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The Oropouche Virus (OROV) has Culicoides paraensis as its main vector in the urban cycle. Diagnoses of acute febrile cases and entomological collections were performed between January and April 2024 in the urban setting of Porto Velho, a city located in the western Brazilian Amazon. A total of 904 human samples were evaluated using RT-qPCR, of which 328 were positive for OROV. In addition, 3,950 insects were captured and Culex quinquefasciatus was the most abundant and predominant species. Seven mosquito samples showed amplification for OROV from pools of Cx. quinquefasciatus, Limatus durhamii and Aedes albopictus. Due to the first record of OROV in the two latter species, our findings can support new hypotheses about potential or bridge vectors. In turn, the high density of Cx. quinquefasciatus reinforces the supposition of this species as the main urban vector in the Amazon in the absence of C. paraensis. Complementary studies of vector competence and capacity are necessary for confirmation., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024. Published by Elsevier B.V.)

This study aimed to assess the diversity patterns of sand fly fauna across different strata and detect Leishmania DNA in these insects in the Mapinguari National Park in Rondônia and Amazonas states, Northern Brazil. Sand flies were collected with "HP" light traps in the canopy (15 m) and at ground level (1 m) on two trails, during August and November 2021 and March and August 2022. Polymerase chain reaction and DNA sequencing were used to identify Leishmania species. A total of 8,040 individuals (2,303♂ - 28.64 %, 5,737♀ - 71.36 %) were collected and 53 species and 13 genera were identified. The most abundant species were Psychodopygus chagasi (21.00 %), Trichophoromyia ubiquitalis (13.61 %) Psychodopygus davisi (11.69 %), Nyssomyia fraihai (10.58 %), Nyssomyia antunesi (6.68 %) and Nyssomyia richardwardi (5.55 %). Species diversity was greater in the canopy (Shannon index H' = 10.8 common species) when compared to ground level (H' = 10.5 common species). We observed a minimum infection rate of 0.45 % (22/4,868 females), in which Leishmania braziliensis DNA was found in Ps. chagasi and Le. lainsoni DNA in Ps. chagasi, Ny. richardwardi, Ps. ayrozai and Th. ubiquitalis. The minimum infection rate of Leishmania in the canopy was 0.47 % (19/4,031) and in the ground was 0.52 % (3/567). In the present study, we observed Le. lainsoni DNA in females of Ny. richardwardi for the first time. The data presented in this study contribute to understanding sand fly diversity and its distribution between the states of Rondônia and Amazonas. They may be useful for implementing targeted control measures to reduce the spread of leishmaniasis and implement entomological surveillance strategies., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Objectives: To evaluate the short and long-term effects of anodal tDCS (a-tDCS) targeting the left temporoparietal area (LTA) on tinnitus severity, annoyance, and loudness., Methods: This is a double-blind, randomized, sham-controlled, and parallel-group clinical trial. A total of 42 individuals with tinnitus were randomized to a-tDCS (n = 24) or sham tDCS (n = 18). The a-tDCS group received tDCS over the LTA during five consecutive day sessions (2 mA, 20 min). The sham group received a placebo current with the same characteristics as the a-tDCS group. Participants were assessed at baseline, after the fifth session, and at the 30-day follow-up, using hearing assessments and symptom questionnaires., Results: There was no effect of comparison between groups or interaction effect (time x group) in all hearing assessments and symptom questionnaires. There was only a main effect of time for Tinnitus Handicap Inventory - THI [F(1.642, 45.988) = 5.128; p = 0.014; η 2 = 0.155]. Bonferroni post hoc showed that there was a significant difference in THI in the sham group between pre and post-treatment [CI (0.107, 14.643; p = 0.046)]. However, there was no difference between pre-treatment and follow-up THI, or between post-treatment and follow-up THI. There was no treatment effect on tinnitus severity (assessed by Tinnitus Functional Inventory - TFI), tinnitus annoyance or loudness (assessed by Visual Analogue Scale - VAS), or tinnitus pitch, loudness or minimum masking level (assessed by tinnitometry)., Conclusion: Five consecutive sessions of a-tDCS targeting LTA do not improve tinnitus severity, annoyance, and loudness. Future studies should investigate if other tDCS protocols are effective or a combination of tDCS with other forms of treatment., Competing Interests: Declaration of competing interest The authors declare that they have no competing interests., (Copyright © 2024 Elsevier Masson SAS. All rights reserved.)

Malaria control and elimination efforts would benefit from the identification and validation of new malaria chemotherapeutics. Recently, a transgenic Plasmodium berghei line was used to perform a series of high-throughput in vitro screens for new antimalarials acting against the parasite sexual stages. The screens identified pyrimidine azepine chemotypes with potent activity. Here, we validate the activity of PyAz90 , the most potent pyrimidine azepine chemotype identified, against P. falciparum and P. vivax in the asexual and sexual stages. PyAz90 blocked parasite transmission to the mosquito vector at nanomolar concentrations and inhibited in vitro asexual parasite multiplication with a fast-action profile. Through the generation of P. falciparum PyAz90- resistant parasites and in vitro assays of mitochondrial activity, we identified cytochrome b as a molecular target of PyAz90 . This work characterizes a promising chemotype that can be explored for the future development of new antimalarials targeting the Plasmodium cytochrome bc 1 complex., Competing Interests: The authors declare no competing financial interest., (© 2024 The Authors. Published by American Chemical Society.)

Sand flies play a crucial role as vectors of bacteria, viruses, and protists, with Leishmania being the most notable among them, transmitted to vertebrate hosts during blood feeding. Understanding the feeding behaviours of sand flies is imperative for gaining insights into their eco-epidemiological roles in the transmission of these infectious agents. This systematic review aimed to answer the question 'What are the blood-feeding sources identified in Brazilian sand flies?' to provide an analysis of their blood-feeding habits. The diverse range of at least 16 vertebrate orders identified as blood sources for 54 sand fly species across different geographic regions was summarised, and the factors potentially associated with the risk of bias in the included studies were analysed. The findings broaden the discussion concerning methods used to identify blood meal sources and shed light on the implications of sand fly feeding behaviours for the transmission dynamics of Leishmania.

