Khabarovsk, Russia
Khabarovsk, Russia

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Rar V.A.,Russian Academy of Sciences | Epikhina T.I.,Russian Academy of Sciences | Pukhovskaya N.M.,Khabarovsk Antiplague Station | Vysochina N.P.,Khabarovsk Antiplague Station | And 2 more authors.
Infection, Genetics and Evolution | Year: 2014

To study Babesia diversity in Ixodid ticks in Russia, Ixodes persulcatus, Haemaphysalis japonica, Haemaphysalis concinna, Dermacentor silvarum, and Dermacentor nuttalli ticks collected in the Far East and Baikal region were assayed for the presence of Babesia spp. using nested PCR. In total, Babesia DNA was detected in 30 of the 1125 (2.7%) I. persulcatus, 17 of the 573 (3.0%) H. concinna, and 12 of the 543 (2.2%) H. japonica but was undetectable in any of the 294 analyzed Dermacentor spp. Partial 18S rRNA gene sequences were determined for all of the positive samples. Among the positive ticks, nine I. persulcatus were infected by Babesia microti 'US'-type, five I. persulcatus were infected by Babesia divergens-like parasites, and 11 I. persulcatus were infected by Babesia venatorum. For all three of these species, the determined 18S rRNA gene sequences were identical to those of the Babesia genetic variants found previously in I. persulcatus in Russia. In addition, five I. persulcatus from the Baikal region and all of the positive Haemaphysalis spp. ticks carried 13 different sequence variants of Babesia sensu stricto belonging to distinct phylogenetic clusters. Babesia spp. from 29 ticks of different species collected in distinct locations belonged to the cluster of cattle and ovine parasites (. Babesia crassa, Babesia major, Babesia motasi, Babesia bigemina, etc.). Babesia spp. from four H. japonica ticks in the Far East belonged to the cluster formed by parasites of carnivores. One more Babesia sequence variant detected in an I. persulcatus tick from the Baikal region belonged to the cluster formed by parasites of cattle and wild cervids (. B. divergens, Babesia capreoli, B. venatorum, Babesia odocoilei, etc.). © 2014 Elsevier B.V..


PubMed | Russian Academy of Sciences and Khabarovsk Antiplague Station
Type: | Journal: Infection, genetics and evolution : journal of molecular epidemiology and evolutionary genetics in infectious diseases | Year: 2014

To study Babesia diversity in Ixodid ticks in Russia, Ixodes persulcatus, Haemaphysalis japonica, Haemaphysalisconcinna, Dermacentor silvarum, and Dermacentor nuttalli ticks collected in the Far East and Baikal region were assayed for the presence of Babesia spp. using nested PCR. In total, Babesia DNA was detected in 30 of the 1125 (2.7%) I. persulcatus, 17 of the 573 (3.0%) H. concinna, and 12 of the 543 (2.2%) H. japonica but was undetectable in any of the 294 analyzed Dermacentor spp. Partial 18S rRNA gene sequences were determined for all of the positive samples. Among the positive ticks, nine I. persulcatus were infected by Babesia microti US-type, five I. persulcatus were infected by Babesia divergens-like parasites, and 11 I. persulcatus were infected by Babesia venatorum. For all three of these species, the determined 18S rRNA gene sequences were identical to those of the Babesia genetic variants found previously in I. persulcatus in Russia. In addition, five I. persulcatus from the Baikal region and all of the positive Haemaphysalis spp. ticks carried 13 different sequence variants of Babesia sensu stricto belonging to distinct phylogenetic clusters. Babesia spp. from 29 ticks of different species collected in distinct locations belonged to the cluster of cattle and ovine parasites (Babesia crassa, Babesiamajor, Babesiamotasi, Babesiabigemina, etc.). Babesia spp. from four H. japonica ticks in the Far East belonged to the cluster formed by parasites of carnivores. One more Babesia sequence variant detected in an I. persulcatus tick from the Baikal region belonged to the cluster formed by parasites of cattle and wild cervids (B. divergens, Babesiacapreoli, B. venatorum, Babesiaodocoilei, etc.).


