Morphological identification of species of the Nuneztovari Complex of Anopheles (Diptera: Culicidae) from an area affected by a Brazilian hydroelectric plant

This investigation was financially supported by Norte Energia SA, Instituto Evandro Chagas and CNPq (grant number 302292/2017-9).Ministério da Saúde. Secretaria de Vigilância em Saúde. Instituto Evandro Chagas. Ananindeua, PA, Brasil / Universidade Federal do Pará. Instituto de Ciências Biológicas. Programa de Pós-Graduação em Biologia de Agentes Infecciosos e Parasitários Belém, PA, Brasil.Ministério da Saúde. Secretaria de Vigilância em Saúde. Instituto Evandro Chagas. Ananindeua, PA, BrasilMinistério da Saúde. Secretaria de Vigilância em Saúde. Instituto Evandro Chagas. Ananindeua, PA, BrasilMinistério da Saúde. Secretaria de Vigilância em Saúde. Instituto Evandro Chagas. Ananindeua, PA, BrasilMinistério da Saúde. Secretaria de Vigilância em Saúde. Instituto Evandro Chagas. Ananindeua, PA, BrasilMinistério da Saúde. Secretaria de Vigilância em Saúde. Instituto Evandro Chagas. Ananindeua, PA, Brasil / Universidade Federal do Pará. Instituto de Ciências Biológicas. Programa de Pós-Graduação em Biologia de Agentes Infecciosos e Parasitários Belém, PA, Brasil.The Nuneztovari Complex of Anopheles (Diptera: Culicidae) comprises four species: An. nuneztovari Gabaldon, An. goeldii Rozeboom & Gabaldon, An. dunhami Causey and An. nuneztovari species A. This study aimed to identify morphologically the species of the Nuneztovari Complex that occur in the area of the Belo Monte hydroelectric dam. The morphological identification of adult males and male genitalia (aedeagus and ventral claspette) was performed. A statistical analysis of the difference in aedeagal leaflet length was done using the Mann-Whitney test. Of the 38 male genitalia of specimens of the Nuneztovari Complex examined, 33 were identified as An. goeldii/An. nuneztovari A and five as An. nuneztovari s.s. A statistically significant difference in aedeagal leaflet length was detected between the species: the mean length was 1.23 μm for An. goeldii/An. nuneztovari A and 9.18 μm for An. nuneztovari s.s. This is the first record of An. nuneztovari s.s.in areas of environmental modification in the Brazilian Amazon. This study provides a measurement tool that can identify and differentiate species of the complex in the region, which can be applied to the other species of the complex as well to other anopheline species; thus, fostering the acquisition of information about the role of each species in malaria transmission

Strategy to improve malaria surveillance system preventing transfusion-transmitted malaria in blood banks using molecular diagnostic

Abstract Background Malaria can be transmitted by blood transfusion through donations collected from asymptomatic or parasitic donors. The parasites are released into the bloodstream during its life cycle and will therefore be present in donated blood by infected individuals. All cases of transfusion-transmitted malaria (TTM) notified since 2005 in Brazil were fatal. A good screening tool for Plasmodium spp. detection in blood units must have a high detection threshold, and the prevention of TTM relies entirely on the exclusion of potentially infected donors. However, in Brazilian blood banks, the screening test relies on blood thick smears examination. Methods The molecular diagnostic based on mitochondrial DNA (mtDNA) using real time PCR (mt-qPCR) was improved to detect Plasmodium falciparum, Plasmodium vivax, and standardized for use in Plasmodium malariae. The analytic sensitivity of this mt-qPCR methodology was performed using a sample of P. vivax. Results The mt-qPCR was highly efficient, and the analytic sensitivity for P. vivax was determined (0.000006 parasites/µL). This method was tested to detect P. vivax and P. falciparum in individuals from two malaria-endemic areas in Brazil, Amazon region (Pará and Rondônia states), the samples were collected in 10 reference units of two blood banks (Pará/nine cities and Rondônia/Porto Velho), and parasites mtDNA were detected in 10 of 2224 potential blood donors (0.45%). In all 10 positive samples, only P. vivax was detected. Conclusion Molecular diagnostic using mt-qPCR was effective in revealing infected potential donors with good perspectives to be applied as screening routine of asymptomatic carriers for preventing transfusion-transmitted malaria in blood banks

