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.

Potential and limits to unravel the genetic architecture and predict the variation of Fusarium head blight resistance in European winter wheat (Triticum aestivum L.).

Genome-wide mapping approaches in diverse populations are powerful tools to unravel the genetic architecture of complex traits. The main goals of our study were to investigate the potential and limits to unravel the genetic architecture and to identify the factors determining the accuracy of prediction of the genotypic variation of Fusarium head blight (FHB) resistance in wheat (Triticum aestivum L.) based on data collected with a diverse panel of 372 European varieties. The wheat lines were phenotyped in multi-location field trials for FHB resistance and genotyped with 782 simple sequence repeat (SSR) markers, and 9k and 90k single-nucleotide polymorphism (SNP) arrays. We applied genome-wide association mapping in combination with fivefold cross-validations and observed surprisingly high accuracies of prediction for marker-assisted selection based on the detected quantitative trait loci (QTLs). Using a random sample of markers not selected for marker-trait associations revealed only a slight decrease in prediction accuracy compared with marker-based selection exploiting the QTL information. The same picture was confirmed in a simulation study, suggesting that relatedness is a main driver of the accuracy of prediction in marker-assisted selection of FHB resistance. When the accuracy of prediction of three genomic selection models was contrasted for the three marker data sets, no significant differences in accuracies among marker platforms and genomic selection models were observed. Marker density impacted the accuracy of prediction only marginally. Consequently, genomic selection of FHB resistance can be implemented most cost-efficiently based on low- to medium-density SNP arrays.

[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.

Quantitative trait loci for adult-plant resistance to Mycosphaerella graminicola in two winter wheat populations.

Septoria tritici blotch (STB) is one of the most important leaf spot diseases in wheat worldwide. The goal of this study was to detect chromosomal regions for adult-plant resistance in large winter wheat populations to STB. Inoculation by two isolates with virulence to Stb6 and Stb15, both present in the parents, was performed and STB severity was visually scored plotwise as percent coverage of flag leaves with pycnidia-bearing lesions. 'Florett'/'Biscay' and 'Tuareg'/'Biscay', each comprising a cross of a resistant and a susceptible cultivar, with population sizes of 316 and 269 F(7:8) recombinant inbred lines, respectively, were phenotyped across four and five environments and mapped with amplified fragment length polymorphism, diversity array technology, and simple sequence repeat markers covering polymorphic regions of ≈1,340 centimorgans. Phenotypic data revealed significant (P < 0.01) genotypic differentiation for STB, heading date, and plant height. Entry-mean heritabilities (h(2)) for STB were 0.73 for 'Florett'/'Biscay' and 0.38 for 'Tuareg'/'Biscay'. All correlations between STB and heading date as well as between STB and plant height were low (r = -0.13 to -0.20). In quantitative trait loci (QTL) analysis, nine and six QTL were found for STB ratings explaining, together, 55 and 51% of phenotypic variation in 'Florett'/'Biscay' and 'Tuareg'/'Biscay', respectively. Genotype-environment and QTL-environment interactions had a large impact. Two major QTL were detected consistently across environments on chromosomes 3B and 6D from 'Florett' and chromosomes 4B and 6B from 'Tuareg', each explaining 12 to 17% of normalized adjusted phenotypic variance. These results indicate that adult-plant resistance to STB in both mapping populations was of a quantitative nature.

Molecular marker assisted broadening of the Central European heterotic groups in rye with Eastern European germplasm.

Broadening the genetic base of heterotic pools is a key to ensure continued genetic gains in hybrid breeding and extend hybrid cultivation to new areas. In the present study, two Central European heterotic pools (Carsten and Petkus) and five Eastern European open-pollinated varieties (OPVs, Pop-1 to Pop-5) were studied with the objectives to (1) investigate the genetic diversity in OPVs and the heterotic pools using molecular and field data, (2) evaluate the molecular diversity among OPVs, (3) examine the combining ability for grain yield of the OPVs when crossed with testers in field trials, and (4) develop a strategy for targeted introgression of OPV germplasm into the heterotic pools. In total, 610 S(0) plants, 347 from OPVs and 263 from heterotic pools, were developed. Clones of the S(0) plants of OPVs were crossed with two testers belonging to each heterotic pool, while clones of heterotic pools were crossed with only the opposite tester. Testcrosses were evaluated for grain yield in multi-location trials. In addition, 589 S(0) plants were fingerprinted with 30 SSR markers. The data revealed that the Carsten pool has a narrow genetic base and should be the primary target for broadening the established heterotic pattern. Mean and genetic variance suggested that Pop-2 and Pop-4 are good candidates for introgression in Petkus pool and Pop-5 in Carsten pool. Nevertheless, introgression of Pop-5 in Carsten could reduce the genetic diversity between heterotic pools. Therefore, we suggest that either selected plants of Pop-5 should be introgressed or more Eastern European germplasm should be fingerprinted and field evaluated to identify promising germplasm for broadening the established heterotic pattern.

