[Analysis of the genome of two Getah virus strains (LEIV 16275 Mar and LEIV 17741 MPR) isolated from mosquitoes in the North-Eastern Asia].

The complete nucleotide consequences of two Getah virus strains isolated in Russia (LEIV 16275 Mar) and Mongolia (LEIV 17741 MPR) were first determined and analyzed. The nucleotide sequences of the gene of surface glycoprotein E2 and complete genome were used to establish the phylogenetic relations of the strains with other representatives of the Semliki forest serocomplex. The study and reference strains of Getah virus were shown to form its own cluster that was far away from other representatives of the complex and the sequence of the E2 gene significantly reflected the identification of phylogenetical groups within the Sempliki forest serocomplex.

[Molecular genetic analysis of a diversity of Getah virus strains isolated from mosquitoes in the North-Eastern Asia].

A molecular genetic study was performed of the Getah virus strains isolated in Russia (Eastern Siberia and Far East) and Mongolia. A phylogenetic analysis was made, by examining the nucleotide sequences of genome fragments that had been obtained by RT-PCR and that included the portions of the E1 and E2 surface glycoprotein genes and the 6K gene. A genetic diversity of Getah virus strains in North-Eastern Asia is discussed.

[Isolation of influenza virus A (Orthomyxoviridae, Influenza A virus), Dhori virus (Orthomyxoviridae, Thogotovirus), and Newcastle's disease virus (Paromyxoviridae, Avulavirus) on the Malyi Zhemchuzhnyi Island in the north-western area of the Caspian Sea].

The paper presents the results of the 2003 and 2006 environmental virological monitoring surveys on the Malyi Zhemchuzhnyi Island where a large breeding colony of sea gull (Laridae) is located. In the past several years, expansion of cormorants (Phalacrocorax carbo) has enhanced the intensity of populational interactions. The investigators isolated 13 strains of influenza A virus (Orthomyxoviridae, Influenza A virus) subtype H13N1 (from sea gulls (n = 4), cormorants (n = 9) 1 strain of Dhori virus (Orthomyxoviridae, Thogotovirus) from a cormorantwith clinical symptoms of the disease, 3 strains of Newcastle disease virus (Paramyxoviridae, Avulavirus) from cormorants. RT-PCR revealed influenza A virus subtype H5 in 3.1% of the cloacal lavages from cormorants. Neutralization test indicated that sera from cormorants contained specific antibodies against West Nile (Flaviviridae, Flavivirus) (15.0%), Sindbis (Togaviridae, Alphavirus) (5.0%), Dhori (10.0%), and Tahini (Bunyaviridae, Orthobunyavirus) (5.0%); sera from herring gulls had antibodies against Dhori virus (16.7%); there were no specific antibodies to Inco (Bunyaviridae, Orthobunyavirus) and mountain hare (Lepus timidus) (Bunyaviridae, Orthobunyavirus) virus.

[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.

[Molecular genetic characteristics of the strain A/chicken/Moscow/2/2007 (H5N1) strain from a epizootic focus of highly pathogenic influenza A among agricultural birds in the neear-Moscow region (February 2007)].

Among agricultural birds in the near-Moscow Region (February 2007), local epizootics caused by the highly pathogenic avian influenza A/H5N1 virus seem to be of unintended manual origin. Such a situation may be considered to be model when the source of inoculation is elucidated in cases of potentially possible acts of bioterrorism. Molecular genetic analysis of isolated A/chicken/Moscow/2/2007 strain established its genetic similarity with the highly pathogenic strains detected in the Black-and-Caspian Sea region in 2006. At the same time, comparison of nucleotide sequences of the strain A/chicken/Moscow/2/2007 with the strains of Qinghai-Siberian genotype (CSG) for which the sequences of full-sized genomes are known in the international databases revealed a significant distinction of the near-Moscow strain from the earlier known analogues. The uniqueness of the primary structure of the PB1 gene is shown. The paper discusses the functional value of amino acid substitutions in the proteins of the strain A/chicken/Moscow/2/2007 and in other variants of CSG of the subtype H5N1.

[Complex environmental and virological monitoring in the Primorye Territory in 2003-2006].