Phylogeny of Anopheles darlingi (Diptera:Culicidae) based on the antimicrobial peptide genes cecropin and defensin

This study was financially supported by the National Council for Scientific and Technological Development (CNPq n° 302292/2017–9), the Coordination for the Improvement of Higher Education Personnel (CAPES n° 88882.183965/2018–01), Instituto Evandro Chagas/SVS/MS and Norte Energia SA.Universidade Federal do Pará. Programa de Pós-graduação em Biologia de Agentes Infecciosos e Parasitários. Belém, PA, Brazil / Ministério da Saúde. Secretaria de Vigilância em Saúde. Instituto Evandro Chagas. Laboratório de Entomologia de Malária. Ananindeua, PA, Brasil / Fundação de Vigilância em Saúde do Amazonas - Dra. Rosemary Costa Pinto. Manaus, AM, BrazilUniversidade Federal do Pará. Programa de Pós-graduação em Biologia de Agentes Infecciosos e Parasitários. Belém, PA, Brazil / Ministério da Saúde. Secretaria de Vigilância em Saúde. Instituto Evandro Chagas. Laboratório de Entomologia de Malária. Ananindeua, PA, BrasillMinistério da Saúde. Secretaria de Vigilância em Saúde. Instituto Evandro Chagas. Ananindeua, PA, BrasilMinistério da Saúde. Secretaria de Vigilância em Saúde. Instituto Evandro Chagas. Ananindeua, PA, BrasilMinistério da Saúde. Secretaria de Vigilância em Saúde. Instituto Evandro Chagas. Laboratório de Entomologia de Malária. Ananindeua, PA, BrasilMinistério da Saúde. Secretaria de Vigilância em Saúde. Instituto Evandro Chagas. Ananindeua, PA, BrasilMinistério da Saúde. Secretaria de Vigilância em Saúde. Instituto Evandro Chagas. Ananindeua, PA, BrasilMinistério da Saúde. Secretaria de Vigilância em Saúde. Instituto Evandro Chagas. Laboratório de Entomologia de Malária. Ananindeua, PA, BrasilUniversidade Federal do Pará. Programa de Pós-graduação em Biologia de Agentes Infecciosos e Parasitários. Belém, PA, Brazil / Ministério da Saúde. Secretaria de Vigilância em Saúde. Instituto Evandro Chagas. Ananindeua, PA, Brasil / Universidade do Estado do Pará. Belém, PA, BrazilUniversidade Federal do Pará. Programa de Pós-graduação em Biologia de Agentes Infecciosos e Parasitários. Belém, PA, BrazilUniversidade Federal do Pará. Programa de Pós-graduação em Biologia de Agentes Infecciosos e Parasitários. Belém, PA, Brazil / Ministério da Saúde. Secretaria de Vigilância em Saúde. Instituto Evandro Chagas. Laboratório de Entomologia de Malária. Ananindeua, PA, BrasilCecropins and defensins are the main classes of antimicrobial peptides in the mosquito innate immune system, acting against bacteria, fungi and protozoa. There is a knowledge gap concerning these peptide genes in anopheline mosquitoes from the Brazilian Amazon. Thus, this work aimed to describe molecular techniques for detecting the genes encoding the antimicrobial peptides cecropin A (CecA) and defensin in Anopheles darlingi mosquitoes and to perform molecular phylogeny of the sequenced genes using the maximum likelihood method and Bayesian inference with other species from different geographic areas. Our results show, for the first time, a molecular biology method for detecting CecA and defensin in Anopheles darlingi that allows for the use of these molecular markers for phylogenetic analysis in anopheline species, separating the species into single and monophyletic clades

VK210 and VK247 genotypes of Plasmodium vivax in anopheline mosquitoes from Brazilian Amazon.