Transmission-blocking vaccines interrupting malaria transmission within mosquitoes represent an ideal public health tool to eliminate malaria at the population level. Plasmodium falciparum and P. vivax account for more than 90% of the global malaria burden, co-endemic in many regions of the world. P25 and P48/45 are two leading candidates for both species and have shown promising transmission-blocking activity in preclinical and clinical studies. However, neither of these target antigens as individual vaccines has induced complete transmission inhibition in mosquitoes. In this study, we assessed immunogenicity of combination vaccines based on P25 and P48/45 using a DNA vaccine platform to broaden vaccine specificity against P. falciparum and P. vivax. Individual DNA vaccines encoding Pvs25, Pfs25, Pvs48/45 and Pfs48/45, as well as various combinations including (Pvs25 + Pvs48/45), (Pfs25 + Pfs48/45), (Pvs25 + Pfs25), and (Pvs48/45 + Pfs48/45), were evaluated in mice using in vivo electroporation. Potent antibody responses were induced in mice immunized with individual and combination DNA vaccines, and specific antibody responses were not compromised when combinations of DNA vaccines were evaluated against individual DNA vaccines. The anti-Pvs25 IgG from individual and combination groups revealed concentration-dependent transmission-reducing activity (TRA) in direct membrane feeding assays (DMFA) using blood from P. vivax-infected donors in Brazil and independently in ex vivo MFA using Pvs25-transgenic P. berghei. Similarly, anti-Pfs25 and anti-Pfs48/45 IgGs from mice immunized with Pfs25 and Pfs48/45 DNA vaccines individually and in various combinations revealed antibody dose-dependent TRA in standard membrane feeding assays (SMFA) using culture-derived P. falciparum gametocytes. However, antibodies induced by immunization with Pvs48/45 DNA vaccines were ineffective in DMFA and require further vaccine construct optimization, considering the possibility of induction of both transmission-blocking and transmission-enhancing antibodies revealed by competition ELISA. These studies provide a rationale for combining multiple antigens to simultaneously target transmission of malaria caused by P. falciparum and P. vivax., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Entomological investigations were conducted for the first time in urban forest remnants of Porto Velho, state of Rondônia, Brazil, to explore the transmission dynamics of Leishmania. Sand fly collections were carried out at ten sites, encompassing both canopy and ground strata, from October to December 2021. A total of 1,671 sand flies were collected, representing 42 species within 12 genera. Nyssomyia Antunesi (n = 384) and Psychodopygus davisi (n = 111) were the most abundant species. Molecular analyses targeting the V7V8 region (18S gene) unveiled the presence of sequences 100% identical to Leishmania infantum in females of Bichromomyia flaviscutellata (1), Nyssomyia Antunesi complex (6), Nyssomyia umbratilis (1), Nyssomyia sp. (1), Psychodopygus ayrozai (1), Ps. davisi (3), Psychodopygus paraensis (1), and Sciopemyia sordellii (1). Sequences 100% similar to Trypanosoma minasense were found in two samples of the Nyssomyia Antunesi complex, and two samples of Sc. sordellii presented 100% identity to a Trypanosoma sp. strain, previously identified in this same sand fly in Rondônia. Sequencing of Cytb fragment suggested Homo sapiens, Dasypus novemcinctus and Tamandua tetradactyla as the blood source for distinct sand flies. The identification of sequences similar to L. infantum in sand flies collected in urban forest fragments is noteworthy, correlating with the recent local and regional occurrence of autochthonous cases of human visceral leishmaniasis. However, further studies are imperative to ascertain the presence of hosts/reservoirs and evaluate the risk of L. infantum transmission to humans., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2024 Silva et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

Water reuse for potable purposes can represent a realistic source supply of drinking water in areas with water scarcity. Therefore, combining conventional wastewater treatment technologies with advanced technologies is necessary to remove contaminants and obtain high-quality and safe water. In this study, the pesticides and degradation products, atrazine (ATZ), hydroxyatrazine (ATZOH), deethylatrazine (DEA), deisopropylatrazine (DIA), simazine (SMZ), ametryn (AMT), diuron (DIU), 2,4-D, fipronil (FIP), fipronil sulfide (FIP-SF) and fipronil sulfone (FIP-SN) were evaluated in effluent after membrane bioreactor (MBR), effluent after advanced treatment by multiple barriers (MBR, reverse osmosis, UV/H 2 O 2 and activated carbon), in tap water collected in the urban region of Campinas and in the Atibaia River (water supply source from city of Campinas). The pesticide concentrations in the Atibaia River and the post-MBR effluent ranged between 1 and 434 ng L -1 and 1 and 470 ng L -1 , respectively. Therefore, the Atibaia River and the post-MBR effluent had the same magnitude pesticide concentrations. In the production of potable water reuse, after the multiple barriers processes, only fipronil (1 ng L -1 ) and atrazine (3 ng L -1 ) were quantified in some of the samples. In tap water from Campinas, atrazine, ATZOH, DEA, diuron, and 2,4-D were quantified in concentrations ranging between 3 and 425 ng L -1 . Therefore, when comparing drinking water obtained from conventional treatment with potable water reuse, according to the pesticides studied, it is possible to conclude that the advanced treatment used on a pilot scale is promising for use in a potable water reuse plant. However, studies involving more microbiological and chemical parameters should be conducted to classify potable water reuse as drinking water., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

The maintenance of a highly productive colony of anopheline mosquitoes requires standardized methods in order to obtain a sufficient number of homogeneous individuals for malaria research. In this context, nutritional status may affect survival, fecundity, and the capacity to support pathogen development. Here we assess the effects of carbohydrate sources on fecundity, survival, and susceptibility to Plasmodium vivax infection in colonies of Anopheles darlingi and Anopheles deaneorum mosquitoes. Newly emerged females from each species were fed either 10% sugar or 15% honey solutions until the end of each experiment. The type of carbohydrate meal did not impact any entomological parameters for An. deaneorum , except for survival. For both species, honey meal significantly increased median survival post-emergence by three to four days, probably due to its nutritional value. For An. darlingi fed with honey, a higher mean frequency in stage 5 was observed at 48 h post-blood-meal, which could indicate a delay in the digestion process. However, no effects on fecundity parameters were observed. Regarding susceptibility, An. darlingi fed with sugar exhibited a low intensity of sporozoites, although any negative effects of sucrose on sporozoites invasions in the salivary glands are unknown. Based on the increase in mosquito survival, a carbohydrate source composed of 15% honey solution could be better for maintaining An. darlingi and An. deaneorum in the lab-rearing context.

