Western equine and St. Louis encephalitis in man,California, 1945-1950

A large group of etiologic agents, some known and some unknown, produce in man a clinical syndrome now labeled "infectious encephalitis."The separation, from this larger group, of single disease entities which cause similar clinical symptoms is possible, but calls for diagnostic acumen plus supporting laboratory evidence. Two etiologically specific entities, western equine encephalitis and St. Louis encephalitis, are frequently encountered in rather well-defined areas of California, the Central Valley and Imperial Valley; and there is a definite seasonal pattern of occurrence-June through October. There are certain guides that are helpful in differential diagnosis. Establishing a diagnosis on the basis of clinical evidence is difficult. Laboratory studies are of great importance not only for diagnosis in the individual case but for advancement of etiology. Specimens are worthless unless taken at proper intervals and submitted by methods described. It is probable that encephalitides caused by still unknown agents exist in California. The isolation and identification of new encephalitogenic viral agents will depend in large measure upon the submission by physicians of suitable specimens from patients with central nervous system disease in which the cause is obscure.     

Western equine encephalomyelitis vaccine produced in chick embryo cell cultures

A method was developed for production of a freeze-dried Western equine encephalomyelitis vaccine from virus propagated in chick embryo cell culture monolayers maintained with a serum-free medium. A sufficient concentration of virus accumulated in the cell culture fluids prior to the occurrence of viral cytopathology to permit the production of a vaccine relatively free from serum and cellular proteins. Inoculation with two mouse ld₅₀ doses of virus per 100 tissue culture cells was found to yield reproducible high virus titers at a convenient harvest time. These harvests were inactivated at 22 C by 0.05% formalin within 48 hr. Potency test results, as measured by the protection of immunized guinea pigs against an intracerebral virus challenge, indicated that the vaccine produced from the virus propagated in cell culture was equal in potency to a lot of whole chick embryo vaccine used to immunize laboratory and field workers subject to a high risk of infection.     

Eastern equine encephalomyelitis virus in experimentally infected bats.

Colonial bats (Myotis supp. and Eptesicus sp.) were infected with eastern equine encephalomyelitis virus by subcutaneous inoculation or by the bite of infected mosquitoes. Bats were maintained in an environment simulating conditions encountered in hibernacula or in summer maternal colonies. Virus was detected in the blood of hibernating bats at irregular intervals over a 42-day observation period; viremia perhaps was influenced by the amount of disturbance (arousal) involved in the blood sampling process. Target organs included brown fat, spleen, lung, kidneys, pancreas, and liver. Neutralizing antibody was not detected in sera collected from these bats between days 4 and 42 post-inoculation. In nonhibernating bats, virus was recovered from mammary glands, brown fat, pancreas, lungs, kidneys, and liver, in addition to blood. Attempts to infect bats orally or to transmit virus to suckling mice by the bite of viremic bats were unsuccessful. Virus was transmitted from viremic chickens to E. fuscus by the bite of Culiseta melanura and Aedes aegypti.     

Phylogenetic and evolutionary analyses of St. Louis encephalitis virus genomes

St. Louis encephalitis virus belongs to the Japanese encephalitis virus serocomplex of the genus Flavivirus, family Flaviviridae. Since the first known epidemic in 1933, the virus has been isolated from a variety of geographical, temporal, and host origins. We have sequenced 10,236 nucleotides of the open reading frame (93.6% of the full-length genome) of 23 of these strains, and have used the sequences to conduct phylogenetic analyses, in order to investigate the forces shaping the evolution of St. Louis encephalitis virus. Contrary to previous reports, we found little evidence for recombination in these isolates. Most of the amino acid sites in the SLEV polyprotein appeared to be under negative selection, with some sites evolving neutrally, and a small number under positive selection. The strongest signal for positive selection was evident in the N-linked glycosylation site of the envelope protein. Intra-strain sequence variability within strains was observed at this site, and analyses suggested that it is under selection in vitro. Furthermore, using heterochronous sequence data, we estimated the most recent expansion of St. Louis encephalitis virus in North America to have happened towards the end of the 19th century.