Exposure to Zika and chikungunya viruses impacts aspects of the vectorial capacity of Aedes aegypti and Culex quinquefasciatus.

Zika (ZIKV) and chikungunya (CHIKV) are arboviruses that cause infections in humans and can cause clinical complications, representing a worldwide public health problem. Aedes aegypti is the primary vector of these pathogens and Culex quinquefasciatus may be a potential ZIKV vector. This study aimed to evaluate fecundity, fertility, survival, longevity, and blood feeding activity in Ae. aegypti after exposure to ZIKV and CHIKV and, in Cx. quinquefasciatus exposed to ZIKV. Three colonies were evaluated: AeCamp (Ae. aegypti-field), RecL (Ae. aegypti-laboratory) and CqSLab (Cx. quinquefasciatus-laboratory). Seven to 10 days-old females from these colonies were exposed to artificial blood feeding with CHIKV or ZIKV. CHIKV caused reduction in fecundity and fertility in AeCamp and reduction in survival and fertility in RecL. ZIKV impacted survival in RecL, fertility in AeCamp and, fecundity and fertility in CqSLab. Both viruses had no effect on blood feeding activity. These results show that CHIKV produces a higher biological cost in Ae. aegypti, compared to ZIKV, and ZIKV differently alters the biological performance in colonies of Ae. aegypti and Cx. quinquefasciatus. These results provide a better understanding over the processes of virus-vector interaction and can shed light on the complexity of arbovirus transmission.

A droga como dispositivo de controle social: uma análise das representações sociais do álcool, maconha e crack na imprensa brasileira / Las drogas como dispositivo de control social: un análisis de las representaciones sociales del alcohol, la marihuana y el crack en la prensa brasileña / Drugs as a social control device: an analysis of the social representations of alcohol, marijuana and crack in the brazilian press

RESUMO. As drogas se consolidam como um dos arquétipos culturais predominantes no cotidiano das sociedades urbanas, sendo sua presença ubíqua em praticamente todas as culturas. Os registros históricos apresentam ampla variabilidade de substâncias que em dado momento eram classificadas como o perigo social da época e que em outro se tornavam banalizadas ou tipificadas como inofensivas. Assim, esse estudo teve como objetivo analisar como dispositivo droga que se consolida em diferentes períodos históricos. Para isso, foram coletadas 4.227 matérias dos jornais Folha da Manhã, Folha da Noite e Folha de São Paulo, que abordassem questões relativas ao álcool (década de 1920), maconha (décadas de 1930 a 1960) e crack (década de 1980 a 2005) e realizada Análise Temática de Conteúdo. Os resultados permitem afirmar que a característica central que define todas as substâncias analisadas nos distintos momentos históricos é o risco social que ela apresenta. A droga se constitui como um risco aos usuários ao mesmo tempo que os institui enquanto uma figura de ameaça social. Ao se referenciar uma substância como uma droga, são ativados sentidos que remetem a um quadro de decadência e criminalidade.

RESUMEN. Las drogas se consolidan como uno de los arquetipos culturales predominantes en la vida cotidiana de las sociedades urbanas, y su presencia ubicua en prácticamente todas las culturas. Los registros históricos muestran una amplia variabilidad de sustancias que en un momento se clasificaron como el peligro social de la época y en otro momento se trivializaron o tipificaron como inofensivas. Así que este estudio tuvo como objetivo analizar cómo diferentes sustancias psicoactivas se encuentran en la prensa como un riesgo social en diferentes momentos. Para ello, se recogieron 4.227 artículos del periódico Folha da Manhã, Folha da Noite y Folha de São Paulo, que abordasen temas relacionados con el alcohol (1920), marihuana (1930 a 1960) y el crack (1980 a 2005) y se realizó un Análisis Temático de Contenido. Los resultados muestran que la característica definitoria de todas las drogas examinadas en los diferentes momentos históricos es el riesgo social que presenta. La droga se constituye como un riesgo para los usuarios mientras los establece como una figura de amenaza social. Al hacer referencia a una sustancia como droga, se activan sentidos que conducen a un marco de decadencia y criminalidad.