Gamete surface protein P48/45 has been shown to be important for male gamete fertility and a strong candidate for the development of a malaria transmission-blocking vaccine (TBV). However, TBV development for Plasmodium vivax homolog Pvs48/45 has been slow because of a number of challenges: availability of conformationally suitable recombinant protein; the lack of an in vivo challenge model; and the inability to produce P. vivax gametocytes in culture to test transmission-blocking activity of antibodies. To support ongoing efforts to develop Pvs48/45 as a potential vaccine candidate, we initiated efforts to develop much needed reagents to move the field forward. We generated monoclonal antibodies (mAbs) directed against Pvs48/45 and characterized putative functional domains in Pvs48/45 using recombinant fragments corresponding to domains D1-D3 and their biological functionality through ex vivo direct membrane feeding assays (DMFAs) using P. vivax parasites from patients in a field setting in Brazil. While some mAbs partially blocked oocyst development in the DMFA, one mAb caused a significant enhancement of the infectivity of gametocytes in the mosquitoes. Individual mAbs exhibiting blocking and enhancing activities recognized non-overlapping epitopes in Pvs48/45. Further characterization of precise epitopes recognized by transmission-reducing and -enhancing antibodies will be crucial to design an effective immunogen with optimum transmission-reducing potential., Competing Interests: The authors declare no conflict of interest.

In this study, we analysed the molecular and morphometric differences of several populations of the putative sand fly vector Psychodopygus davisi (Root, 1934) (Diptera, Psychodidae, Phlebotominae) in Brazil. We amplified the 658 base pair fragments of the DNA barcoding region-cytochrome c oxidase subunit 1 (COI) gene-for 57 specimens of P. davisi and three specimens of Psychodopygus claustrei (Abonnenc, Léger & Fauran, 1979). We merged our data with public sequences of the same species available from GenBank. Then, the combined dataset-87 sequences and 20 localities-was analysed using population structure analysis and different species delimitation approaches. Geometric morphometry of wings was performed for 155 specimens of P. davisi populations from the North, Midwest and Southeast Brazilian regions, analysing the differences in centroid sizes and canonical variates. Molecular analysis indicated high intraspecific genetic distance values for P. davisi (maximum p distance = 5.52%). All algorithms identified P. davisi and P. claustrei as distinct molecular taxonomic units, despite the low interspecific distance (p distance to the nearest neighbour = 4.79%). P. davisi sequences were split into four genetic clusters by population structure analysis and at least five genetic lineages using intermediate scenarios of the species delimitation algorithms. The species validation analysis of BPP strongly supported the five-species model in our dataset. We found high genetic diversity in this taxon, which is in agreement with its wide geographic distribution in Brazil. Furthermore, the wing analysis showed that specimens from the Southeast Region of Brazil are different from those in the North and the Midwest. The evolutionary patterns of P. davisi populations in Brazil suggest the presence of candidate species, which need to be validated in future studies using a more comprehensive approach with both genomic data and morphological characters., (© 2023 Royal Entomological Society.)

Background: In the Amazon region, several species of triatomines occur in the natural environments. Among them, species of the genus Rhodnius are a risk to human populations due to their high rates of infection with Trypanosoma cruzi. The aim of this study was to identify the T. cruzi genotypes in Rhodnius specimens and their relationship with sylvatic hosts from different environments in the Brazilian Amazon. Methods: A total of 492 triatomines were collected from the municipalities of Monte Negro, Rondônia state, and Humaitá, Amazonas state, 382 of them being nymphs and 110 adults. Genotyping of T. cruzi in six discrete typing units (DTUs) was performed using conventional multilocus PCR. The triatomines that were positive for T. cruzi and engorged with blood were also targeted for amplification of the cytochrome B (cytB) gene to identify bloodmeal sources. Results: Of the 162 positive samples, the identified DTUs were TcI (87.65%) and TcIV (12.35%). It was observed that 102 specimens were engorged with a variety of bloodmeals. Triatomines infected with TcI were associated with DNA of all identified vertebrates, except Plecturocebus brunneus. TcIV was detected in triatomines that fed on Coendou prehensilis , Didelphis marsupialis , Mabuya nigropunctata , P. brunneus , Pithecia irrorata , Sapajus apella , and Tamandua tetradactyla. Conclusion: Results highlight the need to understand the patterns of T. cruzi genotypes in Rhodnius spp. and their association with sylvatic hosts to better elucidate their role in the transmission of Chagas disease in the Amazon region.

Circadian behavioral patterns in mosquitoes can be observed through their locomotor activity, which includes fundamental behaviors such as foraging, mating, and oviposition. These habits, which are fundamental to the life cycle of Anopheles mosquitoes, are closely related to pathogen transmission to humans. While rhythmic cycles of locomotor activity have been described in Anopheles species, no studies have been conducted on Anopheles darlingi species, the main malaria vector in the Amazon region. The aim of this study was to investigate how insemination status, blood meal, and Plasmodium vivax infection affect the locomotor activity of An. darlingi. The experiments were performed with 3- to 10-day-old An. darlingi females, which had been fed with 15% honey solution. These mosquitoes were obtained from the Malaria Vector Production and Infection Platform (PIVEM)/FIOCRUZ-RO. The experimental groups were divided into four categories: virgin vs. inseminated, unfed virgin vs. blood-fed virgin, unfed inseminated vs. blood-fed inseminated, and infected blood vs. uninfected blood. Locomotor activity was monitored using the Flybox equipment, capturing images that were subsequently converted into video to measure the insect activity, using PySoLo software. The periodicity and rhythmicity of mosquito locomotor activity were analyzed using MatLab® software. The locomotor activity of An. darlingi females showed a nocturnal and bimodal pattern under LD conditions. When comparing the insemination states and blood meal, there was a reduction in the locomotor activity in inseminated and blood-fed females. However, the P. vivax + infection did not increase locomotor activity of An. darlingi species., (© 2023. The Author(s).)

