Investigation of Yellow Fever Virus at the Human-Animal Interface after a Zika Virus Outbreak in Midwest Brazil.

Following the first report of zika virus in March 2015, Brazil experienced its largest sylvatic yellow fever outbreak between 2016 and 2019. This study aimed to investigate the circulation of yellow fever virus (YFV) in non-human primates (NHPs) and mosquitoes collected in urban parks and other metropolitan areas of midwest Brazil between 2017 and 2018. Whole blood samples from 80 NHPs, including 48 black-tailed marmosets (Mico melanurus) and 2332 mosquitoes from six different genera, were collected in the states of Mato Grosso (MT) and Mato Grosso do Sul (MS) and then tested for YFV by RT-qPCR. Additionally, 23 plasma samples of NHPs were tested for neutralizing antibodies for YFV by a plaque reduction neutralization test (PRNT). No YFV RNA or neutralizing antibodies for YFV were detected in NHPs and mosquitoes from MT and MS. The continuous monitoring of YFV circulation in different species of NHPs and vectors in urban areas is instrumental to quickly assess potentially unknown maintenance cycles of yellow fever at the human-animal interface in Brazil.

Avanços no entendimento da patogenia da covid-19: uma revisão / Advances in understanding covid-19 pathogenesis: a review / Avances en la comprensión de la patogenia de la covid-19: una revisión

Introdução: O SARS-CoV-2, causador da pandemia por Doença por Coronavirus 2019 (COVID-19) gerou desafios à saúde pública, principalmente pelo conhecimento escasso de sua patogênese, e de estratégias terapêuticas e preventivas eficazes. Diversas lacunas de conhecimento sobre a doença envolvem a contribuição de fatores de risco, as doenças concomitantes, os fatores genéticos e imunogenéticos, no direcionamento da resposta imune, bem como a hiperinflamação, a imunidade antiviral e os alelos dos antígenos leucocitários humanos (HLAs) dos pacientes, que estão relacionados ao desfecho da doença. Objetivo: Esta revisão integrativa objetivou aprofundar os conhecimentos sobre os mecanismos de imunidade relacionada aos fatores de risco, da hiperinflamação e da tempestade de citocinas decorrente da infecção por SARS-CoV-2, bem como os avanços de associação dos HLAs nos agravos da doença. Metodologia: A revisão integrativa foi realizada utilizando os descritores nas bases de dados PubMed, SCIELO, CINAHL, SCOPUS, Web of Science, MedNar, Portal periódicos CAPES e Open Journal System, sem limites de janela cronológica. Resultados: Os efeitos da pandemia e os esforços na busca do conhecimento sobre a patogênese da COVID-19 resultaram em avanço no combate da doença. Conseguiu-se relacionar fatores de risco como obesidade, hipertensão, diabetes, doenças renais crônicas, idade, gênero, e outras condições predisponentes ao agravo da doença. Sob o aspecto da patogênese, também houveram progressos no entendimento dos mecanismos celulares e humorais em resposta à doença, bem como conseguiu vincular a resposta da hiperinflamação e o perfil dos HLAs dos pacientes à evolução da doença ao óbito ou à convalescência. Conclusão: Os esforços científicos conjuntos e os avanços na compreensão dos mecanismos da doença conseguiram estabelecer estratégias de combate a COVID-19, resultando no fim da pandemia, porém ainda há avanços que devem ser alcançados para o combate das sequelas dos pacientes convalescentes e para minimização da COVID longa e seus prejuízos.

