Activity Patterns of St. Louis Encephalitis and West Nile Viruses in Free Ranging Birds during a Human Encephalitis Outbreak in Argentina.

St. Louis encephalitis virus (SLEV) (Flavivirus) is a reemerging arbovirus in the southern cone of South America. In 2005, an outbreak of SLEV in central Argentina resulted in 47 human cases with 9 deaths. In Argentina, the ecology of SLEV is poorly understood. Because certain birds are the primary amplifiers in North America, we hypothesized that birds amplify SLEV in Argentina as well. We compared avian SLEV seroprevalence in a variety of ecosystems in and around Córdoba city from 2004 (before the epidemic) and 2005 (during the epidemic). We also explored spatial patterns to better understand the local ecology of SLEV transmission. Because West Nile virus (WNV) was also detected in Argentina in 2005, all analyses were also conducted for WNV. A total of 980 birds were sampled for detection of SLEV and WNV neutralizing antibodies. SLEV seroprevalence in birds increased 11-fold from 2004 to 2005. Our study demonstrated that a high proportion (99.3%) of local birds were susceptible to SLEV infection immediately prior to the 2005 outbreak, indicating that the vertebrate host population was primed to amplify SLEV. SLEV was found distributed in a variety of environments throughout the city of Córdoba. However, the force of viral transmission varied among sites. Fine scale differences in populations of vectors and vertebrate hosts would explain this variation. In summary, we showed that in 2005, both SLEV and to a lesser extent WNV circulated in the avian population. Eared Dove, Picui Ground-Dove and Great Kiskadee are strong candidates to amplify SLEV because of their exposure to the pathogen at the population level, and their widespread abundance. For the same reasons, Rufous Hornero may be an important maintenance host for WNV in central Argentina. Competence studies and vector feeding studies are needed to confirm these relationships.

Continuous myocloni and tonic spasms in a 2-month-old infant with enterovirus 71 brain stem encephalitis.

Brain stem encephalitis is a cardinal presentation of central nervous system involvement in enterovirus 71 infection, and manifests as myoclonus, ataxia, tremor, and autonomic dysfunction. A 2-month-old infant with enterovirus 71 brain stem encephalitis demonstrated continuous myocloni and tonic spasms. On admission, the patient's myoclonus, which mainly involved the shoulders and the arms, was considerably worse during wakefulness and occurred once or twice a minute. Several hours after admission, the myoclonic jerks steadily worsened, appeared ceaselessly every 1 to 2 seconds, and were intermixed with tonic spasms of all four extremities accompanied by crying. Video electroencephalography revealed a normal background without epileptiform discharges and no ictal electroencephalographic changes during the myoclonic jerks and tonic spasms. Complete remission was achieved without complications after completion of a 3-day immunoglobulin therapy. This case suggests that the brain stem may be a major origin site for not only myoclonus but also tonic spasm.

Liver-specific mitochondrial respiratory chain complex I deficiency in fatal influenza encephalopathy.

We report on a 4-year-old boy who died from influenza encephalopathy. The clinical course and microscopic findings of the autopsied liver were compatible with Reye's syndrome. We examined the mitochondrial respiratory chain function by blue native polyacrylamide gel electrophoresis (BN-PAGE), western blotting, and respiratory chain enzyme activity assays. The activity of liver respiratory chain complex (CO) I was markedly decreased (7.2% of the respective control activity); whereas, the other respiratory chain complex activities were substantially normal (CO II, 57.9%; CO III, 122.3%; CO IV, 161.0%). The activities of CO I-IV in fibroblasts were normal (CO I, 82.0%; CO II, 83.1%; CO III, 72.9%; CO IV, 97.3%). The patient was diagnosed with liver-specific complex I deficiency. This inborn disorder may have contributed to the fatal outcome. We propose that relying only on fibroblast respiratory chain complex activities may lead to the misdiagnosis of liver-specific complex I deficiency.

Japanese encephalitis virus down-regulates thioredoxin and induces ROS-mediated ASK1-ERK/p38 MAPK activation in human promonocyte cells.