ABSTRACT Drugs are one of the predominant cultural archetypes in the daily life of urban societies, and their ubiquitous presence in almost all cultures. Historical records show a wide variability of substances that at one point were classified as the social danger of the time and at another time trivialized or typified as harmless. Thus, this study aimed to analyze how different psychoactive substances are constituted in the press as a social risk at different times. For this, we collected 4,227 articles of newspapers Folha da Manhã, Folha da Noite and Folha de São Paulo, that addressed issues related to alcohol (1920), marijuana (1930s to 1960) and crack (1980s to 2005) and performed a Thematic Content Analysis. The results indicate that the central defining characteristic of all drugs examined in the different historical moments is the social risk it has. The drug is constituted as a risk to users while establishing them as a figure of social threat. When referring a substance as a drug, senses are activated that denote to a situation of decadence and crime.

Sequencing of ZIKV genomes directly from Ae. aegypti and Cx. quinquefasciatus mosquitoes collected during the 2015-16 epidemics in Recife.

Zika virus (ZIKV) is a negative sense RNA virus from the Flaviviridae family, which was relatively unknown until the first human epidemic in Micronesia, in 2007. Since then, it spread to French Polynesia and the Americas. Recife, the capital of Pernambuco state and epicenter of the Zika epidemic in Brazil, experienced a large number of microcephaly cases and other congenital abnormalities associated to the ZIKV infection from, 2015 to 16. Evidences suggest that both Aedes aegypti and Culex quinquefasciatus mosquitoes from Recife are capable of replicating and transmitting the virus. Here, we conducted high throughput sequencing of ZIKV genomes directly from Ae. aegypti and Cx. quinquefasciatus mosquitoes collected during the ZIKV epidemics in Recife, in order to investigate the variability and evolution of the virus. We obtained 11 draft ZIKV genomes derived from 5 pools from each Ae. aegypti and Cx. quinquefasciatus species. Genome coverage breadth ranged from 16 to 100% and average depth from 45 to 46,584×. Two of these genomes were obtained from pools of Cx. quinquefasciatus females with no sign of blood in the abdomen. Amino acid substitutions found here were not species-specific. In addition, molecular clock dating estimated that ZIKV draft genomes obtained here were co-circulating in other regions of the country during the epidemics. Overall results highlight that viral mutations and even minor variants can be detected in genomes directly sequenced from mosquito samples and insights about natural viral genomic variability and viral evolution can be useful when designing tools for mosquito control programs.

Persistence of chikungunya ECSA genotype and local outbreak in an upper medium class neighborhood in Northeast Brazil.

The chikungunya East/Central/South/Africa virus lineage (CHIKV-ECSA) was first detected in Brazil in the municipality of Feira de Santana (FS) by mid 2014. Following that, a large number of CHIKV cases have been notified in FS, which is the second-most populous city in Bahia state, northeastern Brazil, and plays an important role on the spread to other Brazilian states due to climate conditions and the abundance of competent vectors. To better understand CHIKV dynamics in Bahia state, we generated 5 complete genome sequences from a local outbreak raised in Serraria Brasil, a neighbourhood in FS, by next-generation sequencing using Illumina approach. Phylogenetic reconstructions revealed that the new FS genomes belongs to the ECSA genotype and falls within a single strongly supported monophyletic clade that includes other older CHIKV sequences from the same location, suggesting the persistence of the virus during distinct epidemic seasons. We also performed minor variants analysis and found a small number of SNPs per sample (b_29L and e_45SR = 16 SNPs, c_29SR = 29 and d_45PL and f_45FL = 21 SNPs). Out of the 93 SNPs found, 71 are synonymous, 21 are non-synonymous and one generated a stop codon. Although those mutations are not related to the increase of virus replication and/or infectivity, some SNPs were found in non-structural proteins which may have an effect on viral evasion from the mammal immunological system. These findings reinforce the needing of further studies on those variants and of continued genomic surveillance strategies to track viral adaptations and to monitor CHIKV epidemics for improved public health control.

