Study of combined foci of mosquito-transmitted arbovirus infections

A total of 5227 serum specimens from humans, horses and swine collected in the seasons of 1968--1976 in 15 administrative areas of the Primorskiy Kray were examined with antigens of a number of mosquito-borne arboviruses: Japanese encephalitis (JE), West Nile (WN), Getah, and Sindbis. Both independent and combined circulation of these viruses in the region was established. Sindbis virus was found to be circulating separately most frequently, West Nile virus the least frequently. According to the results of the serological analysis, the conditions for combined circulation are most closely related for JE and Getah, and JE and WN viruses. The interpretation of the results of examinations of the sera reacting simultaneously with JE and WN virus is most difficult because of close antigenic relationship of these viruses. A set of methods for serological differentiation of bivalent sera to JE and WN viruses and the criteria for the assessment of the results are recommended.     

Anti-arbovirus antibodies positive serum screening in population of Saratov

Seroepidemiological studies revealed the existence of a population stratum immune to West Nile and Sindbis viruses in the Saratov region. Screening of 218 sera showed that the stratum immune to arboviruses increased from 1998 to 2000. The incidence of antibodies to Batai and Sindbis viruses was the highest. Activation of natural foci of these arboviruses can lead to exacerbation of the epidemic situation in the region.     

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.     

Antigenic analysis of West Nile virus strains using monoclonal antibodies

Summary Seventeen monoclonal antibodies (MAbs) were prepared against the flavivirus West Nile strain H442. While the majority of these were specific for the major envelope protein, MAbs directed against the NS1 and ns4a nonstructural proteins were also identified. The MAbs were tested by indirect immunofluorescence against 16 southern African West Nile (WN) isolates, representative strains from the two main WN antigenic groups and several viruses from other flavivirus complexes. The MAb reactivities ranged from WN strain-specific to broadly flavivirus-group reactive. Comparison of the local isolates revealed the presence of several different strains, all of which were antigenically distinct from the representative strains of the two WN antigenic groups.     

Epidemic outbreak of meningitis and meningoencephalitis, caused by West Nile virus, in the Krasnodar territory and Volgograd region (preliminary report)

Sera from 102 inpatients from the Volgograd region (64) and Krasnodar region (38) were tested for antibodies to West Nile (WN) virus in hemagglutination inhibition (HI) test and for IgM and IgG antibodies in enzyme immunoassay (EIA). Diseases etiologically associated with WN virus were diagnosed in 81 patients: in 50 out of 64 in the Volgograd region and in 31 out of 38 in the Krasnodar region, which makes 79.4%. Specificity of antibodies to WN virus was confirmed in HI and EIA with WN antigens, related flaviviruses (Japanese encephalitis and yellow fever), and Sindbis alfavirus. A considerable number and the incidence of WN infection suggest that an epidemic caused by WN virus occurred in the Krasnodar and Volgograd regions in summer 1999.     

Virus neutralizing antibodies to arboviruses in birds of the order Anseriformes in Czechoslovakia.

Sera from birds of the order Anseriformes in Czechoslovakia were examined for virus neutralizing (VN) antibodies to arboviruses. VN antibodies to Sindbis, Calovo and Tahyna viruses were found in 15, 5 and 6 out of 106 greylag goose (Anser anser) sera. Out of 38 ducks, 6 mallards (Anas platyrhynchos) and 1 garganey (Anas querquedula) contained VN antibodies to Sindbis virus, 6 mallards to Calovo virus, 4 mallards and 1 garganey to Tahyna virus, 2 mallards and 1 garganey to tick-borne encephalitis (TE) virus and 1 mallard to West Nile (WN) virus.     

Purification of togavirus haemagglutinins by chromatography on controlled pore glass.

Purified preparations of haemagglutinins from Sindbis, West Nile and tick-borne encephalitis (TBE) viruses could be obtained from infected mouse brains by alkaline extraction, precipitation with protamine and chromatography on polyethylene glycol-coated controlled pore glass with 242 A pore diameter.     

