Murine oncornavirus activation in the pancreas during infection with Venezuelan equine encephalitis virus.

Electron microscopic studies of the pancreases of 3-week-old ICR Swiss mice infected with Venezuelan equine encephalitis virus and killed 6 days post inoculation (at which time they were moribund) indicated significantly more type-C particles than were found in uninfected controls. This phenomenon was apparently possible because of the microanatomy of the pancreas, in which interstital spaces allowed accumulation of virus particles.

Further observations on infections of guinea pigs with Venezuelan encephalitis virus strains.

Guinea pigs from a Guatemalan colony died after subcutaneous inoculating of moderately small doses of equine-benign strains of Venezuelan encephalitis (VE) virus of hemagglutination-inhibition subtype I-E from enzootic habitats in Mexico and Guatemala. Thus these guinea pigs were unlike English short hair and inbred 13 guinea pigs, which usually survive infections with equine-benign VE strains of subtype I-E. We therefore caution others that not all strains of guinea pigs can be used to evaluate the potential equine virulence of VE viruses.

Vector competence of Culex (Melanoconion) taeniopus for allopatric and epizootic Venezuelan equine encephalomyelitis viruses.

The vector competence of Culex (Melanoconion) taeniopus was examined in the laboratory for "enzootic" allopatric and "epizootic" strains of Venezuelan equine encephalomyelitis viruses of hemagglutination inhibition subtypes I, II, III, and IV. Following bloodmeals from viremic hamsters, and extrinsic incubation of 20-22 days, mosquitoes were allowed to refeed for transmission attempts. Infection rates never exceeded 50% with oral doses of less than 10(4) chick embryo cell culture plaque forming units (CECPFU), and approached 100% only after ingestion of greater than or equal to 10(5.5) PFU. Transmission was achieved for some "epizootic" subtype IABC and "enzootic" subtype ID strains after bloodmeals containing greater than or equal to 10(3.4) CECPFU; subtypes II, III, and IV were never transmitted despite oral doses up to 10(5.0) CECPFU. These data contrast sharply with those reported previously for sympatric "enzootic" subtype IE Middle American Venezuelan encephalitis viruses.

Studies on the transmission of Venezuelan equine encephalitis virus by Colombian simuliidae (Diptera).

The ability of Simulium mexicanum and Simulium metallicum to serve as biological or mechanical vectors of an enzootic and an epizootic strain of Venezuelan equine encephalitis (VEE) virus was examined. Guinea pigs were inoculated with the epizootic Cordoba strain or the enzootic RPVP407 strain of VEE virus. Wild-caught adult Simuliidae were fed on the viremic guinea pigs and the virus content of groups of flies was determined at daily intervals post-engorgement to test for viral replication. Flies were refed on suckling mice at greater than or equal to 8 days post-engorgement to test for biological transmission. Other flies were interrupted while feeding on viremic guinea pigs and refed on suckling mice to test for mechanical transmission. Neither S. mexicanum nor S. metallicum appear to be efficient vectors of either strain of VEE virus, although occasional mechanical transmission was obtained. Titers of virus in flies decreased rapidly after engorgement and from 3-12 days post-engorgement virus was detected only in 5%-25% of both species of flies. Although earlier field evidence implicated both S. mexicanum and S. metallicum as vectors of epizootic VEE, we conclude that it is highly unlikely that they play an important role as vectors of the virus in nature.

Responses of guinea pigs to infections with strains of Venezuelan encephalitis virus, and correlations with equine virulence.

Five equine-virulent strains of Venezuelan encephalitis (VE) virus consistently killed guinea pigs (strain English short hair) inoculated subcutaneously, whereas ten equine benign strains did not. Two enzootic eastern Panamanian and Colombian strains of hemagglutination-inhibition antigenic subtype I-D and of undetermined virulence for equine animals also killed guinea pigs. This subtype of VE virus may represent an excpetion to a positive correlation between equine virulence and guinea pig lethality since another strain of ts subtype has been reported to be benign in horses inoculated subcutaneously. Within the strains tested, viruses of subtypes I-ABC and I-D were lethal for guinea pigs, and viruses of other subtypes were benign. There was no correlation between guinea pig and human virulence of VE strains.

Experimental infection of North American mammals with epidemic Venezuelan encephalitis virus.

Eighty-three wild mammals of ten species were inoculated with a first mouse passage level Venezuelan encephalitis (VE) virus isolated from south Texas in 1971 during the VE outbreak. Rodents were highly susceptible to VE infection and circulated VE virus at levels ranging from 10(7.7) to 10(11.5) suckling mouse intracranial lethal doses per ml for 2 to 4 days. These levels of virus could easily infect vector mosquitoes. Mortality was high in all species of rodents except Sigmodon hispidus, adult Neotoma micropus, and adult Peromyscus leucopus. Lagomorphs were susceptible to VE infection but circulated VE virus at or near mosquito threshold levels for 1 day only. Raccoons and opossums were relatively resistant to VE infection or circulated low levels of virus in the blood. Juvenile animals of four species were more susceptible and experienced higher viremias than did adults. In two of the four, mortality was higher in the juveniles than in adults. A consideration of results obtained during this study and those of other investigators led to the conclusion that equines were of major importance, that canines, lagomorphs and rodents were of minor importance, and that other species tested were of no importance as virus amplifiers in VE epizootics.

