Vol 86, No 5 (2009)

Aim . To study rate of detection of bacteria and viruses from dif ferent ta xonomic groups and their associations in children with pyelonephritis. Materials and methods . Two hundred seventy-four patients aged 5 — 10 years divided on two groups were studied: 1 st group — 240 children with chronic secondary pyelonephritis, 2 n d group — children with nephrectomy due to terminal stage of renal obstructive process. Qualitative and quantitative composition of bacterial flora in urine and renal biopsy samples was studied by bacteriological methods as well as presence of viruses (HSVI, HSVII, CMV, EBV, HPV) by means of PCR. Results . In group 1, 72.2% of children had bacterial mixed infection with associations of coagulase-negative staphylococci, Escherichia , peptococci, and Mycoplasma . Herpesviruses and human papillomaviruses were detected in 50.0% of cases. In group 2, bacterial flora was isolated from preoperative urine in diagnostically-significant titer in 91.2% of cases, whereas in urine obtained from the same patients during operation the microorganisms were detected in 38.2% of cases with predominance of Mycoplasma and Ureaplasma . Bacteriological tests of renal biopsy samples yielded bacteria in 29.5% of cases. Studied viruses were detected in preoperative and intraoperative urine as well as in renal biopsy samples in 52.9%, 44.1%, and 58.8% of cases, respectively. In 32.4% of patients viruses were detected in biopsy samples but not in intraoperative renal pelvis’ urine. There was no difference in HPV and CMV detection rate in the nephrectomy group. Conclusion . Bacterial-viral mixed infection is encountered in children with obstructive pyelonephritis and this should be taken into account during diagnostics and treatment of this condition.

Aim . To study associations between clinical signs and results of serological and molecular biological tests in patients with different clinical variants of chronic HCV-infection. Materials and methods . Two hundred twenty-five patients with various clinical course of chronic hepatitis C was tested on the presence of serological markers of HCV-infection, their combinations and HCV RNA. Results . HCV RNA was detected in 78% of patients. Rate of detection of IgM to cor-antigen was higher in RNA HCV-positive patients, which also had higher detection rate of IgG to non-structural viral protein. Anti-сor IgM antibodies were detected more often than others irrespectively from clinical manifestations of the disease. Conclusion . Association between results of serological as well as molecular biological tests and detection of active viral replication in patients with chronic HCV-infection was established. RNA HCV is significantly more frequently detected in serum samples which contain broad spectrum of antibodies to HCV irrespectively from clinical manifestations of the disease. This is important for optimization of diagnostic and treatment measures in practice.

Aim . To determine rate of infection of protozoa by enteroviruses to assess the potential role of protozoa as a natural reservoir of enteroviruses. Materials and methods . The samples were collected from flowing and stagnant water reservoirs in Orenburg region in summer and autumn. The samples of sewages were taken in all stages of their treatment. Cultures of protozoa were isolated with micromanipulator equipped with micropipette, incubated on Pratt’s medium at 25°C and fed with Pseudomonas fluorescens culture. RNA of enteroviruses was detected by reverse transcription polymerase chain reaction (RT-PCR). Results . Seventy-two protozoan species were found in Ural river, whereas 15 and 38 species were found in lakes and sewages respectively. Enteroviruses were detected by RT-PCR in 61.8% cultures of protozoa belonging to 23 species of flagellates, amoebae and ciliates isolated from natural water bodies undergoing anthropogenic impact as well as from sewages in all stages of their treatment. Predominant localization of enteroviruses in dominant taxons of protozoa ( Paraphysomonas sp., Spumella sp., Petalomonas poosilla , Amoeba sp.) was noted. Conclusion . Obtained data confirm presence of enteroviruses in protozoa living both in flowing and stagnant recreation natural water bodies as well as in sewages and confirm the hypothesis of persistence of enteroviruses in protozoa and the reservoir role of the latter. Contingency of life cycles of viruses and protozoa allows to explain the seasonality of aseptic meningitis incidence caused by enteroviruses, which peaks in summer and autumn when protozoa massively multiply in water bodies.

