Monitoramento de vetores do gênero Aedes durante o inverno no município de Barbacena, Minas Gerais, Brasil ­ 2018 / Monitoring of vectors of the genus Aedes during winter in the municipality of Barbacena, Minas Gerais, Brazil ‒ 2018

Objetivo: estimar a oviposição e distribuição espacial de vetores Aedes durante a estação de inverno e correlacionar essas estimativas com dados climáticos do mesmo período. Métodos: estudo de campo conduzido no município de Barbacena-MG, em 2018. O monitoramento, a coleta de ovos e a estimativa de índices estegômicos de vetores Aedes foram obtidos por meio de ovitrampas. Os Índices de Densidade de Ovos (IDO) e de Positividade de Ovitrampas (IPO%) foram estimados conforme estações climáticas e semanas epidemiológicas. A correlação entre parâmetros meteorológicos (temperatura/pluviometria) e índices estegômicos (IDO/IPO) foi determinada pelo coeficiente de Spearman. Resultados: um total de 1.080 ovitrampas, instaladas em 39 bairros, foi analisado durante 10 semanas epidemiológicas, fornecendo um total de 970 ovos. Nas estações de outono, inverno e primavera, foram obtidos, respectivamente, 421, 470 e 70 ovos. Durante o outono, houve variação do IDO entre 14,2 e 34,2. O IPO manteve-se constante em 4,5%. Durante o inverno, houve variação do IDO entre 0,00 e 47,50 e do IPO entre 0,00% e 8,25%. Houve correlação negativa significativa tanto entre aumento da precipitação mensal e diminuição do número de ovos coletados (rho=-0.673) quanto entre aumento da precipitação mensal e diminuição do IPO (rho=-0.612). O valor geral do IDO e do IPO nas 10 semanas foi, respectivamente, 22,04 e 4,17% e, na estação de inverno, foi, respectivamente, 23,50 e 3,73%. Conclusão: os achados corroboram a presença de ovos e vetores do gênero Aedes mesmo em condições climáticas adversas para essas espécies e sustentam ações de manejo sanitário durante todo o ano.

Objective: to estimate oviposition and spatial distribution of Aedes vectors during the winter season and correlate these estimates with climate data from the same period. Methods: field study conducted in the municipality of Barbacena-MG in 2018. Monitoring, egg collection, and estimation of stegomic indices of Aedes were obtained using ovitraps. The Indices of Egg Density (EDI) and Positive Ovitrap (POI%) were estimated according to climatic seasons and epidemiological weeks. The correlation between meteorological parameters (temperature/rainfall) and stegomic indices (EDI/POI) was determined by the Spearman coefficient. Results: a total of 1,080 ovitraps installed in 39 neighborhoods were analyzed during ten (10) epidemiological weeks, providing a total of 970 eggs. In the autumn, winter, and spring seasons, 421, 470, and 70 eggs were obtained, respectively. During the autumn, there was a variation in EDI between 14.2­34.2. The POI remained constant at 4.5%. During the winter, the EDI varied between 0.00­47.50, and the POI varied between 0.00%­8.25%. There was a significant negative correlation, respectively, between an increase in monthly precipitation and a decrease in the number of eggs collected (rho=-0.673) and between an increase in monthly precipitation and a decrease in POI (rho=-0.612). The overall value of EDI and POI in the ten (10) weeks was 22.04 and 4.17%, and in the winter season, they were 23.50 and 3.73%, respectively. Conclusion: the findings corroborate the presence of eggs and vectors of the genus Aedes even in adverse climatic conditions for these species and support health management actions throughout the year.

The Schistosomiasis SpleenOME: Unveiling the Proteomic Landscape of Splenomegaly Using Label-Free Mass Spectrometry.

