[Monitoring of enterovirus circulation in Irkutsk region].

AIM: Monitoring of circulation of enteroviruses (EVI) in Irkutsk Region and study of regional specter of circulating enteroviruses. MATERIALS AND METHODS: 1419 samples from patients with suspected EVI, contact in foci ofenterovirus infection, acute intestine infections and 964 samples of sewage water were studied in total. In 2011 isolation of viral agents from 97 samples positive on enterovirus by RT-PCR from patients with preliminary EVI diagnosis and 5 samples of sewage water of Irkutsk city was carried out. Transplantable line of human rhabdomyosarcoma RD cell culture was used for isolation of enteroviruses. Infection of cells and 2 serial passages of the studied material were carried out. The isolates were typed in neutralization reaction (NR) with a set of 32 diagnostic type-specific immune sera against viral poliomyelitis I-III; Coxsackie B1-6; Coxsackie A2, A4, A7, A9, A10; ECHO 68 - 71; ECHO 2, 4, 7, 8, 9, 12, 16, 20, 25, 26, 27, 29, 31, 33. RESULTS: In 2011 circulation of enterovirus serotypes that were previously absent on the territory of the region was established: ECHO 68, ECHO 70, ECHO 71. These strains were isolated from patients, circulation of ECHO 70 serotype was established also in samples of sewage water. The analysis of enterovirus landscape carried out showed the possibility of complication of epidemic situation on the territory of the region due to change of serovariants of causative agents of non-polioenterovirus infections and detection ofepidemically significant enteroviruses - ECHO 68, 70 and 71 serotypes. CONCLUSION: Determination of specter ofenterovirus serotypes, detection of serotypes that had not previously circulated in Irkutsk Region allows to prognose epidemic situation on morbidity of enterovirus infections and timely develop and make decisions for ensuring epidemiologic welfare of the population.

[Polytypic strains in the genofund of tick-borne encephalitis virus].

Eighteen polytypic tick-borne encephalitis virus (TBEV) strains containing the fragments of E and NS1 protein genes of Siberian and Far Eastern, occasionally Siberian and European subtypes were isolated in the European and Asian parts of the tick-borne encephalitis (TBE) area. They were identified using real-time polymerase chain reaction, hybridization-fluorescence detection with genotype-specific probes, restriction fragment length polymorphism analysis, and E protein sequencing. The polytypic strains were isolated from individual Ixodes persulcatus ticks, their pools, from the blood of patients and the brain of dead patients. The isolation rates of the polytypic strains in the sympathry area of different TBEV subtypes ranged from 4.4% (the Irkutsk Region) to 15.1% (the Yaroslavl Region). In addition to 2 polytypic strains, a strain similar to the TBEV 886-84 strain was isolated. The TBEV subtypes entering into the composition of the polytypic strains show nongenetic interactions, such as neutral replication or competition. The polytypic strains are stable during passages in the cultured pig embryo kidney epithelial cells and on cloning. Mouse brain passage promotes dissociation of polytypic strains. The conditions for the formation of polytypic strains and their role in the etiology of TBE are discussed.

[Antigenic characteristics of morbillivirus isolates from Baikal seals]. / Antigennaia kharakteristika izoliatov morbillivirusa baikal'skoi nerpy.

Four morbillivirus strains were isolated from the Baikal seals Phoca sibirica. The antigen difference between these strains was studied using ELISA with different immune sera and monoclonal antibodies. The assay data indicated three different antigen variants of morbillivirus. The four strains were found to circulate over the Baikal seal population during epizootic.

[Dynamics of expression of Marburg and Ebola virus antigens in infected Vero cells ]. / Dinamika ékspressii antigenov virusov Marburg i Ebola v infitsirovannykh kletakh Vero.

Time course of Marburg and Ebola virus antigens expression in Vero cells was studied by indirect immunofluorescence test. The maximum accumulation of virus specific antigens in Vero cells infected with a high dose was observed after 48-54 h of incubation. It is essential for laboratory diagnosis that virus specific antigens can present as incorporations of different shape and size, starting from small hardly discernible granules (immediately after the virus adsorption) to large lumps, cords, accumulations, and diffuse fluorescence.

