Tick-borne encephalitis virus isolates from natural foci of the Irkutsk region: clarification of the genotype landscape.

The Irkutsk region is the unique territory where all known subtypes of tick-borne encephalitis virus (TBEV) circulate. In the last years, the phenomenon of changes in TBEV subtypes (substitution of the Far-Eastern subtype by the Siberian one) was noted in some regions of the Russian Federation. The results of individual investigation of 11522 Ixodes persulcatus ticks and brain specimens from 81 small mammals collected in natural foci of the Irkutsk region during 2006-2014 are presented in the article. More than 60 TBEV strains have been isolated and studied by virological methods; E gene fragments (1193 b.p.) of 68 isolates have been typed. The majority of the strains (irrespective of subtype) were of high virulence for laboratory mice (LM) in case of both intracerebral and subcutaneous inoculation of virus. All isolates from warm-blooded small mammals and humans were of high virulence for LM, but placed in the same clusters of the phylogenetic tree with ticks collected in the same area. Tick-borne strains of different virulence also did not form separate clusters on the tree. Phylogenetic analysis showed that modern TBEV genotypic landscape of the studied territory is changing toward absolute predominance of the Siberian subtype (94.1%). This subtype is represented by two groups with prototype strains "Zausaev" and "Vasilchenko". The "Vasilchenko" group of strains is spread on the whole territory under study; the strains of "Zausaev" group were isolated previously in the Irkutsk suburbs. The European subtype of TBEV circulates in natural foci of Pribaikalie permanently (at least 5% of the random sampling); the strains are of high virulence for LM. The Far-Eastern TBEV subtype was not found within the group of isolates collected in 20062014. The phylogenetic relationship of the strains under study had a higher correlation with the place of isolation than with the year or source.

[A NATURAL PLAGUE FOCUS. IN GORNYI ALTAI: FORMATION, DEVELOPMENT, AND FUNCTIONING].

The paper gives the results of analyzing the data of long-term studies of the natural focal pattern of plague in the Gornyi Altai natural focus. It describes a wide range of biological processes occurring in the focus and shows the most important patterns of its functioning as a complex multilevel ecological system. The key features of the formation of the focus have been revealed. The plague focus in South-Western Altai has formed relatively, recently, about half a century ago, then it has intensively developed and its enzootic area and the activity of epizootic manifestations have considerably increased. This process is due to the space-time transformations of the basic ecological and population characteristics of Pallas' pika (Ochotoma pallasi), the principal vector of the pathogen of plague and fleas parasitizing the mammal, which is in turn related to the aridization of mountain steppes in South-Western Altai.

[Cholera Vibrio biofilm: production, characterization and role in reservation of causative agent in water environment].

AIM: Experimental production, characterization and evaluation of the role of cholera vibrio biofilm. MATERIALS AND METHODS: 33 strains of Vibrio cholerae eltor O1 and V. cholerae O139 of various epidemic significance and origin were studied in a series of experiments by bacteriologic, microscopic (light-optic, luminescent, scanning electron microscopy), molecular genetics, spectrophotometric and statistical methods. RESULTS: Formation of a biofilm involving inter-cellular bonds, pili and extracellular material and variability of the microorganism (RO-phenotype and transition into uncultivable forms) was shown at various temperature and substrate conditions. A more pronounced ability to form biofilms was detected for strains isolated from environmental samples compared with isolated from clinical material regardless of their epidemic significance. Toxigenic strains of eltor biovar (from surface reservoirs during cholera outbreaks) have demonstrated the highest parameters of optical density compared with toxigenic clinical isolates and non-toxigenic O1 and O139 serogroup cultures. The presence of mbaA1 and mbaA2, vpsR, toxR, hapA genes is common for strains that form a biofilm. CONCLUSION: The data obtained confirm the role of biofilm in reservation of cholera vibrio strains of various epidemic significance in saprophytic phase of microorganism existence.

[The involvement of some flea species in the epizootic process in the Gorno-Altai natural plague focus: spatial and temporary characteristics].

