[Study of the heterogeneity of 16s rRNA gene and groESL operone in the dna samples of Anaplasma phagocytophilum, Ehrlichia muris, and "Candidatus Neoehrlichia mikurensis" determined in the Ixodes persulcatus ticks in the area of Urals, Siberia, and far east of Russia].

A total of 3552 Ixodes persulcatus from Sverdlovsk, Chelyabinsk, Novosibirsk, Irkutsk regions and Khabarovsk Territory were examined on the Ehrlichia and Anaplasma presence by nested PCR based on the 16S rRNA gene. Both Anaplasma phagocytophilum and Ehrlichia muris DNA were found in I. persulcatus in all studied regions. A. Phagocytophilum was detected in 1.3-6.3% of ticks and E. muris - in 2.0-14.1% of ticks. Moreover, "Candidatus Neoehrlichia mikurensis" DNA was found in 8 ticks collected in Novosibirsk, Irkutsk Regions and Khabarovsk Territory. Partial nucleotide sequences of 16S rRNA gene and groESL operone (1240-1300 bp) were determined for 65 samples of A. Phagocytophilum, 17 samples of E. muris and 4 samples of "Candidatus Neoehrlichia mikurensis". Nucleotide sequences of 16S rRNA gene and groESL operone of E. muris and "Candidatus Neoehrlichia mikurensis" were shown to be highly conservative, and nucleotide sequences of groESL operone of both E. muris and "Candidatus Neoehrlichia mikurensis" differed from the sequences found previously in other species of Ixodid tick. On the basis of analysis of the 16S rRNA gene and groESL operone sequences it was concluded that all revealed samples A. Phagocytophilum could be divided into 2 groups. GroESL operone sequences of A. Phagocytophilum from the first group were identical to each other but significantly differed from the known groESL operone sequences (less than 98.2% of similarity), whereas their 16S rRNA gene sequences were identical to the sequence of widely distributed and pathogenic for human A. Phagocytophilum genetic variant (CAHU-HGEl, GenBank AF093788) or differed from it by a single nucleotide substitution. The nucleotide sequences of groESL operone of A. Phagocytophilum from the second group differed from each other by 1-4 nucleotides and were closely related (99.2-99.4% of similarity) to the sequences of groESL operone ofA. phagocytophilum isolates found in Europe in Ixodes ricinus and roe deer. The nucleotide sequences of the 16S rRNA gene of A. Phagocytophilum from the second group were most similar to the sequence of the rare A. Phagocytophilum genetic variant previously found only in China (GenBank DQ342324).

[Detection of Babesia spp. DNA in small mammals and ixodic ticks on the territory of north Ural, west Siberia and far east of Russia].

Totally, 932 small mammals and 458 questing adult Ixodes persulcatus from Sverdlovsk and Novosibirsk regions and Khabarovsk Territory, as well as 128 Haemaphysalis japonica, 34 H. concinna and 29 Dermacentor silvarum from Khabarovsk Territory were examined for the presence of Babesia by nested PCR based on the 18S rRNA gene. Babesia microti DNA was found in samples of small mammals from all the studied regions--in 36.2% of samples from Sverdlovsk region, 5.3% of samples from Novosibirsk region, and 6.7% of samples from Khabarovsk Territory. The determined B. microti 18S rRNA gene sequences from Novosibirsk region (6 sequences) and from Khabarovsk Territory (10 sequences) were identical to each other and to the sequences of pathogenic for human B. microti US-type, while the determined B. microti 18S rRNA gene sequences from Sverdlovsk region (12 sequences) were identical to those of B. microti strain Munich. B. microti were found most frequently in samples of Myodes spp., they were found also in Microtus spp., Apodemus spp., Sorer spp., and Sicista betulinav. It was shown that one of 347 analyzed I. persulcatus from Novosibirsk region and one of 77 I. persulcatus from Khabarovsk Territory contained B. microti US-type DNA. One I. persulcatus from Novosibirsk region contained B. divergens DNA. In this work B. divergens was for the first time determined in I. persulcatus and B. microti in I. persulcatus in Asian part of Russia. Three different genetic variants of Babesia sensu stricto were found in three H. japonica from Khabarovsk Territory. The first genetic variant was closely related to Babesia sp. revealed in a feral raccoon in Japan (99.9% similarity on the basis of 18S rRNA gene sequences). Two others Babesia genetic variants were most similar to the ovine pathogen Babesia crassa (97.1-97.6% similarity on the basis of 18S rRNA gene sequences).

