Vol 89, No 5 (2012)

Aim. Evaluate microbial specter and factors of persistence in facultative anaerobic bacteria isolated from urine during infection of lower urinary tract (ILUT) and secretion of prostate gland (SPG) during chronic bacterial prostatitis (CBP). Materials and methods. Bacteriologic study of urine from 144 women (group I) during exacerbation of uncomplicated ILUT and SPG of 105 patients with CBP (group II) was carried out. Quantitative and qualitative composition of microflora as well as adhesive (AA) and anti-lysozyme (ALA) activity for entero-, coryneformic bacteria and hemolytic staphylococci was determined. Results. In groups I and II aerobic-anaerobic associations with domination of non-clostridial anaerobic bacteria, coagulasenegative staphylococci (CNS), coryneformic bacteria, for urine - enterobacteria were isolated from urine and SPG. S. haemolyticus strains predominated in the CNS group. In group I the frequency of detection of strains of enterobacteria and S. haemolyticus with high AA and ALA were higher (p<0.05) compared with corynebacteria. In group II cultures of S. haemolyticus more frequently (p<0.05) had AA and ALA compared with entero- and corynebacteria. Conclusion. In etiologic structure of ILUT and CBP a tendency of shift of gram-negative flora to gram-positive is observed. Detection of AA and ALA in most of the S. haemolyticus and coryneformic bacteria gives evidence of their pathogenic and persistence potential.

Aim. Evaluation of the role of hemocirculation of a bacterial endotoxin in pregnancy complicated with gestosis. Materials and methods. A complex clinical-laboratory examination of 74 pregnant women at the II-III trimester of gestational period including determination of serum cytokine levels (TNFƒ, IL-1ƒ, IL-2, IL-4, IL-6, IL-8, IL-10, IFNƒ), C-reactive protein, LPS, LPS-binding protein and IgG against R-core LPS in blood plasma was performed. Results. In pregnant women with gestosis an increase in endotoxinemia level (p<0.0001), twofold increase in IgG titers against R-core LPS (p<0.01) and an increase in LBP concentration by 14% (p<0.05) without an increase in concentration of C-reactive protein were shown. Cytokine profile in gestosis was characterized by a twofold statistically significant increase of IL-1ƒ (p<0.0001), IL-8 (p<0.0001) pro-inflammatory cytokine levels, a tendency of IL-6 (p=0.063), TNFƒ (p=0.13) increase and a decrease of Th1 cytokine concentrations: IL-2 - by 6.5 times (p<0.0001), IFNƒ - by 2 times (p<0.0001). Conclusion. The results allow to consider gestoses as a manifestation of increased endotoxin translocation into systemic blood flow during gestation, that determines the necessity to improve therapeutic measures in this pathology.

Aim. Study the presence of genetic determinants of pks pathogenicity island containing clb (colibactin) genes in bacteria of the E. coli M-17 production strain by using PCR. Materials and methods. E. coli M-17 cultures isolated from biopreparations bificol (Microgen) and colibacterin (Biomed) and control strain obtained from Tarasevich State Institute on Standardization and Control (Moscow) were studied. Detection of genetic markers of colibacterin was performed by using multiplex PCR with 4 pairs of primers amplifying the main clb genes: clbB, clbN, clbA and clbQ, generating 575, 711, 981 and 820 bp amplicons, respectively. Results. In genome of all the studied E. coli M-17 strains сlbB, clbN, clbA and clbQ genes associated with the formation of genotoxic colibacterin were detected. Conclusion. Genome of E. coli M-17 bacteria used for production of probiotic preparations colibacterin and bificol contains genetic determinants of genotoxin that require further studies in terms of evidence of harmlessness of production bacteria.

Aim. Carrying out analysis of epizootologic-epidemiologic situation on anthrax that had emerged in Omsk region in 2010 when horse meat from epizootic focus of anthrax was used in production of meat semi-finished products. Materials and methods. Study of samples for detection of anthrax causative agents and strain identification was performed according to guidelines 1.3.2569-09. Strain genotyping was performed by MLVA method. Results. The epizootologic-epidemiologic investigation performed allowed to detect the causes of emergence of anthrax outbreak, its routes and factors of transmission. MLVA genotyping results gave evidence on the single origin of Bacillus anthracis strains isolated from sick animals, humans and food substances. Conclusion. Timely execution of a complex of epizootic and epidemic control measures allowed to localize epizootic and epidemic focus of anthrax as well as prevent a possible large scale development of epidemic complications.