Evandro Chagas Institute/SVS/MS, CNPq (Grant: n° 302292/2017-9), and CAPES (Grant: n° 88882.183965/2018-01).Federal University of Pará. Pós Graduação em Biologia de Agentes Infecciosos e Parasitários. Belém, PA / Ministério da Saúde. Secretaria de Vigilância em Saúde. Instituto Evandro Chagas. Laboratório de Pesquisa Básica em Malária - Entomologia. Ananindeua, PA, Brasil.Ministério da Saúde. Secretaria de Vigilância em Saúde. Instituto Evandro Chagas. Laboratório de Pesquisa Básica em Malária - Entomologia. Ananindeua, PA, Brasil.Ministério da Saúde. Secretaria de Vigilância em Saúde. Instituto Evandro Chagas. Laboratório de Pesquisa Básica em Malária - Entomologia. Ananindeua, PA, Brasil.Ministério da Saúde. Secretaria de Vigilância em Saúde. Instituto Evandro Chagas. Laboratório de Geoprocessamento. Ananindeua, PA, Brasil.Ministério da Saúde. Secretaria de Vigilância em Saúde. Instituto Evandro Chagas. Laboratório de Geoprocessamento. Ananindeua, PA, Brasil.Ministério da Saúde. Secretaria de Vigilância em Saúde. Instituto Evandro Chagas. Laboratório de Pesquisa Básica em Malária - Entomologia. Ananindeua, PA, Brasil.Federal University of Pará. Pós Graduação em Biologia de Agentes Infecciosos e Parasitários. Belém, PA / Ministério da Saúde. Secretaria de Vigilância em Saúde. Instituto Evandro Chagas. Laboratório de Pesquisa Básica em Malária - Entomologia. Ananindeua, PA, Brasil.Federal University of Pará. Pós Graduação em Biologia de Agentes Infecciosos e Parasitários. Belém, PA / Ministério da Saúde. Secretaria de Vigilância em Saúde. Instituto Evandro Chagas. Laboratório de Pesquisa Básica em Malária - Entomologia. Ananindeua, PA, Brasil.Plasmodium vivax sporozoites are differenced by circumsporozoite protein. Studies on the circulation of P. vivax VK210 and P. vivax VK247 in anopheline mosquitoes are important to verify the adaptability of these parasites on mosquitoes in different locations and periods. This study aimed to describe and compare the distribution of these genotypes in anopheline mosquitoes from four states of the Brazilian Amazon. Epidemiological databases about CSP infections on mosquitoes from Pará (2000-2015), Amapá (2000-2010), Roraima (2000-2003 and 2009-2011) and Acre States (2012-2015) were used for analysis. A total of 895 specimens were found infected mainly by P. vivax VK210. We showed that the distribution of P. vivax VK247 changed over time in the main malaria vectors on the Brazilian Amazon. We note that A. darlingi was abundant in certain localities while A. albitarsis s.l. in anothers, which highlights the importance of entomological studies for the control of human malaria

Morphological atypia and molecular profile of

This study aimed to perform morphological and molecular analyses of parasites isolated from the blood of malaria-infected individuals during an outbreak in the Microregion of Cametá, State of Pará, Brazilian Amazon. A total of 260 positive samples were identified by microscopy as Plasmodium vivax; however, in three samples, forms considered unusual for the species were found and defined as morphological atypia of P. vivax. Single P. vivax infection was confirmed by qPCR in all samples. Among 256 genotyped samples, the VK247 genotype alone was identified in 255 samples, and the VK210 genotype was found in only one. The study showed that this malaria outbreak was caused by the etiological agent P. vivax, and for the first time, morphological atypia was described in isolates circulating in Brazil. Likewise, for the first time, the VK247 genotype was detected predominantly in single infections in an area of the State of Pará, which may suggest a greater circulation of the genotype in the region