Virulence determinants in genetically heterogeneous populations of Aeromonads recovered from an urban lagoon.

The diversity and distribution of Aeromonas spp. associated with virulence profiles from the Rodrigo de Freitas Lagoon were investigated using phylogenetic analysis of gyrB/rpoB gene sequences for speciation. The concatenated gyrB/rpoB gene sequences clustered into five species: Aeromonas punctata/caviae (n = 37), A. hydrophila (n = 10), A. dhakensis (n = 16), A. jandaei (n = 1) and A. enteropelogenes/trota (n = 3). The virulence genes (atc/aerA/hlyA/asp/amp) resulted in 19 virulence profiles, distributed heterogeneously among the five Aeromonas species. Out of the 67 isolates, 16% presented five distinct profiles carrying four virulence genes and 7% showed all genes investigated. The hemolytic genes were detected as follows: act 54% (37/67), aerA 36% (24/67), hlyA 26% (18/67) and proteolytic genes such as asp 36% (24/57) and amp in 85% (57/67) were widely distributed in lagoon sampling stations. Meanwhile, 88% (59/67) and 92% (62/67) of the isolates showed hemolytic and proteolytic activity, respectively. Our results demonstrated that concatenated sequences of the gyrB and rpoB genes showed to be an adequate approach for the Aeromonas speciation and prevalence. The high heterogeneity of virulence genes among the species resulted in several virulence profiles, as well as high percentages of hemolytic and proteolytic activity, demonstrating the necessity of further epidemiological surveys of Aeromonas species pathogenicity in an aquatic recreational lagoon.

Genome sequencing reveals coinfection by multiple chikungunya virus genotypes in a recent outbreak in Brazil.

Chikungunya virus (CHIKV) is an RNA virus from the Togaviridae family transmitted by mosquitoes in both sylvatic and urban cycles. In humans, CHIKV infection leads to a febrile illness, denominated Chikungunya fever (CHIKF), commonly associated with more intense and debilitating outcomes. CHIKV arrived in Brazil in 2014 through two independent introductions: the Asian/Caribbean genotype entered through the North region and the African ECSA genotype was imported through the Northeast region. Following their initial introduction, both genotypes established their urban cycle among large naive human populations causing several outbreaks in the Americas. Here, we sequenced CHIKV genomes from a recent outbreak in the Northeast region of Brazil, employing an in-house developed Next-Generation Sequencing (NGS) protocol capable of directly detecting multiple known CHIKV genotypes from clinical positive samples. Our results demonstrate that both Asian/Caribbean and ECSA genotypes expanded their ranges, reaching cocirculation in the Northeast region of Brazil. In addition, our NGS data supports the findings of simultaneous infection by these two genotypes, suggesting that coinfection might be more common than previously thought in highly endemic areas. Future efforts to understand CHIKV epidemiology should thus take into consideration the possibility of coinfection by different genotypes in the human population.

Zika virus detection, isolation and genome sequencing through Culicidae sampling during the epidemic in Vitória, Espírito Santo, Brazil.

BACKGROUND: Zika virus (ZIKV) has been isolated from many mosquito species in nature, but it is believed that the main vectors in urban environments are species of the genus Aedes. Here, we detected and isolated ZIKV in samples from Aedes aegypti, Aedes taeniorhynchus and Culex quinquefasciatus, collected during the Zika epidemic in Vitória, southeast Brazil. Using quantitative real-time polymerase chain reaction, ZIKV detection was performed in mosquito samples collected from February to April 2016. RESULTS: Overall, six pools of mosquitoes were positive for ZIKV: four of Cx. quinquefasciatus, one of Ae. aegypti and one of Ae. taeniorhynchus. Their genomes were sequenced. CONCLUSIONS: These results support and strengthen the hypothesis that other mosquito species can also be involved in ZIKV transmission.