Aim. Evaluate the effect of mass vaccination against hepatitis B implemented within the framework of national project Health on the rate of development of unfavorable outcomes of infection and on average lifetime of the population of Russia. Materials and methods. Hepatitis B markers (НВsAg, anti-НВс, HBeAg, anti-HBe) were determined by enzyme immunoassay in 6211 blood serum samples from conditionally healthy population of 6 regions of the Russian Federation. Analysis of damage to health of the population caused by hepatitis B and favorable effect of mass vaccination against hepatitis B was performed by using mathematical model by Goldstein S.T. et al. (2005), that used results of detection of hepatitis B markers in discrete groups (women of childbearing age, children up to 5 years of age, adults aged 30 years and older). A formula developed by us was used to evaluate the effect of vaccination on the average lifespan of the population of the Russian Federation. Results. The rate of HBsAg detection in the conditionally healthy population of the Tuva Republic was significantly higher (8.1%) than in other studied regions of the Russian Federation (p<0.05): in Moscow Region - 1.6%, in Rostov Region - 1.6%, in Sverdlovsk Region - 1.2%, Khabarosvk Region - 2.0%, in Sakha (Yakutia) Republic - 2.5%. The rate of detection of anti-HBc was significantly higher in the Tuva Republic and Sakha (Yakutia) Republic (46.2% and 42.3%, respectively); in other regions: Moscow Region - 13.6%, Rostov Region - 18.9%, Sverdlovsk Region - 17.5%, Khabarovsk Region - 21.0%. HBeAg in women of childbearing age was detected only in the Tuva Republic (0.4%). The mathematical analysis performed demonstrated that the level of coverage of vaccination of newborn children that in the studied regions is 96.1-99.6% may lead to the reduction of morbidity and mortality from hepatitis B and its outcomes by 91-95% that exceeds the similar parameter obtained by using our model, described in the literature for Africa, Asia and Europe. The calculated level of decrease of morbidity and mortality from hepatitis B and its outcomes may lead to the increase of average lifespan of the population of the Russian Federation by 1.44 - 4.08 months. Conclusion. The strategy of vaccination of newborn children against hepatitis B employed in the Russian Federation significantly reduces the risk of this disease and development of unfavorable outcomes of the disease especially in hyper endemic regions and promotes the increase of average lifespan of the population of Russia.

Aim. Development of technological stages of preparation of experimental influenza whole-virion inactivated adsorbed vaccine based on recombinant influenza virus strains NIBRG-14 and А/Аstana/RG/6:2/2009. Materials and methods. 2 recombinant vaccines influenza strains were used in the study - NIBRG-14 and А/Аstana/RG/6:2/2009. Purification of native virus-containing allantoic fluid was performed by ion-exchange chromatography. The virus was inactivated by formaldehyde. Merthiolate at concentration of 0.1 mg/ml was added to the vaccine as a preserving substance. Aluminium hydroxide was used as an adjuvant. Harmlessness and immunogenicity (HI) of the constructed preparation are determining. Results. Virus-containing materials from recombinant strains with biological activity of 8.5 - 9.0 lg EID50/cm3 and hemagglutination activity of 1:256 - 1:1024 in chicken embryos were obtained. Optimal inactivation regimen of non-purified suspensions by formaldehyde was established and combined scheme of purification and concentration of influenza virus was selected that provide harmlessness and immunogenicity of experimental samples of inactivated vaccines against highly pathogenic influenza A/H5N1 in experiments in mice. Conclusion. The data obtained on quality parameters of intermediate products and final vaccine give evidence on their compliance with normative parameters for whole-virion influenza purified vaccine.

Aim. Study of the apoptogenic effect of Corynebacterium diphtheriae toxigenic strains on mice peritoneal macrophages in vitro. Materials and methods. Evaluation of apoptosis induced by Corynebacterium diphtheriae, Сorynebacterium pseudodiphtheriticum, Staphylococcus aureus, Streptococcus pyogenes strains was performed by characteristic morphological changes in macrophages in smears stained by azure eosin by Romanovsky- Giemsa. Results. Apoptogenic activity of diphtheria infectious agent was established to be determined by diphtheria exotoxin at early (after 1 hour) and surface structures and pathogenicity enzymes at later (3 hours) stages of effect. Conclusion. The ability of diphtheria infectious agent to cause macrophage apoptosis is one of the mechanisms of realization of its pathogenic properties determined by the effect of diphtheria exotoxin, as well as its surface structures and pathogenicity enzymes. The increase of apoptogenic activity of toxigenic strains of С. diphtheriae in association with S. pyogenes may be a pathogenetic base of formation of prolonged forms of bacteria carriage against the background of chronic ENT pathology.

Aim. Study the effect of 1270 nm wavelength laser exposure on the viability of Staphylocoсcus aureus and S. epidermidis cells in vitro. Materials and methods. Reference strain S. aureus 209P and clinical isolate S. epidermidis 26/193 that form bacterial biofilm were used in the study. Ica gene presence in S. epidermidis 26/193 that controls biofilm formation was confirmed in the study by PCR with a primer that generates a specific 814 n.p. amplicon. Experimental device by «ew surgical technologies, Ltd.»was used as a source of emission. The device has a continuous emission mode of laser semiconductor diodes with 1270 nm wavelength. Maximum regulated power of the emission is up to 3 W. Emission power in the studies performed was 150 mW. The time of exposition was 5, 10, 15 and 30 minutes. The amount of viable cells in the experiment and control was determined by calculating CFU/ml. Evaluation of bacterial biofilm formation was performed by the method described in OToole G.A. et al. (2000). Results. A decrease of the number of viable forms of S. aureus 209P by a mean of 52±.0% and 76±.0%, and of S. epidermidis 26/193 by a mean of 48±% and 64±% for 15 and 30 min exposition, respectively, and a significant suppression of biofilm formation by staphylococci was shown. Conclusion. Exposure to 1270 nm laser renders a moderate bactericidal effect on staphylococci cells and significantly suppresses their bacterial biofilm formation.

