[Microbial associations defecting in children with chronic pyelonephritis].

AIM: To study rate of detection of bacteria and viruses from different taxonomic 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: 1st group--240 children with chronic secondary pyelonephritis, 2nd 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.

[Vntr-genotyping of Francisella tularensis strains isolated in the former USSR territory and some European countries during epizootics in 1988 - 1989].

Retrospective VNTR-analysis of 159 Francisella tularensis subsp. holarctica strains isolated in December 1988 - February 1989 in former USSR and some European countries was carried out. Analysis of heterogenic genotypes of strains allow to subdivide them into 30 groups of variants by individual genotypes, while cluster analysis--to subdivide them in 7 clusters with different number of compositions. The predominance of genotype C1 strains isolated on the Rostov and Archangelsk regions and the Crimea was established. F. tularensis strains isolated in winter time 1988 - 1989 in different geographic regions were supposed to be resident cultures typical for their biotope in natural focus of disease.

[The multi-locus VNTR-analysis in studies of the population structure of Yersinia pestis in natural foci]. / Mul'tilokusnyi VNTR-analiz v izuchenii populiatsionnoi struktury Yersinia pestis v prirodnykh ochagakh.

A collection of Yersinia pestis strains was investigated by the multi-locus VNTR analysis. All 9 used locuses were diverse, although they differed between themselves by the quantity of genotypes displaying 4 to 13 variations in the sample. The diversity index (DI) ranged from 0.18 (ms21) to 0.86 (ms46); 8 locuses had DI > 0.5. The statistical processing showed 55 individual genotypes in a group of 81 examined strains, which denoted a high discriminative potentiality of the typing system (DP = 0.98). On the basis of the cluster analysis, the genotypes were shared between 11 main groups. The strains belonging to one genotype group were found to originate, as a rule, from one natural focus. The suggested scheme of typing and of creating the databases of genotypes of plaque agent can be used to establish, with a high probability degree, the source of strains.

[Retrospective VNTR-analysis of genotypes of Vibrio cholerae 01 strains isolated, during the 7th cholera pandemic, in Rostov Region]. / Retrospektivnyi VNTR-analiz genotipov shtammov Vibrio cholerae 01, vydelennykh na territorii Rostovskoi oblasti v gody VII pandemii kholery.

Antiplague Research Institute, Rostov-on-Don, Russia Retrospective multi-locus VNTR-analysis was made for 166 Vibrio cholerae strains isolated, 1967-2001, in Rostov Region from clinical samples (82 strains) and from water samples (84 strains). On the basis of cluster analysis of heterogeneous identification strain genotypes, 45 variations of individual strains were shared between 11 separate clusters, among which the F cluster vibrios were predominant. Having emerged, 1970, in the region, they were widely spread during the 1973-1975 cholera pandemic and were registered, among the isolated strains, till 1992 indicating the possibility of long persistence of V. cholerae 01 in the natural aquatic environment. Presumably, the ecosystem specificity contributed to the long-term vibrio persistence.

[Genotyping of the Francisella tularensis strains isolated from natural foci of tularemia in the Rostov region by multilocus VNTR analysis]. / Genotipirovanie metodom mul'tilokusnogo VNTR-analiza shtammov Francisella tularensis, vydelennykh iz prirodnykh ochagov tuliaremii Rostovskoi oblasti.

On the basis of an analysis of the VNTR alleles' distribution in 109 strains of F. tularensis it was established that 19 genotypes of the disease causative agent circulated in the Rostov Region from 1945 to 2002. The microbe-provoked infection episodes can be divided into polyclonal, monoclonal and cluster ones. A retrospective analysis of the genotypes' distribution is indicative of that strains of similar or of closely-related genotypes circulate simultaneously in the studied territory. All investigated F. tularensis strains could be differentiated into two groups; strains, whose genotypes are encountered almost evenly within the entire Region's territory, belong to group 1; and strains of group 2 displayed a trend towards being geographically bound. Isolations of cultures with similar (close) genotypic features made in prolonged time periods suggest that a part of F. tularensis clones can persist for a long time in environmental foci. A set of strains described by genotype can provide a foundation for a database of the tularemic microbe culture within the geo-information system of the South Federative Okrug of Russia.

[Variable nucleotide tandem repeats (VNTR analysis) in Vibrio cholerae 0139 isolated from humans and from water of surface reservoirs in Russia]. / Variabel'nye tandemnye nukleotidnye povtory (VNTR-analiz) u Vibrio cholerae 0139, vydelennykh ot liudei i uz vody poverkhnostnykh vodoemov Rossii.

The comparative study of variable tandem repeats (VNTR analysis) in genomes of V. cholerae 0139 isolated from humans and from water samples taken from surface reservoirs was carried out. The results of the study of the allele state of 5 loci of tandem repeats in 50 strains of vibrios, carried out in the double-primer polymerase chain reaction (PCR), as well as the earlier comparison of the same isolates in the single-primer PCR, showed essential differences and the absence of clonality in the cultures of the clinical and aqueous origin. The suggestion was made that vibrios with individual VNTR genotypes and having no genes ctx and tcpA, isolated from water samples, were epidemic unimportant representatives of the autochthonous microflora of water reservoirs.

[Variable tandem repeat VcA of Vibrio cholerae]. / Variabel'nyi tandemnyi povtor VcA Vibrio cholerae.

Computer analysis revealed seven potential variable-number tandem-repeat (VNTR) loci in the Vibrio cholerae genome. Specific primers were designed to amplify locus VcA located on chromosome 2 and containing a TGCTGT repeat. The locus was found in all tested strains from a V. cholerae strain collection, the repeat number varying 3 to 23. In total, 14 VcA alleles were observed. The VcA locus was proposed as a marker for the molecular typing of V. cholerae strains.