Anthropogenic activities such as agriculture and industry increase contaminants that reach the water bodies, potentially threatening the biota. Most likely, these pollutants occur in complex mixtures. The effects on organisms can be potentiated (synergism) or reduced (antagonism) according to the interaction between the stressors or the species. Cadmium (Cd) is a toxic metal present in phosphate fertilizers, and fipronil is an insecticide broadly used in sugarcane crops. Copepods are important energy transfer links in aquatic environments, and effects on this group impact the whole trophic chain. In this study, we evaluated the responses of the freshwater Calanoida copepod Notodiaptomus iheringi, naturally present in water bodies that can be affected by sugarcane cultures in Brazil. The organisms were exposed to environmental concentrations of Cd and fipronil, isolated and in the mixture, in acute (48 h) and sub-chronic (8 d) tests. Our data indicate that both contaminants affect the survival of the organisms in acute or sub-chronic exposures. Cadmium did not affect egg production or hatching, while fipronil impacted these endpoints negatively. The Cd-fipronil combination resulted in antagonistic responses in survival (acute and sub-chronic) and egg production. A synergistic response was observed in egg hatching. Our results suggest that Cd presents a protective effect in the mixture with fipronil; however, it is not enough to prevent egg-hatching inhibition. These responses highlight how tricky it is to deal with pollutants' interaction in environmental concentrations since synergism is the most common response to metal-pesticide mixtures. Our data point out N. iheringi as a sensitive organism in the presence of contaminants and reflects the threat of chemical mixtures in concentrations found in water bodies close to sugarcane crops in Brazil., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 The Authors. Published by Elsevier Ltd.. All rights reserved.)

In the study undertaken by Souza et al. [Primates 64(1):153-159, 2022; https://doi.org/10.1007/s10329-022-01038-5 ], published in the most recent volume of this journal, the blood samples of two Alouatta guariba clamitans (Primates, Atelidae) from two municipalities in the state of Rio Grande do Sul, southern Brazil were reported to be positive for Mansonella perstans. This is the first reported finding of M. perstans in A. guariba clamitans, as well as the first time that M. perstans has been recorded in Brazil outside the Amazon region. We would like to express our concern about this finding, specifically with respect to the geographical distribution of M. perstans in Brazil, as, up until this study, this filaria had only been found in the upper Rio Negro region in São Gabriel da Cachoeira, Amazonas, Brazil. Moreover, species identification was performed using partial sequences of three gene fragments, namely internal transcribed spacer 2, 12S, and 18S, yet neither the phylogenetic trees nor the BLAST alignments of these sequences provided supporting evidence that they belong to M. perstans., (© 2023. The Author(s), under exclusive licence to Japan Monkey Centre.)

Obtaining Plasmodium vivax sporozoites is essential for in vitro culture of liver stage parasites, not only to understand fundamental aspects of parasite biology, but also for drug and vaccine development. A major impediment to establish high-throughput in vitro P. vivax liver stage assays for drug development is obtaining sufficient numbers of sporozoites. To do so, female anopheline mosquitoes have to be fed on blood from P. vivax-infected patients through an artificial membrane-feeding system, which in turns requires a well-established Anopheles colony. In this study we established conditions to provide a robust supply of P. vivax sporozoites. Adding a combination of serum replacement and antibiotics to the membrane-feeding protocol was found to best improve sporozoite production. A simple centrifugation method appears to be a possible tool for rapidly obtaining purified sporozoites with a minimal loss of yield. However, this method needs to be better defined since sporozoite viability and hepatocyte infection were not evaluated., (© 2023. Springer Nature Limited.)

Water scarcity is one of the main abiotic factors that limit agricultural production. In this sense, the identification of genotypes tolerant to water deficit associated with irrigation management strategies is extremely important. In this context, the objective of this study was to evaluate the morphology, production, water consumption, and water use efficiency of colored fiber cotton genotypes submitted to irrigation strategies with a water deficit in the phenological phases. Two experiments were conducted in succession. In the first experiment, a randomized block design was used in a 3 × 7 factorial scheme, corresponding to three colored cotton genotypes (BRS Rubi, BRS Jade, and BRS Safira) in seven irrigation management strategies with 40% of the real evapotranspiration (ETr) varying the phenological stages. In the second experiment, the same design was used in a 3 × 10 factorial arrangement (genotypes × irrigation management strategies). The water deficit in the vegetative phase can be used in the first year of cotton cultivation. Among the genotypes, 'BRS Jade' is the most tolerant to water deficit in terms of phytomass accumulation and fiber production.

Aedes aegypti are vector insects of arboviruses such as dengue, Zika, and chikungunya. All available vector control methods have limited efficacy, highlighting the urgent need to find alternative ones. Evidence shows that arachnids like ticks are sources of biologically active compounds. Moreover, chemical modulation of the locomotor and immune systems of vector insects can be used to control arbovirus transmission. The present study evaluated the effectiveness of crude saliva of female Amblyomma cajennense sensu stricto (s.s.) ticks in reducing locomotor activity and inducing an immune response in Ae. aegypti females. Additionally, the study evaluated the protein constitution of tick saliva. For this purpose, the crude saliva obtained from several semi-engorged A. cajennense females was used. A volume of 0.2 nL of crude tick saliva was administered to mosquitoes by direct intrathoracic microinjection. The effect of the tick's saliva on the locomotor activity of the mosquito was observed using Flybox, a video-automated monitoring system, and the hemolymph hemocyte levels were quantified by reading slides under a light microscope. The protein concentration of the crude tick saliva was 1.27 μg/μL, and its electrophoretic profile indicates the presence of proteins with a molecular weight ranging between ∼17 and 95 kDa. Microplusins, ixodegrins, cystatin, actins, beta-actin, calponin, albumin, alpha-globulins, and hemoglobin were the main proteins identified by proteomics in the saliva of A. cajennense. The microinjected saliva had low toxicity for Ae. aegypti females and significantly reduced their locomotor activity, especially in the transition between the light and dark phases. The crude tick saliva did not change the period and rhythmicity of the circadian cycle. The tick saliva significantly increased the number of hemocytes two days after injection and reduced it after five days. These results suggest that further evaluation of the biological properties of tick saliva proteins against Ae. aegypti would be of interest., Competing Interests: Declaration of competing interest All authors disclose some financial and personal relationships with other people or organizations that could inappropriately influence in this work. The authors declare no conflict of interest., (Published by Elsevier Inc.)

