[Identification of Ixodes persulcatus and Ixodes pavlovskyi occidentalis (Ixodidae) by the analysis of the gene fragment COXI (cytochrome oxidase subunit I)].

Ticks of the genus Ixodes were collected in 2010 in the lowland part of Toguchinsk district of Novosibirsk Province (Russia) and in the forest-park area of Novosibirsk Scientific Centre and its outskirts (Sovetskiy district of Novosibirsk), and identified as Ixodes persulcatus (Schulze, 1930) (18 females and 13 males) and Ixodes pavlovskyi (13 females and 10 males). Ten specimens of each sex from each collecting site were examined. The following nine characters were used: the length and width of the scutum (conscutum) and of the gnathosoma in ventral view; the length of palpal segments II-III; the width of the hypostome; the length of idiosoma with scapula, of leg I, of the medial spur on fore coxa (Taiga..., 1985; Filippova, Musatov, 1996; Filippova, Panova, 1998). According to morphometric characters, specimens of Ixodes pavlovskyi collected in the forest-park area of the Novosibirsk Scientific Centre were identified as the subspecies I. p. occidentalis Filippova et Panova, 1998. Nucleotide sequences of the COI mitochondrial gene fragment were determined for 56 ticks. Phylogenetic analysis of the COI gene fragment in representatives of the persulcatus-ricinus species-group dwelling in Asia demonstrated high degree of conservatism. Molecular-genetic methods allow reliable identification of morphologically similar species I. pavlovskyi and I. persulcatus, pathogenic for humans.

[Age changes in Ixodes pavlovskyi (Ixodinae) natural populations from Novosibirsk and its outskirts].

The age structure of I. pavlovskyi natural populations from Novosibirsk and its outskirts was studied with the use of the fat reserves in the midgut and in the fat body as the age index.

[Characteristics of the distribution of ticks Ixodes persulcatus and Ixodes pavlovskyi at the border between the forest and forest-steppe zones in the territory near Ob River].

Field investigations performed in 2009 and 2010 in the Novosibirskiy, Toguchinskiy, and Sovetskiy districts of Novosibirsk Province showed, that at present, Ixodes persulcatus, I. pavlovskyi (subspecies I. pavlovskyi occidentals Filippova et Panova, 1998), and Dermacentor reticulatus (Fabricius, 1794) permanently cohabitate in the woodland park of the Novosibirsk Akademgorodok, and I. pavlovskyi is the dominating species there. The highest abundance of I. pavlovskyi was recorded in pine forests subjected to intensive recreational load. At the same time, I. pavlovskyi was not found in the Novosibirskiy District and in the plane part of Toguchinskiy District, while the abundance of I. persulcatus is almost three times higher in the above mentioned territories, than in the Akademgorodok.

[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).

Genetic diversity of Babesia in Ixodes persulcatus and small mammals from North Ural and West Siberia, Russia.

OBJECTIVE: The aim of this work was to study the prevalence and genetic diversity of Babesia in Ixodes persulcatus ticks and small mammals from Ural and Siberia in Russia. METHODS: In total, 481 small mammals and 922 questing adult I. persulcatus from North Ural (Sverdlovsk region) and West Siberia (Novosibirsk region) were examined for the presence of Babesia by nested PCR based on the 18S rRNA gene. RESULTS: Babesia microti of the 'Munich'-type was found in 36.2% of blood samples of the small mammals from the Sverdlovsk region and B. microti of the 'US'-type in 5.3% of the animals from the Novosibirsk region. Babesia DNA was not detected in 133 analysed I. persulcatus from the Sverdlovsk region; however, it was found in 24 of 789 ticks from the Novosibirsk region. Three distinct Babesia species were detected in I. persulcatus. B. microti 'US'-type was identified in 10 ticks, Babesia closely related to B. divergens/B. capreoli in 2 ticks, and Babesia closely related to B. venatorum (EU1) in 12 ticks. CONCLUSION: To our knowledge, this is the first detection of Babesia sensu stricto in I. persulcatus ticks and of B. microti in I. persulcatus in the Asian part of Russia.

[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.

[The abundance and distribution of the Ixodes persulcatus (Acari: Ixodidae) near its northern spreading limit in the Ural Mountains].

A count of the tick species Ixodes persulcatus Schulze, 1930 was carried out in the "Denezhkin Kamen" Nature Reserve and adjacent territories (the Severoural'sk and Ivdel' Districts of the Sverdlovsk Region, the Northern Urals geographical province) in the 2005. The abundance and distribution of unengorged adults has been evaluated on an area of 22.5 square kilometers (N 60 degrees 27'-60 degrees 30' E 059 degrees 38'-059 degrees 42'). The area includes proportionally main landscape and vegetation elements of the region studied, from mountain analogues of the middle and northern taiga up to tundra. One tick species, I. persulcatus, has been collected by flagging with the abundance from 0.4 up to 6.8 (average 1.6 +/- 0.9) specimens per flag-hour. The observed values of abundance are classified into three classes (I - ticks are absent, II - 1-2 specimens, and III - 3-7 specimens per flag-hour). The class I amounts 20, II - 75, and III - 5% of the area examined. It has been revealed by the expert evaluation of the 2003-2004 and counts of the 2005 that ticks occur stably in the Northern Ural, reaching N 61 degrees and 400 m above sea level. The level of the species abundance remained constant till the middle of summer. In this period the activity of ticks dependent on the weather optimum only.

[Detection of DNA of borrelia circulating in Novosibirsk region].

Detection of DNA of Borrelia burgdorferi sensu lato was performed by PCR in taiga ticks Ixodes persulcatus, in blood samples and skin bioptates of small forest mammals, and in blood and urine samples of humans after attaching of ticks events. In Novosibirsk region both in natural reservoir and in patients with Ixodes ticks-borne borreliosis DNA of Borrelia garinii and Borrelia afzelii are detected. DNA of these borrelia were detected in 8 from 72 of taiga ticks, in 36 from 298 of blood and skin samples of small forest mammals, and in 32 from 102 of human blood and urine samples. In all studied samples DNA of B. garinii from NT29 subgroup was predominated. Borrelia DNA in which sequence of intergenic spacer region was homologous to sequence Chy13p first detected in China has been detected in one blood sample from red-backed vole (Clethrionomys rutilus).