[Experimental infection caused by influenza A (H5N1) virus in common gull (Larus canus)].

The influenza A/common gull/Chany/P/2006 (HSN1) virus strain Isolated from a clinically healthy common gull (Larus canus) caused no death of Its natural host (a common gull). The virus was shown to be capable for effective replication in the tissues of the lung, spleen, and upper respiratory tract and in the intestinal mucosal cells of the common gull with further environmental virus liberation elimination along with mucinous discharges from the cloaca and fauces for 2 weeks. The potential role of this bird species in the circulation of influenza virus is discussed.

[Molecular-biological and antigenic features of H5N1 subtype highly pathogenic influenza virus strains isolated in southern Siberia in 2005-2009].

AIM: Study of molecular-biological and antigenic features of H5N1 subtype virus strains isolated in southern Siberia in 2005-2009. MATERIALS AND METHODS: Study was performed by using standard procedures according to WHO recommendations. RESULTS: Hemagglutinin gene of H5N1 subtype virus strain isolated in Siberia belongs to 2 genetical lineages: 2.2 and 2.3.2. Strains of these 2 lineages have antigenic differences. All of the strains are highly pathogenic for chicken and white mice. CONCLUSION: H5N1 subtype highly pathogenic influenza virus variants of 2 different antigenic lineages that have antigenic differences circulated in Siberia in 2005-2009. A possible role of antigenic drift in evolution of H5N1 subtype influenza virus is discussed.

[Characterization of the H5N1 influenza virus isolated during an outbreak among wild birds in Russia (Tuva Republic) in 2010].

The study of basic biological properties of H5N1 subtype strain isolated during an outbreak among wild birds in Russia in 2010 was presented. The study was carried out using conventional methods according to the WHO recommendations. H5N1 influenza virus isolated in Siberia belonged to clade 2.3.2 of the hemagglutinin gene; the phylogenetic analysis was performed. The antigenic characteristics and the basic genetic markers of biological properties were studied. It was shown that all strains were highly pathogenic for chickens and white mice. Thus, it was shown that in Russia in the 2010 H5N1 virus phylogenetically closely related to Asian variants caused epizootic among wild birds. The potential danger of this variant of the virus for humans was confirmed by different methods. We discussed the possibility of formation of H5N1 influenza natural focus.

[Detection of antibodies to A/H5N1 influenza virus in citizens of Russian Federation].

AIM: To determine levels of antibodies to influenza virus A/H5N1 in serum samples of people living in different regions of Russia in order to assess the risk of infection with avian influenza H5N1. MATERIALS AND METHODS: Two thousand one hundred sixty-eight serum samples were tested by hemagglutination inhibition assay for the presence of antibodies to influenza virus A/H5N1. RESULTS: Twenty-six serum samples obtained from residents of Khanty-Mansiysk Autonomous Area and 2 samples from residents of Novosibirsk region were positive for antibodies to serotype A/H5. There were no clinical cases of avain influenza A/H5N1 infection in medical history of studied persons. CONCLUSION: Since cases of asymptomatic carriage of A/H5N1 influenza virus in water birds are described and ability of the virus to survive in water environment for a long time is shown, it seems logical to detect antibodies to influenza virus A/H5 in sera of subjects living in Russian Federation taking into account that influenza virus A/H5N1 is isolated from wild fowl and poultry since 2005.

[Characteristic of high pathogenic avian influenza virus subtype H5N1 isolated from common gull (Larus canus)].

AIM: To study biological characteristics of H5N1 influenza virus isolated from common gull on south of West Siberia in 2006. MATERIALS AND METHODS: Isolation and characterization of biological characteristics performed according to recommendations of World Health Organization. RESULTS: Influenza virus A (H5N1) was first isolated from common gull (Larus canus) in Russia. Antigen of isolated virus had significant affinity to polyclonal sera obtained against high pathogenic avian influenza viruses H5N1 circulating in South-East Asia. Phylogenetic analysis of isolated strain revealed its belonging to group of Qinghai-related variants of H5N1 influenza virus. Aminoacid structure of hemagglutinin proteolytic cleavage site is characteristic for type A high pathogenicity avian influenza viruses. Experimental infection of chickens demonstrated high pathogenicity of the isolated virus. CONCLUSION: Involvement of common gulls in circulation of subtype H5N1 influenza virus is demonstrated for the first time. Important role of species from Laridae family in unprecedented spreading of H5N1 influenza virus started in 2005 is discussed.