Introduction: SARS-CoV-2, which causes the Coronavirus Disease 2019 (COVID-19) pandemic, has generated challenges to public health, mainly due to the limited knowledge of its pathogenesis and effective therapeutic and preventive strategies. Several gaps in knowledge about the disease involve the contribution of risk factors, concomitant diseases, genetic and immunogenetic factors, in directing the immune response, as well as hyperinflammation, antiviral immunity and human leukocyte antigen (HLAs) alleles of patients, which are related to the outcome of the disease. Aim: This integrative review aimed to deepen knowledge about the mechanisms of immunity related to risk factors, hyperinflammation and the cytokine storm resulting from SARS-CoV-2 infection, as well as advances in the association of HLAs in diseases. Methodology: The integrative review was carried out using the descriptors in the databases PubMed, SCIELO, CINAHL, SCOPUS, Web of Science, MedNar, CAPES periodical portal and Open Journal System, without chronological window limits. Results: The effects of the pandemic and efforts to seek knowledge about the pathogenesis of COVID-19 resulted in progress in combating the disease. It was possible to relate risk factors such as obesity, hypertension, diabetes, chronic kidney disease, age, gender, and other conditions predisposing to the worsening of the disease. From the aspect of pathogenesis, progress has also been made in understanding the cellular and humoral mechanisms in response to the disease, as well as linking the hyperinflammation response and the patients' HLA profile to the progression of the disease to death or convalescence. Conclusion: Joint scientific efforts and advances in understanding the mechanisms of the disease managed to establish strategies to combat COVID-19, resulting in the end of the pandemic, but there are still advances that must be achieved to combat the sequelae of convalescent patients and to minimize of long COVID and its losses.

Introducción: El SARS-CoV-2, causante de la pandemia de la Enfermedad del Coronavirus 2019 (COVID-19), ha generado desafíos a la salud pública, principalmente por el limitado conocimiento de su patogénesis y estrategias terapéuticas y preventivas efectivas. Varios vacíos en el conocimiento sobre la enfermedad involucran la contribución de factores de riesgo, enfermedades concomitantes, factores genéticos e inmunogenéticos, en la dirección de la respuesta inmune, así como la hiperinflamación, la inmunidad antiviral y los alelos del antígeno leucocitario humano (HLA) de los pacientes, que están relacionados con el resultado de la enfermedad. Objetivo: Esta revisión integradora tuvo como objetivo profundizar el conocimiento sobre los mecanismos de inmunidad relacionados con los factores de riesgo, la hiperinflamación y la tormenta de citocinas resultante de la infección por SARS-CoV-2, así como los avances en la asociación de los HLA en las complicaciones de la enfermedad. Metodología: La revisión integradora se realizó utilizando los descriptores de las bases de datos PubMed, SCIELO, CINAHL, SCOPUS, Web of Science, MedNar, portal periódico CAPES y Open Journal System, sin límites de ventana cronológica. Resultados: Los efectos de la pandemia y los esfuerzos por buscar conocimiento sobre la patogénesis de la COVID-19 resultaron en avances en el combate de la enfermedad. Se logró relacionar factores de riesgo como obesidad, hipertensión, diabetes, enfermedad renal crónica, edad, sexo y otras condiciones que predisponen al agravamiento de la enfermedad. Desde el punto de vista de la patogénesis, también se ha avanzado en la comprensión de los mecanismos celulares y humorales de respuesta a la enfermedad, así como en la vinculación de la respuesta de hiperinflamación y el perfil HLA de los pacientes con la progresión de la enfermedad hasta la muerte o la convalecencia. Conclusión: Los esfuerzos científicos conjuntos y los avances en la comprensión de los mecanismos de la enfermedad lograron establecer estrategias para combatir el COVID-19, teniendo como resultado el fin de la pandemia, pero aún quedan avances que se deben lograr para combatir las secuelas de los pacientes convalecientes y para Minimizar el COVID prolongado y sus pérdidas.

Inflammation, fibrosis and E1 glycoprotein persistence in joint tissue of patients with post-Chikungunya chronic articular disease.

