Transcriptomics analysis highlights potential ways in human pathogenesis in Leishmania braziliensis infected with the viral endosymbiont LRV1.

The parasite Leishmania (Viannia) braziliensis is widely distributed in Brazil and is one of the main species associated with human cases of different forms of tegumentary leishmaniasis (TL) such as cutaneous leishmaniasis (CL) and mucosal leishmaniasis (ML). The mechanisms underlying the pathogenesis of TL are still not fully understood, but it is known that factors related to the host and the parasite act in a synergistic and relevant way to direct the response to the infection. In the host, macrophages have a central connection with the parasite and play a fundamental role in the defense of the organism due to their ability to destroy intracellular parasites and present antigens. In the parasite, some intrinsic factors related to the species or even the strain analyzed are fundamental for the outcome of the disease. One of them is the presence of Leishmania RNA Virus 1 (LRV1), an endosymbiont virus that parasitizes some species of Leishmania that triggers a cascade of signals leading to a more severe TL phenotype, such as ML. One of the strategies for understanding factors associated with the immune response generated after Leishmania/host interaction is through the analysis of molecular patterns after infection. Thus, the gene expression profile in human monocyte-derived macrophages obtained from healthy donors infected in vitro with L. braziliensis positive (LbLRV1+) and negative (LbLRV1-) for LRV1 was evaluated. For this, the microarray assay was used and 162 differentially expressed genes were identified in the comparison LbLRV1+ vs. LbLRV1-, 126 upregulated genes for the type I and II interferons (IFN) signaling pathway, oligoadenylate synthase OAS/RNAse L, non-genomic actions of vitamin D3 and RIG-I type receptors, and 36 down-regulated. The top 10 downregulated genes along with the top 10 upregulated genes were considered for analysis. Type I interferon (IFNI)- and OAS-related pathways results were validated by RT-qPCR and Th1/Th2/Th17 cytokines were analyzed by Cytometric Bead Array (CBA) and enzyme-linked immunosorbent assay (ELISA). The microarray results validated by RT-qPCR showed differential expression of genes related to IFNI-mediated pathways with overexpression of different genes in cells infected with LbLRV1+ compared to LbLRV1- and to the control. No significant differences were found in cytokine levels between LbLRV1+ vs. LbLRV1- and control. The data suggest the activation of gene signaling pathways associated with the presence of LRV1 has not yet been reported so far. This study demonstrates, for the first time, the activation of the OAS/RNase L signaling pathway and the non-genomic actions of vitamin D3 when comparing infections with LbLRV1+ versus LbLRV1- and the control. This finding emphasizes the role of LRV1 in directing the host's immune response after infection, underlining the importance of identifying LRV1 in patients with TL to assess disease progression.

Rhodnius spp. infestation in palm trees and natural infection by Trypanosoma cruzi and Trypanosoma rangeli in periurban and rural areas of state of the Rondônia, in the Brazilian Amazon.

The state of Rondônia in the Brazilian Amazon is prone to diseases transmitted by insect vectors because of the environmental and population changes resulting from large hydroelectric projects and the expansion of agricultural and livestock industries. The first case of Chagas disease by vectorial transmission was recorded in 2019 in a rural area in Rondônia, reinforcing the need for entomological surveillance. Hence, our goal was to estimate the abundance of Rhodnius spp. in palm trees located in rural and periurban areas and in Brazil-Bolivia border regions, perform domiciliary searches, and check for possible associations between triatomines and the presence/absence of palm-inhabiting fauna and outdoor farming, domestic animals, and buildings. The sampling took place in five municipalities of Rondônia in 2014 (June to August) and 2015 (April to June). Triatomines were collected by active searches during the selective pruning of palm tree crowns. Domiciliary inspections lasted from 30 to 60 min. A set of captured triatomines was analyzed for Trypanosoma cruzi and T. rangeli infection by PCR. Overall, 496 insects were captured during sampling of 150 palms in rural areas and 150 in periurban areas. No triatomine was found during active searches of 59 dwelling either indoors or outdoors. The majority of triatomines caught in the palm trees were identified as Rhodnius robustus (98.6%), and seven specimens were R. pictipes. Triatomine infestation was observed in only 20% of the sampled palms (61/300) in the vicinity of 26/59 households. Nearly half of the infested palm trees had only one or two triatomines, and few palms presented more than 15 triatomines. The municipality of Buritis had the highest triatomine abundance and percentage of infested palms; however, the highest triatomine density per infested palm was observed in Alvorada D'Oeste, where a quarter of the palms were infested. Ants, arachnids, termites, reptiles, and rodents were frequently found in palm trees. Dogs were the predominant domestic animals in households, whereas hens and cattle were the main farming animals. Model estimates showed that the number of triatomines was affected by the presence of henhouses and varied strongly between localities. No relationships were detected between the average number of triatomines and palm fauna and/or palm height. Overall, approximately half of the triatomines were infected with T. cruzi (51.4%) and/or T. rangeli (47.2%), reinforcing the need for continuous entomological surveillance and implementation of community-based approaches because the Brazilian state of Rondônia borders areas experiencing reinfestation by domiciled species and potential colonization of animal shelters by triatomines.

Draft Whole-Genome Sequence of Leishmania (Viannia) braziliensis Presenting Leishmania RNA Virus 1, from Western Amazon, Brazil.

Leishmania (Viannia) braziliensis is the main etiological agent of tegumentary leishmaniasis in the neotropics. Here, we report a draft genome sequence (31.2 Mb) of an L. braziliensis strain from the western Amazon region of Brazil. This genome sequence will complement those available for other Leishmania species and contribute to further studies focusing on this parasite and the neglected diseases associated with it.