Japanese encephalitis virus (JEV), a mosquito-borne flavivirus, causes severe neurological disease with high mortality. Molecular mechanisms of JEV pathogenesis such as upstream apoptotic processes and pathways are not yet completely resolved or understood. In this study, JEV replication in human promonocyte cells induced time-dependent apoptosis and activated virus dose-dependent caspases 3, 8 and 9. Proteomic analysis demonstrated up- and down-regulated (more or less than 1.5-fold) proteins in JEV-infected promonocyte cells. Biological process categorization showed processes of antioxidation, free radical removal, and sulfur redox metabolism entailed many identified up- and down-regulated proteins. Down-regulation of thioredoxin, confirmed by using Western blotting, was involved in the apoptosis process of the oxidative stress response pathway. JEV infection caused increased intracellular ROS production and activation of ASK1-ERK/p38 MAPK signaling. ERK/p38 MAPK inhibitor PD98059 treatment definitely suppressed this apoptosis. Down-regulation of thioredoxin, increased intracellular ROS, and activation of ASK1-ERK/p38 MAPK signaling all were associated with JEV-induced apoptosis. These results are suggestive of an oxidative stress-pathway as a key element of JE pathogenesis.

Emerging viral encephalitides in dogs and cats.

Few viral pathogens resulting in encephalitis in dogs and cats have emerged over the past decade or so. All are the result of penetration through presumed species barriers and all are considered zoonoses or possible zoonotic pathogens. In all cases, encephalitis is a rare event that has low morbidity but high mortality. More viruses are likely to emerge as pathogenic in our domesticated carnivorous companions as our habitats continue to overlap with the shrinking wildlife habitats. Hopefully, however, none reach the level of distinction that was once held by rabies virus.

[Viral encephalitis virus, a new bioterrorist menace]. / Les virus des encéphalites virales, une nouvelle menace bioterroriste.

Often responsible for little known infections, today viral encephalitis viruses appear as a new bioterrorist menace, because of their easy production and their great pathogenic potential. Spraying is the best way to permit the rapid diffusion of certain encephalitis viruses. Diagnosis of viral encephalitis, predominating in tropical surroundings, is difficult. In the majority of cases, symptoms differ little from those of common flu. With supplementary examinations, the biological abnormalities are usually non-specific. There are no characteristic images on scans or MRI. Identification of the virus in the nasopharynx, blood or cerebrospinal fluid, in serology, PCR or RT-PCR permits confirmation of the virus. Treatment is essentially symptomatic and relies on appropriate reanimation measures. Ribavirin can be indicated in some cases such as the Rift Valley fever, but is formally contraindicated in West Nile encephalitis. The aim of terrorist groups who would use this type of weapon is more to provoke panic and disorganisation than to kill as many people as possible.

Vaccines and animal models for arboviral encephalitides.

Arthropod-borne viruses ("arboviruses") cause significant human illness ranging from mild, asymptomatic infection to fatal encephalitis or hemorrhagic fever. The most significant arboviruses causing human illness belong to genera in three viral families, Togaviridae, Flaviviridae, and Bunyaviridae. These viruses represent a significant public health threat to many parts of the world, and, as evidenced by the recent introduction of the West Nile virus (WNV) to the Western Hemisphere, they can no longer be considered specific to any one country or region of the world. Like most viral diseases, there are no specific therapies for the arboviral encephalitides; therefore, effective vaccines remain the front line of defense for these diseases. With this in mind, the development of new, more effective vaccines and the appropriate animal models in which to test them become paramount. In fact, for many important arboviruses (e.g. California serogroup and St. Louis encephalitis viruses), there are currently no approved vaccines available for human use. For others, such as the alphaviruses, human vaccines are available only as Investigational New Drugs, and thus are not in widespread use. On the other hand, safe and effective vaccines against tick-borne encephalitis virus (TBEV) and Japanese encephalitis virus (JEV) have been in use for decades. New challenges in vaccine development have been met with new technologies in vaccine research. Many of the newer vaccines are now being developed by recombinant DNA technology. For example, chimeric virus vaccines have been developed using infectious clone technology for many of the arboviruses including, WNV, JEV, and TBEV. Other successful approaches have involved the use of naked DNA encoding and subsequently expressing the desired protective epitopes. Naked DNA vaccines have been used for TBEV and JEV and are currently under development for use against WNV. The development of less expensive, more authentic animal models to evaluate new vaccines against arboviral diseases will become increasingly important as these new approaches in vaccine research are realized. This article reviews the current status of vaccines, both approved for use and those in developmental stages, against the major arboviral encephalitides causing human disease. In addition, research on animal models, both past and present, for these diseases are discussed.