Arboviruses in the Iberian Peninsula

The information concerning the presence and activity of arboviruses in the Iberian Peninsula is very scanty. In Portugal serological studies have been done with sera from animals, wild birds and human population. The results have shown that some arboviruses have been active in some areas of the territory. West Nile (WN) virus was isolated from mosquitoes and Dhori and Thogoto viruses have been isolated from ticks. In Spain, in spite of the large territory, it's geographical contiguity to Africa and the existence of different kinds of climates and ecologic conditions, the information is reduced to some surveys done in some restricted regions of the country. In both countries African swine fever was frequent and recently African horsesickness has been introduced to Southern Spain. A straight liaison and cooperation among virology laboratories of both countries and also Morocco should be emphasized in order to establish epidemiological surveillance in Southern Europe.     

Influence of tick-borne encephalitis and West Nile viruses on the chromosomes of pig kidney cells

Changes of the mitotic index (MI) were studied in pig kidney (PS) cells infected either with tick-borne encephalitis (TBE) virus strains Hypr and Ir-32, or with West Nile (WN) virus prototype strain, strains K-99 and E-13. The cell division was arrested by the infection (metaphase barrier). The effect of TBE virus strains was manifested by the appearance of lagging chromosomes while the action of WN virus strains by colchicine--like metaphases. The prototype TBE virus strain Hypr affected the chromosomes and mitotic apparatus of PS cells less markedly than did the freshly isolated Ir-32 strain.     

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

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.     

Comparison of an antibody-capture IgM enzyme-linked immunosorbent assay with IgM-indirect immunofluorescence for the diagnosis of acute Sindbis and West Nile infections

The development and evaluation of an antibody-capture ELISA for the detection of IgM antibodies to Sindbis (SIN) and West Nile (WN) viruses are described. Comparison of the ELISA results with those obtained by indirect immunofluorescence (IIF) antibody tests using both fluorescein and biotinylated anti-human IgM conjugates, showed that the former technique was both more sensitive and specific than the IIF methods. There were no false positives by the ELISA whereas with the IgM-IIF assays a high percentage of false positives were obtained. These were due to rheumatoid factor and also to an interfering factor which was not detected by the RF latex agglutination test. Absorption of the sera with anti-IgG was necessary to eliminate this interference in the IgM-IIF tests.     

Susceptibility of dogs to West Nile virus: a survey and pathogenicity trial

A serological survey of dogs from the highveld region of South Africa showed that 37 per cent (138 of 377) had neutralizing antibodies to West Nile (WN) virus and only 2.7 per cent (10 of 377) had antibodies to Sindbis virus. WN virus was isolated from one of the WN-antibody negative sera. Because these results suggested that dogs may play an important part in the epidemiology of WN virus, a pathogenicity trial was carried out. Two of three dogs infected with WN virus had a mild recurrent myopathy, but no other abnormalities were detected in the biochemical or haematological tests performed on any of the dogs. All three dogs developed antibodies but a low titre-viraemia was detected in only one dog. It was concluded that dogs do not play an important part in the epidemiology of WN virus but they may play a small part in the maintenance of the virus.     

Polykaryocytosis induced by certain arboviruses in monolayers of BHK-21-528 cells

Multinucleated giant cell formation in a clone of BHK-21 cells, BHK-21-528, was tested with certain arboviruses. Eleven out of 19 viruses tested, Chikungunya, Getah, Sagiyama, Sindbis, Western equine encephalitis, St. Louis encephalitis, Japanese encephalitis, West Nile, Bunyamwera, Germiston and California encephalitis virus induced cell fusion after infection. All cases where giant cells were observed during the experiments, involved the B type cell fusion (fusion from within).     

Antigenic analysis of tick-borne encephalitis complex viruses by time-resolved fluoroimmunoassay with monoclonal antibodies.

The antigenic structure of 5 strains of tick-borne encephalitis (TBE) virus and 7 other viruses of the TBE complex was examined by the highly sensitive and specific technique of time-resolved fluoroimmunoassay (TR-FIA). A collection of 8 monoclonal antibodies to the Austrian strain. Neud?rfl, was used in this study. The findings demonstrate the uniformity of the antigenic structure of TBE viruses from different geographic regions of the USSR. In addition, an epitope was detected which is characteristic of western variants of TBE virus, and another epitope was detected which permits the differentiation of the east-Siberian strain, Aina, from other TBE virus strains. The unique nature of Skalica virus was confirmed, and its similarity, but not identity, to Langat TP-21 virus was shown. Substantial variability in the antigenic structure of some TBE complex viruses was also demonstrated.     

Interspecies interactions of arboviruses. I. Competition phenomenon in mixed populations of Japanese encephalitis and West Nile viruses.

