[The first break-through of the genotype 2.3.2 of high-virulence influenza A virus A/HSN1, which is new for Russia, in the Far East].

The epizootic etiologically associated with highly pathogenic avian influenza H5N1 genotype 2.3.2 that is new for Russia among wild and domestic birds in the south of the Primorye Territory during spring migration in April 2008 has been decoded. About 25% of the wild birds of a water complex, which include European teals (Anas crecca), mallard ducks (Anas platyrhynchos), great-crested grebes (Podiceps cristatus), are involved in viral circulation in the area of the Suifun-Khankai plain. Chicken embryos and the cell lines MDCK, SPEV, BHK-21, SW-13 were used to isolate 3 strains from recently deceased hens (A/chicken/Primorje/1/08, A/chicken/Primorje/11/08, and A/chicken/Primorje/12/08) and one strain from a European teal (A/Anas crecca/Primorje/8/08). The strains were deposited in the State Collection of Viruses of the Russian Federation, D. I. Ivanovsky Research Institute of Virology, Russian Academy of Medical Sciences. The nucleotide sequences of the full-sized genomes of A/chicken/Primorje/1/08 and A/Anas crecca/Primorje/8/08 were sent to the International databank GenBank. The strains from domestic and wild birds were shown to be identical. The isolated strains are most close to the strains Alchicken/Viet Nam/10/05, A/chicken/Guangdong/178/04, and A/duck/Viet Nam/12/05. Molecular genetic analysis has indicated that the strains isolated are susceptible to rimantadine and ozeltamivir and less adapted to mammalian cells (particularly, they contain E627 in RV2, which agrees with the biological properties of these strains in vitro). Penetration of the newly isolated virus into the Far East ecosystem provides in the foreseeable future a way for infecting the birds wintering in America and Australia in the nesting places, with further carriage of viral populations there in the period of autumn migrations.

[Interpretation of the epizootic outbreak among wild and domestic birds in the south of the European part of Russia in December 2007].

The paper presents the results of interpreting the epizootic outbreak etiologically associated with high-virulent influenza virus A/H5N1 among domestic and wild birds in the Zernogradsky and Tselinsky districts of the Rostov Region. Epizooty was characterized by a high infection rate in the synanthropic birds of a ground-based complex. RT-PCT revealed influenza virus A/H5 in 60% of pigeons and crows and in around 20% of starlings, and in 10% of tree sparrows. Fifteen viral strains from chickens (Gallus gallus domesticus), Indian ducks (Cairina moschata), rooks (Corvus frugilegus), rock pigeons (Columba livia), tree sparrows (Passer montanus), common starlings (Sturnus vulgaris), and great white herons (Egretta alba) were isolated and deposited in the State Collection of Viruses of the Russian Federation. Full-sized genomes of 5 strains were sequenced and deposited in the international database GenBank. The isolated strains belong to the Quinhai-Siberian (2.2) genotype, an Iranian-Northern Caucasian subgroup, they are phylogenetically closest to the strain A/chicken/Moscow/2/2007 (inducing epizooty among poultry in the near-Moscow Region in February 2007) and have 13 unique amino acid replacements as the consensus of the Quinhai-Siberian genotypes in the proteins PB2, PA, HA, NP, NA, and M2, by preserving thereby 4 unique replacements first describes for the strain A/chicken/Moscow/2/2007. The findings are indicative of a different mechanism that is responsible for bringing the virus into the northeastern part of the Azov Sea area in September 2007 (during the fall migration of wild birds) and in December 2007 in the south-western Rostov Region where a human factor cannot be excluded. Mass infection of synanthropic birds endangers the further spread of epizooty, including that in the central regions of the Russian Federation in spring after near migrants return after wintering.

[Epizooty caused by high-virulent influenza virus A/H5N1 of genotype 2.2 (Qinghai-Siberian) among wild and domestic birds on the paths of fall migrations to the north-western part of the Azov Sea basin (Krasnodar Territory)].

Isolation, followed by the sequencing the full-size genome of strains of A/chicken/Krasnodarl300/07 and A/Cygnus cygnus/Krasnodar/329/07, has shown that they belong to genotype 2.2 (Qinghai-Siberian). The strains were deposited at the State Virus Collection of the Russian Federation and nucleotide consequences were at the International databank GenBank. The strains contained 10 unique amino acid replacements in reference to the consensus of the Qinghai-Siberian genotype in the PB2, PA, HA, NA, and NS1, which suggests that regional variants may form in different parts of an area.