The paper presents the results of monitoring of viruses of Western Nile (WN), Japanese encephalitis (JE), tick-borne encephalitis (TBE), Geta, Influenza A, as well as avian paramicroviruses type I (virus of Newcastle disease (ND)) and type 6 (APMV-6) in the Primorye Territory in 2003-2006. Totally throughout the period, specific antibodies to the viruses were detected by neutralization test in wild birds (7.3%, WN; 8.0%, Geta; 0.7% Batai; 2.8%, Alpine hare (Lepus timidus); by hemagglutination-inhibition test in cattle (11.4% WN; 5.9%, JE; j 3.0%, TBE; 11.6%, Geta), horses (6.1, 6.8, 0, and 25.3%, respectively), and pigs (5.4, 1.5, 0, and 5.9%, respectively) by enzyme immunoassay (IgG) in human beings (0.8, 0.5, 6.8, and 3.2%, respectively. Reverse-transcription polymerase chain reaction (RT-PCR) was used to reveal RNA of the NP segment of influenza A virus in 57.9 and 65% of the cloacal swabs from wild and domestic birds, respectively; and the HA-segment of subtype HH was not detected in 2005. HA/H5 RNA was recorded in 5.5 and 6.7% of the swabs from wild and domestic birds, respectively; 6% of the specimens from domestic birds were M-segment positive in 2006. RNA of influenza A virus NA/H7 and RNA was not detected throughout the years. In 2004, the cloacal swabs 8 isolated influenza A strains: two H3N8 and two H4N8 strains from European teals (Anas crecca), two (H3N8 and H6N2) strains from Baikal teals (A. formosa), one (H10N4) strain from shovelers (A. clypeata), and one (H4N8) from garganeys (A. querquedula). In 2004, one ND virus strain was isolated from the cloacal swabs from European teals (A. crecca). RT-PCR revealed RNA of this virus in some 8 more cloacal swabs from black ducks (A. poecilorhyncha) (3 positive specimens), pheasants (Phasianus colchicus) (n = 2), garganeys (A. querquedula) (n = 1), gadwalls (A. strepera) (n = 1), and geese (Anser anser domesticus) (n = 1). Sequencing of the 374-member fragment of the ND virus F gene, which included a proteolytic cleavage site, could assign two samples to the weakly pathogenetic variants of genotype 1, one sample to highly pathogenic variants of genotype 3a, five to highly pathogenic ones of genotype 5b. Isolation of APMV-6 (2003) from common egrets (Egretta alba) and geese (Ans. anser domesticus) is first described.

[Organization of the Karshi virus strain Leiv-2247 Uz genome and its phylogenetic relationships with other representatives of the Flavivirus genus].

For the first time, the Karshi virus complete genome sequence has been determined and analyzed in this work. The Karshi virus was attributed to the Tick-borne encephalitis group on the basis of phylogenetic analysis of polyprotein sequencing, as well as partial NS5 of the Flavivirus genus. Comprehensive analysis of the longed to its own cluster, which includes also the Royal Farm virus.

[Highly pathogenic influenza A/H5N1 virus-caused epizooty among mute swans (Cygnus olor) in the lower estuary of the Volga River (November 2005)].

Molecular virological studies of the field material collected in the epicenter of epizooty with high mortality among mute swans (Cygnus olor) in the area of the lower estuary of the Volga River (November 2005) could establish the etiological role of highly pathogenic influenza A (HPAI) virus of the subtype H5N1. Ten HPAI/H5N1 strains deposited at the State Collection of Viruses of the Russian Federation with the priority dated December 1, 2005 were isolated from the cloacal/tracheal swabs and viscera of sick and freshly died mute swans. Complete nucleotide sequences of all fragments of the genome of 6 strains have been deposited in the Gene Bank. The paper discusses the molecular genetic characteristics of isolated strains.

[The role of ecovirological zoning in prediction of the influence of climatic changes on arbovirus habitats].

The paper contains methodical basis of ecovirological zoning in the analysis of the problem of interrelation between climatic changes and the activity of natural nidi of arboviral infection. The authors offer an original hieroglyphic system of cartographic signs of arboviruses and a map of prevalence of arboviruses in various landscape zones of Noth Eurasia (within the boundaries of the former USSR.) They analyze the three main cartographic approaches to ecovirological zoning of natural territorial complexes, which imply analysis of a ralation to a landscape, conjugated component analysis, and paleographic reconstruction analysis.

[Detection of the circulation of Crimean-Congo hemorrhagic fever virus in the piedmont steppes of the North Caucasus].

The laboratory verified cases of Crimean-Congo hemorrhagic fever (CCHF) in the piedmont steppes of the North Caucasus (Malgobeksky District, Republic of Ingushetia) are first described. The source of the first infection was Ixodidae ticks; three subsequent sources were contacts with the bloody discharges from patients. CCHF virus genome was detected in the blood of the cattle from an epidemic focus and in the pools of the Ixodes ticks Haemaphysalis parva Neum., 1897 and Boophilus annulatus Say, 1821, taken from cattle. The problem of including the piedmont steppes of the North Caucasus into the CCHF nosological area is discussed.

[Khurdun virus, a presumably new RNA-containing virus associated with coots (Fulica atra), isolated in the Volga river delta].