Aim . To study structure of sak0192 gene in S.agalactiae strains isolated from animals and to assess potential for using it as a molecular epidemiologic marker. Materials and methods . One hundred and fourteen strains of S.agalactiae isolated from cow milk were used. Presence of sak0192 gene as well as pathogenicity genes bac, bca, scpB was determined by PCR. Alleles of sak0192 gene were identified by SSCP method, and their nucleotide sequences — by sequencing. Results . In sak0192 gene of S.agalactiae strains direct repeats and spacers were revealed as well as heterogeneity in its nucleotide sequence. Eight different alleles of sak0192 gene were identified in 114 strains, five of which were characteristic only for strains isolated from animals. Complex analysis of sak0192 gene allele and presence of bac, bca, and scpB revealed 16 different genetic variants, only 3 of which were characteristic for strains isolated from animals and from humans. Conclusion . Polymorphism of sak0192 gene could be used for diagnostics and differentiation of S.agalactiae strains and, together with combination of pathogenicity genes bac, bca, and scpB, for identification of epidemically significant clones — agents of infectious disease in humans and animals.

Aim. To reveal features of rubella epidemic process during start of mass immunization and to determine rubella virus genotypes circulating in Saint-Petersburg. Materials and methods. Official data on rubella morbidity during 1995 — 2007 and number of vaccinated against rubella children and adults were used in this study. During 2006 — 2008 males aged 17 — 20 years with rubella diagnosis were eligible for laboratory test on rubella. Nasopharyngeal swabs and blood specimens were tested by PCR and virus isolation on cell culture (PK13). Genotyping of isolates was performed on the basis of 600 nucleotide sequence of E1 gene from 8731 to 9653 n.p. Results. It was shown that mass vaccination of children and young women against rubella during 4 years resulted in 3-fold drop of rubella incidence in whole population, which diminishes the probability of infection in pregnant women and born of children with congenital rubella syndrome. In age structure of rubella morbidity the proportion of children aged 3 — 6 and 7 — 14 years decreased by 1.5-fold. Epidemic process loss the features of autoregulating system (periodicity and seasonal incidence peaks). Results of genotyping showed that isolates belonged to genotype 1E. High degree of homology (97.7 — 99,6%) to isolates from Barnaul and Belorussia was demonstrated. Conclusion. Issues on isolates’ origin and success of measures on elimination of endemic rubella could be resolved by further studies on isolation and genotyping of rubella virus strains in Saint-Petersburg and North-East region of Russian Federation in the whole.

Aim. To assess the level of endotoxinemia and state of antiendotoxin immunity compared with parameters of humoral immunity in patients with bacterial vaginosis. Materials and methods. Blood sera from 28 women with bacterial vaginosis (main group) and 24 clinically healthy women (control group). Concentration of lypopolysaccharide (LPS) in sera was measured by semi-quantitative gel-clot test using LAL-reagent Endosafe. Concentration of LBP, IgG EndoCAb as well as levels of serum IgA, IgM and IgG were measured by enzyme immunoassay. Results. Conditionally higher level of LPS, 2-fold increase of LBP concentration, 1.7-fold increase of IgG EndoCAb as well as tendency to decrease of IgA level were revealed in sera of women from the main group. Dysbiotic shift in vaginal microecological system of non-pregnant women of child-bearing age was associated with breakthrough of LPS in systemic bloodstream. Registration of vaginosis during pregnancy associated with detection of Chlamydia , possessing LPS similar to enterobacteria, revealed presence of unfavorable variants of prolonged torpid course of dysbiotic signs. Conclusion. During torpid forms of bacterial vaginosis, entotoxinemia and abnormalities in anti-endotoxin immunity are revealed, which the need of optimization of its therapy.

