Low incidence of human coronavirus among hospitalized children in Novosibirsk city, Russia during pre-pandemic period (2013-2020).

We investigated the incidence of 15 respiratory viruses among 2991 children with acute respiratory infections in Novosibirsk city, Russia, prior to the COVID-19 pandemic (2013-2020). Viral infections were detected in 72.5% cases. The incidence of human coronavirus was 2% (Alphacoronaviruses, 63%; Betacoronaviruses, 37%).

Development of a Purified Viral Preparation for Studies of COVID-19 (SARS-CoV-2) Biology.

Different methods for producing bulk quantities of concentrated and purified SARS-CoV-2 for the use as antigens and for the research into COVID-19 biology were tested.

Study of Antiviral Efficiency of Oxidized Dextrans In Vitro and In Vivo.

Antiviral efficiency of oxidized dextrans (OD) with different molecular weights and oxidation degree (OD40min, OD70min, OD40max, and OD70 max) was studied in vitro and in vivo. Dextrans OD40max and OD70max prevented the development of the cytopathic effect of influenza A(H1N1)pdm09 virus in more than 50% MDCK cells vs. control (no OD). Four intranasal doses of OD40min, OD40max, and OD70min and one intranasal dose of OD70max before infection of BALB/c mice with A(H1N1)pdm09 influenza virus significantly reduced mortality and prolonged life span in comparison with controls receiving saline. These and our previous data attest to clear-cut preventive effect of OD in influenza infection.

Pathogenicity assessment of wild-type and mouse-adapted influenza A(H1N1) pdm09 viruses in comparison with highly pathogenic influenza A(H5N1) virus.

Here we compare the results of pathological and virological examinations of mice experimentally infected with either wild-type or mouse-adapted pandemic A(H1N1) pdm09 viruses and highly pathogenic avian influenza (HPAI) virus A(H5N1). Mice were sacrificed on days 1, 3, 6, and 10 post infection or whenever morbidity was severe enough to justify euthanasia. Morbidity rates were calculated on the basis of clinical signs (weight loss, poor hair coat, hunched posture and paresis); virus-induced disease was characterised by the histopathology of lung; virus dissemination was determined by virus isolation on organ samples of lung, brain, liver, kidney and spleen. All mice infected with mouse-adapted A(H1N1) pdm09 died in the course of the experiment, whereas 20% of animals survived the infection with A(H5N1). Echinocyte formation changed the rheological properties of blood in animals infected with either mouse-adapted A(H1N1) pdm09 or A(H5N1). To sum up, the adaptation of pandemic A(H1N1) pdm09 virus can confer an enhanced virulence similar to or even exceeding that of HPAI A(H5N1) virus.

Molecular characterization and phylogenetics of a reassortant H13N8 influenza virus isolated from gulls in Mongolia.

Double reassortant H13N8 influenza A virus was isolated from gull in Mongolia. The basic virological characteristics were studied. Complete genome sequence analysis indicated the complicated evolutionary history. The PA gene belongs to classical Avian-like lineage and more likely originated from non-gull avian virus pool. Data confirm the state of extensive geographic mosaicism in AIV from gulls in the Northern Hemisphere.

Monitoring of influenza viruses in Western Siberia in 2008-2012.

Western Siberia is of great importance in ecology and epidemiology of influenza. This territory is nesting area for great amount of bird species. Territorial relations of Western Siberian birds that are established during seasonal migration are extremely wide since this region is an intersection point of bird migration flows wintering in different regions of the world: Europe, Africa, Middle East, Central Asia, Hindustan, and South East Asia. Reassortant influenza viruses that can cause outbreak among population may emerge in Western Siberia with high probability. Thus, it is extremely important to carry out widespread study of circulated viruses, their molecular biological properties, phylogenetic links in this region, as well as herd immunity to influenza virus serotypes with epidemic potential.

[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.

Ecology of influenza virus in wild bird populations in Central Asia.

The study provides the results of avian influenza virus surveillance in Central Asia during 2003-2009. We have analyzed 2604 samples from wild birds. These samples were collected in Kazakhstan (279), Mongolia (650), and Russia (1675). Isolated viruses from samples collected in Mongolia (13 isolates) and in Russia (4 isolates) were described. Virological analysis has shown that six isolates belong to the H3N6 subtype and five isolates belong to the H4N6 subtype. Two H1N1 influenza viruses, one H10N7 virus, two H3N8 viruses, and an H13N8 virus that is new for Central Asia have been also isolated. Samples were taken from birds of six orders, including several species preferring water and semiaquatic biotopes, one species preferring dry plain regions, and one more species that can inhabit both dry and water biotopes.

[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.

[Genetical features of influenza virus A (H1N1) strain that caused the 2009 pandemic].

Genetical features of the A(H1N1) influenza virus strain that caused the 2009 pandemic are analyzed in the review. Mutations typical for this strain, unique and similar to influenza viruses of swine, avian and seasonal types, and phenotypic (pathologic) features associated with them, that are experimentally confirmed, are described. A possibility of reassortation of avian and swine influenza viruses and possible epidemiologic consequences are discussed.

[Studies of properties of pandemic influenza virus A/H1N1 circulated in Russian Federation in 2009 - 2010].

