Renato Pereira de Souza

Probable transfusion-transmitted Zika virus in Brazil

Zika virus (ZIKV) is an emerging arthropod‐borne flavivirus transmitted by Aedes mosquitoes. Recent commentaries regarding ZIKV routes of transmission describe a potential transmission by transfusion. Herein, we report a probable case of transfusion‐transmitted ZIKV infection through a platelet transfusion that was detected from postdonation information.

Identifying Rodent Hantavirus Reservoirs, Brazil

Bolomys lasiurus and Oligoryzomys nigripes are rodent reservoirs of Araraquara-like and Juquitiba-like hantaviruses, which cause HPS in Brazil.

Hantavirus Pulmonary Syndrome, Central Plateau, Southeastern, and Southern Brazil

This syndrome is an increasing health problem because of human encroachment into habitats of rodent reservoirs.

St. Louis encephalitis vírus: first isolation from a human in São Paulo state, Brasil

This paper reports the isolation of St. Louis encephalitis virus (SLEV) from a febrile human case suspected to be dengue, in Sao Pedro, Sao Paulo State. A MAC-ELISA done on the patients acute and convalescent sera was inconclusive and hemagglutination inhibition test detected IgG antibody for flaviviruses. An indirect immunofluorescent assay done on the C6/36 cell culture inoculated with the acute serum was positive for flaviviruses but negative when tested with dengue monoclonal antibodies. RNA extracted from the infected cell culture supernatant was amplified by RT-PCR in the presence of NS5 universal flavivirus primers and directly sequenced. Results of BLAST search indicated that this sequence shares 93% nucleotide similarity with the sequence of SLEV (strain-MSI.7), confirmed by RT-PCR performed with SLEV specific primers. Since SLEV was identified as the cause of human disease, it is necessary to improve surveillance in order to achieve early detection of this agent in the state of Sao Paulo and in Brazil. This finding is also an alert to health professionals about the need for more complete clinical and epidemiological investigations of febrile illnesses as in the reported case. SLEV infections can be unrecognized or confused with other ones caused by an arbovirus, such as dengue.

Detection of a new yellow fever virus lineage within the South American genotype I in Brazil.

Nucleotide sequences of two regions of the genomes of 11 yellow fever virus (YFV) samples isolated from monkeys or humans with symptomatic yellow fever (YF) in Brazil in 2000, 2004, and 2008 were determined with the objective of establishing the genotypes and studying the genetic variation. Results of the Bayesian phylogenetic analysis showed that sequences generated from strains from 2004 and 2008 formed a new subclade within the clade 1 of the South American genotype I. The new subgroup is here designated as 1E. Sequences of YFV strains recovered in 2000 belong to the subclade 1D, which comprises previously characterized YFV strains from Brazil. Molecular dating analyses suggested that the new subclade 1E started diversifying from 1D about 1975 and that the most recent 2004–2008 isolates arose about 1985. J. Med. Virol. 82:175–185, 2010.

Dengue Virus Type 4 Phylogenetics in Brazil 2011: Looking beyond the Veil

Dengue Fever and Dengue Hemorrhagic Fever are diseases affecting approximately 100 million people/year and are a major concern in developing countries. In the present study, the phylogenetic relationship of six strains of the first autochthonous cases of DENV-4 infection occurred in Sao Paulo State, Parana State and Rio Grande do Sul State, Brazil, 2011 were studied. Nucleotide sequences of the envelope gene were determined and compared with sequences representative of the genotypes I, II, III and Sylvatic for DEN4 retrieved from GenBank. We employed a Bayesian phylogenetic approach to reconstruct the phylogenetic relationships of Brazilian DENV-4 and we estimated evolutionary rates and dates of divergence for DENV-4 found in Brazil in 2011. All samples sequenced in this study were located in Genotype II. The studied strains are monophyletic and our data suggest that they have been evolving separately for at least 4 to 6 years. Our data suggest that the virus might have been present in the region for some time, without being noticed by Health Surveillance Services due to a low level of circulation and a higher prevalence of DENV-1 and DENV- 2.

Yellow fever outbreak affecting Alouatta populations in southern Brazil (Rio Grande do Sul State), 2008-2009.

The natural transmission cycle of Yellow Fever (YF) involves tree hole breeding mosquitoes and a wide array of nonhuman primates (NHP), including monkeys and apes. Some Neotropical monkeys (howler monkeys, genus Alouatta) develop fatal YF virus (YFV) infections similar to those reported in humans, even with minimum exposure to the infection. Epizootics in wild primates may be indicating YFV circulation, and the surveillance of such outbreaks in wildlife is an important tool to help prevent human infection. In 2001, surveillance activities successfully identified YF‐related death in a black‐and‐gold howler monkey (Alouatta caraya), Rio Grande do Sul State (RGS) in southern Brazil, and the YFV was isolated from a species of forest‐dwelling mosquito (Haemagogus leucocelaenus). These findings led the State Secretariat of Health to initiate a monitoring program for YF and other 18 arboviral infections in Alouatta monkeys. The monitoring program included monkey captures, reporting of monkey casualties by municipalities, and subsequent investigations. If monkey carcasses were found in forests, samples were collected in a standardized manner and this practice resulted in increased reporting of outbreaks. In October 2008, a single howler monkey in a northwestern RGS municipality was confirmed to have died from YF. From October 2008 to June 2009, 2,013 monkey deaths were reported (830 A. caraya and 1,183 A. guariba clamitans). Viruses isolation in blood, viscera, and/or immunohistochemistry led to the detection of YF in 204 of 297 (69%) (154 A. g. clamitans and 50 A. caraya) dead Alouatta monkeys tested. The number of municipalities with confirmed YFV circulation in howlers increased from 2 to 67 and 21 confirmed human cases occurred. This surveillance system was successful in identifying the largest YF outbreak affecting wild NHP ever recorded. Am. J. Primatol. 74:68–76, 2012.

