[Use of RT-PCR and enzyme immunoassays for the specific diagnosis of Crimean hemorrhagic fever].

The objective of the investigation was to evaluate the efficiency of the RT-PCR kit "AmplySens CHF" produced by InterlabService of the Central Research Institute of Epidemiology and that of the ELISA kits made by the D. I. Ivanovsky Research Institute of Virology for the specific diagnosis of Crimean hemorrhagic fever (CHF). Examination of sera from CHF patients from the Astrakhan Region showed that positive RT-PCR results were observed in 95.2 and 37.5% on days 4-8 and 9-13 after disease onset, respectively; but they were absent on days 13-17. Positive ELISA-IgM results were found in 93% on disease days 6 to 16. A high percentage (78.9%) of positive IgG samples was seen only on days 9-16. Thus, RT-PCR has a marked efficiency in diagnosing CHF until day 8 of illness while ELISA-IgM has it on day 8 or later. ELISA-IgG can be considered to be a confirming rather than compulsory test. The findings suggest that the RT-PCR kit "AmplySens CHF" produced by InterlabService of the Central Research Institute of Epidemiology and that of the ELISA kits made by the D. I. Ivanovsky Research Institute of Virology have a pronounced sensitivity and specificity and a high efficiency when concurrently used to verify CHF in patients.

[Pathogenicity of West Nile virus: molecular markers].

The review describes the phenotypic properties, structure, and expression pattern of West Nile virus genome (Flaviviridae, Flavivirus, Japanese encephalitis antigenic complex), as well as the clinical picture and pathogenesis of its etiologically related disease West Nile fever. It also analyzes the available data on the impact of genetic mutations in the genome on the biological properties of the virus.

[Population interactions of West Nile virus (Flaviviridae, Flavivirus) with arthropode vectors, vertebrates, humans in the middle and low belts of Volga delta in 2001-2006].

The population interactions of West Nile virus in the middle and lower regions of the Volga delta in 2001-2006 were studied. The authors established major species of the mosquitoes Anopheles messeae, An. hyrcanus, Culex piplens, Cx modestus, Coquilletidia richiardii in anthropogenic biocenoses and An. hyrcanus, Coq. richiardii in the natural ones; vertebrates - crows (Corvidae) and domestic animals in the anthropogenic biocenoses and cormorants in the natural ones. The ticks Hyalomma marginatum were found to play an active role in the circulation of the virus and in the stability of infection foci. The epicenter of an endemic area is situated in the anthropogenic biocenoses of the middle belt where high infection rates were found in humans (20-40% immune stratum), mosquitoes, and birds. A complete sequence of 14 strains was detected in 44% from 1 to 5 nucleotide replacements to a genome, most frequently at E154 positions Asn --> Ser and Ser --> Pro, which prevents glycosylation and reduces neuroinvasion. Analysis of 100 RT-PCR-positive samples revealed the absolute predominance of genotypes 1 with single findings of genotype 2 and 4.

[Molecular virological monitoring of Newcastle disease virus strains (Paramyxoviridae, Avulavirus) in the populations of wild birds in the Volga estuary (the 2001 data)].

The paper analyzes the results of isolation of Newcastle disease virus (NDV) strains from 336 swaps of 31 wild bird species collected in the 2001 summer in the Volga estuary (Astrakhan Region). Twenty-seven NDV strains were isolated from little terns (Sterna albifrons) (n=11; infection rate, 24.4%), great cormorants (Phalacrocorax carbo) (n=6; 11.1%), coots (Fulica atra) (n=8; 6.5%), sandwich terns (Sterna sandvicensis) (n=1; 100%), and common redshanks (Tringa totanus) (n=1; 50.0%). Four strains were sequenced by the 374 n. a. residue fragment from the beginning of the F gene, one of them was by the full F gene, and another (Stemal/Astrakhan/2755/2001) was by the full genome. Nucleotide sequences have allowed the authors to classify corresponding NDV strains as 5b genotype and the analysis of the amino acid sequence of the F-protein cleavage site has shown them to belong to a non-pathogenic group.

[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.

[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.

[Ecology and evolution of influenza viruses in Russia (1979-2002)]. / Ekologiia i évoliutsiia virusov grippa v Rossii (1979-2002 gg.).