[Genotyping of Yersinia pestis: variability of locus (CAAA)n in natural strains isolated in territory of former SSSR]. / Genotipirovanie Yersinia pestis: variabel'nost' lokusa (CAAA)n u prirodnykh shtammov, vydelennykh na territorii byvshego SSSR.

Genome polymorphism by the locus (CAAA)n was studied in 69 strains of Yersinia pestis isolated from natural foci of the former Soviet Union. The polymorphism was found to be represented by ten alleles in chromosomes, which could be regarded as evidence of variability of this VNTR-locus (diversity index, DI = 0.86). The value of DI was found to vary substantially: from 0.24 in a group of vole strains from seven isolates from the Transcaucasian highlands to 0.77 in nine strains from the Central Asia desert focus. The allele polymorphism of the variable locus (CAAA)n in natural strains of Y. pestis was suggested to be used as a possible genetic marker of the strain. It was concluded that the oligonucleotide primers used in polymerase chain reaction should be upgraded to the genotyping accuracy.

[Viral-bacterial-fungal associations in chronic tonsillitis in children]. / Viruso-bakterial'no-gribkovye assotsiatsii pri khronicheskom tonzillite u detei.

The microflora of palatal tonsils was studied in 84 children with chronic tonsillitis in comparison with that in the control group of 38 healthy children. In most of the sick children viral-bacterial and less frequently viral-bacterial-fungal associations were detected with the prevalence of reo- and adenoviruses, Epstein-Barr viruses, coagulase negative staphylococci and Staphylococcus aureus, as well as peptostreptococci. Adhesive activity and persistence factors among the main bacterial pathogens were shown to be widely prevalent. The depth of the lesion of tonsillar tissue by the infective agents of bacterial and fungal nature, as well as their persistence potential, depended on the taxonomic position of these microorganisms.

[Vibrio cholerae O139 isolated from humans and the water from open reservoirs: a comparative genotyping]. / Vibrio cholerae O139, vydelennykh ot liudei i iz vody otkrytykh vodoemov: sravnitel'noe genotipirovanie.

The comparative study of the genomes of V. cholerae O139 isolated from humans and from water of surface reservoirs was carried out with the use of single- and double-primer polymerase chain reaction (PCR). The profiles of polymorphic DNA fragments obtained in this study made it possible to find out differences between groups of strains, as well as the individual features of some of them. The comparison of strains isolated from humans and from water in single-primer PCR revealed that they, in spite of the general similarity of their genomes, essentially differed, which was probably due to changes in the genome of this infective agent. Strains of aqueous origin lacked genes ctx and tcpA, which made them epidemiologically unimportant.

[Database on nucleotide sequences used as primers of microorganisms]. / Baza dannykh o nukleotidnykh posledovatel'nostiakh, ispol'zuemykh v kachestve praimerov mikroorganizmov.

The data base (DB) "Primers of microorganisms" for the accumulation and systematization of information on oligonucleotide sequences, used as primers in polymerase chain reaction, has been created. This DB includes data on primers for the laboratory diagnostics of 20 bacterial genera (Aerococcus, Aeromonas, Bartonella, Borrelia, Burkholderia, Chlamydia, Clostridium, Corynebacterium, Escherichia, Francisella, Helicobacter, Legionella, Listeria, Mycobacterium, Mycoplasma, Salmonella, Shigella, Staphylococcus, Vibrio, Yersinia) and 6 viral families (Arenaviridae, Flaviviridae, Hepadnaviridae, Herpesviridae, Picornaviridae, Retroviridae). DB contains data on 145 pairs of primers and 530 bibliographic sources. The retrospective depth of DB is 10 years (1987-1996), and it is replenished as new Russian and foreign documented sources of information arrive.

[Cloning and study of the function of recA-like gene of Yersinia pestis in Escherichia coli cells]. / Klonirovanie i issledovnie funktsii recA-podobnogo gena Yersinia pestis v kletkakh kishechnoi palochki.

A 2 kb fragment of Yersinia pestis genome cloned in Escherichia coli cells of the strain HB101 contains a gene able to complement the recA-dependent deficiency of E. coli cells in UV-resistance, resistance to alkylating agents, UV- and MNNG-induced mutability. Cellular capability for homologous recombination in crosses with HfrH donor, derepressed synthesis of bacteriocins (colicin E1 and pesticin 1) is also complemented by the fragment in E. coli recA-strains. The obtained data suggest the functional homology of the cloned recA-like gene product with the product of E. coli recA-gene.

[Genetic determination of the vibriocinogenicity trait in Vibrio cholerae of the El Tor biotype]. / Issledovanie geneticheskoi determinirovannosti priznaka vibriotsinogennosti u kholernykh vibrionov biotipa El-Tor.

In two different strains of cholera vibrios two recA-dependent plasmids, pVib I (1.9-2.2 Md) and pVib II (5.2-5.8 Md), have been detected. These plasmids determine the synthesis of vibriocin, coagulase and fibrinolysin, which has been established by the cotransformation of the DNA of plasmids pVib I and pBR322 and by the transfer mobilization with the use of plasmid RP4.

[Isolation, purification and physicochemical properties of pesticin I from cells of Yersinia pestis strain EV]. / Vydelenie, ochistka i nekotorye fiziko-khimicheskie svoistva pestitsina I iz kletok shtamma EV Yersinia pestis.

Pesticin I has been isolated and purified from Y. pestis strain EV. The homogeneous preparation of Pesticin has been shown to be monomer protein with a molecular weight of 65000 daltons, having three immunologically identical alpha-, beta- and gamma-forms with different isoelectric points. The amino acid composition of Pesticin I is presented. Rabbit anti-serum to the beta-form of the preparation of Pesticin has been obtained.