Survival of arboviruses in trypanosome-infected triatomines.

The potential of triatomines to maintain arboviruses was demonstrated by the ability of Rhodinius prolixus with experimentally punctured abdomen to harbor Venezuelan equine encephalitis (VEE) virus for at least 4 months and St. Louis encephalitis virus for 1 month. At 30 days after infection VEE virus was found at low titers in Trypanosoma cruzi-infected R. prolixus and at moderate titers in T. rangeli-infected R. neglectus. Transmission of VEE virus by bite of punctured bugs was successful 2 weeks after virus ingestion; attempts at 30 days failed.

Arbovirus investigations in Argentina, 1977-1980. I. Historical aspects and description of study sites.

This is the introductory paper to a series on the ecology of arboviruses in Argentina. Epizootics of equine encephalitis have occurred since at least 1908, principally in the Pampa and Espinal biogeographic zones, with significant economic losses; human cases of encephalitis have been rare or absent. Both western equine and eastern equine encephalitis viruses have been isolated from horses during these epizootics, but the mosquitoes responsible for transmission have not been identified. A number of isolations of Venezuelan equine encephalitis (VEE) virus were reported between 1936 and 1958 in Argentina, but the validity of these findings has been seriously questioned. Nevertheless, serological evidence exists for human infections with a member of the VEE virus complex. Serological surveys conducted in the 1960s indicate a high prevalence of infection of humans and domestic animals with St. Louis encephalitis (SLE), and 2 SLE virus strains have been isolated from rodents. Human disease, however, has rarely been associated with SLE infection. Only 7 isolations of other arboviruses have been described (3 of Maguari, 1 of Aura, 2 of Una, and 1 of an untyped Bunyamwera group virus). In 1977, we began longitudinal field studies in Santa Fe Province, the epicenter of previous equine epizootics, and in 1980 we extended these studies to Chaco and Corrientes provinces. The study sites are described in this paper.

Comparative infections of epizootic and enzootic strains of Venezuelan equine encephalomyelitis virus in Amblyomma cajennense (Acari: Ixodidae).

To compare the potential for an enzootic or an epizootic strain of Venezuelan equine encephalomyelitis (VEE) virus to infect Amblyomma cajennense (F.), larval ticks were fed on guinea pigs (strain 13) inoculated with an enzootic viral strain of variant I-E (68U201) or on guinea pigs inoculated with an epizootic strain of variant I-A (Trinidad donkey). Peak viremias were 10(5.2) plaque-forming units (PFU)/ml and 10(7.3) PFU/ml in guinea pigs infected with enzootic and epizootic viral strains, respectively. Ticks feeding on enzootic- and epizootic-infected hosts had viral titers of 10(2.5) and 10(3.9) PFU per tick, respectively, at drop-off. Although epizootic virus was recovered from 98% (127 of 130) of larval ticks up to 16 d after drop-off, enzootic virus was recovered from 95% (19 of 20) at drop-off (mean titer, 10(2.5) PFU per tick), with recovery rates declining rapidly to 2 of 10 (mean titer, 10(1.4) PFU per tick) by 16 d after drop-off. Transstadially transmitted epizootic virus was found in 0.4% (12 of 2,950) of unfed nymphs (mean titer, 10(2.8) PFU per tick) 63 d after drop-off, 1% (5 of 521) fed nymphs 69 d after drop-off, and 1% (4 of 400) of unfed adults (mean titer, 10(3.6) PFU per tick) 106 d after drop-off. No enzootic virus was recovered from 4,600 unfed nymphs tested 63 d after drop-off.

Modification of Venezuelan equine encephalomyelitis virus infection in mice by X radiation.

A highly virulent strain of Venezuelan equine encephalomyelitis (VEE) virus produced less severe histopathologic changes in brain tissues of mice previously exposed to sublethal total-body x-irradiation than it caused in nonirradiated mice. Prior exposure to 600 R of x-irradiation virtually eliminated the lesions of vasculitis and encephalitis that were found in the infected nonirradiated control mice. Mean peak brain lesion scores generally decreased as radiation exposure dose was increased. Irradiation of mice before inoculation often decreased median time to death, whereas the severity of pathologic changes in brain tissues from inoculated irradiated mice was often reduced, without significantly altering ultimate host survival. The inflammatory response did not appear to have a significant role in clearance of this virus from the brain. There was no evidence that participation of the immune response contributed to total mortality from VEE virus encephalitis, as indicated by the failure of radiation immunosuppression to reduce mortality. Death apparently was caused by the direct cytocidal effects of VEE virus replication.

[Morphological study of a mixed alphavirus infection]. / Morfologicheskoe izuchenie smeshannoi infektsii al'favirusami.