INTRODUCTION: Chikungunya chronic joint disease causes debilitating arthralgia, significantly impacting the quality of life of affected individuals. METHODS: In this study, patients underwent clinical follow-ups, joint biopsies, and pre-biopsy and 24 months post-biopsy serum dosage of cytokines. RESULTS: All participants were female and had pain in 12 joints on average, with 41.17% exhibiting moderate disease activity. Histopathological analysis revealed collagen deposition. Indirect immunofluorescence detected the CHIKV glycoprotein E1 antigen, and an increase in cytokines. CONCLUSIONS: Persistent inflammation and ineffective antiviral immune responses leading to antigen persistence may contribute to chronic CHIKV arthritis.

Dengue virus serotype 2 genotype III evolution during the 2019 outbreak in Mato Grosso, Midwestern Brazil.

DENV-2 was the main responsible for a 70% increase in dengue incidence in Brazil during 2019. That year, our metagenomic study by Illumina NextSeq on serum samples from acute febrile patients (n = 92) with suspected arbovirus infection, sampled in 22 cities of the state of Mato Grosso (MT), in the middle west of Brazil, revealed eight complete genomes and two near-complete sequences of DENV-2 genotype III, one Human parvovirus B19 genotype I (5,391 nt) and one Coxsackievirus A6 lineage D (4,514 nt). These DENV-2 sequences share the aminoacidic identities of BR4 lineage on E protein domains I, II and III, and were included in a clade with sequences of the same lineage circulating in the southeast of Brazil in the same year. Nevertheless, 11/34 non-synonymous mutations are unique to three strains inthis study, distributed in the E (n = 6), NS3 (n = 2) and NS5 (n = 3) proteins. Other 14 aa changes on C (n = 1), E (n = 3), NS1 (n = 2), NS2A (n = 1) and NS5 (n = 7) were first reported in a genotype III lineage, having been already reported only in other DENV-2 genotypes. All 10 sequences have mutations in the NS5 protein (14 different aa changes). Nine E protein aa changes found in two sequences, six of which are unique, are in the ectodomain; where the E:M272T change is on the hinge of the E protein at domain II, in a region critical for the anchoring to the host cell receptor. The NS5:G81R mutation, in the methyltransferase domain, was found in one strain of this study. Altogether, these data points to an important evolution of DENV-2 genotype III lineage BR4 during this outbreak in 2019 in MT. Genomic surveillance is essential to detect virus etiology and evolution, possibly related to immune evasion and viral fitness changes leading to future novel outbreaks.

Diagnostic and vaccine potential of Zika virus envelope protein (E) derivates produced in bacterial and insect cells.

Introduction: In the present study we evaluated the features of different recombinant forms of Zika virus (ZIKV) proteins produced in either bacterial (Eschericha coli) or insect cells (Drosophila melanogaster). The ZIKV-envelope glycoprotein (EZIKV) is responsible for virus entry into host cells, is the main target of neutralizing antibodies and has been used as a target antigen either for serological tests or for the development of subunit vaccines. The EZIKV is composed of three structural and functional domains (EDI, EDII, and EDIII), which share extensive sequence conservation with the corresponding counterparts expressed by other flaviviruses, particularly the different dengue virus (DENV) subtypes. Methods: In this study, we carried out a systematic comparison of the antigenicity and immunogenicity of recombinant EZIKV, EDI/IIZIKV and EDIIIZIKV produced in E. coli BL21 and Drosophila S2 cells. For the antigenicity analysis we collected 88 serum samples from ZIKV-infected participants and 57 serum samples from DENV-infected. For immunogenicity, C57BL/6 mice were immunized with two doses of EZIKV, EDI/IIZIKV and EDIIIZIKV produced in E. coli BL21 and Drosophila S2 cells to evaluate humoral and cellular immune response. In addition, AG129 mice were immunized with EZIKV and then challenge with ZIKV. Results: Testing of samples collected from ZIKV-infected and DENV-infected participants demonstrated that the EZIKV and EDIIIZIKV produced in BL21 cells presented better sensitivity and specificity compared to proteins produced in S2 cells. In vivo analyses were carried out with C57BL/6 mice and the results indicated that, despite similar immunogenicity, antigens produced in S2 cells, particularly EZIKV and EDIIIZIKV, induced higher ZIKV-neutralizing antibody levels in vaccinated mice. In addition, immunization with EZIKV expressed in S2 cells delayed the onset of symptoms and increased survival rates in immunocompromised mice. All recombinant antigens, either produced in bacteria or insect cells, induced antigen-specific CD4+ and CD8+ T cell responses. Conclusion: In conclusion, the present study highlights the differences in antigenicity and immunogenicity of recombinant ZIKV antigens produced in two heterologous protein expression systems.