[Detection of Borrelia miyamotoi in ticks Ixodes persulcatus from Russia].

Unfed adult Ixodes persulcatus ticks from five regions of Russia were examined to analyze the distribution and diversity of Borrelia miyamotoi. DNA of B. miyamotoi was found in 1.8% of ticks from Leningrad Oblast, 2.9% from Sverdlovsk, 4.5% from Novosibirsk, 2.3% from Irkutsk Oblast, and 2.5% from Khabarovsk Krai. The molecular typing of the B. miyamotoi DNA was based on the partial sequencing of the 16S rRNA, p66, and glpQ genes. The only genetic variant of B. miyamotoi was detected in all samples of ticks collected from these five territories.

[Trends in the spread of pandemic influenza A(H1N1) swl in the Far East in 2009].

The paper describes the trend in the spread of pandemic influenza A(H1N1) swl virus in the Far East, which started in this region 2-3 months later than that in the European part of Russia. By mid-October seasonal epidemic influenza was practically displaced by pandemic one.

[PCR detection of the causative agents of infections transmitted by ticks on the Kamchatka Peninsula].

There has been recently a rise in referrals for Ixodes tick bites in the spring and summer periods in the Kamchatka Territory. Among the dominant tick species, there has been the taiga tick Ixodes persulcatus habiting the extensive areas of the southern and central parts of the peninsula. Examination of 84 I. persulcatus females collected from human beings and domestic animals in 2003 to 2007 detected DNA of the pathogens of tick-borne borreliosis (B. burgdorferi sensu lato), rickettsiasis (R. tarasevichiae and R. helvetica), and Ehrlichiosis/anaplasmosis (A. phagocytophilum). Tick-borne encephalitis RNA and antigens and babesiasis DNA were not found in the study samples. Despite the small number of taiga ticks in Kamchatka, the detection of the pathogens of various infectious diseases in the ticks suggests that there may be a risk for contamination of the peninsula's population with these pathogens.

[Differentiantion of Listeria monocytogenes strains isolated in the Far East and European part of Russia on the basis of polymorphism of genes encoding invasion factors].

Forty Listeria monocytogenes isolates obtained in European and Far East regions of Russia were differentiated on the basis of polymorphism of 5 markergenes. Total length of concatemers obtained after sequencing of internal fragments of genes inlA, inlB, inlC, inlE and prs was 3029 b.p. Comparative analysis of concatemers' sequences revealed 237 variable nucleotides. Totally, 25 sequence types were revealed, and isolates from European and Far East regions belonged to different types. On the dendrogram isolates split on 2 clusters, which correspond to early described phylogenetic lines of L. monocytogenes specie. Isolates obtained in European and Far East regions formed independent subclusters within main clusters. Fifteen clinical isolates of L. monocytogenes belonged to 7 different types. Analysis of epidemiologic data on time and place of isolates obtaining suggested that isolates of the same sequence type are epidemiologically related and might represent one strain; index of discrimination for proposed typing method was calculated as 0.982.

[Natural foci of tick-borne encephalitis on the southwestern coast of Sakhalin]. / Prirodnye ochagi kleshchevogo éntsefalita na iugo-zapadnom poberezh'e Sakahlina.

Nucleic acids of Ixodes persulcatus were studied by molecular hybridization in the natural focus of tick-borne encephalitis in Kholmsk District of Sakhalin Province. The studies have shown wide dissemination of viral RNA in the focus. The infectivity of ticks in various sites of habitation varied from 3.5 to 18.5%, their number fluctuating from 0.4 to 300 and more imago per flag-hour. The most active part of the natural focus has been determined using zoological-viral indexes. The viral strain of tick-borne encephalitis has been isolated.

