Species and Genetic Diversity of Representatives of the <i>Anaplasmataceae</i> Family Found in the Sympatry Zone of the <i>Ixodes, Dermacentor</i> and <i>Haemaphysalis</i> Genera Ticks

Introduction.On the territory of the Ekhirit-Bulagatsky district of the Irkutsk region zones of sympatry of four Ixodes ticks species are found, where the species and genetic diversity of infectious agents transmitted through tick bites may be more pronounced than in foci with a mono-dominant type of ticks’ population. In this connection, the study of the species and genetic diversity of representatives of the Anaplasmataceae family in the sympatry zone of the Ixodes ticks of closely related species was of scientific interest.Objective: To study the species and genetic diversity of members of the Anaplasmataceae family in the zones of sympatry of Ixodes ticks Ixodes persulcatus, Dermacentor silvarum, D. nuttalli and Haemaphysalis concinna, to identify the main carriers and potential reservoir hosts of ehrlichia and anaplasma.Methods. In the course of the study, 1106 specimens of adult ticks and 49 samples of small mammalian livers from the Ekhirit-Bulagatsky area were analyzed. Anaplasma and ehrlichia DNA were detected by two-round PCR in the presence of genus- and species-specific primers from the 16S rRNA gene region. The nucleotide sequences of the 16S rRNA gene and the fragment of the groESL operon were identified in some samples. Sequencing was carried out according to the Sanger method. Comparative analysis was performed using the BLASTN program and ClustalW method. Epidemiological data analysis was performed using parametric methods of statistical processing of the material.Results. The DNA of Ehrlichia muris and Anaplasma phagocytophilum were detected in all studied species of ticks in their sympatry area. However, the rate of infection of taiga ticks was significantly higher than that of H. concinna and Dermacentor spp. Potential reservoir hosts of the Anaplasmataceae family members can be classified as Microtus oeconomus, M. gregalis, Myodes rutilus and Sorex spp. When analyzing the nucleotide sequences of the 16S rRNA gene, three genetic variants of anaplasma were detected. The nucleotide sequences of the A. phagocytophilum groESL operon belonged to two genetic groups

Epizootic Situation on Anaplasmosis of Small Ruminants in the Irkutsk Region

Anaplasmosis of ruminants is a group of natural focal infections caused by bacteria from the genus Anaplasma of the Anaplasmataceae family. The main etiological agent of anaplasmosis in sheep, goats, and wild ruminants is Anaplasma ovis, which parasitizes in the erythrocytes of these animals. The purpose of this study was the finding and identification of Anaplasma spp. in the blood of small ruminants using genetic methods and obtaining data on the distribution of anaplasmosis in the Irkutsk region. 20 goat blood samples, 611 sheep blood samples and 209 Dermacentor nuttalli ticks from 12 districts of the Irkutsk region were examined for the presence of Anaplasma spp. Only one type of anaplasma, A. ovis, was found among the genotyped samples. A. ovis was found in the blood of sheep and goats in all of the studied districts of the Irkutsk region. The proportion of sheep blood samples containing anaplasma DNA varied from 30 % to 85 %, in goats – from 10 % to 100 % in different districts, and averaged 57.8 % in sheep and 55,0 % in goats. Frequency of infection of D. nuttalli ticks with A. ovis was 5.7 %. The nucleotide sequences of the samples detected in the blood of small ruminants on the territory of the Irkutsk region differed from each other by a single nucleotide substitution and were identical to the sequences of the type strain Haibei, as well as the sequences of A. ovis previously found in the blood of sheep from Mongolia, deer from China, and Dermacentor niveus and Dermacentor nuttalli ticks from China. These sequences were also identical to the sequences previously found in the blood of sheep from Altai and in Dermacentor nuttalli ticks from Tuva, which indicates the wide distribution of these A. ovis genovariants in Siberia and the probable role of D. nuttalli as a carrier of the agent of anaplasmosis of small ruminants in the Irkutsk region