Pre-West Nile virus outbreak: perceptions and practices to prevent mosquito bites and viral encephalitis in the United States.

Mosquitoes can transmit over 100 of the viruses that can cause encephalitis, meningitis, and hemorrhagic disease in humans (Chin 2000; Gubler 1996; Monath 1989). While much is known about the ecology, epidemiology, and clinical manifestations of the arboviral encephalitides (Campbell et al. 2002; Centers for Disease Control and Prevention 1997; Gubler 1998; Hayes 1989; Hubálek and Halouzka 1999), little empirical research exists regarding the U.S. population's knowledge of mosquitoes and arboviral encephalitis, particularly prior to the U.S. outbreak of West Nile virus (WNV) in 1999. A nationally representative 55-item survey instrument was successfully administered to 1,500 adults in the United States and an additional 250 adults in six states in the Northeast (Connecticut, Delaware, New Jersey, New York, Pennsylvania, and Rhode Island) regarding mosquitoes and mosquito-borne viral encephalitis. A summary outcome measure for mosquito bite prevention was created. Analyses revealed that the following were statistically significant predictors of behaviors taken to prevent mosquito bites: being concerned about being bitten by mosquitoes, perceived effectiveness of staying indoors in late afternoon and early evening was protective, perceived effectiveness that mosquito repellent is not harmful to health, owning dogs and/or cats as pets, being married, and being > or = 18-44 years old. Being concerned about being bitten by mosquitoes was the most robust predictor of behavioral action to prevent mosquito bites (OR = 7.3; 95% CI = 4.3, 12.2). Observed misperceptions and inadequate knowledge regarding insect repellents suggest increased promotion of the safety and efficacy of DEET-containing insect repellents is warranted.

Induction of nodavirus disease in seabass, Dicentrarchus labrax, using different infection models.

The aim of this study was to investigate the susceptibility of juvenile and adult seabass, which are generally thought to be refractory to nodavirus. Moreover, preliminary immunological studies were performed to examine the immune response of adult seabass. Successful transmission of the disease was experimentally demonstrated in juvenile and adult seabass as ascertained by the presence of the clinical signs of the disease, re-isolation of the virus in the SSN-1 cell line and subsequent confirmation by histopathology and immunohistochemistry. Bigger seabass not only developed the clinical disease but also suffered mortalities. Serum neutralisation titres were considered low in this study.

A new tick-borne encephalitis-like virus infecting New England deer ticks, Ixodes dammini.

To determine if eastern North American Ixodes dammini, like related ticks in Eurasia, maintain tick-borne encephalitis group viruses, we analyzed ticks collected from sites where the agent of Lyme disease is zoonotic. Two viral isolates were obtained by inoculating mice with homogenates from tick salivary glands. The virus, which was described by reverse transcriptase polymerase chain reaction and direct sequencing of the amplification products, was similar to, but distinct from, Powassan virus and is provisionally named "deer tick virus." Enzootic tick-borne encephalitis group viruses accompany the agents of Lyme disease, babesiosis, and granulocytic ehrlichiosis in a Holarctic assemblage of emergent deer tick pathogens.

Cold stress-induced neuroinvasiveness of attenuated arboviruses is not solely mediated by corticosterone.

In previous studies we have shown that various stress paradigms can induce the penetration of noninvasive, attenuated viruses into the central nervous system (CNS). Since glucocorticoids levels are elevated during stress, we compared the effect of cold stress and corticosterone (CS) injection on neuroinvasiveness of a non-invasive encephalitic virus, WN-25 (West Nile). Exposure of inoculated mice to cold stress or CS resulted in high viremia and a marked increase in mortality when compared to control untreated mice. Exposure of WN-25 inoculated mice to cold treatment or CS injection led to high blood virus levels as compared to nontreated mice (3.2 and 3.1 vs > 1 log 10 PFU/ml). Cold stress or CS (5000 ng/mouse) treatment caused a mortality rate of 70% and 50% of the WN-25 inoculated mice respectively. No mortality was recorded in control inoculated groups (p < 0.05). Passive transfer serum from uninfected cold stressed mice to WN-25 inoculated nonstressed mice, resulted in similar mortality. The levels of CS in passive transferred serum from cold stressed animals was 500 ng/ml, only 2% (100 vs. 5000 ng) of the CS dose required to obtain a similar effect on viral penetration and mortality when CS was injected directly. Therefore, we concluded that CS was not the sole factor responsible for the cold stress effect on the viral infection outcome.