Inflammation, fibrosis and E1 glycoprotein persistence in joint tissue of patients with post-Chikungunya chronic articular disease

ABSTRACT Introduction: Chikungunya chronic joint disease causes debilitating arthralgia, significantly impacting the quality of life of affected individuals. Methods: In this study, patients underwent clinical follow-ups, joint biopsies, and pre-biopsy and 24 months post-biopsy serum dosage of cytokines. Results: All participants were female and had pain in 12 joints on average, with 41.17% exhibiting moderate disease activity. Histopathological analysis revealed collagen deposition. Indirect immunofluorescence detected the CHIKV glycoprotein E1 antigen, and an increase in cytokines. Conclusions: Persistent inflammation and ineffective antiviral immune responses leading to antigen persistence may contribute to chronic CHIKV arthritis.

Regional mutations in CHIKV-ECSA genomes and detection of other viruses in the serum of acute febrile patients by a metagenomic approach in Mato Grosso, Central-Western Brazil, 2018.

Mato Grosso (MT) State is part of central western Brazil and has a tropical permissive environment that favors arbovirus outbreaks. A metagenomic approach was used to identify viral genomes in seven pools of serum from patients (n=65) with acute febrile disease. Seven chikungunya virus (CHIKV) genomes were determined, showing four amino acid changes found only in CHIKV genomes obtained in MT since 2018: nsP2:T31I, nsP3: A388V, E3:T201I and E3:H57R, in addition to other mutations in E1, nsP2 and nsP4. Six parvovirus B19 (B19V) genotype I genomes (4771-5131 nt) showed four aa alterations (NS1:N473D, R579Q; VP1:I716T; and 11 kDa:V44A) compared to most similar B19V from the USA. Coinfection between CHIKV and B19V was evidenced in 22/65 (33.8%) patients by RT‒PCR and PCR, respectively. Other viruses found in these pools include human pegivirus C, torque teno virus 3, an unclassified TTV and torque teno mini virus. Metagenomics represents a useful approach to detect viruses in the serum of acute febrile patients suspected of arbovirus disease.

Persistent SARS-CoV-2 antigen presence in multiple organs of a naturally infected cat from Brazil.

Background: Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is the etiological agent of the disease coronavirus 2019 (COVID-19) in humans. SARS-CoV-2 has been identified in cats with or without clinical signs. Case presentation: We describe the pathological and molecular findings in a six-month-old asymptomatic cat with SARS-CoV-2 infection from Brazil, belonging to a human family with COVID-19 cases. The pool of nasopharynx and oropharynx swabs at day zero tested positive by RT-qPCR for SARS-CoV-2. No amplification resulted from molecular testing performed on days 7 and 14. The cat was hit by a car and died 43 days after the molecular diagnosis. Immunohistochemistry at post-mortem examination demonstrated nucleocapsid protein in samples from the lungs, kidneys, nasal conchae, trachea, intestine, brain and spleen. Conclusion: The present study has highlighted the possibility that viral antigens can be detected by immunohistochemistry in multiple organs six weeks after infection, although the same tissues tested negative by RT-PCR.

Clinical and genomic data of sars-cov-2 detected in maternal-fetal interface during the first wave of infection in Brazil.