MODERN EPIDEMIOLOGY OF VECTOR-BORNE TICK-BORNE INFECTIONS IN NORTHERN DISTRICTS OF THE IRKUTSK REGION

Проведен вирусолого-микробиологический мониторинг семи северных районов Иркутской области. Полученные в ходе исследования данные свидетельствуют об изменении эпидемиологической ситуации в отношении клещевых инфекций на территории данных районов. Отмечается продвижение природных очагов на север. На территории северных районов выявлены сочетанные очаги, клещевых инфекций, в том числе обнаружена циркуляция новых для Восточной Сибири патогенов - E. muris, A. phagocytophilum, B. miyamotoi

IZOENZYME STRUCTURE OF POPULATIONS OF TICKS IXODES PERSULCATUS SCHULZE (ACARI: IXODIDAE) IN RECREATION AREAS OF IRKUTSK CITY

Allozyme structure of the population of ticks Ixodes persulcatus in 8 enzyme systems is shown in two samples of ticks from the recreational areas in suburbs of Irkutsk. From the total of 13 loci, which encode these enzymes, eight were polymorphic. Based on allozyme analysis of populations of mites, it is found that most of them show intrapopulation heterogeneity and. the lack of difference between the two populations from different foci. A similar pattern may indicate a large rate of migration of genes between populations and. their close relatives

MOLECULAR GENETIC ANALYSIS OF INFECTION AGENTS OF FARM ANIMALS ANAPLASMOSIS ON THE TERRITORY OF WESTERN AND EASTERN SIBERIA

A total of 452 blood samples of cattle, sheep and goat collected in different regions of Altai Republic, Altai region, Novosibirsk and Irkutsk regions were examined on the presence of Anaplasma DNA using nested PCR with subsequent sequencing of PCR fragments. Anaplasma DNA was found in all examined blood samples of goat, 75.2 % samples of sheep, and 49.3 % samples of cattle. A molecular genetic analysis has demonstrated that intraerythrocytic Anaplasma ovis circulates in goat and sheep blood, while an intraerythrocytic Anaplasma sp. Omsk and a new intraerythrocytic Anaplasma sp. Sibl22, which cannot be attributed to any known species, circulate in cattle blood. In addition to intraerythrocytic Anaplasma, DNA of monocytic Anaplasma bovis was found in one blood sample of cattle

Chimeric Antibody 14D5 Protects Mice against the Far-Eastern, Siberian, and European Tick-borne Encephalitis Virus

Tick-borne encephalitis virus (TBEV), belonging to the Flaviviridae family, is the most significant pathogen transmitted by Ixodes ticks and causing one of the most severe human neuroinfections. In Russia, serum immunoglobulin produced from the donor blood is currently used for post-exposure prophylactic and therapy of tick-borne encephalitis virus. However, it is known that preparations obtained from donated blood have certain disadvantages, and therefore development of novel preparations for post exposure prophylaxis and therapy of tick-borne encephalitis is required. To develop an alternative preparation, which does not include donor blood, a chimeric antibody ch14D5 against glycoprotein E of TBEV was constructed.This study was aimed to investigate protective efficacy of the chimeric antibody ch14D5 against the Far-Eastern, Siberian, and European subtypes of TBEV in in vivo experiments.A peripheral mouse model of tick-borne encephalitis was used in this study: the chimeric antibody ch14D5 was administrated intravenously in mice one day after their intraperitoneal infection with TBEV strains Sofjin, Vasilchenko, and Absettarov. Anti-TBEV serum immunoglobulin was used as a control preparation, which was administered in the same way. Protective efficacy of the chimeric antibodies 14D5 was assessed using the log-rank test. In the study, the presence or absence of antibody-dependent enhancement of infection (ADE) was examined when mice, infected with different subtypes of the TBEV, got the antibody ch14d5.Obtained results demonstrated high efficacy of the ch14D5 antibody in post-exposure prophylaxis of the disease in mice infected with any of the used TBEV strains, as well as the absence of ADE.It was shown that protective efficacy of antibody ch14D5 is higher than that of the anti-TBEV serum immunoglobulin, and antibody ch14D5 could be used for development of a therapeutic preparation for post-exposure prophylaxis