Schistosomiasis is a neglected parasitic disease that affects millions of people worldwide and is caused by helminth parasites from the genus Schistosoma. When caused by S. mansoni, it is associated with the development of a hepatosplenic disease caused by an intense immune response to the important antigenic contribution of adult worms and to the presence of eggs trapped in liver tissue. Although the importance of the spleen for the establishment of immune pathology is widely accepted, it has received little attention in terms of the molecular mechanisms operating in response to the infection. Here, we interrogated the spleen proteome using a label-free shotgun approach for the potential discovery of molecular mechanisms associated to the peak of the acute phase of inflammation and the development of splenomegaly in the murine model. Over fifteen hundred proteins were identified in both infected and control individuals and 325 of those proteins were differentially expressed. Two hundred and forty-two proteins were found upregulated in infected individuals while 83 were downregulated. Functional enrichment analyses for differentially expressed proteins showed that most of them were categorized within pathways of innate and adaptive immunity, DNA replication, vesicle transport and catabolic metabolism. There was an important contribution of granulocyte proteins and antigen processing and presentation pathways were augmented, with the increased expression of MHC class II molecules but the negative regulation of cysteine and serine proteases. Several proteins related to RNA processing were upregulated, including splicing factors. We also found indications of metabolic reprogramming in spleen cells with downregulation of proteins related to mitochondrial metabolism. Ex-vivo imunophenotyping of spleen cells allowed us to attribute the higher abundance of MHC II detected by mass spectrometry to increased number of macrophages (F4/80+/MHC II+ cells) in the infected condition. We believe these findings add novel insights for the understanding of the immune mechanisms associated with the establishment of schistosomiasis and the processes of immune modulation implied in the host-parasite interactions.

Understanding global changes of the liver proteome during murine schistosomiasis using a label-free shotgun approach.

Schistosomiasis is an endemic disease affecting over 207 million people worldwide caused by helminth parasites of the genus Schistosoma. In Brazil the disease is responsible for the loss of up to 800 lives annually, resulting from the desabilitating effects of this chronic condition. In this study, we infected Balb/c mice with Schistosoma mansoni and analysed global changes in the proteomic profile of soluble liver proteins. Our shotgun analyses revealed predominance of up-regulation of proteins at 5weeks of infection, coinciding with the onset of egg laying, and a remarkable down-regulation of liver constituents at 7weeks, when severe tissue damage is installed. Representatives of glycolytic enzymes and stress response (in particular at the endoplasmic reticulum) were among the most differentially expressed molecules found in the infected liver. Collectively, our data contribute over 70 molecules not previously reported to be found at altered levels in murine schistosomiasis to further exploration of their potential as biomarkers of the disease. Moreover, understanding their intricate interaction using bioinformatics approach can potentially bring clarity to unknown mechanisms linked to the establishment of this condition in the vertebrate host. SIGNIFICANCE: To our knowledge, this study refers to the first shotgun proteomic analysis to provide an inventory of the global changes in the liver soluble proteome caused by Schistosoma mansoni in the Balb/c model. It also innovates by yielding data on quantification of the identified molecules as a manner to clarify and give insights into the underlying mechanisms for establishment of Schistosomiasis, a neglected tropical disease with historical prevalence in Brazil.

Shotgun proteomics to unravel the complexity of the Leishmania infantum exoproteome and the relative abundance of its constituents.

The exoproteome of some Leishmania species has revealed important insights into host-parasite interaction, paving the way for the proposal of novel disease-oriented interventions. The focus of the present investigation constituted the molecular profile of the L. infantum exoproteome revealed by a shotgun proteomic approach. Promastigotes under logarithmic phase of growth were obtained and harvested by centrifugation at different time points. Cell integrity was evaluated through the counting of viable parasites using propidium iodide labeling, followed by flow cytometry analysis. The 6h culture supernatant, operationally defined here as exoproteome, was then conditioned to in solution digestion and the resulting peptides submitted to mass spectrometry. A total of 102 proteins were identified and categorized according to their cellular function. Their relative abundance index (emPAI) allowed inference that the L. infantum exoproteome is a complex mixture dominated by molecules particularly involved in nucleotide metabolism and antioxidant activity. Bioinformatic analyses support that approximately 60% of the identified proteins are secreted, of which, 85% possibly reach the extracellular milieu by means of non-classic pathways. At last, sera from naturally infected animals, carriers of differing clinical forms of Canine Visceral Leishmaniasis (CVL), were used to test the immunogenicity associated to the L. infantum exoproteome. Western blotting experiments revealed that this sub-proteome was useful at discriminating symptomatic animals from those exhibiting other clinical forms of the disease. Collectively, the molecular characterization of the L. infantum exoproteome and the preliminary immunoproteomic assays opened up new research avenues related to treatment, prognosis and diagnosis of CVL.