Persistent detection of Zika virus RNA from an infant with severe microcephaly - a case report.

BACKGROUND: Zika virus (ZIKV) is a recently emerged arbovirus, which infection during pregnancy is associated with a series of congenital malformations, collectively denominated Congenital Zika Syndrome (CZS). Following infection, ZIKV RNA has a median duration period of 10 days in plasma and up to 6 months in semen in immunocompetent adult individuals. Moreover, ZIKV is able to replicate and persist in fetal brains and placentas, consequently, infection is associated with pregnancy loss, albeit the pathogenic mechanisms are still unknown. CASE PRESENTATION: Here we report a CZS case of an infant born during the ZIKV outbreak in northeast Brazil, the child presented recurrent episodes of seizures with prolonged presence of ZIKV RNA on the central nervous system (CNS) and blood. ZIKV RNA was identified and partially sequenced from a sample of cerebrospinal fluid (CSF) obtained from the infant with 6 months of life, and later from another sample after the infant completed 17 months of life. Commonly congenital infections were discarded based on STORCH (syphilis, toxoplasmosis, rubella, cytomegalovirus and herpes simplex virus) negative laboratory results. Presence of specific ZIKV antibodies on both mother and children confirmed the association of severe microcephaly and ZIKV infection, diagnosed after birth. CONCLUSIONS: Altogether, our data raise the possibility that CZS cases may result in prolonged viral presence, these findings could be useful for therapy and diagnostic recommendations.

Perfil de resistência aos antibióticos e prevalência dos genes qacEΔ1 e sul1 em Pseudomonas aeruginosa de efluente hospitalar / Antibiotic resistance pattern and prevalence of qacEΔ1 and sul1 genes in Pseudomonas aeruginosa from hospital wastewater

Introdução: Efluentes hospitalares representam riscos à saúde pública e ambiental devido à presença de microrganismos patogênicos, drogas e produtos químicos. Pseudomonas aeruginosa é um patógeno oportunista frequentemente encontrado no ambiente hospitalar. Objetivo: Avaliar o resistoma de isolados de P. aeruginosa da estação de tratamento de esgoto hospitalar (ETEH) de um complexo hospitalar na cidade do Rio de Janeiro. Método: Vinte isolados dos cinco estágios da ETEH foram identificados como P. aeruginosa pelo sequenciamento do gene 16S rRNA. A suscetibilidade aos antibióticos foi determinada segundo o CLSI e os genes qacEΔ1 e sul1 foram detectados pela PCR. Resíduos de sulfonamidas foram pesquisados por cromatografia líquida de alta eficiência acoplada à espectrometria de massas sequencial. Resultados: Foi demonstrada a presença de sulfametoxazol em nível inferior a 50 ng∙L−1, resistência às sulfonamidas (80%) seguida pelas quinolonas (50%) e 13 perfis de suscetibilidade aos antimicrobianos. Os genes qacEΔ1-sul1 foram detectados em 100% dos isolados, sugerindo a presença de integrons de classe 1 em toda a ETEH. Conclusões: Os resultados sinalizaram limitações no tratamento e a propagação de genes de resistência nas etapas da ETEH. Esses dados contribuem com órgãos competentes no desenho de ações preventivas frente aos impactos negativos à saúde pública.

Introduction: Hospital effluents may pose great environmental risk due to the presence of pathogenic microorganisms, drugs and chemical components. Pseudomonas aeruginosa is an opportunistic pathogen frequently found in hospital environment. Objective: To evaluate the resistome of P. aeruginosa from the hospital wastewater treatment plant (HWTP) in a hospital complex of Rio de Janeiro city. Method: Twenty isolates from the five stages of the HWTP were identified as P. aeruginosa by 16S rRNA gene sequencing analysis. Susceptibility to antibiotics was determined according to CLSI and qacEΔ1 and sul1 genes were detected by PCR. Sulphonamide residues were investigated by high performance liquid chromatography coupled to sequential mass spectrometry. Results: The sulfamethoxazole has been demonstrated at a level below 50 ng L-1. Sulfonamide resistance (80%) has been demonstrated followed by quinolone class (50%) and 13 susceptibility patterns to antimicrobials. The qacEΔ1-sul1 genes were detected in 100% of isolates suggesting the presence of class 1 integrons in the whole HWTP. Conclusions: The results signalized limitations of HWTP and propagation of resistance genes in all stages of the HWTP. These data also contribute to the environmental sanitary surveillance in the design of prevention actions against negative impact on the public health.

