Equine leukoencephalomalacia associated with ingestion of corn contaminated with fumonisin B1

This article describes clinical, etiologic and pathologic diagnosis of an outbreak of equine leukoencephalomalacia. Two samples of the corn consumed by the affected horses contained fumonisin B1 at levels of 46 and 53 ug/g and Fusarium moniliforme, a good in vitro mycotoxin producer.

In-situ hybridization for demonstration of equine herpesvirus type 1 DNA in paraffin wax-embedded tissues and its use in horses with disseminated necrotizing myeloencephalitis.

The detection of equine herpesvirus type 1 (EHV-1) in infected cell cultures, and in tissues taken at necropsy, by the in-situ hybridization technique is described. A 4.9 kb Bam HI fragment of EHV-1 vaccine strain RacH was used as a probe after labelling with [alpha-32P] thymidine 5'-triphosphate ([32P]TTP) or digoxigenin-deoxyuridine 5'-triphosphate (dUTP). Both probes specifically detected EHV-1 DNA in either cytospin or paraffin wax-embedded preparations of infected cells. The digoxigenin-labelled probe was further used to examine tissue sections of equine fetuses which had been aborted due to EHV-1 infection. In all cases positive hybridization signals were mainly associated with the nuclei. Positive results were confirmed by immunostaining of EHV-1 antigen in adjacent sections. However, both methods failed to detect EHV-1 in spinal cord sections of six horses suffering from disseminated necrotizing myeloencephalitis (DNM). These results support the hypothesis that DNM is not caused by a productive viral infection of parenchyma of the nervous system but is immunologically mediated.

Evolution of alphaviruses in the eastern equine encephalomyelitis complex.

Evolution of viruses in the eastern equine encephalomyelitis (EEE) complex was studied by analyzing RNA sequences and oligonucleotide fingerprints from isolates representing the North and South American antigenic varieties. By using homologous sequences of Venezuelan equine encephalomyelitis virus as an outgroup, phylogenetic trees revealed three main EEE virus monophyletic groups. A North American variety group included all isolates from North America and the Caribbean. One South American variety group included isolates from the Amazon basin in Brazil and Peru, while the other included strains from Argentina, Guyana, Ecuador, Panama, Trinidad, and Venezuela. No evidence of heterologous recombination was obtained when three separate regions of the EEE virus genome were analyzed independently. Estimates of the overall rate of EEE virus evolution (nucleotide substitution) were 1.6 x 10(-4) substitution per nucleotide per year for the North American group and 4.3 x 10(-4) for the Argentina-Panama South American group. Evolutionary rate estimates for the North American group increased over 10-fold (from about 2 x 10(-5) to 4 x 10(-4)) concurrent with divergence of two monophyletic groups during the early 1970s. The North and South American antigenic varieties diverged roughly 1,000 years ago, while the two main South American groups diverged about 450 years ago. Analysis of multiple strains isolated from an upstate New York transmission focus during the same years suggested that, in certain locations, EEE virus may be relatively isolated for short time periods.

Western equine encephalomyelitis: virulence markers and their epidemiologic significance.

Comparative studies are described on the virulence of the western equine encephalomyelitis (WEE) complex viruses for mice. Three epizootic WEE virus strains (McMillan, Cba 87, and Cba CIV 180) and five enzootic WEE complex viruses (Highlands J [HJ], Y62-33, Aura, Fort Morgan [FM], and WEE AG80-646) were examined. The neurovirulence and the neuroinvasiveness of these viruses for adult mice were established and correlated with viremia and virus replication in brain tissue. Adult mice inoculated intraperitoneally showed differential responses that corresponded to the epidemiologic attributes of WEE viruses. Viruses associated with equine epizootics were neurovirulent and neuroinvasive, whereas enzootic viruses were neither neuroinvasive nor neurovirulent. In North America, HJ virus appears to be an antigenic link with an intermediate virulence between epizootic WEE virus and the enzootic FM virus. The HJ virus has been associated with rare cases of sporadic equine and human diseases. In South America, no virus with intermediate virulence characteristics has been described. We speculate that epizootics may arise from nonpathogenic strains such as AG80-646 maintained in enzootic transmission cycles.