Igolkina Y.,Russian Academy of Sciences | Bondarenko E.,Join stock company Vector Best | Rar V.,Russian Academy of Sciences | Epikhina T.,Russian Academy of Sciences | And 7 more authors.
Ticks and Tick-borne Diseases | Year: 2016

Rickettsia spp. are intracellular Gram-negative bacteria transmitted by arthropods. Two potentially pathogenic rickettsiae, . Candidatus Rickettsia tarasevichiae and . Rickettsia helvetica, have been found in unfed adult . Ixodes persulcatus ticks. The aim of this study was to assess the prevalence and genetic variability of . Rickettsia spp. in . I. persulcatus ticks collected from different locations in the Russian Far East. In total, 604 adult . I. persulcatus ticks collected from four sites in the Khabarovsk Territory (continental area) and one site in Sakhalin Island were examined for the presence of . Rickettsia spp. by real-time PCR. Nested PCR with species-specific primers and sequencing were used for genotyping of revealed rickettsiae. The overall prevalence of . Rickettsia spp. in ticks collected in different sites varied from 67.9 to 90.7%. However, the proportion of different . Rickettsia species observed in ticks from Sakhalin Island significantly differed from that in ticks from the Khabarovsk Territory. In Sakhalin Island, . R. helvetica prevailed in examined ticks, while . Candidatus R. tarasevichiae was predominant in the Khabarovsk Territory. For . gltA and . ompB gene fragments, the sequences obtained for . Candidatus R. tarasevichiae from all studied sites were identical to each other and to the known sequences of this species. According to sequence analysis of . gltA, . o(cyrillic)mpB and . sca4 genes, . R. helvetica isolates from Sakhalin Island and the Khabarovsk Territory were identical to each other, but they differed from . R. helvetica from other regions and from those found in other tick species. For the first time, DNA of pathogenic . Rickettsia heilongjiangensis was detected in . I. persulcatus ticks in two sites from the Khabarovsk Territory. The . gltA, . ompA and . o(cyrillic)mpB gene sequences of . R. heilongjiangensis were identical to or had solitary mismatches with the corresponding sequences of . R. heilongjiangensis found in other tick species. © 2016 Elsevier GmbH.


PubMed | Russian Academy of Sciences, Institute of Chemical Biology and Fundamental Medicine, Join stock company Vector Best and Khabarovsk Antiplague Station
Type: Journal Article | Journal: Ticks and tick-borne diseases | Year: 2016

Rickettsia spp. are intracellular Gram-negative bacteria transmitted by arthropods. Two potentially pathogenic rickettsiae, Candidatus Rickettsia tarasevichiae and Rickettsia helvetica, have been found in unfed adult Ixodes persulcatus ticks. The aim of this study was to assess the prevalence and genetic variability of Rickettsia spp. in I. persulcatus ticks collected from different locations in the Russian Far East. In total, 604 adult I. persulcatus ticks collected from four sites in the Khabarovsk Territory (continental area) and one site in Sakhalin Island were examined for the presence of Rickettsia spp. by real-time PCR. Nested PCR with species-specific primers and sequencing were used for genotyping of revealed rickettsiae. The overall prevalence of Rickettsia spp. in ticks collected in different sites varied from 67.9 to 90.7%. However, the proportion of different Rickettsia species observed in ticks from Sakhalin Island significantly differed from that in ticks from the Khabarovsk Territory. In Sakhalin Island, R. helvetica prevailed in examined ticks, while Candidatus R. tarasevichiae was predominant in the Khabarovsk Territory. For gltA and ompB gene fragments, the sequences obtained for Candidatus R. tarasevichiae from all studied sites were identical to each other and to the known sequences of this species. According to sequence analysis of gltA, mpB and sca4 genes, R. helvetica isolates from Sakhalin Island and the Khabarovsk Territory were identical to each other, but they differed from R. helvetica from other regions and from those found in other tick species. For the first time, DNA of pathogenic Rickettsia heilongjiangensis was detected in I. persulcatus ticks in two sites from the Khabarovsk Territory. The gltA, ompA and mpB gene sequences of R. heilongjiangensis were identical to or had solitary mismatches with the corresponding sequences of R. heilongjiangensis found in other tick species.


Rar V.A.,Russian Academy of Sciences | Livanova N.N.,Russian Academy of Sciences | Panov V.V.,Russian Academy of Sciences | Doroschenko E.K.,Gamaleya Institute of Epidemiology and Microbiology | And 3 more authors.
Ticks and Tick-borne Diseases | Year: 2010