The relative number of plague pathogen strains isolated from some flea species in different Gorno-Altai natural plague foci substantially varies; this indicator also varies with time. These patterns are due to the difference in the structure of multispecies communities of ectoparasites in these areas and their long-term transformation. As of now, the three species Paradoxopsyllus scorodumovi, Ctenophyllus hirticrus, and Amphalius runatus are widely involved in pathogen transmission in all three foci. These ectoparasites should be referred to as main plague vectors. In each focus, they are joined by other flea species, such as Rhadinopsylla dahurica and Amphipsylla primaris in the Ulandryk focus, Frontopsylla hetera, R. dahurica, Paradoxopsyllus kalabukhovi, and Paramonopsyllus scalodae in the Tarkhatin focus, and P. scalonae and P. kalabukhovi in the Kurai focus, which should be classified as an additional vector.

[Change in the habitat of Yersinia pestis in the Gorno-Altaisk natural focus of plague].

The paper analyzes the change that occurred in the habitat of the causative agent of plague in its Gorno-Altaisk natural focus in 1961 to 2012. Since 1961 when the plague microbe was found to come from the southern slopes of the Saylyugem mountain range, which are located in Mongolia, to the northern slopes situated in Russia, a gradual expansion of the habitat of Yersenia pestis subsp. altaica had commenced in South-Eastern Altai. During the considered period, the area where epizootic manifestations were registered showed an 11-fold increase. In most cases, the spread of the plague pathogen within the focus was natural and occurred in the successive and closely related settlements of Mongolian pikas (Ochotona pallasi). By now, the plague microbe has been widely distributed in three populations of this small animal, which inhabit the territory of South-Eastern Altai.

[Ratio of the numbers of Ixodes persulcatus imagoes in the near-Baikal area with the use of different recording methods].

The data of estimating the number of Ixodes perculcatus by two methods (the number of ticks per flag-hour and that per flag-kilometer) in the near-Baikal area were analyzed. According to these values, the abundance of taiga ticks in individual calculations ranged from 0 to 394. The association between the time of passage and the number of collected ticks was positive and high significant. The correction factors were calculated to obtain the abundance values expressed in per flag-kilometer units from the abundance values per flag-hour and conversely. The value of these factors depended on the number of ticks.

[The genotypy structure of a natural Daphnia population based on the phenotypic response on change in the amount of food].

Natural populations of Daphnia are genetically heterogeneous with regard to phenotypic response to modification in the food quantity (a set of metric quantitative traits). In the majority of the traits, three genetically determined response types to the environmental factor studied have been detected: arithmetic means decrease (R1), increase (R2) or remain constant (St) as a result of a decrease in the food quantity. This phenotypic response was shown to occur in the whole set of the metric traits studied, irrespective of physiological specialization. The problems of assessment of genetic population structure based on phenotypic response of the individuals reflected in quantitative traits are discussed.

[Dispersal of the flea Ctenophyllus hirticrus and spreading of plague epizooties in Gorny Altai].

Gradual dispersion of an abundant flea species Ctenophyllus hirticrus specific to the Pallas's pika (the main plague carrier), is revealed in the Gorno-Altai natural plague focus on the territory, occupied by two populations of this lagomorph. Spreading of Yersinia pestis in these areas took place a short time later the rise of this ectoparasite's abundance. It is supposed that the colonization of these areas by C. hirticrus was one of the factors determined epizooties spreading within the focus and formation of new sites of stable Y. pestis preservation.

[Genotypic structure of a Drosophila population by motor activity of the imago]. / Genotipicheskaia struktura populiatsii drozofily po dvigatel'noi aktivnosti imago.

Analysis of the variation of adult locomotor activity in four samples taken at different times from a natural population of Drosophila melanogaster showed that the total variation of this trait is relatively stable in time and has a substantial genetic component. Genotypic structure of the population for locomotor activity is characterized by the presence of large groups of genotypes with high and low values of this trait. A possible explanation for the presence of such groups in a population is cyclic density-dependent selection.

[Direction of selection in experimental populations of Drosophila during cyclic change in their density]. / Napravlenie otbora v éksperimental'nykh populiatsiiakh Drozofily pri tsiklicheskom izmenenii ikh plotnosti.

The influence of cyclic fluctuations of Drosophila population density, which result from varying exploitation pressure and feeding conditions, on reorganization of the genotypic composition, evaluated for individual reproductive capacity, was studied. It was found that productivity, which was assessed in individual cultures by the number of adult offsprings, was higher for individuals that lived in low density conditions as compared to those in dense populations. These differences persisted in offsprings of compared individuals in two successive generations, thus testifying to their genetic determination. The genotypic composition of populations may change in one or two generations because of the selective pressure of density.