Brazil has the highest SARS-CoV-2 case-fatality rate in pregnant women in the Americas. In this study, clinical and virological findings of five mildly symptomatic pregnant women and their infected fetuses/newborns treated at a referral hospital for COVID19-pregnant women in Midwestern Brazil are reported. Mother and fetal samples were tested by RT-qPCR, ECLIA and Illumina MiSeq sequencing. From the five cases, one resulted in spontaneous abortion, one was stillborn, two were preterm births and one full-term birth. Maternal and fetal placenta, newborn and stillborn secretions were SARS-CoV-2+; one neonate developed ground-glass opacities in his lungs. One neonate's umbilical cord was IgG+ and all were IgM negative upon hospital discharge. Genomes recovered from two placentas belong to the B.1.1.28 and B.1.1.33 lineages and present nonsynonymous mutations associated with virus fitness and infectivity; other not frequently reported mutations (B.1.1.33: NSP3 V2090G, M A2S and ORF3ab S253P and Y264N; B.1.1.28: NSP3 E995D, NSP12 R240K, NSP14H1897Y and in ORF7b V21F) were found in proteins involved in viral replication, viral induction of apoptosis, viral interference on interferon and on NF-Κß pathways. Phylogeny indicates the south of Brazil as the possible origin of these lineages circulating in Mato Grosso State. These findings contribute to describe SARS-CoV-2 infection and outcomes in pregnant women and their fetuses, at any stage of gestation and even in mild symptomatic cases.

A Novel Host of an Emerging Disease: SARS-CoV-2 Infection in a Giant Anteater (Myrmecophaga tridactyla) Kept Under Clinical Care in Brazil.

A young male free-ranging giant anteater (Myrmecophaga tridactyla) was found with paralysis of pelvic limbs on a highway and kept under human care. Radiographs confirmed multiple incomplete fractures in the thoracolumbar vertebrae. Due to the poor prognosis, euthanasia was chosen. The infection was established by viral SARS-CoV-2 RNA detection in the rectal swab, spleen and kidney samples. Immunohistochemistry detected the viral nucleocapsid protein in sections of the lungs, liver, spleen, lymph nodes, and large intestine sections, and spike protein antigen in the lung tissue. Pilosa order species should be included as potential hosts of natural infection of SARS-CoV-2.

Neurological infection by chikungunya and a triple Arbovirus co-infection in Mato Grosso, Central Western Brazil during 2019.

BACKGROUND: Neurological viral infection is frequently associated to enterovirus, herpesvirus and arboviruses. These infections may cause severe clinical outcomes, long lasting sequelae or death. Few studies have addressed viral neurological infections etiology in Brazil. OBJECTIVES: Identification of viruses in the cerebral spinal fluid (CSF) of human neurological infections suspected of viral etiology during January and May 2019 in Midwestern Brazil. MATERIALS AND METHODS: Clinical, laboratory and epidemiological information was gathered from medical records. In addition, an aliquot of the sampled CSF was subjected to viral RNA/DNA extraction, randomic dscDNA amplification by PCR, DNA purification and Ilumina HiSeq 2500 sequencing. RESULTS: Six viral genomes belonging to Chikungunya virus (CHIKV) East-Central-South African (ECSA) genotype (10.834-11.804 nt in length) confirmed lately by RT-PCR for CHIKV envelope were present in all six liquor samples. These genomes present two mutations, nsP2:T31I and nsP3:A388V, shared with other Mato Grosso State strains from 2019, not present in sequences of the virus from previous years obtained in the State. One case was a triple co-infection also confirmed through RT-PCR, with Dengue virus serotype 4 genotype II (NS5; 874 nt) and Oropouche virus genotype IA (segment S; 302 nt). CSF was clear and colorless (5/6 patients), with >10% of lymphomononuclear cells (6/6), 1-99 erythrocytes/mm3 (5/6), glucose levels >50 mg/dl (4/5) e > 10 mg/dl of proteins (4/4). One patient evolved to death, and another, a newborn, presented sequelae after recovery. CONCLUSIONS: Despite herpesviruses and enteroviruses are frequent etiologies of neurological infections, the casuistic here reported was associated to arboviruses already known to be responsible for acute febrile illness outbreaks in the state of Mato Grosso, Midwestern Brazil.