The research results on ecology and evolution of influenza A viruses, which has been conducted by the Center of Ecology and Evolution of influenza Viruses of Ivanovsky's Institute of Virology, Russian Academy of Medical Sciences, for more than 30 years, are summarized in the paper. A gene pool of influenza A viruses circulating in Russia's territory was defined. Foci of influenza A viruses were detected in natural biocenosis. Issues conditioned by the population interrelations of influenza viruses, i.e. between the populations of wild and home animals and the populations of people, are also under discussion.

[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.

West Nile virus and other zoonotic viruses in Russia: examples of emerging-reemerging situations.

Studies of the interactions of vertebrates, viruses and arthropod vectors of these viruses were monitored in terms of different ecological groups of viruses transmitted by mosquitoes and ticks in Northern Eurasia in an area encompassing more than 15 million km2. About 90 viruses were isolated, including 24 new to science. Newly recognized infections of vertebrates, including humans, were described. Many unusual epidemic situations were analysed. Permanent efforts were established to prevent bioterrorist activities and their consequences. Extensive epidemic outbreaks of West Nile fever (WNF; i.e., fever caused by West Nile virus) and Crimean-Congo hemorrhagic fever (CCHF) with unusual high mortality appeared in the last four years in southern Russia. We determined infection rates in humans, domestic and wild animals, mosquitoes and ticks from natural and synanthropic biocenoses [Editorial note: "synanthropic" means, roughly, all species living with (c.f. lice, fleas) or near people, such as in houses (c.f. house mice), parks (c.f. Rattus spp.), and the like, rather like "peridomestic", but not strictly so; "biocenosis" is the biome, the "totality of living populations in a particular habitat, which itself is only a part of the ecosystem".]. CCHF virus strains were phylogenetically similar to strains isolated in this area 35 years ago but different from Central-South-Asian and African strains. Before the outset of the current emergence of epidemic WNF, three genetic variants of this virus had been isolated in USSR, two African and one Indian. Phylogenetic analysis of complete genome sequences of epidemic strains demonstrated considerable similarity to strains from USA and Israel and differences from strains isolated in the same USSR areas 20-30 years before. In addition to strains of genotype 1, we isolated strains of second and third lineages and a strain of a fourth genetic variant. Nucleotide differences of these strains from all three genotypes was about 30%. The emerging WNF situation in Russia for the last 4 years probably has been the result of not only natural and social factors, but also to introduction of more virulent strains or by evolution of the virus.

[ West Nile fever among residents of the Astrakhan region in 2002]. / Zabolevaemost' i zarazhaemost' naseleniia Astrakhanskoi oblasti likhoradkoi Zapadnogo Nila v 2002 g.

Thirty-three persons infected with West Nile fever were detected in 2002 in the Astrakhan Region; the diagnosis was confirmed serologically and the maximal number of the infected was registered in August, same year. The indices of the specific humoral immunity varied from 3.3% to 27.1%. A monitoring determined the highest infection risk among the residents of the Volga middle delta.

[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.

[The study of immune structure of Astrakhan region population to West Nile virus in 1998 and 1999]. / Izuchenie immunostruktury naseleniia Astrakhanskoi oblasti k virusu Zapadnogo Nila v 1998 i 1999 gg.

Immunostructure of the Astrakhan Region population to West Nile fever (WNF) was studied in the preepidemic period (1998) and after the outbreak (1999). Among the sera obtained in 1998, 63 (26.3%) were positive in neutralization reaction, 84 (27.1%) in enzyme immunoassay IgG and 20 (7.8%) in HAIR. IgM-antibodies were found in none of 142 samples. Overall number of donors having antibodies to WNF virus by three reactions reached 97 (31.6%). In the sera obtained in 1999, virus neutralizing antibodies were detected in 72(44.4%) cases, specific IgM antibodies detected by EIA_in 5(3.1%), IgG_in 44(27.1%), antihemagglutinins_in 11 (6.9%). The number of positive findings in 4 reactions in 1999 was 81(50.0%). The results of examination of the sera collected for two years (1998 and 1999) were the following: of 402 samples examined in NR positive were 135(33.6%), of 304 five (1.6%) were IgM positive, 128(27.1%) of 472 were IgG positive, and 31(7.4%) of 417 responded in HAIR. Overall index of humoral immunity for 2 years was 37.9% (in males and females 39.8 and 32.8%, respectively. In persons aged 20-29 years_36.9%, 50-59 years_42.9%. Thus, by 2-year results, antibodies to WNF virus occurred in 51.9% rural citizens and 35.0% city population.

[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.