Circulation of eastern equine encephalitis, western equine encephalitis, Ilhéus, Maguari and Tacaiuma viruses in equines of the Brazilian Pantanal, South America.

Neutralizing antibodies to EEE (6.7%), WEE (1.2%), ILH (26.6%), MAG (28.2%) and TCM (15.7%) viruses were found in sera of 432 equines of the Brazilian Pantanal, area where undiagnosed horse deaths are frequently observed. A 4-fold rise in CF titer to EEE virus was detected in acute and convalescent sera of an encephalitis horse sacrificed in 1992. Antibodies to EEE, ILH, MAG and TCM viruses were detected in horses less than 2 years old indicating recent circulation of these viruses in the Pantanal. The evidence of recent equine encephalitis associated with rising CF titer to EEE warrants a more intensive study with attempts to isolate virus from horses with clinical manifestations of encephalitis.

[Epidemiology of encephalitis caused by arbovirus in the Brazilian Amazonia]. / Epidemiologia das encefalities por arbovírus na Amazônia brasileira.

An overview of ecological, epidemiological and clinical findings of potential arthropod-borne encephalitis viruses circulating in the Amazon Region of Brazil are discussed. These viruses are the Eastern Equine Encephalitis (EEE), Western Equine Encephalitis (WEE), St. Louis Encephalitis (SLE), Mucambo (MUC) and Pixuna (PIX). These last two are subtypes (III and IV) of Venezuelan Equine Encephalitis virus. The areas of study were the highways and projects of development, as well as places where outbreaks of human diseases caused by arboviruses had been detected. These viruses are widespread in all Amazonia, and at least four of them, EEE, WEE, SLE and MUC are pathogenic to man. EEE and WEE infections were detected by serology, while SLE and MUC by either serology and virus isolation. The PIX virus has the lowest prevalence and, it was isolated in only a few cases, one being from a laboratory infection. Wild birds are the main hosts for all these viruses, except MUC, whose major hosts are rodents. The symptoms presented by infected people were generally a mild febrile illness. Although, jaundice was observed in two individuals from whom SLE was isolated. A comparison of the clinical symptoms presented by the patients in the Amazon Region and other areas of America, especially in the USA is made. In Brazilian Amazon region epidemics have not been detected although, at least, one EEE epizootic was recorded in Bragança, Para State, in 1960. At that time, of 500 horses that were examined 61% were positive to EEE by HI and of them 8.2% died. On the other hand, SLE has caused four epizootics in a forest near Belem. Wild birds and sentinel monkeys were infected, but no human cases were reported.

Viral encephalitis.

Nonherpetic encephalitis outside the newborn period is usually a self-limited disease. The majority of patients will recover without significant sequelae, and require only supportive therapy during the acute illness. Though the underlying viral etiology frequently will escape detection, identification of the infecting agent has considerable prognostic value which can complement clinical measures of severity of disease. The most important initial task of the clinician faced with a case of presumptive viral encephalitis is to eliminate the possibility of a treatable illness. Once this has been done, the diagnosis of viral encephalitis can be supported by documenting the characteristic slow-wave background activity on EEG, and a mild lymphocellular pleocytosis in the CSF. Because viral encephalitis can be caused by such a large number of organisms, the search for an etiology can be daunting. Realizing that all the agents described above can, at times, cause encephalitis without any clue to their identity, one nevertheless may use several pieces of historical information to narrow the possibilities. Travel history, animal exposures, immunization history, and seasonality all may help to steer the search in a particular direction and, indeed, may point to a nonvirologic cause as well. In addition, detection of extraneurological signs and symptoms may strongly indicate a specific virologic diagnosis. Finally, knowledge of concurrent community epidemic patterns, and of surveillance data routinely collected by local and state health departments, can help to increase or decrease the likelihood of a given pathogen. The causative viral agent usually can be identified by serological testing and viral culture. Occasionally, single serological determinations are diagnostic: in rabies (when the patient has not received immune prophylaxis), eastern equine encephalitis, and HIV, since seropositivity is strongly associated with symptomatic illness; and in Epstein-Barr virus, if a panel of antibody determinations which can time the infection is available. In addition, high CSF: serum titers for antibody against any neurotropic agent is usually diagnostic, though the absence of a high central nervous system antibody titer does not eliminate any potential viral pathogen. With these few exceptions, a single serological determination for a given pathogen is almost always impossible to interpret; paired sera (one obtained upon diagnosis, and one obtained 10 to 14 days later, either just prior to hospital discharge or at a follow-up visit) are far more helpful. Many viruses that directly infect the central nervous system are difficult to recover from the CSF; therefore, viral isolation from the nasopharynx and stool also should be sought.(ABSTRACT TRUNCATED AT 400 WORDS)