Chikungunya, Zika, Mayaro, and Equine Encephalitis virus detection in adult Culicinae from South Central Mato Grosso, Brazil, during the rainy season of 2018.

INTRODUCTION: Several arboviruses causing human disease have been reported in Brazil. In nature, arboviruses maintain a lifecycle involving vertebrates and vectors, which may contribute for periodical reemergence of those of public health concern in tropical regions, as Mato Grosso State (MT). In this study, we searched for arboviruses in mosquito body pools sampled during the rainy season of 2018 in 21 bird watching points of Cuiabá and Varzea Grande, South Central MT. METHODS: In total, 2873 (57%) males and 2167 (43%) females belonging to six urban and sylvatic mosquito genera allocated to 398 pools were subjected to RNA extraction and RT-PCR for arboviruses. Positive pools were subjected to virus isolation in C6/36 cells. RESULTS: A total of 102/398 pools, 66/233 (29.6%) of females, and 36/165 (21.8%) of males, mostly sampled in May (31/102), were positive for arboviruses. Chikungunya virus (CHIKV) was distributed in 19 points, Zika virus (ZIKV) was found in 14 points, Mayaro virus (MAYV) in 10 points, and East Equine encephalitis virus (EEEV) in three points. Culex quinquefasciatus pools (39/89 of females and 24/99 of males) were positive for CHIKV, ZIKV, and MAYV; Aedes (Stg) aegypti pools (11/46 of females and 12/33 of males) for CHIKV, ZIKV, MAYV, and EEEV; Aedes albopictus female pools (8/29) for CHIKV, ZIKV, and EEEV; and Psorophora albigenu (2/12) and Psorophora ferox female pools (4/16) for CHIKV. CONCLUSIONS: Arbovirus molecular detection in mosquito populations varies considerable between geographical regions and epidemics, influenced by genetic characteristics and microbiome interference on virus replication. Although infected females are responsible for the transmission to vertebrates during bloodfeeding, male infection by CHIKV, ZIKV, and MAYV resultant from vertical route could lead to interepidemic maintenance of these arboviruses in their natural reservoirs.

Persistent SARS-CoV-2 antigen presence in multiple organs of a naturally infected cat from Brazil

Background: Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is the etiological agent of the disease coronavirus 2019 (COVID-19) in humans. SARS-CoV-2 has been identified in cats with or without clinical signs. Case presentation: We describe the pathological and molecular findings in a six-month-old asymptomatic cat with SARS-CoV-2 infection from Brazil, belonging to a human family with COVID-19 cases. The pool of nasopharynx and oropharynx swabs at day zero tested positive by RT-qPCR for SARS-CoV-2. No amplification resulted from molecular testing performed on days 7 and 14. The cat was hit by a car and died 43 days after the molecular diagnosis. Immunohistochemistry at post-mortem examination demonstrated nucleocapsid protein in samples from the lungs, kidneys, nasal conchae, trachea, intestine, brain and spleen. Conclusion: The present study has highlighted the possibility that viral antigens can be detected by immunohistochemistry in multiple organs six weeks after infection, although the same tissues tested negative by RT-PCR.(AU)

Persistent SARS-CoV-2 antigen presence in multiple organs of a naturally infected cat from Brazil

Abstract Background: Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is the etiological agent of the disease coronavirus 2019 (COVID-19) in humans. SARS-CoV-2 has been identified in cats with or without clinical signs. Case presentation: We describe the pathological and molecular findings in a six-month-old asymptomatic cat with SARS-CoV-2 infection from Brazil, belonging to a human family with COVID-19 cases. The pool of nasopharynx and oropharynx swabs at day zero tested positive by RT-qPCR for SARS-CoV-2. No amplification resulted from molecular testing performed on days 7 and 14. The cat was hit by a car and died 43 days after the molecular diagnosis. Immunohistochemistry at post-mortem examination demonstrated nucleocapsid protein in samples from the lungs, kidneys, nasal conchae, trachea, intestine, brain and spleen. Conclusion: The present study has highlighted the possibility that viral antigens can be detected by immunohistochemistry in multiple organs six weeks after infection, although the same tissues tested negative by RT-PCR.