The magnitude of copepods' responses to pesticides, individually and in mixture, is little understood. The aims of this study were to evaluate: (i) the effects of the pesticides fipronil and 2,4-D, individually and in mixture, on the freshwater copepod Notodiaptomus iheringi; and (ii) the survival and the feeding rate of copepods after the exposure. Acute toxicity tests using the commercial formulations of fipronil and 2,4-D, individually and in mixture, were performed. The LC 10-48h , LC 20-48h , and LC 50-48h of fipronil to N. iheringi were 2.38 ± 0.48, 3.08 ± 1.14, and 4.97 ± 3.30 μg L -1 , respectively. For 2,4-D the LC 10-48h , LC 20-48h , and LC 50-48h were 371.18 ± 29.20, 406.93 ± 53.77, and 478.24 ± 107.77 mg L -1 , respectively. Morphological damages on the copepods exposed to pesticides were observed at all concentrations. Fungal filaments covering dead organisms were presented at the treatment highest concentration (R5:7.43 ± 2.78 μg L -1 fipronil). The mixture of the pesticides presented synergistic effects on the mortality of N. iheringi. Post-exposure tests showed no difference between the treatments and the control on the mortality and on the feeding rate for 4 h. However, since delayed toxicity of pesticides can occur, longer post-exposure tests using N. iheringi should be tested. N. iheringi is a key species in the aquatic Brazilian ecosystem and showed sensitivity to fipronil and 2,4-D; thus, more studies with this species assessing other responses are recommended., (© 2023. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

Biting midges of the genus Culicoides are insects of proven medical and veterinary importance, because of their role in the transmission of viruses, bacteria, protozoa and nematodes. Culicoides paraensis has been considered the main vector of the Oropouche Virus (OROV) in the urban cycle of the disease in the neotropics. Due to the great abundance of Culicoides spp. in the State of Rondônia and its epidemiological history of OROV, we investigated the biting activity in humans, the abundance as a function of meteorological parameters and seasonality, and the detection of OROV. Entomological collections occurred in three municipalities from Brazilian State of Rondônia: Porto Velho, Ariquemes, and Ouro Preto do Oeste. GLMM's were used to determine if Culicoides spp. abundance was predicted by seasonal, diurnal, and meteorological factors. Total RNA was extracted from insects and viral RNA detection was performed using the S segment as the target region of OROV via RT-qPCR. In total, 7315 individuals were captured and identified as C. paraensis. In the dry season, 1488 individuals (24.5%) were recorded, 4591 (75.5%) in the rainy season, with peaks of biting activity between 4pm and 6pm. All variables showed a significative effect on the midge abundance. The rainy season, temperature between 30 °C and 32 °C and relative air humidity between 75% and 85% were the main predictive parameters for capturing the highest average number of insects. Our results confirm diurnal activity of C. paraensis and its greatest abundance in rainy periods. No sample was positive for the OROV, which could be explained by the virus absence in local human populations, C. paraensis as a minor vector species in the sampled localities, and probable low rate of infection of biting midges. Our findings on hourly and seasonal biting activities can provide support to intervention actions regarding vector control and surveillance of this species. This was the first study to collect and analyze biting midges in a region where human OROV cases had already been detected, but without previous information on entomovirological surveillance., Competing Interests: Declaration of Competing Interest The authors declare no competing interests., (Copyright © 2023 Elsevier B.V. All rights reserved.)

Malaria is caused by parasite of the genus Plasmodium and is still one of the most important infectious diseases in the world. Several biological characteristics of Plasmodium vivax contribute to the resilience of this species, including early gametocyte production, both of which lead to efficient malaria transmission to mosquitoes. This study evaluated the impact of currently used drugs on the transmission of P. vivax. Participants received one of the following treatments for malaria: i) chloroquine [10 mg/kg on day 1 and 7.5 mg/kg on day 2 and 3] co-administered with Primaquine [0.5 mg/kg/day for 7 days]; ii) Chloroquine [10 mg/kg on day 1 and 7.5 mg/kg on day 2 and 3] co-administered with one-dose of Tafenoquine [300 mg on day 1]; and iii) Artesunate and Mefloquine [100 mg and 200 mg on day 1, 2 and 3] co-administered with Primaquine [0.5 mg/kg/day for 14 days]. Patient blood was collected before treatment and 4 h, 24 h, 48 h and 72 h after treatment. The blood was used to perform a direct membrane feeding assay (DMFA) using Anopheles darlingi mosquitoes. The results showed 100% inhibition of the mosquito infection after 4 h using ASMQ+PQ, after 24 h for the combination of CQ+PQ and 48 h using CQ+TQ. The density of gametocytes declined over time in all treatment groups, although the decline was more rapid in the ASMQ+PQ group. In conclusion, it was possible to demonstrate the transmission-blocking efficacy of the malaria vivax treatment and that ASMQ+PQ acts faster than the two other treatments., Competing Interests: The authors have declared that no competing interests exist., (Copyright: This is an open access article, free of all copyright, and may be freely reproduced, distributed, transmitted, modified, built upon, or otherwise used by anyone for any lawful purpose. The work is made available under the Creative Commons CC0 public domain dedication.)