Application of universal primers for identification of Foot-and-mouth disease virus and Swine vesicular disease virus by PCR and PCR-ELISA.

Two approaches for simultaneous identification of both Foot-and-mouth disease virus (FMDV) and Swine vesicular disease virus (SVDV) are described: (1) a single-step reverse transcription-PCR with three primers and (2) a PCR-ELISA assay with two universal primers for genome amplification and two virus-specific probes for identification. These methods are based on the use of 3D gene universal PCR primers, the structure of which was optimized and refined due to the close relationship between the two viruses belonging to different genera of the Picornaviridae family. In procedure (1), a three-primer PCR containing one universal antisense primer and two virus-specific primers was shown to differentiate between FMDV and SVDV in one reaction, due to the different length of the amplified DNA fragments (600 and 340 base pairs, respectively). In procedure (2), the two viruses were identified by PCR-ELISA, i.e. PCR for the 3D gene followed by two parallel hybridizations with FMDV and SVDV-specific probes in microplate wells and ELISA detection. The application of universal primers could halve the number of PCR experiments in both cases, as compared to the usual virus-specific PCR procedures. Also, we investigated the 3D gene structure of several SVDV strains isolated at different times. No essential changes were detected in the regions coding for conserved motifs of the RNA-dependent RNA polymerase recognized by our universal primers. The multi-primer PCR was successfully tested on 38 FMDV and 15 SVDV strains, and the PCR-ELISA on 32 FMDV and 16 SVDV strains including clinical material from disease cases.

[Characteristics of field isolates of Newcastle disease virus isolated in the course of outbreaks in the poultry plant in the Leningrad region in 2000]. / Kharakteristika polevogo izoliata virusa n'iukaslskoi bolezni, vydelennogo vo vremia vspyshki na ptitsefabrike v Leningradskoi oblasti v 2000 g.

A field isolate of Newcastle disease virus (NDV) was isolated in the Russko-Vysotskaya poultry farm, Leningrad region. Within four days after infection, the isolate caused 100% mortality in 60-day-old susceptible chickens. The HA titer of the allantoic fluid samples collected after one passage in SPF-chicken embryos was 1:512, and it reacted only with the NDV specific antiserum in HI test. Intracerebral pathogenicity index and mean embryo death time were 1.97 and 49 hours, respectively. The isolate has the amino acid sequence of the protease cleavage site of the fusion protein F0 (112R-R-Q-R-R-F117), which is similar to that in the velogenic strains of NDV. Therefore, it was concluded that the virus isolated in this work was an ethiological agent of the ND outbreak in this poultry farm.

[Adenovirus KR95, isolated from chickens during an outbreak of hydropericarditis, is the pathogen of this disease]. / Adenovirus KR95, vydelennyi ot tsypliat vo vremia vspyshki gidroperikardita, iavliaetsiia vozbuditelem étogo zabolevaniia.

Virus agent KR95 was isolated from the liver of dieoff chickens during an outbreak of hydropericarditis syndrome at a poultry farm in Russia. Electron microscopic examination of the virus morphology, comparative restriction cleavage map construction, DNA-DNA hybridization, and analysis of structural proteins from purified and disrupted virions showed that the agent is to be classified as type 1 avian adenovirus.

[Comparative analysis of the VP2 variable region of the gene from infectious bursal disease virus isolates]. / Sravnitel'nyi analiz variabel'noi oblasti gena VP2 izoliatov virusa infektsionnoi bursal'noi bolezni.

Variable cDNA regions in the VP2 gene of 24 isolates of infectious bursal disease virus (IBDV) isolated in Russia in 1993-1996 were amplified by the "nested" PCR and sequenced. The primary structure analysis of the VP2 gene variable region revealed 2 major groups of IBDV isolates. The first group consisted of the isolates with the structure identical or closely related to the highly virulent European strains CS89, 74/89A, 661, JY86, and DV86, the second group included the isolates with a high level of homology to the vaccine strains PBG98 and Cu-1. In addition, two isolates with original structure were identified, which differed from previously studied strains.