Ecological aspects and molecular detection of Leishmania DNA Ross (Kinetoplastida: Trypanosomatidae) in phlebotomine sandflies (Diptera: Psychodidae) in terra firme and várzea environments in the Middle Solimões Region, Amazonas State, Brazil

Background: Phlebotomine sand flies (Diptera: Psychodidae) are insects of medical importance due to the role that some species play in the transmission of leishmaniasis. This work aimed to study some ecological aspects among sand flies fauna inhabiting two different environments: the várzea (lowland Amazonian forest) and terra firme (upland Amazonian forest), both located in Tefé Municipality, Amazonas State, Braziland to detect Leishmania infection in those phlebotomine populations. Methods: Sand flies were collected using HP light traps. Collection took place over the course of six months: January, February, April, August, September, and October of 2013. To detect natural infection by Leishmania, DNA samples were extracted from female sand flies and submitted to Polymerase Chain Reaction (PCR) targeting the kDNA gene; Leishmania species were identified by PCR-RFLP targeting the hsp70 gene and genetic sequencing. Results: In all, 5,716 individuals were collected, and 46 species were identified. Trichophoromyia ubiquitalis (3,330 - 58.26%) and Nyssomyia antunesi (661 - 11.26%) were the most abundant species. Species richness was greater in terra firme environments (42 species) than in the várzea environments (22 species), and forests ecotopes (43 species) were richer than peridomiciles (28 species). DNA of Leishmania was found in Th. ubiquitalis and Psychodopygus davisi, both of which inhabit the terra firme environment and sequencing analysis confirmed the presence of Leishmania (Viannia) lainsoni DNA in Th. ubiquitalis in Tefé Municipality. Conclusions: The high abundance of Th. ubiquitalis and Ps. davisi and detection of DNA of Leishmania sp. may indicate that both species could be putative vectors for American Cutaneous Leishmaniasis (ACL) in the terra firme environment of Tefé. The sand fly fauna found in várzea is rich and diverse, exhibiting several species, nevertheless the seasonal hydric stress during part of the year that could influence the local diversity, if compared with other studies. This is the first report in Amazonas State of Th. ubiquitalis with presence of L. (V.) lainsoni DNA. © 2015 Pereira et al.; licensee BioMed Central

Bionomia de Migonemyia migonei (Diptera, Psychodidae, Phlebotominae) em condições experimentais