Identification of an antigenic subtype of eastern equine encephalitis virus isolated from a human.

Eastern equine encephalitis (EEE) virus was isolated from the cerebrospinal fluid of a 6-year-old male who had clinically diagnosed aseptic meningitis and subsequently died. Several standard serologic tests that use polyclonal antibody and indirect immunofluorescence and hemagglutination inhibition tests that use monoclonal antibody provided evidence that the isolate was an antigenic subtype of prototype North American EEE virus. We believe that this is the first evidence of an antigenic subtype of EEE virus.

Characterization of the mesenteronal infection with Western equine encephalomyelitis virus in an incompetent strain of Culex tarsalis.

Interactions of western equine encephalomyelitis (WEE) virus are compared with mesenterons of 2 genetically selected susceptible (WS) and refractory (WR) lines of Culex tarsalis. Both WS and WR females had similar susceptibility when parenterally inoculated with virus, thus it was initially thought that resistance in WR Cx. tarsalis was associated with a mesenteronal infection barrier. Present data on viral growth in mesenterons dissected from females fed on virus-soaked pledgets suggest that virus infected and multiplied in some WR mesenterons, but to significantly lower titers than in WS mesenterons. The proportion of WR females with infected mesenterons varied depending on the time after feeding, incubation temperature, and whether mesenterons were incubated with WEE viral antibody before viral assay. The percentage of WR mesenterons infected did not increase significantly when diethylaminoethyl dextran was added to the infectious bloodmeal, the pH of the infectious bloodmeal was altered, or virus was introduced by intrathoracic inoculation into the hemocoel. It was concluded that the low titers of virus detected in both WR mesenterons and whole mosquitoes were influenced by the genetic ability of WR Cx. tarsalis to modulate WEE viral titers to low or undetectable levels after peroral or parenteral infection. These findings make it difficult to determine what proportion of the WR mesenterons are resistant to infection with WEE virus. WS and WR Cx. tarsalis were equally susceptible to peroral infection with the flavivirus St. Louis encephalitis and the bunyavirus Turlock.

Eastern equine encephalitis in Massachusetts: a report of 16 cases, 1970-1984.

We studied the case records of 16 patients with eastern equine encephalitis (EEE) in Massachusetts from 1970 to 1984 and compared their presentations, courses, and outcomes with the data available from previous epidemics. In recent years, there has been a greater frequency of EEE in adults, whereas in the past it was considered a disease of children. Also, prognosis for a good functional recovery seems to be correlated with age over 40 years, a long prodromal course (5 to 7 days) of constitutional symptoms, and the absence of coma. Previous reports did not mention these significant correlations. We also stress the positive and negative diagnostic correlations, in order to distinguish between EEE and herpes simplex encephalitis.

The ecology of western equine encephalomyelitis virus in the Central Valley of California, 1945-1985.