[Neutralizing morbillivirus antibodies in the population of the Baikal seal Phoca sibirica]. / Neitralizuiushchie morbillivirus antitela v populiatsii baikal'skikh nerp Phoca sibirica.

The antibodies neutralizing Baikal seal morbillivirus (BSMV) were studied in sera from 148 normal Baikal seals (Phoca sibirica). The virus-neutralizing antibodies were demonstrated in sera of 61.5% of the animals examined. A comparative analysis of antibodies in NT and cell-ELISA method is presented. The role of BSMV in morbidity and mortality of seals (Phoca sibirica) is discussed.

[Ebola virus reproduction in cell cultures]. / Reproduktsiia virusa Ebola v kletochnykh kul'turakh.

Ebola-Zaire virus production in Vero and BGM cells was studied. The CPE developed in both cell cultures. The cell monolayer destruction by 80-90% was seen at a low multiplicity of infection in 7-8 days after virus inoculation. An overlay composition was developed for virus titration using plaque assay. The plaque production was shown to be directly proportional to the virus dose. The curve of Ebola virus production in Vero cell culture fluid was determined. At a multiplicity of infection of 0.01 PFU/cell, the maximum virus titer of 10(6.4) PFU/ml was reached in 7 days postinfection. Specific antisera were generated by inoculation of guinea pigs. Indirect immunofluorescent assay was used for testing of virus-specific antigen and antibody.

[An experimental infection of the seal Phoca sibirica with the morbilli virus]. / Eksperimental'noe infitsirovanie nerpy Phoca sibirica morbillivirusom.

A Baikal seal (Phoca sibirica) was experimentally infected with Baikal seal morbillivirus (BSMV) isolated from Baikal seals during an epizootic in 1987-1988. The seal was infected with BSMV with an infectious titer of 10(7.0) TCD50/ml, and daily observations of the animal clinical condition were made. The virus-specific antibodies in the seal serum were determined by ELISA and virus neutralization test. The clinical picture showed a mild infection. The ELISA-active antibodies were detected 10 days postinfection and reached the maximum in 20 days. Virus-neutralizing antibodies were detected in 16 days after infection, reached the maximum titer of 1:640 in 20 days and remained at this level for 39 days (the observation period). These data indicate that BSMV can induce a disease in the natural host with production of virus-neutralizing antibodies. The results of this work and the earlier reports show that the epizootic in Baikal seals was induced by BSMV.

[The isolation of the morbilli virus from the Baikal seal Phoca sibirica and its preliminary characterization]. / Vydelenie morbillivirusa ot baikal'skoi nerpy Phoca sibirica i ego predvaritel'naia kharakteristika.

A morbillivirus was isolated from the organs of a seal (Phoca sibirica) which had died during 1987-1988 epizootic in Baikal. This Baikal seal morbillivirus (BSM) was adapted to Vero cell cultures in which it induced a cytopathic effect developing to complete destruction of the monolayer. Typing of BSM was done by indirect immunofluorescence test and enzyme immunoassay using antibodies to distemper and measles viruses. A method for virus concentration and purification was developed. According to electron microscopic examinations, the virus virions were spherical particles of heterogeneous sizes over 100 nm in diameter. The clinical picture of seal infection, pathological anatomy and histopathology are described. A possible role of BSM in the epizootics of Baikal seals is discussed.

[Antibodies to morbilli virus of the Baikal seal in its natural host]. / Antitela k morbillivirusu baikal'skoi nerpy u prirodnogo khoziaina.

Morbillivirus of Baikal seal (BSM) was isolated from organs of a dead animal during 1987-1988 epizootic of Baikal seal (Phoca sibirica). A method of cellular enzyme immunoassay for testing for virus-specific antibodies was developed using BSM. The method was used for antibody detection in sera of 115 apparently normal seals collected in the spring of 1989. Antibody to BSM were found in sera from 75 animals. Examinations of seropositive animals of different age and sex were carried out. The results obtained indicate a possible role of BSM in the epizootic.