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The genus Orthopodomyia Theobald, 1904 (Diptera: Culicidae) comprises 36 wild mosquito species, with distribution largely restricted to tropical and temperate areas, most of which are not recognized as vectors of epidemiological importance due to the lack of information related to their bionomy and involvement in the cycle transmission of infectious agents. Furthermore, their evolutionary relationships are not completely understood, reflecting the scarcity of genetic information about the genus. Therefore, in this study, we report the first complete description of the mitochondrial genome of a Neotropical species representing the genus, Orthopodomyia fascipes Coquillet, 1906, collected in the Brazilian Amazon region. Using High Throughput Sequencing, we obtained a mitochondrial sequence of 15,598 bp, with an average coverage of 418.5×, comprising 37 functional subunits and a final portion rich in A + T, corresponding to the control region. The phylogenetic analysis, using Maximum Likelihood and Bayesian Inference based on the 13 protein-coding genes, corroborated the monophyly of Culicidae and its two subfamilies, supporting the proximity between the tribes Orthopodomyiini and Mansoniini, partially disagreeing with previous studies based on the use of molecular and morphological markers. The information generated in this study contributes to a better understanding of the taxonomy and evolutionary history of the genus and other groups of Culicidae.
Assuntos
Culicidae , Genoma Mitocondrial , Filogenia , Animais , Culicidae/genética , Culicidae/classificação , Brasil , Sequenciamento de Nucleotídeos em Larga EscalaRESUMO
The Orthoflavivirus ilheusense (ILHV) is an arbovirus that was first isolated in Brazil in 1944 during an epidemiologic investigation of yellow fever. Is a member of the Flaviviridae family and it belongs to the antigenic complex of the Ntaya virus group. Psorophora ferox is the primary vector of ILHV and this study presents the isolation and phylogenetic analysis of ILHV in a pool of Ps. ferox collected in the state of Goiás in 2021. Viral isolation tests were performed on Vero cells and C6/36 clones. The indirect immunofluorescence test (IFI) was used to confirm the positivity of the sample. The positive sample underwent RT-qPCR, sequencing, and phylogenetic analysis. This is the first report of ILHV circulation in this municipality and presented close relationship between this isolate and another ILHV isolate collected in the city of Belém (PA).
Assuntos
Culicidae , Filogenia , Animais , Brasil , Células Vero , Culicidae/virologia , Chlorocebus aethiops , Flavivirus/genética , Flavivirus/isolamento & purificação , Flavivirus/classificação , Mosquitos Vetores/virologiaRESUMO
Bats are widely distributed in Brazil, including the Amazon region, and their association with viral pathogens is well-known. This work aimed to evaluate the metavirome in samples of Molossus sp. bats captured in the Brazilian Amazon from 2019 to 2021. Lung samples from 58 bats were divided into 13 pools for RNA isolation and sequencing followed by bioinformatic analysis. The Retroviridae family showed the highest abundance of viral reads. Although no complete genome could be recovered, the Paramyxoviridae and Dicistroviridae families showed the formation of contigs with satisfactory identity and size characteristics for further analysis. One contig of the Paramyxoviridae family was characterized as belonging to the genus Morbillivirus, being grouped most closely phylogenetically to Porcine morbillivirus. The contig related to the Dicistroviridae family was identified within the Cripavirus genus, with 94%, 91%, and 42% amino acid identity with Culex dicistrovirus 2, Rhopalosiphum padi, and Aphid lethal paralysis, respectively. The presence of viruses in bats needs constant updating since the study was able to identify viral sequences related to families or genera still poorly described in the literature in association with bats.
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Aedes aegypti is a mosquito native to the African continent, which is now widespread in the tropical and subtropical regions of the world. In many regions, it represents a major challenge to public health, given its role in the cycle of transmission of important arboviruses, such as Dengue, Zika, and Chikungunya. Considering the epidemiological importance of Ae. aegypti, the present study sequenced the partial mitochondrial genome of a sample collected in the municipality of Balsas, in the Brazilian state of Maranhão, followed by High Throughput Sequencing and phylogenetic analyses. The mitochondrial sequence obtained here was 15,863 bp long, and contained 37 functional subunits (thirteen PCGs, twenty-two tRNAs and two rRNAs) in addition to a partial final portion rich in A+T. The data obtained here contribute to the enrichment of our knowledge of the taxonomy and evolutionary biology of this prominent disease vector. These findings represent an important advancement in the understanding of the characteristics of the populations of northeastern Brazil and provide valuable insights into the taxonomy and evolutionary biology of this prominent disease vector.