[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.

A microsatellite map of wheat.

Hexaploid bread wheat (Triticum aestivum L. em. Thell) is one of the world's most important crop plants and displays a very low level of intraspecific polymorphism. We report the development of highly polymorphic microsatellite markers using procedures optimized for the large wheat genome. The isolation of microsatellite-containing clones from hypomethylated regions of the wheat genome increased the proportion of useful markers almost twofold. The majority (80%) of primer sets developed are genome-specific and detect only a single locus in one of the three genomes of bread wheat (A, B, or D). Only 20% of the markers detect more than one locus. A total of 279 loci amplified by 230 primer sets were placed onto a genetic framework map composed of RFLPs previously mapped in the reference population of the International Triticeae Mapping Initiative (ITMI) Opata 85 x W7984. Sixty-five microsatellites were mapped at a LOD >2.5, and 214 microsatellites were assigned to the most likely intervals. Ninety-three loci were mapped to the A genome, 115 to the B genome, and 71 to the D genome. The markers are randomly distributed along the linkage map, with clustering in several centromeric regions.

Genetic mapping of 66 new microsatellite (SSR) loci in bread wheat.

In hexaploid bread wheat ( Triticum aestivum L. em. Thell), ten members of the IWMMN ( International Wheat Microsatellites Mapping Network) collaborated in extending the microsatellite (SSR = simple sequence repeat) genetic map. Among a much larger number of microsatellite primer pairs developed as a part of the WMC ( Wheat Microsatellite Consortium), 58 out of 176 primer pairs tested were found to be polymorphic between the parents of the ITMI ( International Triticeae Mapping Initiative) mapping population W7984 x Opata 85 (ITMI pop). This population was used earlier for the construction of RFLP ( Restriction Fragment Length Polymorphism) maps in bread wheat (ITMI map). Using the ITMI pop and a framework map (having 266 anchor markers) prepared for this purpose, a total of 66 microsatellite loci were mapped, which were distributed on 20 of the 21 chromosomes (no marker on chromosome 6D). These 66 mapped microsatellite (SSR) loci add to the existing 384 microsatellite loci earlier mapped in bread wheat.

Chromosomal location and genetic mapping of the mismatch repair gene homologs MSH2, MSH3, and MSH6 in rye and wheat

The efficiency of homeologous recombination is influenced by mismatch repair genes in bacteria, yeast, and mammals. To elucidate a possible role of these genes in homeologous pairing and cross-compatibility in plants, gene probes of wheat (Triticum aestivum) specific for the mismatch repair gene homologues MSH2, MSH3, and MSH6 were used to map them to their genomic positions in rye (Secale cereale). Whereas MSH2 was mapped to the short arm of chromosome 1R, MSH3 was mapped to the long arm of chromosome 2R and MSH6 to the long arm of chromosome 5R. Southern blots with nullisomic-tetrasomic (NT) lines of wheat indicated the presence of the sequences on the respective homeologous group of wheat chromosomes. Additionally, an MSH6-specific homologue could also be detected on homoeologous group 3 of wheat. However, in the well-known, highly homoeologous pairing wheat mutant ph1b the MSH6-specific sequence is not within the deleted part of chromosome 5BL, indicating that the pairing phenotype is not due to a loss of one of the mismatch repair genes tested.

Daily intakes of 134Cs, 137Cs, 40K, 232Th, and 238U in Ukrainian adult males.

Total diet samples were collected to estimate dietary intakes of several radionuclides for Ukrainian males by duplicate portion studies. The samples were collected in the Kiev, Rovno and Volynsky regions in autumn of 1994. Some foodstuffs, including milk and potato, were also collected in the same regions. Daily intakes of 232Th and 238U were estimated to be 2.0 mBq and 7.5 mBq per person, respectively. The estimated daily intake of 40K, 85 Bq per person, was equivalent to that of Reference Man (the intake of a West European and a North American). Daily intakes of 137Cs were found to be quite different between some regions. Daily intakes were 1.8 Bq per person in the Kiev region and 12 Bq per person in the Rovno and Volynsky regions. Daily intake of 134Cs was below 1 Bq per person.

[Effect of alginic acid and its salts on the dynamic of 85Sr and 137Cs accumulation in rats]. / Vliianie al'ginovoi kisloty i ee solei na dinamiku nakopleniia 85Sr i 137Cs v organizme krys.

Accumulation of 85Sr and 137Cs radionuclide blend in rats was studied. Effect of alginic acid, its salts as well as foods containing the polysaccharides on dynamics of accumulation was established. It was shown that efficiency of compounds studied depends on type of salt of alginic acid and presence of guluronic acid in it.

[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.

[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.

[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.