BRIEF CHARACTERISTIC OF EUROPEAN GENOTYPE TICK-BORNE ENCEPHALITIS VIRUS STRAINS IDENTIFIED IN SIBERIAN REGION

The molecular-genetic analysis of 13 strains of Western genotype TBEV isolated in Western and Eastern Siberia demonstrated two groups of strains differed geneticallyfrom each other and had a high level of E gene sequences homology within each group. Nevertheless, the heterogeneity of biological propertiesfor some strains within a group was observed

The distribution of tick-borne encephalitis virus genotypes in different types of landscapes of Eastern Siberia

To find out what is the role of some environmental factors in the formation of a heterogeneous viral population, the analysis of the distribution of tick-borne encephalitis virus (TBEV) genotypes in different types of landscapes of Eastern Siberia has been carried out. In our study, it was shown that TBEV area in all investigated landscapes of Eastern Siberia is represented not only by TBEV of genotype 3, absolute dominance of which is noted in the studied regions, regardless the landscapes where the foci are located, but also by strains of other genotypes. The relationship between genotypes 1,3 and "polytypic" strains with a certain type of landscape and the degree of their development by man has not been established. The prevailing number of strains of TBEV genotype 5 is isolated from the material collected in the Transbaikalia, in the landscapes of the mountain taiga. They, as well as the strain of genotype 4, are found in the foci, where joint circulation of several TBEV genotypes is noted. The strains of TBEV genotype 2 were revealed in the landscapes of plains and plateaus taiga and insular steppes and forest steppes of the southern part of the Baikal region. Moreover, the detection of RNA of TBEV genotype 2 in the "polytipic" 763-87 and 765-87 strains isolated in the floodplain and forest-steppe landscapes of the Barguzin hollow does not exclude the possibility of circulation of TBEV genotype 2 in the Transbaikalian foci. The greatest genetic heterogeneity of TBEV is observed in natural foci located along Lake Baikal and in the southeastern part of Transbaikalia

Characteristics of genetic and phenotypic properties of tick-borne encephalitis virus strains isolated from various source on the territory of Eastern Siberia

Since ixodid ticks and their feeders (vertebrates) form the basis of the tick-borne encephalitis virus (TBEV) population, it was interesting to obtain a genetic characteristic of strains isolated from various sources in the territory of Eastern Siberia. In our study, it was found that the circulation of TBEV of genotypes 1, 2, 3, 5 and "polytypic" strains in the territory of Eastern Siberia is maintained both by the main vectors (I. persulcatus ticks) and by their feeders (small and large mammals, birds). The distribution of genotypes in strains groups from ixodid ticks and vertebrates varied. TBEV strains of genotypes 1 and 2 were significantly more often isolated from rodents, and genotype 3 from ixodid ticks (p < 0.05). Isolates of genotype 3 (85.5 %) prevailed in the group of strains isolated from ixodid ticks, of which 29.6 % belonged to "Vasilchenko", and 25.6 % to "Zausaev"subgenotypes. Genotypes 1 and 2 were represented by single isolates (3.3 % and 1.3 %, respectively). Among strains isolated from warm-blooded hosts, the proportion of genotype 1 was 35.3 %, genotype 2 -11.8 %, genotype 3 - 35.3 %. More than half of genotype 3 strains (58.3 %) were related to "Vasilchenko" subgenotype, and the strains of "Zausaev" subgenotype were not detected. The obtained data suggest that ixodid ticks and warm-blooded animals are amplifiers of TBEV of genotypes 3 and 1, respectively. The certain differences in the phenotypic characteristics of strains isolated from ticks and warm-blooded animals have been revealed. It has been found that strains isolated from ticks are more homogeneous in their antigenic characteristics, less heterogeneous in S-feature than strains isolated from warm-blooded sources. The strains isolated from warm-blooded animals demonstrate somewhat greater resistance to warming but reducing the reproduction of the virus at 42 °C, and have greater virulence compared to strains isolated from ticks