Anopheles darlingi is a major malaria vector in the Amazon region and, like other vectors, harbors a community of microorganisms with which it shares a network of interactions. Here, we describe the diversity and bacterial composition from the midguts and salivary glands of lab-reared and field-captured An. darlingi using metagenome sequencing of the 16S rRNA gene. The libraries were built using the amplification of the region V3-V4 16S rRNA gene. The bacterial community from the salivary glands was more diverse and richer than the community from the midguts. However, the salivary glands and midguts only showed dissimilarities in beta diversity between lab-reared mosquitoes. Despite that, intra-variability was observed in the samples. Acinetobacter and Pseudomonas were dominant in the tissues of lab-reared mosquitoes. Sequences of Wolbachia and Asaia were both found in the tissue of lab-reared mosquitoes; however, only Asaia was found in field-captured An. darlingi , but in low abundance. This is the first report on the characterization of microbiota composition from the salivary glands of An. darlingi from lab-reared and field-captured individuals. This study can provide invaluable insights for future investigations regarding mosquito development and interaction between mosquito microbiota and Plasmodium sp.

Safe and effective malaria transmission-blocking chemotherapeutics would allow a community-level approach to malaria control and eradication efforts by targeting the mosquito sexual stage of the parasite life cycle. However, only a single drug, primaquine, is currently approved for use in reducing transmission, and drug toxicity limits its widespread implementation. To address this limitation in antimalarial chemotherapeutics, we used a recently developed transgenic Plasmodium berghei line, Ookluc, to perform a series of high-throughput in vitro screens for compounds that inhibit parasite fertilization, the initial step of parasite development within the mosquito. Screens of antimalarial compounds, approved drug collections, and drug-like molecule libraries identified 185 compounds that inhibit parasite maturation to the zygote form. Seven compounds were further characterized to block gametocyte activation or to be cytotoxic to formed zygotes. These were further validated in mosquito membrane-feeding assays using Plasmodium falciparum and P. vivax. This work demonstrates that high-throughput screens using the Ookluc line can identify compounds that are active against the two most relevant human Plasmodium species and provides a list of compounds that can be explored for the development of new antimalarials to block transmission.

Objective:  To evaluate the seroprevalence of toxoplasmosis among puerperal women cared for at a tertiary university hospital and the level of understanding of these puerperal women about toxoplasmosis, vertical transmission, and its prophylaxis., Methods:  For this cross-sectional study, we evaluated 225 patients using presential interviews, prenatal documentation, and electronic medical records. Data were stored using Research Electronic Data Capture (REDCap) software. Prevalence rates were estimated by the presence of reactive IgG antibodies against Toxoplasma gondii . Data analysis was performed using the chi-square test and calculation of the odds ratio (OR). Seroreactivity to T. gondii and exposure variables (age, educational level, and parity) were analyzed using a confidence interval (95%CI) and a significance level of 5% (p < 0.05)., Results:  The seropositivity rate for T. gondii was 40%. There was no association between seroprevalence and age. Primiparity was a protective factor against seropositivity and low education was a risk factor., Conclusion:  Knowledge of T. gondii infection and its transmission forms was significantly limited, presenting a risk for acute maternal toxoplasmosis and vertical transmission of this protozoan. Increasing the education level regarding the risk of toxoplasmosis during pregnancy could reduce the rates of infection and vertical transmission of this parasite., Competing Interests: The authors have no conflict of interest to declare., (Federação Brasileira de Ginecologia e Obstetrícia. This is an open access article published by Thieme under the terms of the Creative Commons Attribution License, permitting unrestricted use, distribution, and reproduction so long as the original work is properly cited. (https://creativecommons.org/licenses/by/4.0/).)

Culicoides biting midges are capable to transmit Oropouche virus, Bluetongue virus and Mansonella spp. This study aimed to assess the utility of DNA barcode as an alternative method in the Culicoides species identification. The study was conducted in Jamari National Forest. Biting midges were collected using HP light traps during four months, February, April, August and October 2018. Insects were morphologically identified to the species level, and rest of the body were subjected to DNA extraction and PCR targeting a fragment of the cytochrome c oxidase subunit I (COI) gene, which were analyzed and deposited in GenBank. A phylogenetic gene tree was reconstructed using RAxML software, and the sequences were assigned at Molecular Operational Taxonomic Unit (MOTU) level by species delimitation algorithms. According to morphological approach, 18 species of 2 subgenera and 7 species groups were identified. A total of 191 new COI barcodes from 18 species were generated. Of these, fifteen species have been deposited for the first time in all datasets in the world. These sequences allowed the correct identification of 188 and 187 specimens according to the BM and BCM criteria, respectively. The intraspecific genetic distances ranged from 0 to 16.5%, while the interspecific ones ranged from 2.1 to 27.1%. The nominal species Culicoides glabellus and C. tetrathyris splitted into three and two MOTUs, respectively, except for mPTP, indicating a cryptic diversity in these species. Also, sequences of C. pseudodiabolicus formed two MOTUs using all algorithms, except for PTP and ABGD, suggesting the existence of two potential species. In contrast, some barcodes of C. quasiparaensis and C. paraensis merged into a single MOTU, which can be explained by the complex characteristics of the paraensis group, since these species have similar morphological characters. Here, we provided the first COI barcodes for biting midges in Rondônia and Brazil, and demonstrates that these are sufficient to discriminate between some species., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2022. Published by Elsevier B.V.)

Salt stress during pre-harvest limits the shelf life and post-harvest quality of produce; however, silicon nutrition can mitigate salt stress in plants. Thus, we evaluated the effects of salinity and fertilization with Si, in pre-harvest, on the morpho-physiological characteristics of onion bulbs during shelf life. The experiment was set up in randomized complete blocks, with treatments arranged in split-split plots. The plots had four levels of electrical conductivity of irrigation water (0.65, 1.7, 2.8, and 4.1 dS m -1 ). The subplots had five fertilization levels with Si (0, 41.6, 83.2, 124.8, and 166.4 kg ha -1 ). The sub-sub plots had four shelf times (0, 20, 40, and 60 days after harvest). Irrigation water salinity and shelf time reduced firmness and increased the mass loss of onion bulbs during shelf life. Salt stress reduced the contents of sugars and total soluble solids of onion bulbs during storage; however, Si supply improved the contents of these variables. Salinity, Si supply, and shelf time increased the concentrations of pyruvic and ascorbic acids in onion bulbs during shelf life. Si doses between 121.8 and 127.0 kg ha -1 attenuated the impacts caused by moderate salinity, increasing the synthesis of metabolites and prolonging the onion bulbs' shelf life.