Sialovirome of Brazilian tropical anophelines.

Anophelinae is a widely dispersed Culicidae subfamily that may carry a unique virome. Here we herein report the set of viruses found in 323 salivary glands of 16 anopheline species sampled at Upper Pantanal, Chapada dos Guimarães National Park and Coxipó river basin, South Central Mato Grosso, Brazil, pooled (n = 11) and subjected to high throughput sequencing. Metagenomics revealed the presence of nine viral sequences belonging to novel viruses from seven viral families: Purunga is a putative novel orbivirus sharing 74% and 65% aa identity, respectively, with the VP1 and VP3 segments of Changuinola serogroup, Jaracatiá flavivirus shares 60% amino-acid (aa) identity with Aedes flavivirus. Coxipó dielmovirus and Chapada dielmovirus shared 51% and 39% aa identity with Merida virus. Coloiado-orthomyxo like virus is 57.1-64.8% identical at aa level to Aedes albonnulatus orthomyxo-like virus. Mujica picorna-like virus shares 49% aa identity with Flen picorna-like virus and Chiquitos virus is 50% similar to Ista virus, both from Picornavirales order. Cerrado partiti-like-virus shares 75-86% aa identity with Atrato partiti-like virus 2. We also found the S and L segments of Anopheles triannulatus orthophasmavirus (92% identity) in Anopheles lutzi from Chapada dos Guimarães. The identification of these putative novel viruses underscore the wide dispersion of viruses in culicid hosts contributing to extensions on mosquito virome descriptions.

Insect-specific viruses and arboviruses in adult male culicids from Midwestern Brazil.

Viruses were identified from male anthropophilic mosquitoes from Mato Grosso (MT) State, Midwest Brazil from February 2017 to January 2018. Mosquitoes tested included Aedes (Stegomyia) aegypti (1139 males; 84 pools), Culex quinquefasciatus (9426 males; 179 pools), Culex sp. (3 males; 3 pools) and Psorophora albigenu (1 male; 1 pool) collected from four cities of MT. Pools were subjected to viral RNA extraction followed by RT-PCRs specific for ten flaviviruses, five alphaviruses and Simbu serogroup of orthobunyaviruses. Positive pools were passaged three times in VERO cells (alphavirus and orthobunyavirus) or C6/36 cells (flavivirus), with isolates confirmed through RT-PCR and nucleotide sequencing. We detected pools positive for Ilhéus (1 pool), dengue serotype 4 (1), Mayaro (12), equine encephalitis virus (1) yellow fever (1), Oropouche (2), Zika (4) and chikungunya (12) viruses. High throughput sequencing of arbovirus positive pools identified 35 insect-specific viruses (ISVs) from the families Circoviridae (2), Parvoviridae (2), Totiviridae (1), Flaviviridae (1), Iflaviridae (2), Mesoniviridae (4), Nodaviridae (2), Luteoviridae (1), Phasmaviridae (1) Phenuiviridae (2), Rhabdoviridae (2), Orthomyxoviridae (1), Xinmoviridae (1), and unclassified Bunyavirales (1), unclassified Picornavirales (3), unclassified Riboviria (4) and taxon Negevirus (5). From these, five novel viruses were tentatively named Mojica circovirus, Kuia iflavirus, Muxirum negevirus, Lambada picorna-like virus and Tacuru picorna-like virus. Our findings underscore the diversity and wide geographical distribution of ISVs and arboviruses infecting male culicids.

