Human seroprevalence data indicate other factors than climatic conditions influencing dirofilariosis transmission in the Russian Federation.

The species of the genus Dirofilaria are filarial parasites causing zoonotic infections in humans with an increasing incidence in temperate and tropical areas of the world. Due to its classification as a vector-borne disease, the most important factors influencing dirofilariosis transmission are those related to climate, such as temperature and humidity. However, other factors linked with human behaviour can influence the distribution of the parasite. Although the Russian Federation could be considered as a non-suitable area for Dirofilaria spp. transmission due to its climatic conditions, one third of the human cases of dirofilariosis have been declared in this country. Here, seroepidemiological data on human dirofilariosis for five different regions distributed throughout the Russian Federation (Rostov, Moscow, Ekaterinburg, Yakutia and Khabarovsk) were obtained. A total of 940 serum samples from totally random donors living in these areas were analysed by enzyme-linked immunosorbent assay for the detection of anti-Dirofilaria immitis immunoglobulin G antibodies. Similar seroprevalence data ranging from 3.41% in Yakutia to 6.95% in Khabarovsk, with no significant correlation with climatic data of yearly average temperature and rainfall from these regions were found. These results suggest that other factors probably related to human behaviour, and not only climatic conditions, might be facilitating the spread of human dirofilariosis in these areas.

[Specific genetic features of the Russian forms of bovine leukemia virus].

Genetic peculiarities of bovine leukemia virus isolates (BLV) spread throughout Russia and Ukraine (BLV-1, BLV-2, and BLV-4) have been characterized based on pol gene polymorphism. Seven viral forms have been detected. The variability of BLV isolates did not exceed 1% within one form. Despite the recent inhabitation of BLV in Russia in the middle of 20th century, Russian BLV variants are characterized by several specific nucleotide substitutions. Point mutations that result in the changes in the aminoacid sequence of reverse transcriptase of BLV specific to distinct viral forms were observed. C --> G transition at the 2752 position (relatively to the reference genome AF033818), which results in the substitution of glutamic to asparaginic acid (GAG --> GAC), is specific to form BLV-2. This mutation was demonstrated in BLV isolates from Ukraine. The T --> G substitution at the 2758 position, which results in the substitution of isoleucine for methionine (ATT --> ATG), is specific to BLV-4 and BLV-7 forms. The BLV-4 form was only detected in Russia and Ukraine. The present study also includes a review of the published data concerning BLV variability. The existing classifications of BLV forms have been critically conceived and the new optimal classification of BLV forms with the maximal resolution has been suggested.

[Identification and classification of bovine leukemia virus isolates in Russia and Ukraine based on the pol viral gene polymorphism].

Bovine leukemia virus (BLV) is a widespread specific pathogen of cattle. Analysis of the pol viral gene polymorphism has been used to characterize the polymorphism of BLV isolates at stock-breeding farms in Russia and Ukraine. The fragments of the pol gene corresponding to the reverse transcriptase and integrase 494 and 233 bp in size, respectively, have been used for analysis. Phylogenetic analysis has revealed several variants of BLV clustered with a high bootstrap support in Russia and Ukraine. A new classification of BLV variants is suggested. Comparison of phylograms based on the polymorphism of the nucleotide sequences of the integrase and reverse transcriptase domains did not show topological conflicts. Therefore, recombination between BLV variants has not been found.

[Comparison of ISSR polymorphism among cattle breeds].

Polymorphism analysis of DNA fragments flanked by (TC)9G and (CT)9G inverted dinucleotide microsatellite repeats in 766 animals of 19 cattle breeds and one breeding type revealed 66 fragments, of which 64 were polymorphic. The breeds proved to differ in the frequency and presence or absence of amplified DNA fragments at the genomic level, indicating that ISSR fingerprinting is informative for differentiating the PCR product spectra and cattle breeds. Multilocus ISSR polymorphism analysis identified the group of fragments that can be used as Bos taurus and B. indicus species markers to describe the standards of breeds, their genetic profiles, and breed-specific patterns. Based on ISSR polymorphism, a prototypal gene pool of cattle was constructed and the breeds closest to it were identified. Genetic diversity analysis made it possible to assume that an optimal mean heterozygosity is characteristic of cattle breeds and that deviations from this optimum are indicative of various processes occurring in the population (breed).

[Polymorphism of the BoLA-DRB3 gene in the Mongolian, Kalmyk, and Yakut cattle breeds].

Polymorphism of the BoLA-DRB3 gene was studied with the use of the PCR-RFLP technique in three cattle breeds (Mongolian, Kalmyk, and Yakut) representing the Bos taurus turano-mongolicus group. 35 BoLA-DRB3.2 alleles were detected in the Mongolian breed and 34 alleles in the Kalmyk breed. The frequencies of alleles in both populations are distributed rather evenly: the frequencies of the most widely represented alleles (*18, *20, and *28) in the Mongolian cattle varied from 7.75 to 8.45%. The most frequent alleles in the Kalmyk cattle were *28(14.52%), *24(7.26%), and *12(6.45%). Only five alleles were identified in the Yakut cattle breed. The prevailing allele was *29 (77.3%); a relatively frequent allele was *1(13.1%), and the remaining three alleles constituted only 9.6%. Such a low level of diversity of BoLA-DRB3 gene alleles was not observed earlier in any other cattle breed. The Mongolian and Kalmyk breeds showed a wide diversity of BoLA-DRB3 genotypes (56 and 51 genotypes, respectively) and a high level of expected heterozygosity (He = 0.953 and 0.946, respectively). Both breeds had a deficiency of heterozygotes (Mongolian cattle: Ho = 0.775, D = -0.187; Kalmyk cattle: Ho = 0.708, D= -0.252). A low level ofgenotypic diversity for the BoLA-DRB3 locus (only seven genotypes; the frequency for the genotype *29/*29 is 71.4%) and a very low level of observed heterozygosity (Ho = 0.12) were revealed in the Yakut breed. BoLA-DRB3.2 alleles associated with resistance to persistent lymphocytosis caused by the bovine leukemia virus (total frequencies 15.49 and 24.19%) and to various forms of mastitis (total frequencies 12.68 and 20.96%, respectively) were identified in the Mongolian and Kalmyk animals. In the Yakut breed, alleles associated with resistance to diseases are represented only by the BoLA-DRB3.2 allele *7 (1.2%). Thus, the Mongolian and Kalmyk cattle breeds are characterized by a wide diversity of alleles and genotypes for the BoLA-DRB3 gene. In contrast, the population of Yakut cattle from the Verkhoyanskii raion of the Republic of Sakha has a poor diversity of alleles and genotypes for the BoLA-DRB3 gene and a very low level of heterozygosity, suggesting an unfavorable state of the population that is probably caused by inbreeding depression due to a long-term isolation and a small number of animals.

[Analysis of the genetic structure of Tuvinian short-fat-tailed sheep populations with the use of the ISSR-PCR method].

The genetic structure of populations of the Tuvinian short-fat-tailed sheep was studied with the use of the ISSR-PCR (Inter Simple Sequence Repeats) method in 18 farms of Tyva. Data on the spectrum of ISSR fragments of DNA were obtained using the (AG)9C primer. Analysis of intermicrosatellite polymorphism permitted us to determine genomic characteristics of the populations, their genealogical relations, and the parameters of genetic diversity within the populations and the breed as a whole. Three genetic notions were considered on the basis of the results of this analysis: gene pool profile, gene pool standard, and breed-specific pattern. The data obtained can be used to carry out population genetic monitoring, to develop a breeding strategy, and to conserve in situ the Tuvinian sheep breed and breeds of other domesticated species.