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As part of a continuous effort to investigate the viral communities associated with wild mammals at the human-animal interface in an Amazonian metropolitan region, this study describes the detection of a novel rodent-borne arterivirus. A sample containing pooled organs of Oecomys paricola was submitted to RNA sequencing, and four sequences taxonomically assigned as related to the Arteriviridae family were recovered, corresponding to an almost complete genome of nearly 13 kb summed. In the phylogenetic analysis with the standard domains used for taxa demarcation in the family, the tentatively named Oecomys arterivirus 1 (OAV-1) was placed within the clade of rodent- and porcine-associated viruses, corresponding to the Variarterivirinae subfamily. The divergence analysis, based on the same amino acid alignment, corroborated the hypothesis that the virus may represent a new genus within the subfamily. These findings contribute to the expansion of the current knowledge about the diversity, host and geographical range of the viral family. Arterivirids are non-human pathogens and are usually species-specific, but the susceptibility of cell lines derived from different organisms should be conducted to confirm these statements for this proposed new genus in an initial attempt to assess its spillover potential.
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Arteriviridae , Arterivirus , Animais , Suínos , Filogenia , Brasil , Arterivirus/genética , Mamíferos , RoedoresRESUMO
Mycobacterium tuberculosis (Mtb) Central Asian Strain (CAS) Lineage 3 (L3) genotype is predominantly found in East-Africa, Central-Asia, Western-Asia, and South-Asia; however, a new spoligotyping CAS/SIT2545 was found in northern regions of Brazil. We aimed to characterize and describe the genetic diversity and perform a phylogenetic assessment of this novel genotype. We performed 24-MIRU-VNTR loci and Whole-genome sequencing (WGS) of six Brazilian isolates previously spoligotyped. The libraries were prepared using a Nextera-XT kit and sequenced in a NextSeq 550 Illumina instrument. We performed lineage assignment and genomic characterization. From publicly available genomes of Mtb L3 and other lineages, we created a robust dataset to run the MTBSeq pipeline and perform a phylogenetic analysis. MIRU-VNTR and WGS confirmed CAS/SIT2545 belongs to L3. Out of 1691 genomes, 1350 (79.83%) passed in quality control (genomic coverage > 95%). Strain 431 differed in 52 single nucleotide variants (SNV), confirming it does not belong to the same transmission chain. The eight genomes from a global dataset clustered closer to Brazilian strains differed in >52 SNVs. We hypothesized L3 and L1 were introduced in Brazilian Northern in the same historical event; however, there is a need for additional studies exploring the genetic diversity of Mtb Brazilian Northern.
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The viral genus Henipavirus includes two highly virulent zoonotic viruses of serious public health concern. Hendra henipavirus and Nipah henipavirus outbreaks are restricted to Australia and Southeast Asia, respectively. The Henipavirus genus comprises mostly bat-borne viruses, but exceptions have already been described as novel viruses with rodents and shrews as reservoir animals. In the Americas, scarce evidence supports the circulation of these viruses. In this communication, we report a novel henipa-like virus from opossums (Marmosa demerarae) from a forest fragment area in the Peixe-Boi municipality, Brazil, after which the virus was named the Peixe-Boi virus (PBV). The application of next-generation sequencing and metagenomic approach led us to discover the original evidence of a henipa-like virus genome in Brazil and South America and the original description of a henipa-like virus in marsupial species. These findings emphasize the importance of further studies to characterize PBV and clarify its ecology, impact on public health, and its relationship with didelphid marsupials and henipaviruses.
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Quirópteros , Vírus Hendra , Infecções por Henipavirus , Vírus Nipah , Animais , Infecções por Henipavirus/epidemiologia , Brasil/epidemiologia , GenômicaRESUMO
Lack of routine surveillance in countries endemic for bovine tuberculosis (TB) and limited laboratory support contributes to the inability to differentiate the Mycobacterium tuberculosis Complex species, leading to an underestimated burden of the disease. Here, Whole-Genome Sequencing of Mycobacterium bovis isolated from tissues with TB-like lesions obtained from cattle and buffalos at Marajó Island, Brazil, demonstrates that recent transmission of M. bovis is ongoing at distinct sites. Moreover, the M. bovis epidemiology in this setting is herein found to be dominated by an endemic and unique clade composed of strains evolved from a common ancestor that are now genetically differentiated from other M. bovis clades. Additionally, envisioning a rapid strain differentiation and tracing across multiple settings, 28 globally validated strain-specific SNPs were identified, three of which considered as robust markers for the M. bovis Marajó strain. In conclusion, this study contributes with data regarding the identification of a novel M. bovis phylogenetic clade responsible for ongoing transmission events in both cattle and buffalo species in Brazil, provides a framework to investigate the dissemination of this highly prevalent strain and, holds the potential to inform TB control strategies that may help to prevent the spread of bovine and zoonotic TB.