To assess sorption properties of Spherocelle beads consisting of particles of macroporous cellulose with various charges in relation to bacterial cells of manufacturing probiotic strains from different taxonomic groups. Materials and methods . The following manufacturing strains: Bifidobacterium bifidum 1, Lactobacillus plantarum 8РА-3 and Escherichia coli M-17, as well as 3 variants of Spherocelles’ matrix: neutral, with positive and negative charges, were used. Results . Spherocelle globules DEAE with a positive charge of the matrix were successively used for designing of immobilized probiotic preparations. Efficacy of sorbent is determined by sorption of >1000 viable cells as well as bacterial metabolites interacting in conditions of sorbent-regulated pH on each globule with diameter 100 — 180 μm. It provides, on the one hand, prolonged viability of probiotic bacteria in culture fluid within 6 months and, on the other hand, optimal pharmacokinetics of preparation due to gradual desorption of metabolites from sorbent globules. Conclusion . Sorbent Spherocell DEAE is biocompatible with cells of manufacturing strains of lactobacilli, bifidobacteria and E.coli and recommended for designing of immobilized probiotics.

Aim . To compare usage of native and formalinized erythrocytes from different animal species in hemagglutination inhibition (HI) test for detection of level of specific antibodies to H5N1 influenza virus in sera of mammals. Materials and methods . Level of anti-H5 antibodies to influenza H5 control antigen and to influenza viruses A/Common gull/ Chany/2006 (H5N1), A/duck/Tuva/01/06 (H5N1), A/Anas platyrhynchos/Chany Lake/9/03 (H5N3) was determined by hemagglutination inhibition test in two influenza A (H5) reference antisera as well as in ferret antisera to native strains of avian influenza virus. Equine, rhesus macaque, sheep, guinea pig, goose, and chicken erythrocytes were used. Results . Using reference antisera, H5 hemagglutinin was detected in all tested antigens with all used erythrocytes. While testing ferret antisera in HI test with reference antigen, anti-H5 antibodies were not detected or detected in extremely low titre (1/80) and only with equine erythrocytes. In most cases, titers of anti-H5 antibodies in HI test with formalinized erythrocytes were higher than with native ones. Conclusion . During monitoring for antibodies to H5N1 avian influenza virus in human population it is necessary to use native strains of A/ H5N1 along with reference antigen. It is possible to use formalinized equine, rhesus macaque, goose, and chicken erythrocytes.

Aim. To study effect of lectin L II of Paenibacillus polymyxa 1460 on cytokine status of mice during modeling of infections caused by Staphylococcus, Escherichia and Yersinia . Materials and methods. Lectin LII of P.polymyxa 1460, bacterial cultures Staphylococcus aureus 209-P, Escherichia coli O1, and Yersinia enterocolitica 12 were used. Mice were inoculated intraperitoneally with 0.2 ml of suspension of bacterial culture grown during 24 hours (5,000 m.c./ml). Lectin (0.4 mcg/ml) in dose 0.2 ml was administered 24 h before and 1 h after inoculation. Serum samples from sacrificed animals were obtained 1, 6, and 24 hours after inoculation and concentrations of IL-1, IL-6, and TNF-α were measured in them using optical density values. Results. It was established that level of TNF-α in serum decreased during staphylococcal infection, whereas levels of IL-1 and IL-6 decreased during all modeled infections. Ability for correction of cytokine balance in organism of experimental animals by administration of lectin 24 h before and 1 h after inoculation with bacteria was demonstrated. Conclusion. Presented studies testify to the effect of bacterial lectin on cytokine-production activity of macrophages during phagocytosis of bacteria and infectious process.

Chitosan biopolymer obtained by deacetylation of chitin has antibacterial and antimycotic action. Known data about mechanisms of biocide effect of chitosan are described in the review. Role of chemical structure — molecular weight, level of deacetylation and presence of nanoparticles — in the expression of antibacterial and antimycotic activity is discussed.

Possible mechanisms of persistence on the example of Chlamydia trachomatis in conditions of herpes simplex virus type 2 (HSV-2) superinfection in vitro and in vivo are described. Emergence of persisting forms of Chlamydia as well as factors influencing on this process are considered. Contemporary views on pathogenesis of viralbacterial infection with HSV-2 and C.trachomatis as well as interactions of the agents with local immunity factors are described. It was suggested that there are signaling pathways through which HSV-2 changes life cycle of Chlamydia .