Trypanosoma brucei EIF4E2 cap-binding protein binds a homolog of the histone-mRNA stem-loop-binding protein.

Trypanosomatids are parasitic protozoans characterized by several unique structural and metabolic processes that include exquisite mechanisms associated with gene expression and regulation. During the initiation of protein synthesis, for instance, mRNA selection for translation seems to be mediated by different eIF4F-like complexes, which may play a significant role in parasite adaptation to different hosts. In eukaryotes, the heterotrimeric eIF4F complex (formed by eIF4E, eIF4G, and eIF4A) mediates mRNA recognition and ribosome binding and participates in various translation regulatory events. Six eIF4Es and five eIF4Gs have been described in trypanosomatids with several of these forming different eIF4F-like complexes. This has raised questions about their role in differential mRNA translation. Here we have studied further TbEIF4E2, the least known eIF4E homologue from Trypanosoma brucei, and found that it is not associated with an eIF4G homolog. It is, however, associated with mature mRNAs and binds to a histone mRNA stem-loop-binding protein (SLBP), one of two Trypanosoma SLBP homologs (TbSLBP1 and TbSLBP2). TbSLBP1 is more similar to the mammalian counterpart while TbSLBP2 is exclusive to trypanosomatids and related organisms. TbSLBP2 binds to TbEIF4E2 through a conserved central region missing in other SLBP homologs. Both SLBPs, as well as TbEIF4E2, were found to localize to the cytoplasm. TbEIF4E2 and TbSLBP2 are differentially expressed during cell culture, being more abundant in early-log phase, with TbSLBP2 also showing cell-cycle dependent expression. The new data reinforce unique aspects of the trypanosomatid eIF4Es, with the TbEIF4E2-TbSLBP complex possibly having a role in differential selection of mRNAs containing stem-loop structures.

ΔNp73 overexpression promotes resistance to apoptosis but does not cooperate with PML/RARA in the induction of an APL-leukemic phenotype.

Here, we evaluated whether the overexpression of transcriptionally inactive ΔNp73 cooperates with PML/RARA fusion protein in the induction of an APL-leukemic phenotype, as well as its role in vitro in proliferation, myeloid differentiation, and drug-induced apoptosis. Using lentiviral gene transfer, we showed in vitro that ΔNp73 overexpression resulted in increased proliferation in murine bone marrow (BM) cells from hCG-PML/RARA transgenic mice and their wild-type (WT) counterpart, with no accumulation of cells at G2/M or S phases; instead, ΔNp73-expressing cells had a lower rate of induced apoptosis. Next, we evaluated the effect of ΔNp73 on stem-cell self-renewal and myeloid differentiation. Primary BM cells lentivirally infected with human ΔNp73 were not immortalized in culture and did not present significant changes in the percentage of CD11b. Finally, we assessed the impact of ΔNp73 on leukemogenesis or its possible cooperation with PML/RARA fusion protein in the induction of an APL-leukemic phenotype. After 120 days of follow-up, all transplanted mice were clinically healthy and, no evidence of leukemia/myelodysplasia was apparent. Taken together, our data suggest that ΔNp73 had no leukemic transformation capacity by itself and apparently did not cooperate with the PML/RARA fusion protein to induce a leukemic phenotype in a murine BM transplantation model. In addition, the forced expression of ΔNp73 in murine BM progenitors did not alter the ATRA-induced differentiation rate in vitro or induce aberrant cell proliferation, but exerted an important role in cell survival, providing resistance to drug-induced apoptosis.