Venezuelan equine: encephalomyelitis virus: structural components / Componentes estructurales: virus encefalomielitis equina venezolana

Venezuelan equine encephalomyelitis (VEE) virus, purified in sucrose density gradients was examined with the electron microscope before and after sodium desoxycholate (DOC) treatment. The structure of the nucleocapsid revealed 10 to 12 nm ring shaped units organized into an icosahedral symmetry. Density gradied fractions exhibiting hemagglutinating activity after DOC treatment show 12 to 18 nm particles lined by short projections. These isolated hemagglutinating subunits are though to correspond to the superficial spikes of VEE virus

Efectos del pH y la concentración iónica sobre la infecciosidad del virus de la encefalitis equina venezolana / Effecs of pH and ionic concentration on VEE virus infection

El virus de la EEV a una temperatura de 37ºC sufre alteraciones en su nivel de infecciosidad, que varían según la concentración de la solución empleada. En buffer citrato 100 mM a pH 3,4 y 5, hay pérdida notable de la infecciosidad viral; cuando se usan concentraciones de 5 y 10 mM, a los mismos pH, no se observa ninguna variación en su nivel de infecciosidad. En presencia de soluciones diluídas de cloruro de magnesio pH 4 y a 77ºC, el virus es estable en su nivel de infecciosidad; lo que no se observa en las soluciones de elevada concentración (1 ó 2 M). Cuando se emplea una solución de MgSO4, no ocurre ninguna variación del nivel de infecciosidad del virus tratado en las mismas condiciones

Encefalitis equina venezolana: estudio serológico en aves de la región de la Guajira venezolana, durante un brote epidémico / Serological survery in birds during an outbreak of veneuelan equine encephalitis virus in the venezuelan Guajira

Para determinar el papel que algunas aves pudieran tener en el inicio y diseminación del virus de la Encefalitis equina Venezolana en períodos epidémicos, se estudiaron los sueros de 37 aves capturadas en zonas de actividad viral, durante una epidemia causada por este virus en la Guajira venezolana en el mes de octubre de 1973. Las muestras de suero fueron procesadas por el método de inhibición de placas bajo capa de agar, utilizando como antígeno virus de la encefalitis venezolana, cepa Guajira. No se observó reducción de placas en relación al virus control, por lo que se deduce que los sueros probados no poseían anticuerpos contra el virus. Aunque la muestra es muy pequeña, se podría concluir que estas aves podrían accidentalmente estar involucradas en el ciclo epidemiológico del virus, pero no jugarían papel importante en la diseminación

Respuesta de anticuerpos contra encefalitis equina venezolana en equidos vacunados con la vacuna a virus vivo modificado (TC-83) / Evaluation of the attenuated vezezuelan equine encephalitis vaccine (TC-83) in equine

A raíz de la epidemia de Encefalitis Equina Venezolana (EEV) de 1973, se utilizó por primera vez en Venezuela, la vacuna a virus vivo modificado (TC-83). Para evaluar la efectividad de la misma, se estudiaron 49 pares de sueros de équidos, sin antecedentes de vacunación o enfermedad, que fueron vacunados con la vacuna TC-83. Se comprobó que 28 de estos animales poseían anticuerpos contra EEV en el suero prevacunación. De los 21 restantes, sin anticuerpos neutralizantes ni inhibidores de la hemaglutinación, 17 convirtieron al cabo de seis semanas después de la vacunación. A pesar de ser una muestra muy pequeña, creemos que estos resultados apoyan la tesis sustentada por algunos investigadores sobre la efectividad de la vacuna a virus vivo atenuado TC-83, contra Encefalitis Equina Venezolana. Asimismo sustentamos la idea de que la vacunación efectuada a tiempo en las zonas en peligro durante 1973, contribuyó a detener la extensión de la epidemia