30th Brazilian Society for Virology 2019 Annual Meeting-Cuiabá, Mato Grosso, Brazil.

The 30th meeting of the Brazilian Society for Virology (SBV) was held, for the first time in its 30 years of existence, in Cuiabá, the capital of Mato Grosso State, Central Western Brazil, a tropical region between the three richest biomes in the world: Amazon Florest, Cerrado and Pantanal. In recent years, the field of virology has been built in the State. The aim of this report is to support participants and virologists to receive the most up-to-date information about the meeting, which occurred from 16 to 19 October 2019. National and international speakers gave SBV the opportunity to learn about their experience on their virology fields, sharing recent scientific findings, compiling conferences, round table presentations and work presentations in oral and poster sessions. The meeting held over 300 attendants, who were also involved on oral and poster presentations, showing a great variety of recent unpublished studies on environmental, basic, animal, human, plant and invertebrate virology. In addition, SBV offered the Helio Gelli Pereira award for the best research studies in each field presented during the meeting. The 30th meeting of SBV was very productive and has also encouraged scientific partnership and collaboration among virologists worldwide.

Influence of climatic variables on the Aedes aegypti and Culex quinquefasciatus populations in Mato Grosso, Brazil.

INTRODUCTION: Aedes aegypti and Culex quinquefasciatus are vector species responsible for the transmission of important arboviruses. METHODS: Adult mosquitoes were collected in the urban areas of four municipalities in Mato Grosso within 1 year. RESULTS: A total of 19,110 mosquitoes were collected. Among them, 16,578 (86,8%) were C. quinquefasciatus (44% female and 56% male); 2,483 (13%), A. (Stegomyia) aegypti (54% female and 46% male); and 49 (0,30%), from the genus Psorophora, Anopheles, Coquilettidia, and Sabethes. A significant correlation was observed between the number of mosquitoes from all species and dew point (female mosquitoes, p = 0.001; male mosquitoes, p = 0.001). CONCLUSIONS: The results of this study may be used as environmental indicators of mosquito populations.

Influence of climatic variables on the Aedes aegypti and Culex quinquefasciatus populations in Mato Grosso, Brazil

Abstract INTRODUCTION: Aedes aegypti and Culex quinquefasciatus are vector species responsible for the transmission of important arboviruses. METHODS: Adult mosquitoes were collected in the urban areas of four municipalities in Mato Grosso within 1 year. RESULTS: A total of 19,110 mosquitoes were collected. Among them, 16,578 (86,8%) were C. quinquefasciatus (44% female and 56% male); 2,483 (13%), A. (Stegomyia) aegypti (54% female and 46% male); and 49 (0,30%), from the genus Psorophora, Anopheles, Coquilettidia, and Sabethes. A significant correlation was observed between the number of mosquitoes from all species and dew point (female mosquitoes, p = 0.001; male mosquitoes, p = 0.001). CONCLUSIONS: The results of this study may be used as environmental indicators of mosquito populations.

Novel Viruses in Mosquitoes from Brazilian Pantanal.

Viruses are ubiquitous and diverse microorganisms arising as a result of interactions within their vertebrate and invertebrate hosts. Here we report the presence of different viruses in the salivary glands of 1657 mosquitoes classified over 28 culicinae species from the North region of the Brazilian Pantanal wetland through metagenomics, viral isolation, and RT-PCR. In total, 12 viruses were found, eight putative novel viruses with relatively low similarity with pre-existing species of viruses within their families, named Pirizal iflavirus, Furrundu phlebovirus, Pixé phlebovirus, Guampa vesiculovirus, Chacororé flavivirus, Rasqueado orbivirus, Uru chuvirus, and Bororo circovirus. We also found the already described Lobeira dielmorhabdovirus, Sabethes flavivirus, Araticum partitivirus, and Murici totivirus. Therefore, these findings underscore the vast diversity of culicinae and novel viruses yet to be explored in Pantanal, the largest wetland on the planet.