Totally, 2590 questing adult Ixodes persulcatus ticks and 1458 small mammals from Ural, Siberia, and the Far East as well as 53 Haemaphysalis concinna, 136 Haem. japonica, and 43 Dermacentor silvarum ticks - exclusively adults - from the Far East were examined for the presence of Ehrlichia and Anaplasma by nested PCR based on the 16S rRNA gene. Both Anaplasma phagocytophilum and Ehrlichia muris were found in I. persulcatus and small mammals from all the studied regions. Myodes spp., Microtus spp., Sorex araneus, Apodemus peninsulae, and Tamias sibiricus were naturally infected with An. phagocytophilum and E. muris. Five of the examined I. persulcatus and 5 of the examined wild rodents from Siberia and the Far East were infected with 'Candidatus Neoehrlichia mikurensis'. The determined 16S rRNA gene sequences of 'Candidatus Neoehrlichia mikurensis' were identical to the sequences of Japanese isolates, while the determined groESL sequences were unique. A new Ehrlichia sp. variant closely related to the Ehrlichia sp. EHf669 found in Haem. flava from Japan was detected in 11% of Haem. japonica ticks. New Anaplasmataceae bacteria genetically distinct from the known species of this family were found in 3 adult Derm. silvarum from the Far East and in 2 I. persulcatus from Siberia and the Far East. In the Far East, about 15% of the captured small mammals were naturally infected with recently discovered Ehrlichia sp. Khabarovsk. Ehrlichia sp. Khabarovsk was found in about 20% of Myodes spp. and S. araneus but was undetectable in any of the 236 studied Ap. peninsulae. A three-year study has demonstrated that An. phagocytophilum and E. muris were detectable in small mammals from the Far East captured only after the beginning of the tick activity season, from May to November. Ehrlichia sp. Khabarovsk was found in mammals trapped in all the examined periods, from February to November. © 2010 Elsevier GmbH. All rights reserved.


Rar V.A.,Russian Academy of Sciences | Epikhina T.I.,Russian Academy of Sciences | Livanova N.N.,Russian Academy of Sciences | Panov V.V.,Russian Academy of Sciences | And 3 more authors.
Molecular Genetics, Microbiology and Virology | Year: 2010

A total 932 small mammals and 458 questing adult Ixodes persulcatus from Sverdlovsk, Novosibirsk regions and Khabarovsk Territory as well as 128 Haemaphysalis japonica, 34 H. concinna and 29 Dermacentor silvarum from Khabarovsk Territory was examined on the Babesia presence by nested PCR based on the 18S rRNA gene. Babesia microti DNA were found in samples from small mammals in all studied regions - in 36.2% of samples from Sverdlovsk region, in 5.3% of samples from Novosibirsk region and in 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 the sequence of B. microti strain Munich. B. microti were found most frequently in samples from Myodes spp., they were found also in Microtus spp., Apodemus spp., Sorex spp. and Sicista betulina. One from 347 analyzed I. persulcatus from Novosibirsk region and one from 77 I. persulcatus from Khabarovsk Territory were shown to contain B. microti US-type DNA. One I. persulcatus from Novosibirsk region have contained B. divergens DNA. This is the first determination of B. divergens in I. persulcatus and the first determination of B. microti in I. persulcatus in Asian part of Russia. Three novel genetic variants of Babesia sensu stricto were revealed in three H. japonica from Khabarovsk Territory. One novel Babesia 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 Babesia crassa (97.1-97.6% similarity); they clustered together with ruminant pathogens B. crassa and Babesia major. © 2010 Allerton Press, Inc.


Rar V.A.,Russian Academy of Sciences | Epikhina T.I.,Russian Academy of Sciences | Livanova N.N.,Russian Academy of Sciences | Panov V.V.,Russian Academy of Sciences | And 4 more authors.
Vector-Borne and Zoonotic Diseases | Year: 2011

The specimens of 3552 questing adult Ixodes persulcatus and 1698 blood/tissue samples of small mammals collected in Ural, Siberia, and Far East of Russia were assayed for the presence of Anaplasma phagocytophilum by nested PCR based on the 16S rRNA gene. Totally, A. phagocytophilum was detected in 112 tick and 88 mammalian samples. The nucleotide sequences of the 16S rRNA gene and groESL operon (1244-1295 bp) were determined for A. phagocytophilum samples from 65 ticks and 25 small mammals. Six different 16S rRNA gene variants differing by 1-5 nucleotide substitutions were detected, and only one variant matched the sequences deposited in GenBank. Analysis of groESL sequences allowed the A. phagocytophilum samples to be divided into three groups; moreover, the samples from different groups also differed in the 16S rRNA gene sequences. The A. phagocytophilum sequences from group I were detected in 11 Myodes spp. samples from West Siberia and Far East and in 19 I. persulcatus samples from all examined regions; from group II, in 10 samples of Myodes spp. and common shrews (Sorex araneus) from Ural; and from group III, in four samples of Asian chipmunks (Tamias sibiricus) from West Siberia and Far East; and in 46 I. persulcatus samples from all examined regions. The nucleotide sequences of A. phagocytophilum groESL operon from groups I and II were strictly conserved and formed with A. phagocytophilum groESL sequence from a Swiss bank vole (Myodes glareolus) (GenBank accession no. AF192796), a separate cluster on the phylogenetic tree with a strong bootstrap support. The A. phagocytophilum groESL operon sequences from group III differed from one another by 1-4 nucleotides and formed a separate branch in the cluster generated by European A. phagocytophilum strains from roe deer (Capreolus capreolus) and Ixodes ricinus ticks. © Copyright 2011, Mary Ann Liebert, Inc.