Reemergence of yellow fever: detection of transmission in the State of São Paulo, Brazil, 2008

INTRODUCTION Following yellow fever virus (YFV) isolation in monkeys from the São José do Rio Preto region and two fatal human autochthonous cases from the Ribeirão Preto region, State of São Paulo, Brazil, two expeditions for entomological research and eco-epidemiological evaluation were conducted. METHODS A total of 577 samples from humans, 108 from monkeys and 3,049 mosquitoes were analyzed by one or more methods: virus isolation, ELISA-IgM, RT-PCR, histopathology and immunohistochemical. RESULTS Of the 577 human samples, 531 were tested by ELISA-IgM, with 3 positives, and 235 were inoculated into mice and 199 in cell culture, resulting in one virus isolation. One sample was positive by histopathology and immunohistochemical. Using RT-PCR, 25 samples were processed with 4 positive reactions. A total of 108 specimens of monkeys were examined, 108 were inoculated into mice and 45 in cell culture. Four virus strains were isolated from Alouatta caraya. A total of 931 mosquitoes were captured in Sao Jose do Rio Preto and 2,118 in Ribeirão Preto and separated into batches. A single isolation of YFV was derived from a batch of 9 mosquitoes Psorophora ferox, collected in Urupês, Ribeirão Preto region. A serological survey was conducted with 128 samples from the municipalities of São Carlos, Rincão and Ribeirão Preto and 10 samples from contacts of patients from Ribeirão Preto. All samples were negative by ELISA-IgM for YFV. CONCLUSIONS The results confirm the circulation of yellow fever, even though sporadic, in the Sao Paulo State and reinforce the importance of vaccination against yellow fever in areas considered at risk.

Arbovírus Ilheus em aves silvestres (Sporophila caerulescens e Molothrus bonariensis)

OBJETIVO: Relatar o isolamento do virus Ilheus no Estado de Sao Paulo e avaliar o seu impacto para a saude publica. METODOS: O isolamento de virus foi realizado em camundongos albinos Swiss, a partir de sangue de aves silvestres, capturadas com redes de espera tipo mist net, armadas no nivel do solo, no Parque Ecologico do Tiete, Sao Paulo. A identificacao das cepas isoladas foi feita pelos testes de inibicao da hemaglutinacao, fixacao de complemento e neutralizacao em camundongos. Amostras de plasma de aves e de mamiferos silvestres foram submetidas a pesquisa sorologica para deteccao de anticorpos inibidores de hemaglutinacao. RESULTADOS: Foram isoladas duas cepas do virus Ilheus em sangue de aves das especies Sporophila caerulescens e Molothrus bonariensis e detectados anticorpos em aves das especies Columbina talpacoti, Geopelia cuneata, Molothrus bonariensis e Sicalis flaveola, em saguis das especies Callithrix jacchus e Callithrix penicillata e no quati Nasua nasua. CONCLUSOES: O isolamento do virus Ilheus e a deteccao de anticorpos especificos em aves residentes, migratorias e de cativeiro, em saguis e quatis, comprovam a presenca desse agente no Parque Ecologico do Tiete. O comportamento migratorio de aves silvestres pode determinar a introducao do virus em outras regioes. Considerando-se a patogenicidade para o homem e a confirmacao da circulacao desse agente viral em area urbana, frequentada para atividade de lazer e de educacao, o risco de ocorrencia de infeccao na populacao humana nao pode ser descartado.

Experimental Zika virus infection induces spinal cord injury and encephalitis in newborn Swiss mice.

A widespread epidemic of Zika virus (ZIKV) infection was reported in 2015 in South and Central America, with neurological symptons including meningoencephalitis and Guillain-Barré syndrome in adults, besides an apparent increased incidence of microcephaly in infants born to infected mothers. It is becoming a necessity to have a trustworthy animal model to better understand ZIKV infection. In this study we used newborn white Swiss mice as a model to investigate the ZIKV strain recently isolated in Brazil. ZIKV was inoculated via intracerebral and subcutaneous routes and analysed through gross histopathology and immunohistochemistry. Here we demonstrated first that the intracerebral group (ICG) displayed severe cerebral lesions, with neuronal death, presence of apoptotic bodies, white matter degeneration and neutrophil perivascular cuffing. In the subcutaneous group (SCG), we observed moderate cerebral lesions, morphologically similar to that found in ICG and additional myelopathy, with architectural loss, marked by neuronal death and apoptotic bodies. Interestingly, we found an intense astrogliosis in brain of both groups, with increased immunoexpression of GFAP (glial fibrillary acidic protein) and presence of hypertrophic astrocytes. The spinal cord of subcutaneous group (SCG) exhibited reduction of astrocytes, but those positive for GFAP were hypertrophic and presented prolonged cellular processes. Finally significant lesions in the central nervous system (CNS) were present in newborn mice inoculated by both routes, but SCG method led to an important neurological manifestations (including myelopathy), during a longer period of time and appears for us to be a better model for ZIKV infection.