Antibiotic treatment has been used to enhance anopheline susceptibility to Plasmodium infection, because bacterial microbiota play a fundamental role in modulating the vector competence of mosquitoes that transmit Plasmodium parasites. However, few studies have examined the impact of antibiotic treatments on Plasmodium vivax sporogonic development in neotropical anopheline mosquitoes. Herein, we assessed the impact of antibiotic treatment on P. vivax development and survival in Anopheles darlingi , the main vector of malaria in the Amazon region. Female mosquitoes were treated continuously with antibiotics to impact the gut bacterial load and then tested for prevalence, infection intensity, and survival in comparison with untreated mosquitoes. Antibiotic-fed mosquitoes had not dramatic impact on P. vivax development previously observed in P. falciparum . However, antibiotic treatment increases mosquito survival, which is known to increase vectorial capacity. These findings raise questions about the effect of antibiotics on P. vivax development and survival in An. darlingi ., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2022 Santos, Magi, Andrade, Bastos, Pereira, Medeiros and Araujo.)

Background: Sandflies are insects important for the transmission cycles of the leishmaniases. Despite being studied since the 1960s in the State of Rondônia (Brazil), several gaps exist regarding our working knowledge of these insects. This study aimed to construct an up-to-date database of sandflies using complementary information from the speciesLink database and the scientific literature, as well as to elaborate integrated abundance maps. We identified 153,155 records of sandflies captured in Rondônia between 1965-2021; after exclusion, 147,258 reports (speciesLink - 3,408, Rondônia studies - 143,850) associated with 15 genera and 140 species were mapped. The most abundant species observed were Psychodopygusdavisi (Root, 1934) (43,818 records), Nyssomyiawhitmani (Antunes & Coutinho, 1939) (12,594), Psychodopyguscarrerai (Barretto, 1946) (11,840), Psychodopygushirsutus (Mangabeira, 1942) (9,676), Nyssomyiaantunesi (Coutinho, 1939) (8,847), Trichophoromyiaubiquitalis (Mangabeira, 1942) (5,505), Psychodopygusgeniculatus (Mangabeira, 1941) (4,644), Pintomyianevesi (Damasceno & Arouck, 1956) (4,140), Trichophoromyiaauraensis (Mangabeira, 1942) (3,579), Psychodopyguscomplexus (Mangabeira, 1941) (2,659), Nyssomyiafraihai (Martins, Falcão & Silva, 1979) (2,504) and Bichromomyiaflaviscutellata (Mangabeira, 1942) (1,418). A total of 20 records of Leishmania detection corresponded to eight sand fly species. The present dataset provides updated information on the distribution of sandflies of Rondônia, including those considered potential vectors of Leishmania , which should prove useful to guide future studies., New Information: The present study provides an extensive dataset built from all studies reporting phlebotomine sandflies in the Brazilian State of Rondônia. Online distribution maps can aid scientists who wish to consult the updated list of sand fly species and view the distribution of these insects, as well as those considered potential vectors of Leishmania. The results of the present study can serve as the basis for future studies on sandflies conducted in the State., (Antonio Marques Pereira Júnior, Moreno Magalhães de Souza Rodrigues, Jansen Fernandes Medeiros.)

Ticks are obligatory parasites and transmit several pathogens to animals and humans. This study aimed to report the parasitism of ticks on carcasses of animals hunted by hunters from communities near Mapinguari National Park, at the shared border of Amazonas and Rondonia states, northern Brazil. The ticks were collected from heads of carcasses of the following animals: South American tapir (Tapirus terrestris), collared peccary (Pecari tajacu) and long-nosed armadillo (Dasypus beniensis). Six Amblyomma tick species were identified: Amblyomma coelebs, Amblyomma naponense, Amblyomma latepunctatum, Amblyomma oblongoguttatum, Amblyomma scalpturatum, Amblyomma triste and Amblyomma spp. The most abundant tick species were A. oblongoguttatum and A. scalpturatum. DNA extraction and polymerase chain reaction (PCR) were performed on all ticks to test for the presence of rickettsial gltA gene fragments, but no amplification was observed. This study reports, for the first time, the parasitism of A. coelebs in the armadillo D. beniensis and the presence of A. triste in the states of Amazonas and Rondonia, updating the tick fauna to 24 and 39 species for these states, respectively., (Copyright © 2022. Published by Elsevier GmbH.)

One strategy to identify transmission foci is based on vector monitoring, and efficient methods are important for vector control. Our study evaluated different light sources (red, green, ultraviolet, blue, and incandescent light) to capture sand fly in Porto Velho, Brazil. We also evaluated Leishmania and blood meal sources in females. A total of 1,943 individuals were identified in 45 species level-taxa, with Trichophoromyia ubiquitalis (Mangabeira) (n = 364), Nyssomyia antunesi (n = 241), Bichromomyia flaviscutellata (Mangabeira) (n = 222), and Psychodopygus davisi (Root) (n = 148) being the most abundant. Incandescent light captured most individuals (n = 589), followed by blue (n = 471), green (n = 452), ultraviolet (n = 281) and red (n = 150). No significant difference was observed between the species composition and lights (PERMANOVA: Pseudo F = 1.29, p = 0.14, NMDS: Stress 0.18). The Shannon and Simpson indices demonstrated a high diversity captured using all lights. Our data demonstrated that LEDs are alternative devices for sand fly capture, with blue and green LEDs presenting similar results to incandescent light. 53 pools were analyzed, only one pool was positive for kDNA and hsp70 targets [Nyssomyia fraihai (Martins, Falão & Silva)] and identified as Endotrypanum spp., suggesting that other trypanosomatids may circulate in the locality. Choloepus hoffmanni (two-toed sloth) Peters, Homo sapiens Linnaeus, Proechimys gardneri (Gardner's spiny rat) Da Silva, and Tamandua tetradactyla (lesser anteater) (Linnaeus) were blood meal sources identified in females, increasing the knowledge about sources used by these insects., (Copyright © 2022 Elsevier B.V. All rights reserved.)