Reeves' concept of the summer transmission cycle of western equine encephalomyelitis virus in 1945 was that the virus was amplified in a silent transmission cycle involving mosquitoes, domestic chickens, and possibly wild birds, from which it could be transmitted tangentially to and cause disease in human and equine populations. Extensive field and laboratory studies done since 1945 in the Central Valley of California have more clearly defined the specific invertebrate and vertebrate hosts involved in the basic virus transmission cycle, but the overall concept remains unchanged. The basic transmission cycle involves Culex tarsalis as the primary vector mosquito species and house finches and house sparrows as the primary amplifying hosts. Secondary amplifying hosts, upon which Cx. tarsalis frequently feeds, include other passerine species, chickens, and possibly pheasants in areas where they are abundant. Another transmission cycle that most likely is initiated from the Cx. tarsalis-wild bird cycle involves Aedes melanimon and the blacktail jackrabbit. Like humans and horses, California ground squirrels, western tree squirrels, and a few other wild mammal species become infected tangentially with the virus but do not contribute significantly to virus amplification.

A newly recognized vesiculovirus, Calchaqui virus, and subtypes of Melao and Maguari viruses from Argentina, with serologic evidence for infections of humans and horses.

In 1983, 17 virus strains were isolated from mosquitoes collected during an outbreak of western equine encephalitis in Santa Fe Province, Argentina. Strains of western equine encephalitis, Venezuelan equine encephalitis, St. Louis encephalitis, and Antequera viruses were isolated, as were several bunyaviruses of the California and Bunyamwera serogroups and a new vesiculovirus. Complement fixation and neutralization tests were used to identify the California serogroup virus as a subtype of Melao virus, the Bunyamwera serogroup virus as a subtype of both Maguari and Playas viruses, and the vesiculovirus as a newly recognized agent for which the name Calchaqui virus is proposed. A limited serosurvey of horses and humans in Santa Fe Province and horses from the adjacent Santiago del Estero Province was performed to determine the prevalence of neutralizing antibody to the subtypes of Melao and Maguari viruses and to Calchaqui virus. The high prevalence of antibodies to these three agents indicates the need for further studies of their disease potential in horses, because they are closely related to several other viruses that are known equine pathogens.

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.

Human eastern equine encephalitis. Electron microscopic study of a brain biopsy.

An electron microscopic study was done on brain biopsy tissue from an eight-month-old female with acute eastern equine encephalitis diagnosed by indirect immunofluorescence. Rare clusters of round virions were found almost exclusively in the extracellular space. All virions observed had spiked envelopes, and their sizes averaged approximately 55 nm. Also found were rare enveloped virions along with degenerate organelles in a membrane-bound structure in the cytoplasm of macrophages. Intracytoplasmic development of virions was not found. Tubuloreticular complexes were seen in the endothelial cells and macrophages. This is the first report of an electron microscopic study on biopsy material from a case of human eastern equine encephalitis. It will extend the usefulness of brain biopsy in the diagnosis of acute encephalitis.

Isolation and preliminary characterization of mutants of western equine encephalomyelitis virus with altered virulence in chickens.

A small plaque (sp) and four temperature-sensitive (ts) mutants of western equine encephalomyelitis virus (WEEV) were obtained from a highly virulent field isolate (BFS-1703) after mutagenesis with 5-azacytidine or nitrosoguanidine or following undiluted passage in mosquito cell culture. Three ts mutants (158, 214, and 260) expressed stable ts phenotypes as determined both by six-hour leak yield and efficiency of plaquing (EOP) analyses. Progeny of ts mutant 214 grown at the nonpermissive temperature lacked hemagglutinin activity. The sp mutant (148) was as highly virulent as the wild-type virus (WTV) for newly hatched chickens (infected when 12 hours old), whereas the ts mutants were attenuated in their virulence for this host. Attentuation was correlated with lower viremia titers in 24-hour postinfection blood samples. The WTV and those mutants able to infect 21-day-old chickens produced nonlethal infections but the growth of ts 158 was apparently restricted in this high-temperature nonpermissive host. All ts mutants except ts 158 lost their ts phenotype after growth for 24 hours in newly hatched chickens and in duck embryonic cell culture (DECC) maintained at 42 degrees C.