The prototype strain LEIV-Ast 01-5 of the unclassified enveloped RNA-containing Khurdun virus, less than 220 nm in size, which is widely distributed among coots (Fulica atra) in the Volga River delta, was deposited on November 4, 2004, at the State Virus Collection (SVC # 992). The virus was isolated annually (2001-2004) with a frequency of 2.3-8.5% (mean 6.3%) when examining 348 coots caught in the lower and middle zones of the Volga River delta. Virological examinations used mixed pools of the brain and spleen to inoculate neonatal albino mice and the cellular line Vero-E6. One strain was isolated from a pygmy cormorant (Phalacrocorax pygmaeus). The virus could not be isolated from other species of 1080 birds, 20 hares, 140,000 mosquitoes of 5 predominant species, and 6,700 Hyalomma marginatum ticks. Any antigenic relationship of the virus with all the viruses early isolated in the Northern Caspian Sea region has not been found. ELISA has been developed to detect and identify Khurdun virus antigen.

[West Nile virus infection of agricultural animals in the Astrakhan region, as evidenced by the 2001-2004 serological surveys].

Sera sampled from 2,884 farming animals in the Astrakhan region in 2001 to 2004 were investigated by the hemagglutination inhibition test (HIT) in order to indicate specific antibodies to West Nile virus (WNV). HIT-positive samples were investigated by the neutralization test (NT). WNV antibodies were detected in all the examined species of animals: horses (the proportion of positive tests throughout the observation averaged 9.8%; the agreement with NT results was 94.1%), cattle (6,4 and 72.%), camels (5.2 and 41.7%), pigs (3.1 and 75%), and sheep (2.2 and 57.1). Relationships between the environmental features of WNV in different natural zones, the infection rate, and the conditions of keeping farming animals in the Astrakhan region are analyzed.

[Circulation of West Nile virus (Flaviviridae, Flavivirus) and some other arboviruses in the ecosystems of Volga delta, Volga-Akhtuba flood-lands and adjoining arid regions (2000-2002)]. / Osobennosti tsirkuliatsii virusa Zapadnogo Nila (Flaviviridae, Flavivirus) i nekotorykh drugikh arbovirusov v ékosistemakh del'ty Volgi, Volgo-Akhtubinskoi poimy i sopredel'nykh aridnykh landshaftakh (2000-2002 gg.).

Comprehensive virological, serological as well as genetic studies of the ecology of West Nile Virus (WNV) as well as of some other arboviruses were undertaken in different ecosystems in the territories of the Astrakhan Region and of the Kalmyk Republic. The main carriers (mosquitoes, ticks, birds and mammals) were defined as involved in the circulation of viruses within the natural and anthropogenic biocenosis. Phylogenetic examinations of isolated strains and samples, which were positive in RT-PCR, showed an absolute predominance of genotype I virus that was most closely related to American and Israeli strains. At the same time, epidemic strains had up to 6% of nucleotide differences versus the historic strains isolated in the same region 20-30 years ago. Besides, the circulation of genotype IV was discovered; it was characterized by a lower pathogenicity, which, possibly, ensures the shaping of a pronounced immune interlayer bearing no epidemic consequences. An analysis of the study results on the WNV ecology denotes the epicenter of the endemic territory located in the middle part of the Volga delta.

[Detection of West Nile virus in birds in the territories of Baraba and Kulunda lowlands (West Siberian migration way) during summer-autumn of 2002]. / Vyiavlenie virusa Zapadnogo Nila u ptits na territorii Barabinskoi i Kulundinskoi nizmennostei (Zapadno-Sibirskii proletnyi put') v letne-osennii period 2002 g.

West Nile Virus (WNV) was discovered in 3 species of birds collected in summer-autumn, 2002, in the South of Western Siberia. WNV was identified by ELISA and RT-PCR. Three of 5 dead rooks (Corvus frugilegus), which were found in the territory of the Kulunda plain, were WNV-infected. WNV RNA was detected in 2% of samples of internal organs of aquatics birds, i.e. teal (Anas crecca) and garganey (Anas querquedula), caught in the Chany Lake (Baraba plain). Nucleotide sequencing of the 300-472 aa fragment of WNV protein E gene showed the maximum level of homology with strain WNV/LEIV-Vlg99-27889, which was isolated from a patient in Volgograd (1999). A high homology level of nucleotide sequencing denotes the relationship between the WNV circulating in the Northern Caspian Region and in the South of Western Siberia.

[Screening of birds in the Volga delta (Astrakhan region, 2001) for the West Nile virus by reverse transcription-polymerase chain reaction]. / Obsledovanie ptits del'ty Volgi (Astrakhanskaiia oblast', 2001g.) na nalichie virusa likhoradki Zapadnogo Nila metodom obratnoi transkriptsii-polimeraznoi tsepnoi reaktsii.