Two phenomena, competition and independent reproduction, were observed on co-cultivation of Japanese encephalitis (JE) and West Nile (WN) viruses in permissive systems. Competition occurred in populations propagated in mouse brains and led to elimination of JE virus. Only clones of WN virus were isolated from these populations at the 1st-3rd passage level. No competition was observed in mixed infection of primary Syrian hamster kidney (SHK) cell culturess the population had the combined immunological phenotype JE+WN+, and contained discrete JE and WN virus clones and clones with duplex antigenic markers. Upon subcloning, the latter dissociated into JE and WN viruses. The competition phenomenon depended upon 3 factors: co-cultivation system, peculiarities of the virus strains, and virus dose. The capacity of WN virus strains to eliminate JE virus in mixed infection of mice varied.     

Human antibody response to immunization with 17D yellow fever and inactivated TBE vaccine

The antibody response against flaviviruses tick-borne encephalitis (TBE), Kyasanur Forest disease (KFD), Murray Valley encephalitis (MVE), West Nile fever (WNF), Japanese B encephalitis (JE), dengue 2 (DEN-2), and yellow fever (YF) was studied in humans after administration of an inactivated TBE virus vaccine. Individuals were either prevaccinated with 17D yellow fever (experimental group) or without any previous exposure to flaviviruses (control group). The appearance of serum titres of homologous and heterologous haemagglutination inhibition (HI) antibodies, heterotypic DEN-2 neutralizing antibodies, and TBE enzyme-linked immunosorbent assay (ELISA) antibodies were examined. Individuals prevaccinated with the 17D yellow fever developed an antibody pattern that contrasted with that of the control group. This pattern was characterized as follows: (1) Predominantly anti-TBE IgG antibodies appeared earlier and in higher titres than in the control group, (2) heterologous HI antibodies cross-reacting with the WN flavivirus subgroup preceded the appearance of homologous HI antibodies, (3) a broad spectrum HI response was observed against all flaviviruses tested, and (4) low titre heterotypic DEN-2 neutralizing antibodies were formed in about half of the cases. These observations are discussed in the context of cross-reactivity, cross-protection and virus infection enhancement.     

Studies on the glycosylation of flavivirus E proteins and the role of carbohydrate in antigenic structure

The glycosylation pattern of several flavivirus E proteins as well as the role of carbohydrate in biological functions and the antigenic structure of tick-borne encephalitis (TBE) virus were investigated by the use of specific endoglycosidases. Endoglycosidase F digestion revealed the presence of a single asparagine-linked oligosaccharide side chain in TBE virus (Western and Far Eastern subtype), Louping III virus, Murray Valley encephalitis virus, and Rocio virus. Consistent with published sequence data, the E protein of West Nile virus apparently is not glycosylated at all. Evidence derived from digestion experiments using endoglycosidase H indicates that the tick-borne viruses contain high-mannose type N-linked oligosaccharide side chains, whereas that of the mosquito-borne Murray Valley encephalitis virus and Rocio virus is endoglycosidase H resistant. Complete deglycosylation of TBE virus by endoglycosidase F did not impair infectivity and HA activity. Carbohydrate does not seem to play a major role in the antigenic structure of the TBE virus glycoprotein since the reactivity of the native virus and the deglycosylated virus was identical when analyzed with monoclonal as well as polyclonal immune sera.     

Antigenic variation of the viruses belonging to the tick-borne encephalitis complex as revealed by human convalescent serum and monoclonal antibodies

Solid-phase enzyme-linked immunoassay (ELISA) was used for the detection of antigenic relationships and/or differences among the viruses belonging to the tick-borne encephalitis (TBE) complex. Monoclonal antibodies of IgM class with haemagglutination-inhibiting activity to the Skalica strain of TBE virus were used to compare the TBE complex viruses. Antigenic analysis of 9 viruses of the TBE complex, isolated from Eurasia and America showed close relationships among them. Nevertheless, it was possible to differentiate the Skalica strain from Langat, louping-ill and Omsk haemorrhagic fever (OHF) viruses by ELISA when monoclonal antibodies and antigens were diluted 1:10,000. Monoclonal antibodies to the Russian spring-summer encephalitis virus did not react with the Skalica strain in immunofluorescence test. By the use of convalescent serum no reaction was found with louping-ill, Russian spring-summer encephalitis, Powassan and OHF viruses in haemagglutination-inhibition (HI) test.