Putative contributions of the sex chromosome proteins SOX3 and SRY to neurodevelopmental disorders

Coordenação de Aperfeiçoamento de Pessoalde Nível Superior, Grant/Award Number: DSProgramDS-1750212PROEX-1669479PROEX-33002010073P7; Fundaçãode Amparo à Pesquisa do Estado de São Paulo,Grant/Award Number: 2011/04956-62011/14658-22014/00041-12014/00591-12014/10488-32015/06281-7; Universidade Federaldo ABC, Grant/Award Number: InstitutionalScholarship; UFABC; CAPES, Grant/AwardNumber: DS-1750212; FAPESP, Grant/AwardNumbers: 2014/10488-3, 2011/04956-6,2014/00591-1, 2014/00041-1,2015/06281-7, 2011/14658-2.Universidade de São Paulo. Faculdade de Medicina. Instituto de Psiquiatria, Hospital das Clinicas HCFMUSP. Laboratório de Psicopatologia e Terapêutica Psiquiátrica . São Paulo, SP, Brazil.Universidade de São Paulo. Faculdade de Medicina. Instituto de Psiquiatria, Hospital das Clinicas HCFMUSP. Laboratório de Psicopatologia e Terapêutica Psiquiátrica . São Paulo, SP, Brazil / University of São Paulo. Inter-institutional Grad Program on Bioinformatics. São Paulo, SP, Brazil.Federal University of ABC. Center of Mathematics, Computing and Cognition. Santo André, SP, Brazil.Universidade de São Paulo. Faculdade de Medicina. Instituto de Psiquiatria, Hospital das Clinicas HCFMUSP. Laboratório de Psicopatologia e Terapêutica Psiquiátrica . São Paulo, SP, Brazil.Universidade de São Paulo. Faculdade de Medicina. Instituto de Psiquiatria, Hospital das Clinicas HCFMUSP. Laboratório de Psicopatologia e Terapêutica Psiquiátrica . São Paulo, SP, Brazil.Brazilian Center for Research in Energy and Materials. Brazilian Biosciences National Laboratory. Campinas, SP, Brazil.Universidade de São Paulo. Faculdade de Medicina. Instituto de Psiquiatria, Hospital das Clinicas HCFMUSP. Laboratório de Psicopatologia e Terapêutica Psiquiátrica . São Paulo, SP, Brazil.Universidade de São Paulo. Faculdade de Medicina. Instituto de Psiquiatria, Hospital das Clinicas HCFMUSP. Laboratório de Psicopatologia e Terapêutica Psiquiátrica . São Paulo, SP, Brazil.Universidade de São Paulo. Faculdade de Medicina. Instituto de Psiquiatria, Hospital das Clinicas HCFMUSP. Laboratório de Psicopatologia e Terapêutica Psiquiátrica . São Paulo, SP, Brazil / Ministério da Saúde. Secretaria de Vigilância em Saúde. Instituto Evandro Chagas. Laboratório de Pesquisas Básicas em Malária - Entomologia. Ananindeua, PA, Brasil.Brazilian Center for Research in Energy and Materials. Brazilian Biosciences National Laboratory. Campinas, SP, Brazil.Universidade de São Paulo. Faculdade de Medicina. Instituto de Psiquiatria, Hospital das Clinicas HCFMUSP. Laboratório de Psicopatologia e Terapêutica Psiquiátrica . São Paulo, SP, Brazil / University of São Paulo. Inter-institutional Grad Program on Bioinformatics. São Paulo, SP, Brazil / Universidade de São Paulo. Faculdade de Medicina. Instituto de Psiquiatria. Hospital das Clinicas HCFMUSP. São Paulo, SP, Brazil / National Institute of Developmental Psychiatry for Children and Adolescents (INPD). São Paulo, SP, Brazil / Universidade de São Paulo. Faculdade de Medicina FMUSP. São Paulo, SP, Brazil.The male-biased prevalence of certain neurodevelopmental disorders and the sex-biased outcomes associated with stress exposure during gestation have been previously described. Here, we hypothesized that genes distinctively targeted by only one or both homologous proteins highly conserved across therian mammals, SOX3 and SRY, could induce sexual adaptive changes that result in a differential risk for neurodevelopmental disorders. ChIP-seq/chip data showed that SOX3/SRY gene targets were expressed in different brain cell types in mice. We used orthologous human genes in rodent genomes to extend the number of SOX3/SRY set (1,721). These genes were later found to be enriched in five modules of coexpressed genes during the early and mid-gestation periods (FDR < 0.05), independent of sexual hormones. Genes with differential expression (24, p < 0.0001) and methylation (40, p < 0.047) between sexes were overrepresented in this set. Exclusive SOX3 or SRY target genes were more associated with the late gestational and postnatal periods. Using autism as a model sex-biased disorder, the SOX3/SRY set was enriched in autism gene databases (FDR ≤ 0.05), and there were more de novo variations from the male autism spectrum disorder (ASD) samples under the SRY peaks compared to the random peaks (p < 0.024). The comparison of coexpressed networks of SOX3/SRY target genes between male autism and control samples revealed low preservation in gene modules related to stress response (99 genes) and neurogenesis (78 genes). This study provides evidence that while SOX3 is a regulatory mechanism for both sexes, the male-exclusive SRY also plays a role in gene regulation, suggesting a potential mechanism for sex bias in ASD