[Monitoring of antibodies to influenza A virus in populations of different regions of West Siberia].

AIM: To study levels of antibodies to influenza virus in sera of subjects residing in different regions of West Siberia in order to assess the risk of infection with avian influenza virus H5N1. MATERIALS AND METHODS: Two hundred and sixty-five serum samples were tested for the presence of antibodies to influenza virus A/New Caledonia/99 (H1N1), A/New York/55/2005 (H3N2), A/Whooper swan/ Mongolia/244/2005 (H5N1) by hemagglutination inhibition assay (HAI) and reaction of microneutralization. RESULTS: All tested sera were negative for antibodies to H5N1. 14.2% and 44.1% of sera were positive for antibodies to H1N1 in HAI and reaction of microneutralization respectively. In respect of antibodies to H3N2 virus, the proportion of positive sera was higher--40.3% and 76.2%, respectively. CONCLUSION: Results of such studies are very actual, especially during pandemic threat. Furthermore, such information allows to better predict consequences of seasonal influenza epidemics caused by serotypes circulating at the present time.

[Investigation of susceptibility of influenza viruses A (H1N1), the cause of infection in humans in April-May 2009, to antivirals in MDCK cell culture].

Biological properties of influenza viruses A (H1N1), that were the cause of the infection in humans in April - May 2009, and the action of the Russian antivirals on their reproduction were studied in vitro. The nucleotide sequence in the viruses was determined and followed by detection of the mutations responsible for resistance to the antiinfluenza drugs. The experiments showned that arbidol and ribavirin had a selective inhibitory action on reproduction of the viruses in the MDCK cell culture while rimantadine had no affect on their reproduction. The data were confirmed by the results of the genome analysis in influenza viruses A/California/04/2009(H1N1), A/California/07/2009(H1N1) and A/Moscow/01/2009(H1N1)swl, that revealed no replacements defining the resistance to arbidol while the viruses contained a mutation in position 31 of M2 protein, responsible for the resistance to adamantans.

[Hemagglutination inhibition test for retrospective diagnosis of avian influenza in mammals].

AIM: To compare usage of native and formalinized erythrocytes from different animal species in hemagglutination inhibition (HI) test for detection of level of specific antibodies to H5N1 influenza virus in sera of mammals. MATERIALS AND METHODS: Level of anti-H5 antibodies to influenza H5 control antigen and to influenza viruses A/Common gull/Chany/2006 (H5N1), A/duck/Tuva/01/06 (H5N1), A/Anas platyrhynchos/Chany Lake/9/03 (H5N3) was determined by hemagglutination inhibition test in two influenza A (H5) reference antisera as well as in ferret antisera to native strains of avian influenza virus. Equine, rhesus macaque, sheep, guinea pig, goose, and chicken erythrocytes were used. RESULTS: Using reference antisera, H5 hemagglutinin was detected in all tested antigens with all used erythrocytes. While testing ferret antisera in HI test with reference antigen, anti-H5 antibodies were not detected or detected in extremely low titre (1/80) and only with equine erythrocytes. In most cases, titers of anti-H5 antibodies in HI test with formalinized erythrocytes were higher than with native ones. CONCLUSION: During monitoring for antibodies to H5N1 avian influenza virus in human population it is necessary to use native strains of A/H5N1 along with reference antigen. It is possible to use formalinized equine, rhesus macaque, goose, and chicken erythrocytes.

[Isolation and molecular characterization of the influenza virus A/H5N1 strains isolated during outbreak of avian influenza among birds in the European part of Russia in 2005: strain with ozeltamivir-resistance mutation was found].

Isolation and characterization of the influenza virus A/H5N1 strains, isolated from chicken in the Yandovka village (Tula Region) and from wild swan near the orifice of the Volga River that died during an outbreak of avian flu in autumn 2005, were carried out. Genetic and phylogenetic analyses were performed. The goals of the analysis were to determine possible geographical origin of the strain, genetic similarity of isolated strains to earlier sequenced isolates, epidemic potential, existence of pathogenicity markers, and resistance to antiviral drugs. It was shown that the isolated influenza virus belonged to highly pathogenic variants of China origin by a reassortment of viruses genotypes Z and V circulated in poultry and wild birds. A number of molecular markers of pathogenicity to gallinaceous birds and mammals were found out. Mutations in the hemagglutinin gene promoting potentially high rate of replication in humans as well as mutations causing the resistance to amantadine/rimantadine were not found. The strain isolated from wild swan had the mutation causing resistance to tamiflu/ozeltamivir.

