Replication of Rocio virus in primary cultures of mouse neural cells

ABSTRACT Objective This study verified the replication efficiency of the Rocio virus in a primary culture of mouse neural cells. Methods Mixed primary cultures (neurons/glia) obtained from the brains of newborn isogenic BALB/c mice were inoculated with Rocio virus on the 7 th day of culture, and the development of cytopathogenic effects was monitored. The infection was confirmed via immunocytochemistry (anti-ROCV), while viral replication was quantified in infected primary cultures. The titration method used depended on the infection period. Results Rocio virus efficiently infected primary cultured neural cells, with the highest viral titer causing cytopathic changes was observed at 2 days post infection. The virus-infected primary culture survived for up to 7 days post infection, and viral load quantitation showed viral replication kinetics compatible with the cell death kinetics of cultures. Conclusion The findings of this study suggest that mouse neural cell primary cultures support Rocio virus replication and could be used as an alternative system for studying Flavivirus infection in the central nervous system.

Níveis séricos de vitamina D e alterações hepáticas em pacientes com malária na região Amazônica / Serum levels of vitamin d and hepatic changes in patients with malaria in the Amazon region / Niveles séricos de vitamina d y alteraciones hepáticas en pacientes con malaria en la Amazonia

Vários estudos sugerem a importância da vitamina D ­ 25(OH)D ­ na evolução clínica dos pacientes com malária. Entretanto, a prevalência de deficiência de 25(OH)D na população amazônica é pouco conhecida, havendo também poucos estudos com pacientes diagnosticados com malária. Assim, o objetivo deste estudo foi avaliar os níveis séricos de 25(OH)D em pacientes com malária e sua relação com dados epidemiológicos, parasitológico e provas de função hepática. Para tanto, foi realizado um estudo transversal analítico com um grupo de pacientes com malária e um grupo controle no município de Itaituba (PA), Brasil, no período de janeiro de 2018 a outubro de 2019. Elaborou-se um protocolo para avaliação dos dados sociodemográficos, parasitológicos e laboratoriais, adotando-se o nível de significância de 5%. A prevalência de deficiência de 25(OH)D foi observada nos pacientes com malária (28,5%) e no grupo controle (24,6%), sem diferença estatística; porém, entre os residentes no garimpo, os níveis séricos foram estatisticamente menores nos pacientes com malária. Os níveis séricos de transaminase glutâmico-pirúvica (TGP) apresentaram correlação inversa com os de 25(OH)D. As provas de função hepática foram significativamente maiores no grupo com malária. Dessa forma, este estudo evidenciou a deficiência de 25(OH)D em Itaituba. Alterações hepáticas pela infecção plasmodial podem ter contribuído para a correlação inversa observada entre os níveis de TGP e 25(OH)D.

Several studies suggest the importance of vitamin D ­ 25(OH)D ­ in the clinical evolution of patients with malaria. However, the prevalence of 25(OH)D deficiency in the Amazonian population is little known, and studies with patients diagnosed with malaria are scarce. Thus the objective of this study is to evaluate the serum levels of 25(OH)D in patients with malaria and its relationship with epidemiological and parasitological data and liver function tests. To that end, an analytical cross-sectional study was carried out with a group of patients with malaria and a control group in the municipality of Itaituba (PA), Brazil, from January 2018 to October 2019. A protocol was elaborated for the evaluation of sociodemographic, parasitological, and laboratory data, adopting a significance level of 5%. Results: The prevalence of 25(OH)D deficiency was observed in patients with malaria (28.5%) and in the control group (24.6%), with no statistical difference; however, among residents in the mining, serum levels were statistically lower in patients with malaria. The glutamic-pyruvic transaminase (GPT) serum levels showed an inverse correlation with 25(OH)D levels. Liver function tests were significantly higher in the malaria group. Thus, this study evidenced 25(OH)D deficiency in Itaituba. Hepatic changes due to plasmodial infection may have contributed to the inverse correlation observed between GPT and 25(OH)D levels.

Diversos estudios sugieren la importancia de la vitamina D ­[25(OH)D]­ en la evolución clínica de pacientes con malaria. Sin embargo, la prevalencia de la deficiencia de 25(OH)D en la población amazónica es poco conocida y existen pocos estudios en pacientes con malaria. Ante esto, el objetivo de este estudio fue evaluar los niveles séricos de 25(OH)D en pacientes con malaria y su relación con datos epidemiológicos, parasitológicos y pruebas de función hepática. Para ello, se realizó un estudio transversal analítico en el grupo de pacientes con malaria y en un grupo control en el municipio de Itaituba (PA), Brasil, de enero de 2018 a octubre de 2019. Se elaboró un protocolo para la evaluación de datos sociodemográficos, parasitológicos y de laboratorio, adoptando un nivel de significancia del 5%. La prevalencia de deficiencia de 25(OH)D se observó en pacientes con malaria (28,5%) y en el grupo control (24,6%), sin diferencia estadística; sin embargo, entre los residentes en la minería, los niveles séricos fueron estadísticamente inferiores en pacientes con malaria. Los niveles séricos de transaminasa glutámico pirúvica (TGP) mostraron una correlación inversa con los niveles de 25(OH)D. Las pruebas de función hepática fueron significativamente más altas en el grupo de malaria. De esta manera, se evidenció deficiencia de 25(OH)D en la población de Itaituba. Los cambios hepáticos debido a la infección plasmodial pueden haber contribuido a la correlación inversa observada entre los niveles de TGP y 25(OH)D.

First isolation of West Nile virus in Brazil

BACKGROUND Serological evidence of West Nile virus (WNV) infection has been reported in different regions of Brazil from equine and human hosts but the virus had never been isolated in the country. OBJECTIVES We sought to identify the viral etiology of equine encephalitis in Espírito Santo state. METHODS We performed viral culture in C6/36 cells, molecular detection of WNV genome, histopathology and immunohistochemistry from horse cerebral tissue. We also carried out sequencing, phylogenetic analysis and molecular clock. FINDINGS Histopathologic analysis from horse cerebral tissue showed injury related to encephalitis and WNV infection was confirmed by immunohistochemistry. The virus was detected by reverse transcription quantitative polymerase chain reaction (RT-qPCR) from brain tissue and subsequently isolated in C6/36 cells. WNV full-length genome was sequenced showing the isolated strain belongs to lineage 1a. The molecular clock indicated that Brazilian WNV strain share the same common ancestor that were circulating in US during 2002-2005. MAIN CONCLUSIONS Here we report the first isolation of WNV in Brazil from a horse with neurologic disease, which was clustered into lineage 1a with others US WNV strains isolated in beginning of 2000's decade.