Rar V.A.,Russian Academy of Sciences | Epikhina T.I.,Russian Academy of Sciences | Pukhovskaya N.M.,Khabarovsk Antiplague Station | Vysochina N.P.,Khabarovsk Antiplague Station | Ivanov L.I.,Khabarovsk Antiplague Station
Molecular Genetics, Microbiology and Virology | Year: 2013

In total, 484 Haemaphysalis japonica, 359 Haemaphysalis concinna, and 221 Dermacentor silvarum collected in Amur oblast and Khabarovsk krai of the Russian Far East were examined regarding the presence of Anaplasmataceae bacteria using nested PCR. All positive samples were characterized by analysis of the 16S rRNA gene and/or groESL operone nucleotide sequences. Forty-nine H. japonica and three H. concinna were shown to contain DNA of two new Ehrlichia genetic variants. On the basis of 16S rRNA gene and groESL operone nucleotide sequences analysis, these genetic variants were found to be most closely related to Ehrlichia spp. revealed in Haemaphysalis spp. ticks in Japan. Four H. concinna from Amur oblast were shown to contain DNA of a new Anaplasma bovis genetic variant, which corresponded to the A. bovis genetic variant revealed in a red gray-backed vole and a Siberian chipmunk from the Far East. Three H. concinna and nine D. silvarum contained DNA of atypical bacteria that cannot be attributed to any Anaplasmataceae genera based on the determined sequences of the 16S rRNA gene fragments. The revealed atypical bacteria significantly differed from each other and did not form a separate genetic group on the basis of 16S rRNA gene sequences. © 2013 Allerton Press, Inc.


Rar V.A.,Russian Academy of Sciences | Pukhovskaya N.M.,Khabarovsk Antiplague Station | Ryabchikova E.I.,Russian Academy of Sciences | Vysochina N.P.,Khabarovsk Antiplague Station | And 4 more authors.
Ticks and Tick-borne Diseases | Year: 2015

Recently, a new Ehrlichia genetic variant, Ehrlichia sp. Khabarovsk, was identified in tissue samples of small mammals captured in the Russian Far East. To further characterize Ehrlichia sp. Khabarovsk, tissue homogenate from a naturally infected gray red-backed vole (Myodes rufocanus) was passaged three times in newborn laboratory mice. Using nested PCR Ehrlichia sp. Khabarovsk DNA was detected in tissue samples from infected mice at 1-4 weeks post inoculation. Electron microscopic examination revealed morulae containing gram-negative bacterial cells in monocytes of mouse spleen and liver. The size and ultrastructure of these cells corresponded to those described previously and allowed us to identify the bacteria as Ehrlichia sp. The comparison of ehrlichial 16S rRNA, groEL and gltA genes and putative GroEL and GltA amino acid sequences has demonstrated that Ehrlichia sp. Khabarovsk, like Ehrlichia ruminantium, is more distant from all other Ehrlichia species than these species are between themselves. Phylogenetic analysis has shown that Ehrlichia sp. Khabarovsk belongs to the clade formed by Ehrlichia spp. but clusters separately from other Ehrlichia species and genetic variants. These data indicate that Ehrlichia sp. Khabarovsk can be considered as a new candidate species. We propose to designate it as '. Candidatus Ehrlichia khabarensis' according to the territory where this species was found. © 2015 Elsevier GmbH.


PubMed | Russian Academy of Sciences and Khabarovsk Antiplague Station
Type: Journal Article | Journal: Ticks and tick-borne diseases | Year: 2015

Recently, a new Ehrlichia genetic variant, Ehrlichia sp. Khabarovsk, was identified in tissue samples of small mammals captured in the Russian Far East. To further characterize Ehrlichia sp. Khabarovsk, tissue homogenate from a naturally infected gray red-backed vole (Myodes rufocanus) was passaged three times in newborn laboratory mice. Using nested PCR Ehrlichia sp. Khabarovsk DNA was detected in tissue samples from infected mice at 1-4 weeks post inoculation. Electron microscopic examination revealed morulae containing gram-negative bacterial cells in monocytes of mouse spleen and liver. The size and ultrastructure of these cells corresponded to those described previously and allowed us to identify the bacteria as Ehrlichia sp. The comparison of ehrlichial 16S rRNA, groEL and gltA genes and putative GroEL and GltA amino acid sequences has demonstrated that Ehrlichia sp. Khabarovsk, like Ehrlichia ruminantium, is more distant from all other Ehrlichia species than these species are between themselves. Phylogenetic analysis has shown that Ehrlichia sp. Khabarovsk belongs to the clade formed by Ehrlichia spp. but clusters separately from other Ehrlichia species and genetic variants. These data indicate that Ehrlichia sp. Khabarovsk can be considered as a new candidate species. We propose to designate it as Candidatus Ehrlichia khabarensis according to the territory where this species was found.

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