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Mycobacterium bovis/genética , Tuberculose Bovina/microbiologia , Animais , Brasil , Búfalos , Bovinos , Mycobacterium tuberculosis/genética , Filogenia , Polimorfismo de Nucleotídeo Único/genética , Tuberculose/microbiologia , Sequenciamento Completo do Genoma/métodos , Zoonoses/microbiologiaRESUMO
Dengue fever, chikungunya, and Zika are diseases caused by viruses transmitted by Aedes aegypti and Aedes albopictus. In Brazil, the number of human infections is high, but few studies are performed in mosquito vectors. This study aimed to investigate the presence of Zika, Dengue and Chikungunya viruses in Ae. aegypti and Ae. albopictus from the municipalities of Alto Alegre, Caxias, Codó, and São Mateus do Maranhão, located in the state of Maranhão, Northeast Brazil. The mosquitoes were collected with a mechanical aspirator, identified, triturated, and then submitted to RNA extraction and RT-qPCR. The positive samples were confirmed by virus isolation and genome sequencing. Three hundred and forty-eight Ae. aegypti (176 males and 172 females) and 12 Ae. albopictus (eight males and four females) were collected and tested. Ae. aegypti was the only vector positive in two municipalities-Codó, with detection of Chikungunya virus (CHIKV) belonging to the East-Central-South African genotype, and in Caxias, with detection of Dengue virus (DENV)-2 belonging to the Asian/American genotype. The detection of CHIKV and DENV-2 is evidence that those viruses are maintained in arthropod vectors, and shows the epidemiological risk in the area for chikungunya cases and a possible increase of severe dengue cases, associated with the occurrence of dengue hemorrhagic fever.
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Aedes/virologia , Vírus Chikungunya/fisiologia , Vírus da Dengue/fisiologia , Mosquitos Vetores/virologia , Animais , Brasil/epidemiologia , Febre de Chikungunya/epidemiologia , Febre de Chikungunya/transmissão , Febre de Chikungunya/virologia , Vírus Chikungunya/classificação , Vírus Chikungunya/isolamento & purificação , Dengue/epidemiologia , Dengue/transmissão , Dengue/virologia , Vírus da Dengue/classificação , Vírus da Dengue/isolamento & purificação , Feminino , Genótipo , Geografia , Humanos , Masculino , Filogenia , Reação em Cadeia da Polimerase em Tempo RealRESUMO
Parrots (Order Psittaciformes) are globally distributed birds that, together with members of the Columbiformes, represent the most susceptible animals, in regards to infection by Chlamydophila psittaci, which is an obligate, zoonotic, intracellular bacterium that causes chlamydiosis in domestic and wild birds and psittacosis in humans. The aim of the present study was to assess the occurrence of C. psittaci in Brazilian psittacids kept in conservation breeding sites in Pará State, Brazil. Cloacal and oropharynx swab samples were collected from 201 psittacids that were distributed among four breeding sites: Metropolitan Area of Belém (C1 and C2), Northeastern Pará (C3), and Low Amazon (C4). The samples were screened for C. psittaci using semi-nested PCR, and the resulting incidence data were analyzed using proportion and chi-square tests. Chlamydophila infection was confirmed for all the breeding sites, with an overall prevalence of 31.84%, and no species-specific predisposition was observed. Furthermore, 13.93% of the sampled birds eliminated the infectious agent by the cloaca, whereas 11.44% eliminated the agent by the oropharynx, and 6.47% eliminated the agent by both routes. Moreover, there was a significant difference between the incidence of Chlamydophila infection of breeding sites C2 and C3 (p=0.029), which yielded the smallest and largest number of diagnosed cases, respectively. In the present study, most of the birds (27.86%) were considered unapparent carriers of Chlamydophila infection, and only 3.98% of the birds yielded both a positive diagnosis and clinical signs of chlamydiosis.
Psitacídeos são aves distribuídas em todo o mundo e, juntamente com Columbiformes, representam os animais mais suscetíveis a uma infecção causada por Chlamydophila psittaci, uma bactéria intracelular, obrigatória, zoonótica que causa clamídia em aves domésticas e selvagens e psitacose em humanos. O objetivo deste estudo foi avaliar a ocorrência de C. psittaci em diferentes espécies de psitacídeos da fauna brasileira mantidos em criadouros conservacionistas no Estado do Pará, Brasil. Amostras de swabs de cloaca e orofaringe de 201 psitacídeos distribuídos em quatro criadouros nas mesorregiões Metropolitana de Belém (C1 e C2), Nordeste do Pará (C3) e Baixo Amazonas (C4) foram utilizados. As amostras foram submetidas ao teste molecular de semi-nested PCR. As análises estatísticas foram realizadas de acordo com o teste de proposição por R e teste do qui-quadrado (p 0,05). A presença de Chlamydophila sp. foi confirmada em todos os criadouros, com uma prevalência de 31,84% de aves infectadas, com predisposição não específica da espécie encontrada para a infecção entre as aves amostradas. Os resultados da semi-nested PCR mostraram que 13,93% das aves eliminaram o agente infeccioso pela cloaca, 11,44% pela orofaringe e 6,47% por ambas. Além disso, quando aplicado em cada local, este teste mostrou uma diferença estatisticamente significativa entre os criadouros C2 e C3 (p = 0,029), que apresentou o menor e maior número de casos diagnosticados, respectivamente. A maioria dos animais, ou 27,86%, foi considerada como portadora inaparente da infecção e apenas 3,98% das aves com diagnóstico positivo apresentaram algum sinal clínico sugestivo da doença.