Submitted by Ycaro Santos ([email protected]) on 2019-04-11T20:35:46Z No. of bitstreams: 1 Dissertação Eric Marialva.pdf: 2999412 bytes, checksum: 29c5e848de9e9571d6d45a5e39e75840 (MD5)Approved for entry into archive by Ycaro Santos ([email protected]) on 2019-04-11T20:39:10Z (GMT) No. of bitstreams: 1 Dissertação Eric Marialva.pdf: 2999412 bytes, checksum: 29c5e848de9e9571d6d45a5e39e75840 (MD5)Made available in DSpace on 2019-04-11T20:39:10Z (GMT). No. of bitstreams: 1 Dissertação Eric Marialva.pdf: 2999412 bytes, checksum: 29c5e848de9e9571d6d45a5e39e75840 (MD5) Previous issue date: 2019Conselho Nacional de Pesquisa (CNPq) – PVE 2014 / Processo 401262/2014-6Fundação Oswaldo Cruz. Instituto de Pesquisas Leônidas e Maria Deane. Manaus, AM, Brasil.A espécie Migonemyia migonei é vetora de Leishmania braziliensis no Brasil. Estudos recentes demostraram que essa espécie pode estar participando do ciclo de transmissão de Leishmania infantum chagasi no Brasil e Argentina. O presente estudo teve como objetivo estudar a biologia de Mi. migonei, verificando morfologia dos imaturos, aspectos de colonização em massa e desenvolver modelo de transmissão de Le. in.chagasi através da picada da mesma, de indivíduos colonizados. Para alcançar esses objetivos foram feitos vários experimentos, os estágios imaturos (ovo, larva e pupa ) foram analisados por MEV e microscopia de luz; colonização em massa foi verificado ciclo de vida, longevidade, fecundidade, fertilidade e preferência de oviposição em substratos; para transmissão, fêmeas de Mi. migonei foram infectadas com Le. in. chagasi e Le. braziliensis (controle) de isolados em meio de cultura; , posteriormente fêmeas infectadas foram colocadas para picar camundongos. Como resultado da descrição, os ovos de Mi. migonei apresentam um exocório com estruturas poligonais, a quetotaxia das larvas e pupas apresentaram diversas cerdas diferenciadas de outras espécies. O ciclo de vida de ovo até emergência do adulto foi de 62,1 dias em média. A maior fertilidade e fecundidade foi com o sangue de hamster (36,68, p <0.05), seguido de humano (23,28), camundongo (19,63) e pinto (12,20). Fêmeas de Mi. migonei alimentadas com maçã sobreviveram maior tempo (10 dias) seguido de água açucarada a 10% (7 dias) e água (5 dias). Não foi observado diferença na taxa de oviposição dos diferentes substratos. Quanto a transmissão Mi. migonei foi capaz de transmitir Le. in. chagasi, a quantidade de parasita variou de 10 a 1000 em uma única orelha. Estes resultados contribuem para biologia de Mi. migonei e desvendar seu papel como vetora na transmissão de Le. in. chagasi.The species Migonemyia migonei is a vector of Leishmania braziliensis in Brazil. Recent studies have shown that this species may be participating in the transmission cycle of Leishmania infantum chagasi in Brazil and Argentina. The present study had as objective to study the biology of Mi. migonei, verifying the morphology of the immature, aspects of mass colonization and to develop a model of transmission of Le.in. chagasi through the sting of it. In order to reach these objectives, several experiments were carried out, the immature stages (egg, larva, and pupa) were analysed by MEV and light microscopy; colonization in mass was verified life cycle, longevity, fecundity, fertility and preference of oviposition in substrates; for transmission Mi.migonei was infected with L. in. chagasi and Le. braziliensis (control), was later placed to prick mice. The eggs of Mi. migonei present an exochorion with polygonal structures, which chaetotaxy of larvae and pupae presented differentiated bristles of other species. The egg life cycle up to adult emergence was 62.1 days on average. The highest fertility and fecundity was with hamster blood (36.68, p <0.05), followed by the human (23.28), mouse (19.63) and chick (12.20). Mi. migonei females fed with apple survived longer (10 days) followed by 10% water (7 days) and water (5 days). No difference was observed in the oviposition rate of the different substrates. As for transmission Mi.migonei was able to transmit Le. in. chagasi, the amount of parasite varied from 10 to 1000. These results may contribute to the biology of Mi. migonei and its role as a vector in the transmission of Le. in. chagasi

Distribution and diversity of mosquitoes and Oropouche-like virus infection rates in an Amazonian rural settlement.

Mosquito diversity and disease transmission are influenced by landscape modifications, i.e., vectors and pathogens previously found only in forests are now found close to human environments due to anthropic changes. This study determined the diversity and distribution of mosquitoes in forest environments in order to analyze the potential vectors of Amazonian forest arboviruses. Mosquitoes were collected by 1) vertical stratification from forest canopy and ground areas using Hooper Pugedo (HP) light traps and human attraction and 2) horizontal stratification using HP light traps in peridomicile, forest edge, and forest environments near the Rio Pardo rural settlement, Amazonas, Brazil. A total of 3,750 mosquitoes were collected, representing 46 species. 3,139 individuals representing 46 species were sampled by vertical stratification. Both the Shannon-Weaver diversity index (H') and equitability (J') were higher in the canopy than on the ground. 611 individuals representing 13 species were sampled by horizontal stratification. H' decreased in the following order: forest edge > forest > peridomicile, and J' was greater at the forest edge and smaller in the peridomicile environment. Moreover, H' was higher for the human attraction collection method than the HP traps. A total of 671 pools were analyzed by RT-qPCR; three species were positive for Oropouche-like viruses (Ochlerotatus serratus, Psorophora cingulata, and Haemagogus tropicalis) and the minimum infection rate was 0.8%. The composition of mosquito species did not differ significantly between anthropic and forest environments in Rio Pardo. Some mosquito species, due to their abundance, dispersion in the three environments, and record of natural infection, were hypothesized to participate in the arbovirus transmission cycle in this Amazonian rural settlement