[Circulation of type A influenza viruses in poultry during the pre-epizootic period of 2005 in Novosibirsk region].

Work is devoted to studying of circulation of avian flu influenza A (H5N1) virus at poultry in territory of Novosibirsk region in preepizootic period. 1901 sample from clinically healthy poultry is collected and investigated. On chicken embryos it has been isolated 28 virions, including from poultry of the industrial maintenance--16, from the poultry contained in private farmsteads--12. At the analysis of isolates it has been shown, that among poultry in preepizootic period circulated various serovars of influenza viruses namely, HI, H4 and H3 serogroups. The percent of isolates from geese of the industrial maintenance increased to 0.6% depending on area. The percent of isolates from a bird contained in private farmsteads, increased to 7.5%. The greatest percent of isolates was observed from the samples taken from hens--77. Thus, it is shown, that among poultry circulation of not pathogenic influenza viruses of type A is the usual phenomenon, and asymptomatic carriers 7.5% of a livestock. Also it has been established, that in preepizootic period 2005 (January-June) at poultry it is not revealed influenza viruses of H5 serotype, caused epizootia at Summer of 2005 in territory of Western Siberia.

[Study of highly pathogenic H5N1 influenza virus isolated from sick and dead birds in Western Siberia].

The mass destruction of domestic birds has been registered in July, 2005 in Novosibirsk region. Influenza virus H5N1 have been isolated from bodies of the lost birds on developing chicken embryos and identified by serological and molecular biological methods. M-gene and genes coding hemagglutinin and neurominidase were in part sequening. The phylogenetic analysis of hemagglutinin gene has shown, that the isolated viruses are forming a claster with strains, isolated from birds during outbreak of the bird's flu on lake Tsinghai (Qinghai) in China in 2005, in Japan in 2004, and also with H5N1 strains, isolated from the person and birds in the countries of Southeast Asia during ipizootia in 2003-2004. The site of the restriction which associated with pathogenicity of isolated avian influenza viruses H5 serogroup, corresponds to sequence of high pathogenic strains, circulating in the countries of Southeast Asia. The test for pathogenicity with use of chickens has confirmed, that researched strains were high pathogenic for birds.

[Rabies epizootic situation in the Novosibirsk region in 1997 - 2003].

Starting from the end of 1997, a rise in the epizooty of rabies with peaks in 1998 and 2002 was noted. 550 cases of rabies were registered in wild and domestic animals, as well as one lethal case of rabies in man in 2001. Wild carnivorous animals are traditionally regarded as the reservoir and source of rabies virus (38%). In the Novosibirsk region this disease was registered in badgers, wild herbivorous animals, myomorph rodents, hares, marmots. Urban cases of rabies constituted 23%, cases of rabies among agricultural animals constituted 14% of all rabies cases in the Novosibirsk region.

[Molecular genetic characterization of rabies virus, isolated in Western Siberia]. / Molekuliarno-geneticheskaia kharakteristika izoliatov virusa beshenstva, vydelennykh na teritorii zapadnoi Sibiri.

The paper covers a study of four rabies viruses, isolated in 2003 from man, wild foxes, badgers and home cats inhabiting entirely different districts of Novosibirsk suburb, and a sample taken from a bat from Maslaninsky district. The samples were studied by fluorescent antibody microscopy and a biological test on white mice, RT-PCR-analysis of NP-genes and sequencing. The study revealed that related variants of rabies viruses circulated in Novosibirsk region, and those formed two groups in genotype 1. Most of the amino acid replacements found in the study are only characteristic of the viruses circulating in the given territory.

[Lissavirus in bats residing in the South of West Siberia].

We caught, 2002-2003 in caves of Novosibirsk Region and Altai Territory, 52 bats (Myotis daubentoni, Myotis brandtii, Murina leucogaster, Plecotus auritus). Examinations of the animals' brain samples by fluorescence antibody microscopy and RT-PCR revealed lyssaviruses, genotype 1, which were close to strains provoking rabies in the local animals. According to the results, bats can be involved in keeping alive the rabies foci in Western Siberia, which must be taken into account in antiepizootic measures.