Estudio leucocitario en animales de laboratorio infectados con virus de encefalitis equina venezolana / Leucocitary study in laboratory animals infected with venezuelan equine encephalitis virus

Se estudió el número de glóbulos blancos y fórmula leucocitaria en animales de laboratorio tales como ratones, ratas y conejos antes y después de producirles una infección experimental aguda con el virus de la Encefalitis equina Venezolana. Se observó una leucopenia que varió de acuerdo a la sensibilidad de la especie animal. Las fórmulas leucocitarias presentaron variaciones, acompañadas de células atípicas las cuales persistieron hasta el final de la infección. Se observó también una vacuolización celular que alcanzó al 6 por ciento y que no estuvo presente nunca en los controles. Los conejos presentaron una eosinofilia que abarcó hasta un 36 por ciento en su fórmula leucocitaria poco antes de morir. Algunas ratas sobrevivieron y al cabo de 14 días presentaron títulos de anticuerpos inhibidores de la hemaglutinación entre 1/80-1/160, lo cual indica su resistencia a la infección viral

Detección de anticuerpos al virus de la encefalitis equina venezolana en équidos de los Distritos Mara y Páez de la Guajira Venezolana: 1984 evaluación prevacunal / Venezuelan equine encephalitis antibodies detección in equidae from the Guajira venezolana Mara and Paez Districts: 1984 prevacunal evaluation

Con el propósito de conocer el estado inmunológico de la población de équidos de los Distritos Mara y Páez de la Guajira Venezolana en relación al Virus de la Encefalitis Venezolana, fueron obtenidos entre mayo y junio de 1984, muestras de sangre de 236 équidos procedentes de los Distritos Mara (126) y Páez (110). Inmediatamente después fueron inoculados con 2 ml de la vacuna TC-83 (virus modificado de encefalitis equina venezolana (EEV). Vecol/Colombia). De las muestras procesadas (221), el 53 por ciento correspondió a equinos, el 43,6 por ciento asnales y 3,4 por ciento mulares. El 53 por ciento mostró títulos inhibidores de la hemaglutinación (IHA) mayores de 1:20 en la muestra previa a la vacunación a expensas principalmente de los animales con antecedentes de vacunación. La mayoría de los títulos IHA estuvieron entre 1:20 y 1:640. Se demostró que para ese año el 53 por ciento de la población de equidos del Distrito Mara y Páez, presentaban anticuerpos contra EEV

Inoculación experimental del paramixovirus del ojo azul en el gato doméstico (Felis catus) / Experimental inoculation of blue eye paramyxovirus in the cat (Felis catus)

En este trabajo se seleccionó al gato doméstico, debido a que la población felina tiene amplia distribución dentro de las granjas porcinas, principalmente por ser usada como control de roedores. Quince gatos adultos fueron inoculados con 4 ml de paramixovirus de ojo azul (POA) con un título de 10 DICC/ml por vía intranasal aplicado como bomba de aspersión. A los 0, 14 y 21 días (PI) se obtuvieron muestras séricas para detección de anticuerpos contra POA por las pruebas de inhibición de la hemoglutinación beta (IHA) y seroneutralización método beta (SN). Para el aislamiento viral se tomó hisoponasal y ocular a los 4 días, y biopsia de tonsila a los 7 días. Los animales fueron sacrificados a los 21 días posinoculación (PI). Se tomaron muestras de encéfalo, pulmón y tonsila para las prueba de inmunofluorescencia (IF) directa y para estudio histopatológico (HP). Todas las muestras de biopsia de tonsila e hisopo nasal y ocular fueron negativas en cultivo celular (CC) en los tres pases ciegos. En la prueba de IHA se obtuvieron los siguientes resultados: el primer muestreo fue negativo y el segundo y tercero detectaron títulos entre 1:6 y 1:192. En las SN, el primer muestreo fue negativo y en el segundo y tercero se detectaron títulos entre 1:4 y 1:64. La IF directa de órganos fue negativa para pulmón, tonsila y encéfalo. En el estudio HP no hubo cambios significativos. De los resultados obtenidos se concluye que el gato tiene la capacidad de seroconvertir a POA sin ser necesariamente un portador