Aim. Study the specter of low-manifest infections (LMI) and their role in children and adolescents with diseases of central nervous system (CNS) against the background of consequences of perinatal damage of nervous system (PDNS). Materials and methods. Infectologic and neurologic examinations were carried out in 42 patients with consequences of PDNS (17 girls and 25 boys, 3 - 15 years). Detection of LMI resulted in etiotropic therapy with evaluation of clinical and laboratory data in dynamics. Results. In 93% (39/42) of patients causative agents of LMI were diagnosed in various combinations and in various biological materials. Among those: Chlamydia spp. - in 71% of patients, Mycoplasma spp. - in 31%, Ureaplasma urealyticum - in 14% (in total the listed microorganisms were diagnosed in 83% of patients); Herpesviridae family viruses - in 75% (HHV-6 - in 67%, VEB - in 36%, CMV - in 11%, HSV-1,2 - in 11%). Combination of Chlamydia spp. with HHV-6 (R tetr=+0,61) and with VEB (R tet=+0,74) (P<0,05) was detected. None of the patients had typical signs of encephalitis clinically or based on MRT. MRT signs of gliosis-atrophic changes in the CNS were detected in all the patients. Reduction of a number of psycho-neurologic and neurologic syndromes was noted in all the patients during LMI therapy. Conclusion. Most of the patients with consequences of PDNS had low-intensity inflammatory-degenerative process in the CNS determined by LMI, first of all by Chlamydia spp. as well as Mycoplasma spp.

Aim. Study the intensity of anti-endotoxin response in intestine of children with atopical dermatitis and intestine dysbioses. Materials and methods. 110 children with atopic dermatitis of various localization were examined in an outpatient setting. Besides detection of dysbiotic disorders in intestine by using bacteriological seeding on differential-diagnostic media and criteria of evaluation of degree of micro-ecological disorders according to Industry standard of Ministry of Health of Russian Federation «rotocol of patient management. Intestine dysbacteriosis»(2003), content of antibodies against LPS of enterobacteria belonging to various immunoglobulin classes (IgG, IgA, IgM, IgE) by EIA was studied in coprofiltrates. Results. An increase of content of various immunoglobulins, specific LPS of enterobacteria that depended on the degree of intensity of dysbiotic disorders was detected in children. Allergization in children correlated with an increase of level of anti-endotoxin IgE and IgM (0.8±.2) and decrease of IgA content in intestine (-0.8±.2). The higher the level of serum anti-LPS antibodies, the more frequently allergic manifestation occur in children. Conclusion. Correlation between the level of anti-endotoxin serum IgE and frequency of occurrence of allergic manifestations in children was shown.

In the last quarter of century virus of Rift valley fever (RVF) sharply extended its distribution by moving from Africa to Asia and evolving from low- to high pathogenic for humans causing severe hemorrhagic disease, practically equaling in this respect with some members of a group of extremely dangerous pathogens. Morbidity and epidemics of RVF are analyzed. Evolution of epidemic development of the infection is examined. Necessity of development of means and methods for diagnostics, prophylaxis and therapy of RVF is underlined.

Data characterizing the reservation forms of plague infectious agent in Tuva natural focus are presented in the review. Yersinia pestis was shown to persist most of the year in Citellophilus tesquorum altaicus imago - the main carrier, getting into the animal organism only for a short time. An increased ability to aggregate in autumn and accumulate in clumps of C. tesquorum altaicus females that are more adapted to survive the cold season compared with males promote the persistence of the microorganism. The plague infectious agent in the altered form survives in the organism of females not only the winter period but also longer periods of time that is demonstrated by the facts of detection of it after 646 days of staying in the carrier. Moreover Yersinia pestis can persist for more than 400 days in the substrate of the nest of long-tailed ground squirrel infected by excrements and corpses of plague fleas. The substrate of the nest infected in summer-autumn period of the previous year may determine the primary infection of ground squirrels by plague infectious agent in the next epizootic season. On ground squirrels infected during contact with nest substrate, infection of intact fleas may be possible, and so the initiation of a new cycle of transmission of the infectious agent. Adaptation of the plague infectious agent to unfavorable existence conditions in the carrier is expressed in the changes of its morphology and ultrastructure that is evidenced by the facts of isolation of the infectious agent from corpses of fleas situated in the substrate, in the L-form, as well as results of phase-contrast and electron microscopy of the digestive tract of C. tesquorum altaicus.