[Population density as a regulation factor of genetic structure and size of animal populations. Phenotypic variability of reaction to increase in density in Drosophila populations]. / Plotnost' naseleniia kak faktor reguliatsii geneticheskoo struktury i chislennosti populiatsii zhivotnykh. Fenotipicheskaia izmenchivost' po reaktsii osobei na uvelichenie plotnosti v populiatsiiakh drozofily.

The presence of considerable level of phenotypic variability for the nature of fertility change in conditions of imago density increase has been shown in Drosophila natural populations. Genetic peculiarities of insects play the main role in its determination. The genotype groups sensitive and stable to density increase exist in the populations. The former have relatively high fertility at low density and low fertility at overcrowding. The latter are of contrary nature of fertility at given density conditions.

[Fluctuations in numbers in box populations of Drosophila and selective genetic mechanism of their regulation]. / Kolebaniia chislennosti iashchichnykh populiatsii drozofily i selektsionno-geneticheskii mekhanizm ikh reguliatsii.

Box populations of Drosophila melanogaster are characterized by two types of periodical fluctuations of numbers: with low and high frequency. High frequency fluctuations are determined by existence of preimago and imago stages and subsequent delay in density-dependent limitation of imago reproduction, duration of which is determined by time of preimago stage. The period of these fluctuations should be limited within two generation, that is confirmed by experimental data. Low frequency fluctuations with the period of 13-15 generations are the result of ecological density-dependent effect. In this case during pick density one can observe continuous degradation of population (i.e. decrease in fecundity and life time of imago) and following decrease in numbers. Temporary changes in fecundity of females and their offspring of the second generation are positively correlated with low frequency fluctuations in numbers. Such relationships show the possibility of density-dependent, cyclic, genetic changes in fecundity connected with fluctuations in numbers. It means that at the phase of growth in numbers when the density is still low, the selection is directed to the individuals with high fecundity sensible to overpopulation. The phase of decline in numbers is connected with high density and selection directed to the individuals with low fecundity in low density populations. The changes in genetic structure of fluctuating population lead to the weakening of this fluctuations and to the maintaining of population under such conditions.

[Epizootological role of fleas in the Gorno-Altai natural plague focus (a review)]. / Epizootologicheskaia rol' blokh v Gorno-Altaiskom prirodnom ochage chumy (obzor).

Epizootological role of fleas in the Gorno-Altai natural plague focus (Sailugemsk focus) and numerous data on the flea viability are analyzed and generalized. Information concerning the flea natural infectivity with Yersinia pestis altaica is represented. Ecological peculiarities of some flea species parasitizing the main host, Mongolian pika Ochotona pallasi, and nature of their interrelations with Y. pestis are investigated. It is shown that the flea taxocenosis provides the permanent all year-round circulation of Y. pestis in the Gorno-Altai natural focus. Certain combinations of structural elements of the flea taxocenosis have a dominant significance in determination the circulation process at different phases of the annual epizootic cycle.

[Asymmetry of the chaetae in fleas (Citellophilus tesquorum) as a possible marker of their capacity for blocking]. / Asimmetriia shchetinok u blokh (Citellphilus tesquorum) kak vozmozhnyi marker sposobnosti k blokoobrazovaniiu.

The fluctuating asymmetry was studied according to two bilateral numeric features of chaetaxy in groups of female Citellophilus tesquorum fleas. One group included insects with the recorded gizzard block during infection with the virulent plague agent strain. The fleas with the block were characterized by considerably higher levels of fluctuating asymmetry as compared with those without fleas.

[An experimental study of the ability of the hybrid crosses of Citellophillus tesquorum fleas of 2 subspecies to transmit the causative agent of plague]. / Eksperimental'noe izuchenie sposobnosti gibridov ot skreshchivaniia blokhi Citellophillus tesquorum dvukh podvidov peredavat' vozbuditelia chumy.

The ability to transmit of plague bacillus was studied in the first-, third- and fourth-generation hybrids bred from C.t.altaicus and C.t.sungaris. The fleas were inoculated with two virulent strains from the Tuva and Transbaikal natural foci of plague. The bacillus was demonstrated to infect the hybrids, form gizzard block, and be transmitted to white mice, causing the animals' death.