Integrative Variation Analysis Reveals that a Complex Genotype May Specify Phenotype in Siblings with Syndromic Autism Spectrum Disorder

<div><p>It has been proposed that copy number variations (CNVs) are associated with increased risk of autism spectrum disorder (ASD) and, in conjunction with other genetic changes, contribute to the heterogeneity of ASD phenotypes. Array comparative genomic hybridization (aCGH) and exome sequencing, together with systems genetics and network analyses, are being used as tools for the study of complex disorders of unknown etiology, especially those characterized by significant genetic and phenotypic heterogeneity. Therefore, to characterize the complex genotype-phenotype relationship, we performed aCGH and sequenced the exomes of two affected siblings with ASD symptoms, dysmorphic features, and intellectual disability, searching for <i>de novo</i> CNVs, as well as for <i>de novo</i> and rare inherited point variations—single nucleotide variants (SNVs) or small insertions and deletions (indels)—with probable functional impacts. With aCGH, we identified, in both siblings, a duplication in the 4p16.3 region and a deletion at 8p23.3, inherited by a paternal balanced translocation, t(4, 8) (p16; p23). Exome variant analysis found a total of 316 variants, of which 102 were shared by both siblings, 128 were in the male sibling exome data, and 86 were in the female exome data. Our integrative network analysis showed that the siblings’ shared translocation could explain their similar syndromic phenotype, including overgrowth, macrocephaly, and intellectual disability. However, exome data aggregate genes to those already connected from their translocation, which are important to the robustness of the network and contribute to the understanding of the broader spectrum of psychiatric symptoms. This study shows the importance of using an integrative approach to explore genotype-phenotype variability.</p></div

Female sibling STRING network analysis.

<p>Biggest component of the connecting brain-expressed genes present in the unbalanced translocation, altered by the shared and exclusive rare inherited SNV and indels found in the female sibling.</p

Summary of the characteristics observed in our patients, compared with those of other cases described in the literature.

<p>Summary of the characteristics observed in our patients, compared with those of other cases described in the literature.</p

Details of the hemizygous variations found in the exome of the male sibling.

<p>Details of the hemizygous variations found in the exome of the male sibling.</p