on wound infections of general surgery hospitals. A direct and strong correlation (R=0.846, p=0.000132) between hemolytic and phospholipase activity was established during evaluation of virulence properties of the isolated strains. The degree of film-forming ability varied significantly from 0.122 to 1.412 OD; 64.3% of the studied cultures were highly film-forming variants. Statistically significant association between biofilm formation and other studied properties was not found. 4 strains produced VIM2-type metallo-betalactamase and had identical RAPD profiles. Conclusion. Considering that earlier the similar cultures were not detected and all of them were isolated at a short interval of time, we have made a conclusion, that their short-term circulation is probably associated with introduction, which was the reason for patient infection. P. aeruginosa could be the etio-pathogen of both early and later complications of cardio-surgical interventions.

Principles of mass parallel sequencing, otherwise called next generation sequencing (NGS), appeared at the beginning of2000s and were realized in dozens of NGS platforms. High performance and sequencing speed of NGS platforms opened wide horizons for scientists in the field of genomic studies, including metagenomic, first of all related to studies of structure of various microbiocenoses. Dozens of studies dedicated to studies of microbiome and virome of various biotopes of humans in normal state and pathology by using NGS platforms have appeared, forming novel conceptions on pathogenesis and epidemiology of various infectious diseases. Significant cost reduction of the analysis facilitates expansion of sphere of application for NGS technologies not only in the field of fundamental, but also applied microbiologic studies, including etiologic diagnostics of infectious diseases. Due to the increase of the number of cases of infectious diseases, that do not have a typical clinical presentation, use of metagenomic approach is of particular importance, allowing to carry out detection of a wide spectrum of causative agents of bacterial, viral and parasitic infections. Technologic features of mass parallel sequencing platform, main methods of metagenomic studies and bioinformatics approaches, used for the analysis of data obtained, are presented in the review. Studies on healthy human microbiome and in pathology are described; possibilities and perspectives of metagenomic approach application in diagnostics and system of epidemiologic control of infectious diseases are examined.

The analysis of the international experience in providing measures of health and disease safety at the Olympic Games was done. The stages of the formation of bio-security system at public events were considered, including measures to prevent infection outbreaks, the use of computer and information technologies.

To improve the sanitary and epidemiological surveillance at the Olympic Games has developed a system of GIS for monitoring objects and situations in the region of Sochi. The system is based on software package ArcGIS, version 10.2 server, with Web-java.lang.Object, Web-server Apach, and software developed in language java. During the execution of the tasks are solved: the stratification of the region of the Olympic Games for the private and aggregate epidemiological risk OCI various etiologies, ranking epidemiologically important facilities for the sanitary and hygienic conditions, monitoring of infectious diseases (in real time according to the preliminary diagnosis). GIS monitoring has shown its effectiveness. Information received from various sources, but focused on one portal. Information was available in real time all the specialists involved in ensuring epidemiological well-being and use at work during the Olympic Games in Sochi.