Infection of birds, residing in the Volga lower and middle delta, with West Nile Virus (WNV) genome was detected by the RT-PCR method. A total of 315 samples of bird organs, collected in the Astrakhan region in August 2001, were examined. Positive results, with various severity degrees, were found in coots (15.1%) and in cormorants (14.3%) in the lower delta. As for the middle delta, the infection rate among coots, herons, sea-gulls and terms was found to be identical and amounted to 8-13%, it was essentially higher in cormorants--42%. The infection rate of land-based birds did not exceed 5% in synanthropic biocenosis. According to a partial sequencing of the 5'-end region of WNV genome, all positive tests can be described as belonging to the 1st WNV genotype. The obtained results are indicative of a high activity of circulation of WNV among the birds of the water and near-water complexes in the mentioned region.

[The isolation of Dhori viruses (Orthomyxoviridae, Thogotovirus) and Crimean-Congo hemorrhagic fever virus (Bunyaviridae, Nairovirus) from the hare (Lepus europaeus) and its ticks Hyalomma marginatum in the middle zone of the Volga delta, Astrakhan region, 2001]. / Isoliatsiia virusov Dkhori (Orthomyxoviridae, Thogotovirus) i Krymskoi-Kongo gemorragicheskoi likhoradki (Bunyaviridae, Nairovirus) ot zaitsa (Lepus europaeus) i sobrannykh s nego kleshei Hyalomma marginatum v srednei sone del'ty Volgi, Astrakhanskaia oblast', 2001 g.

In August, 2001, in the middle zone of the delta of the Volga River, the Astrakhan region, during investigation of the natural foci of West Nile fever and Crimean--Congo hemorrhagic fever (CCHF), the material from the hare (Lepus europaeus, Pallas, 1778 (Lagomorpha, Leporidae) and collected from it the ticks Hyalomna marginatum Koch 1844, was obtained. 4 strains of Dhori virus (Orthomyxoviridae, Thogotovirus) and 2 strains of CCHF virus (Bunyaviridae, Nairovirus) were isolated. This is the first isolation of Thogotovirus genus virus from the wild vertebrates. Considering the overlap of the Dhori virus and CCHF virus areas, similar ecology and the isolation both viruses from the same pool of the ticks, the necessity for the use of the test-system for indication of the viruses, differential diagnosis and accumulation of the data concerning the role of Dhori virus in the human and farm animals pathology is discussed.

[Analysis of new variants of West Nile fever virus]. / Analiz novykh variantov virusa likhoradki Zapadnogo Nila.

The complete nucleotide sequences for 6 strains of the West Nile fever virus were determined. For the first time the complete nucleotide sequences of the Indian isolate and Krsn190 strain, that is the most far phylogenetically from all isolates known at present time were established. The scheme for separation of virus variants into 4 groups and criteria for determination the group to which the isolate belongs are suggested.

[Isolation of the West Nile fever virus from the great cormorant Phalacrocorax carbo, the crow Corvus corone, and Hyalomma marginatum ticks associated with them in natural and synanthroic biocenosis in the Volga delta (Astrakhan region, 2001)]. / Izoliatsiia virusa likhoradki Zapadnogo Nila ot bol'shogo baklana Phalacrocorax carbo, vorony Corvus corone i sobrannykh s nee kleshchei Hyalomma marginatum v prirodnykh i sinantropnykh biotsenozakh v del'te Volgi (Astrakhanskaia oblast', 2001 g.).

Four strains identified as West Nile fever virus by inhibited hemagglutination and neutralization tests, enzyme immunoassay, and reverse transcription polymerase chain reaction were isolated during a virological examination of birds and their collected ticks in the natural and synanthropic biocenoses of the Volga delta. The strains were isolated from the great cormorant (Phalacrocorax carbo), the crow (Corvus corone) and its collected Hyalomma marginatum nymphs. The types of interpopulational relations in the ecological system wild-birds-virus-mosquitoes-synanthroic birds-ticks are discussed.

[Genetic analysis of West Nile fever virus, isolated in the south of the Russian plain (Volgograd and Astrakhan regions) in 1999]. / Geneticheskii analiz virusov likhoradki Zapadnogo Nikia, vydelennykh na iuge Russkoi ravniny (Volgogradskaia i Astrakhanskaia oblasti) v 1999 g.

Two strains of West Nile virus, Vlg 27889 and Ast 986, were isolated from the brain of a dead man and from the blood of a patient, respectively, during an outbreak of serous meningitis and meningoencephalitis in July-September, 1999, in the Volgograd and Astrakhan regions. Analysis of parts of genome of the strains cloned from cell culture by reverse transcription and polymerase chain reaction demonstrated their identity and appurtenance to group I West Nile viruses.