Dozens of orthobunyaviruses have been isolated in Brazil, and at least thirteen have been associated with human disease. The Oropouche virus has received most attention for having caused explosive epidemics with hundreds of thousands of cases in the north region between the 1960sand the 1980s, and since then has been sporadically detected elsewhere in the country. Despite their importance, little is known about their enzootic cycles of transmission, amplifying hosts and vectors, and biotic and abiotic factors involved in spillover events to humans. This overview aims to combine available data of neglected orthobunyaviruses of several serogroups, namely, Anopheles A, Anopheles B, Bunyamwera, California, Capim, Gamboa, Group C, Guama, Simbu and Turlock, in order to evaluate the current knowledge and identify research gaps in their natural transmission cycles in Brazil to ultimately point to the future direction in which orthobunyavirus research should be guided.

https://www.mdpi.com/1999-4915/14/5/987/pdf?version=1652928714

An Overview of Neglected Orthobunyaviruses in Brazil

During these past years, several studies have provided serological evidence regarding the circulation of West Nile virus (WNV) in Brazil. Despite some reports, much is still unknown regarding the genomic diversity and transmission dynamics of this virus in the country. Recently, genomic monitoring activities in horses revealed the circulation of WNV in several Brazilian regions. These findings on the paucity of genomic data reinforce the need for prompt investigation of WNV infection in horses, which may precede human cases of encephalitis in Brazil. Thus, in this study, we retrospectively screened 54 suspicious WNV samples collected between 2017 and 2020 from the spinal cord and brain of horses with encephalitis and generated three new WNV genomes from the Ceará and Bahia states, located in the northeastern region of Brazil. The Bayesian reconstruction revealed that at least two independent introduction events occurred in Brazil. The first introduction event appears to be likely related to the North American outbreak, and was estimated to have occurred in March 2013.The second introduction event appears to have occurred in September 2017 and appears to be likely related to the South American outbreak. Together, our results reinforce the importance of increasing the priority of WNV genomic monitoring in equines with encephalitis in order to track the dispersion of this emerging pathogen through the country.

/pdf/retrospective-investigation-in-horses-with-encephalitis-r96j1yvr.pdf

Retrospective Investigation in Horses with Encephalitis Reveals Unnoticed Circulation of West Nile Virus in Brazil

West Nile virus is a flavivirus transmitted by mosquitoes, mainly of the genus Culex. In Brazil, serological studies have already indicated the circulation of the virus since 2003, with the first human case detected in 2014. The objective of the present paper is to report the first isolation of WNV in a Culex (Melanoconion) mosquito. Arthropods were collected by protected human attraction and CDC light bait, and taxonomically identified and analyzed by viral isolation, complement fixation and genomic sequencing tests. WNV was isolated from samples of Culex (Melanoconion) mosquitoes, and the sequencing analysis demonstrated that the isolated strain belonged to lineage 1a. The finding of the present study presents the first evidence of the isolation and genome sequencing of WNV in arthropods in Brazil.

https://www.mdpi.com/2414-6366/8/4/237/pdf?version=1681989724

First Isolation and Genome Sequence Analysis of West Nile Virus in Mosquitoes in Brazil

Many human arboviruses are also pathogenic for horses, and some of these have emerged recently. A descriptive cross-sectional observational study was conducted to assess the prevalence of West Nile virus (WNV) and other arboviruses among 77 horses on the rural properties of the Espirito Santo state, Brazil. Serum samples were screened for arbovirus-reactive antibodies using the hemagglutination inhibition technique and subsequently a plaque reduction neutralization test for the confirmation of exposure from sera was used to detect heterotypic immune reactions. Overall, the total antibodies against at least one arbovirus of Alphavirus, Flavivirus, and Orthobunyavirus genera were detected in 39 (50.6%) animals. The antibodies to Phlebovirus were not detected in any sample. When the 24 WNV hemagglutination inhibition (HI)-positive samples were tested by the plaque-reduction neutralization test 90%, 9 (32.1%) were positive for WNV antibodies and 14 (50%) for Saint Louis encephalitis virus. Our findings indicate that the region provides ideal conditions for the emergence of arboviruses, reinforcing the need for further surveillance of mosquito-transmitted diseases in domestic animals.

Serological Evidence of Arboviruses in Horses During West Nile Fever Monitoring Surveillance in Southeastern Brazil

Infections with arboviruses are reported worldwide. Saint Louis encephalitis (SLEV) and West Nile viruses (WNV) are closely related flaviviruses affecting humans and animals. SLEV has been sporadically detected in humans, and corresponding antibodies have been frequently detected in horses throughout Brazil. WNV was first reported in western Brazil over a decade ago, has been associated with neurological disorders in humans and equines and its prevalence is increasing nationwide. Herein, we investigated by molecular and serological methods the presence or evidence of SLEV and WNV in equines from Rio de Janeiro. A total of 435 serum samples were collected from healthy horses and tested for specific neutralizing antibodies by plaque reduction neutralization test (PRNT90). Additionally, serum and central nervous system samples from 72 horses, including horses with neurological disorders resulting in a fatal outcome or horses which had contact with them, were tested by real-time reverse transcription–polymerase chain reaction (RT-qPCR) for both viruses. Adopting the criterion of four-fold antibody titer difference, 89 (20.4%) horses presented neutralizing antibodies for SLEV and five (1.1%) for WNV. No evidence of SLEV and WNV infection was detected by RT-qPCR and, thus, such infection could not be confirmed in the additional samples. Our findings indicate that horses from Rio de Janeiro were exposed to both SLEV and WNV, contributing to the current knowledge on the distribution of these viruses flaviviruses in Brazil.

/pdf/serological-evidence-of-exposure-to-saint-louis-encephalitis-iw106m7f.pdf

Serological Evidence of Exposure to Saint Louis Encephalitis and West Nile Viruses in Horses of Rio de Janeiro, Brazil

In June 2019, a horse with neurological disorder was diagnosed with West Nile virus (WNV) in Boa Viagem, a municipality in the state of Ceara, northeast Brazil. A multi-institutional task force coordinated by the Brazilian Ministry of Health was deployed to the area for case investigation. A total of 513 biological samples from 78 humans, 157 domestic animals and 278 free-ranging wild birds, as well as 853 adult mosquitoes of 22 species were tested for WNV by highly specific serological and/or molecular tests. No active circulation of WNV was detected in vertebrates or mosquitoes by molecular methods. Previous exposure to WNV was confirmed by seroconversion in domestic birds and by the detection of specific neutralizing antibodies in 44% (11/25) of equids, 20.9% (14/67) of domestic birds, 4.7% (13/278) of free-ranging wild birds, 2.6% (2/78) of humans, and 1.5% (1/65) of small ruminants. Results indicate that not only equines but also humans and different species of domestic animals and wild birds were locally exposed to WNV. The detection of neutralizing antibodies for WNV in free-ranging individuals of abundant passerine species suggests that birds commonly found in the region may have been involved as amplifying hosts in local transmission cycles of WNV.

Francisco das Chagas Almeida

Papers

West Nile Virus in the State of Ceará, Northeast Brazil.