[Tick-borne encephalitis in a northern zone of coniferous and broad-leaved forests of a focal region of the Amur River basin]. / Kleshchevoi éntsefalit na severe zony khvoino-shirokolistvennykh lesov ochagovogo regiona Priamur'ia.

The geographical variability of the population of tick-borne encephalitis (TBE) virus in the northern part of the zone of combined coniferous and broad-leaved forests was established; this variability was manifested by higher virulence and homogeneity of TBE virus strains, as shown in experiments on white mice receiving the virus extraneurally, in comparison with the southern part of the zone and by higher virus carriership of the ticks Ixodes persulcatus. With the epizootic situation remaining tense and the danger of TBE virus infection still present, TBE morbidity and mortality rates decreased in the years of the construction of the Baikal-Amur Railroad, which was due to greater attention given to measures for the prophylaxis of TBE during this period.

[Demonstration of the tick-borne encephalitis virus in ticks by means of nucleic acid hybridization]. / Indikatsiia virusa kleshchevogo éntsefalita v kleshchakh metodom gibridizatsii nukleinovykh kislot.

The method of molecular hybridization of nucleic acids (MHNA) is compared to the traditional bioprobe technique in the study of virus carriership of I. persulcatus ticks collected in the South and in the North of the area of coniferous and broad-leaved forests in the Khabarovsk Territory. Higher sensitivity of the MHNA method than that of the bioprobe was demonstrated by studying the tick pools: 4.9% versus 1.6% (1985), 18.2% versus 3.6% (1986). Virus carriership studied by MHNA in isolated ticks comprised 8.6% (1985) and 10.8% (1986) in the south of the region and 20.0% (1985)--in the north; average content of RNA of the tick-borne encephalitis virus comprised 10 pg. MHNA allows massive studies of individual ticks and better indication of the virus to be carried out; the response can be obtained in 24-48 hours.

[Effect of detergents on the hemagglutinating activity of different strains of tick-borne encephalitis virus]. / Vliianie detergentov na gemaggliutiniruiushchuiu aktivnost' razlichnykh shtammov virusa kleshchevogo éntsefalita.

A new genetic marker, sensitivity of hemagglutinins of tick-borne encephalitis virus (TBE) strains to the effect of hyamine 10-X (Hs), was used for characterization of the hemagglutinating component of freshly isolated TBE virus variants with differing neurovirulence and for strain grouping. This marked may also be used for directed search of strains highly active in HI.

[Detection of the natural heterogeneity of natural populations of tick-borne encephalitis viruses and a grouping of the strains]. / Vyiavlenie estestvennoi neodnorodnosti prirodnykh populiatsii virusa kleshchevogo éntsefalita i gruppirovka shtammov.

Eighty strains of tick-borne encephalitis virus isolated from I. persulcatus ticks collected in 1976-1981 in the same biotopes in the south and north of the zone of coniferous-broad-leaved forests, and two biotopes of the southern taiga subzone of the Khabarovsk region and Sakhalin island, as well as from the brains of fatal cases were studied. Significant differences were found between maximal and minimal values of the invasive activity in spatially separated populations of the same zone (t = 2.32, P less than 0.02). Significant interpopulation differences were also found in the value of peripheral activity (t = = 2.77, P less than 0.02) and invasivity index (t = 3.83, P less than 0.01). Natural heterogeneity of the strains was found both by previously known markers (high neurovirulence for mice, invasivity index, plaque size in SPEV cell culture, viremia in mice, stability to thiamine 10X- and a new marker proposed by us: the degree of adsorption of hemagglutinins on an ion-exchanger DEAE-Sephadex A-50. Strains of tick-borne encephalitis virus were grouped in two landscape zones, this grouping indicated the existence of significant interpopulation differences.