In combined paired cultivation of 8 Venezuelan equine encephalomyelitis virus, multiploid virions formed in 17.8% of cases. Five clones with this effect were used in mixed infections with clones of Semliki Forest and Sindbis viruses. In these infections changes in virions of the virus progeny were observed in 20% and 16.6%, respectively. Mixed cultivation of pairs of Sindbis and Semliki Forest viruses resulted in formation of multiploid virions in 42.8%. Besides, in two mixed populations formed upon combined multiplication of the latter viruses virions of unusual shapes were found: rounded, oval, elongated, and triangular designated as polymorphic.

[Alphavirus reproduction in cultures of transplantable human lymphoblastic cells in acute infection]. / Reproduktsiia al'favirusov v kul'turakh perevivaemykh limfoblastoidnykh kletok cheloveka pri ostroi infektsii.

The results of investigations of acute infection of continuous human B- and T-cells with typical members of the alphavirus group--Semliki Forest, Sindbis, and Venezuelan equine encephalomyelitis viruses, are presented. Virus amplification was shown to pass through the typical phases: eclipse, logarithmic growth, plateau. Infectious virus production per one cell was from 10 to 10,000 PFU in various cultures. Cell infection results in interferon production. Replication of the viruses under study in lymphoblastoid cell cultures is not accompanied by the active cytocidal effect. The regularities determining the sensitivity of lymphoblastoid cells to viruses in general and alphaviruses in particular are discussed. Proceeding from the results of the study of alphavirus replication in human continuous B- and T-cells it is suggested that this system be used as a model for the analysis of antiviral activity of interferon, its inducers, and chemopreparations in special cells. Lymphoblastoid and fibroblast interferon are as active in B-cells and show no antiviral activity in continuous T-cells, as interferon inducer.

[Use of continuous human lymphoblastoid cell lines (T- and B-origin) to produce persistent tick-borne encephalitis virus and Venezuelan equine encephalomyelitis virus infections]. / Ispol'zovanie perevivaemykh limfoblastoidnykh linii kletok cheloveka (T- i B-proiskhozhdeniia) dlia polucheniia persistentnoi infektsii virusa kleshchegogo éntsefalita i virusa venesuél'skogo éntsefalomielita loshadei.

Persistent infection with tick-borne encephalitis virus (TBE) was established in experimentally infected continuous lymphoblastoid human cell lines Raji, L-101 (of B-origin) and 1387 (T-origin) and with Venezuelan equine encephalomyelitis (VEE) virus in Raji and 1387 lines. The persistently infected lines produced infectious virus, the cells showed specific fluorescence in immunofluorescent tests, and electron microscopic examinations revealed TBE and VEE virions in sections.

[Variability of attenuated alphavirus strains in virion aggregate infection]. / Izmenchivost' attenuirovannykh shtammov al'favirusov v usloviiakh infektsii agregatami virionov.

A natural process of infection with multiploid virions was stimulated and the genetic effects occurring in this infection were studied in experiments with homogeneous attenuated strain of Venezuelan equine encephalomyelitis virus producing the smallest plaques. The process of infection with multiploid virions was reproduced by infecting the cells with artificially obtained aggregates. Under these conditions, small- and large-plaque virus was produced which in some cases had a higher virulence than the parental strain. A possible mechanism of this phenomenon and its practical implications for the evaluation of the properties of attenuated alphavirus strains is discussed.

[Effect of virulent and anttenuated variants of viruses of American equine encephalomyelitis on the mitotic activity of the cells]. / Vliianie virulentynykh i attenuirovannykh variantov virusov Amerikanskogo entsefalomielita loshadei na mitoticheskii rezhim kletok.

A comparative study of reproduction of Venezuelan and Eastern equine encephalomyelitis (VEE and EEE) viruses in L cell culture showed the disorders of the mitotic activity of the infected cultures to depend on the variant of the inoculated virus and the degree of its attenuation. The highest number of pathological mitosis was observed within 12--24 hours after inoculation of the cultures, the attenuated variants of both the viruses causing lower numbers of pathological mitoses than virulent variants. EEE virus was found to induce accumulation of pyknotized nuclei in the stage of telophase in the culture.

[Antiviral factor isolated from infected cell cultures]. / Antivirusnyi faktor, vydeliaemyi iz infitsirovannykh kul'tur kletok.

An antiviral factor of protein nature inhibiting reproduction of VEE, herpes, vaccinia VSV, fowl plague viruses was isolated from infected cell cultures. The factor has no virucidal or prophylactic effect, stable to low pH values and heating at 100 degrees C for 30 min.

Venezuelan equine encephalitis virus propagation in the olfactory tract of normal and immunized mice.

The role of virus spread in the induction of damage to the central nervous system (CNS) of mice infected with Venezuelan equine encephalitis virus (VEEV) via the respiratory route was studied. The virus concentration in various organs and in the blood, the sensitivity to different doses of virus, and ultrastructural lesions in various tissues were examined. It is concluded that VEEV can enter the CNS of nonimmunized mice both by vascular and by olfactory pathways, whereas in immunized mice the olfactory pathway is the main route.