AIM: Studies of cultural, virologic, antigenic properties of 89 samples of pandemic influenza A(H1N1) 2009 virus isolated in Russian Federation from May 2009 to March 2010. MATERIALS AND METHODS: Properties of isolated samples were compared with those of the reference strain A/ California/04/2009 (H1N1). RESULTS: Studies of biological properties and analysis of genome nucleotide sequences of the isolated samples showed that those strains are closely related to the reference strain. CONCLUSION: Monitoring of genetic, virologic and antigenic properties of pandemic influenza A(H1N1) 2009 virus isolates carried out from May 2009 to March 2010 did not reveal significant changes in the abovementioned properties of the virus or emergence of mutations that can lead to such changes.

Influenza A/H1N1pdm virus in Russian Asia in 2009-2010.

In total 3566 blood sera samples were collected in the Russian Far East and Central and Western Siberia in 2009-2010. The presence of antibodies to influenza A/H1N1pdm, seasonal H1N1 and H3N2 influenza viruses in the sera was tested in the hemagglutination inhibition test. 29.5% of samples from the Far East were positive to pandemic influenza, this value for Central and Western Siberia is 2-fold lower (12.8% and 11%, respectively). Fifty-six influenza A/H1N1pdm viruses were isolated during 2009-2010 from samples collected in Central and Western Siberia and the Russian Far East. Molecular and genetic properties of 23 strains were studied. Nucleotide and amino-acid sequences of pandemic influenza virus strains were identical (99% identity or more) to reference strain A/California/04/2009. Sporadic substitutions in antigenic sites were detected but did not change antigenic characteristics of strains.

[Pandemic influenza virus A (H1N1) in Amur region in autumn 2009].

AIM: Isolation and study of molecular genetic characteristics of pandemic influenza virus A (H1N1) circulated in Amur region in autumn 2009 as well as testing of serum samples taken from citizens of this region during November- December 2009 in order to measure levels of antibodies to socially significant serotypes of influenza A virus. MATERIALS AND METHODS: Strain of pandemic influenza virus A/Blagoveschensk/01/2009 (H1N1) was isolated on MDCK cell culture and nucleotide sequences of all eight segments of viral genome were determined. Five hundred seventy-six serum samples taken in Amur region in autumn 2009 were tested by hemagglutination inhibition assay. RESULTS: Nucleotide sequence of A/Blagovechensk/01/2009 (H1N1) strain was 99.7% identical to reference influenza virus strain A/California/04/2009. Diagnostically significant titers of antibodies to pandemic influenza virus were observed in 46.3% of persons younger 30 years old and in 20.1% older persons. Antibodies to seasonal influenza virus H1N1 and H3N2 were detected in 39.5 and 29.8% of persons respectively. CONCLUSION: Final seroepidemiological picture of distribution of pandemic virus in Amur region matches with the one for seasonal influenza virus A (H1N1): > 60% of seropositive persons were registered in age group < 18 years old, and this proportion increases with increasing age.

[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.

Surveillance and identification of influenza A viruses in wild aquatic birds in the Crimea, Ukraine (2006-2008).

The ecology of avian influenza (AI) viruses in wild aquatic birds of Asia is poorly understood, especially for the H5N1 high pathogenicity AI (HPAI) viruses. From March 2006 through November 2008, 20 AI viruses were isolated in the Crimea region of Ukraine with an overall frequency of virus recovery of 3.3%. All the viruses were isolated from three species of dabbling ducks: mallard (Anas platyrhynchos), wigeon (Anas penelope), and garganey (Anas querquedula), making the frequency of virus recovery for dabbling ducks 6.3%. The viruses were predominantly isolated during the fall sampling period. All viruses were genetically and antigenically characterized. No H5N1 HPAI viruses were isolated, but other HA and NA subtypes were identified including H3N1 (2), H3N6 (3), H3N8 (4), H4N6 (6), H5N2 (3), H7N8 (1), and H10N6 (1) subtypes. All isolates were of low pathogenicity, as determined by the intravenous pathogenicity index of 0.00. For H5N2 and H7N8 isolates, the HA gene was sequenced and the phylogenetic analysis revealed possible ecologic connections of the Crimea region with AI viruses from Siberia and Europe. No influenza A isolates were recovered from other Anseriformes (diving ducks [two species of pochards] and graylag geese), Columbiformes (collared doves), Gruiformes (coot), and Galliformes (gray partridges).

[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.

[Isolation of influenza A virus from wild birds in western part of Mongolia].

AIM: To study circulation of influenza A viruses in western part of Mongolia. MATERIALS AND METHODS: Isolation and characterization of influenza viruses was performed according to recommendations of WHO. RESULTS: Circulation of influenza A viruses subtypes H3N6, H4N6, H1N1, H13N8 in different wild bird species in western part of Mongolia was documented. CONCLUSION: Taxonomic and ecologic heterogeneity of bird species involved in continuous circulation of influenza A viruses was revealed. Subtype H13N8 was isolated for a first time from herring gull on territory of western Mongolia.

[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.

[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.

Pathogenesis of infectious disease of mice caused by H5N1 avian influenza virus.

The pathogenesis of a disease caused by Qinghai-like H5N1 influenza virus in BALB/c mice was studied. Clinical, morphological, and immunological characteristics of the experimental infection caused by highly pathogenic A/duck/Tuva/01/06/ (H5N1) virus are described.