Ribeirão das Pedras, a 10-km-long stream from the source to mouth, is part of a predominantly urban catchment located in Campinas metropolitan area in the state of São Paulo, Brazil, and it is also surrounded by sugarcane farms. Monthly sampling of 31 selected emerging contaminants (ECs) was conducted for 1 year (October 2018 to October 2019) in five points, including the spring, agricultural, and urban areas, to assess the dynamics and impact of ECs on the stream. The ECs were quantified using LC-MS/MS analysis. Out of the 31 ECs monitored in this study, 13 were detected in the Ribeirão das Pedras catchment, which were mainly pesticides and caffeine. Eight ECs (hexazinone, malathion, desethylatrazine (DEA), desisopropylatrazine (DIA), fipronil, ametryn, 2-hidroxyatrazine, and diuron) were detected with risk quotients higher than 1, indicating some level of environmental concern. Statistical analyses showed that caffeine, hexazinone, atrazine, DEA, and DIA were the most statistically important contaminants in temporal analysis, with caffeine concentrations varying randomly. Hexazinone, atrazine, DIA, and DEA concentrations increased from November 2018 to January 2019, and atrazine, hexazinone, and DEA concentrations increased from June 2019 to September 2019. Spatial analysis indicates that the spring of Ribeirão das Pedras is the only statistically different sampling point, with lower concentrations of EC. Points 3 and 5, both located in urban areas next to the stream's mouth, differ from each other due to the possible dilution of caffeine downstream of point 3 and domestic sewage discharge upstream of point 5., (© 2022. The Author(s), under exclusive licence to Springer Nature Switzerland AG.)

Chagas disease is a zoonotic disease caused by the hemoflagellate Trypanosoma cruzi and transmitted primarily by triatomine vectors. Triatomines are hematophagous insects that feed on a variety of vertebrate hosts. The Chagas disease transmission cycle is closely related to the interactions between vectors, parasites, and vertebrate hosts. Knowledge of triatomine food sources is critical to understanding Chagas disease transmission dynamics. The aim of this study was to identify blood meal sources used by triatomines from different environments in the Brazilian Amazon. A total of 25 captures were conducted in four environments. Triatomine specimens were captured on palm trees and were identified by morphological and morphometric characters. Blood meal sources identification was conducted using a traditional PCR followed by Sanger sequencing of mtDNA cytb gene. Sequencing was successful in 167 specimens and a total of 21 blood meal sources were identified: two reptilians, six birds, and 13 mammals. Among these 21 species, three (Tamandua tetradactyla, Didelphis marsupialis and Rattus rattus) are considered reservoir of T. cruzi. Knowledge of the relationship between triatomines and possible reservoirs can help to elucidate the enzootic cycle of T. cruzi in the Amazon region and guide control strategies for Chagas disease transmission in that region., (Copyright © 2022 Elsevier B.V. All rights reserved.)

Background: The colonization of mosquitoes susceptible to Plasmodium vivax via direct membrane feeding assay (DMFA) has the potential to significantly advance our knowledge of P. vivax biology, vector-parasite interaction and transmission-blocking vaccine research. Anopheles darlingi and Anopheles deaneorum are important vectors of malaria in the Western Brazilian Amazon. Since 2018, well-established colonies of these species have been maintained in order to mass produce mosquitoes destined for P. vivax infection. Plasmodium susceptibility was confirmed when the colonies were established, but susceptibility needs to be maintained for these colonies to remain good models for pathogen transmission. Thus, the susceptibility was assessed of colonized mosquitoes to P. vivax isolates circulating in the Western Amazon., Methods: Laboratory-reared mosquitoes from F10-F25 generations were fed on P. vivax blood isolates via DMFA. Susceptibility was determined by prevalence and intensity of infection as represented by oocyst load seven days after blood feeding, and sporozoite load 14 days after blood feeding. The effect of infection on mosquito survival was evaluated from initial blood feeding until sporogonic development and survival rates were compared between mosquitoes fed on infected and uninfected blood. Correlation was calculated between gametocytaemia and prevalence/intensity of infection, and between oocyst and sporozoite load., Results: Significant differences were found in prevalence and intensity of infection between species. Anopheles darlingi showed a higher proportion of infected mosquitoes and higher oocyst and sporozoite intensity than An. deaneorum. Survival analysis showed that An. deaneorum survival decreased drastically until 14 days post infection (dpi). Plasmodium vivax infection decreased survival in both species relative to uninfected mosquitoes. No correlation was observed between gametocytaemia and prevalence/intensity of infection, but oocyst and sporozoite load had a moderate to strong correlation., Conclusions: Colonized An. darlingi make excellent subjects for modelling pathogen transmission. On the other hand, An. deaneorum could serve as a model for immunity studies due the low susceptibility under current colonized conditions. In the application of DMFA, gametocyte density is not a reliable parameter for predicting mosquito infection by P. vivax, but oocyst intensity should be used to schedule sporozoite experiments., (© 2022. The Author(s).)

Objectives: To evaluate the effect of tDCS on tinnitus distress, loudness and psychiatric symptoms., Methods: A systematic literature search of PubMed, Web of Science, Cochrane Library, VHL, EMBASE, PsycINFO, OVID, and CINAHL databases was carried out on articles published until July 2021. Inclusion criteria were published controlled trials using tDCS intervention with tinnitus patients, using a sham/control group, and measuring tinnitus loudness, distress and/or psychiatric symptoms. A meta-analysis was performed for the overall effect as well as to compare subgroups according to tDCS target (left temporoparietal area (LTA) and dorsolateral prefrontal cortex (DLPFC))., Results: Fourteen articles with 1031 participants were included. Six studies applied tDCS over the DLPFC, six over the LTA and two over both areas. Although the overall meta-analysis showed that tDCS significantly decreased tinnitus loudness (SMD=-0.35; 95%CI=-0.62 to -0.08, p = 0.01) and distress (SMD=-0.50, 95%CI=-0.91 to -0.10, p = 0.02).The subgroup analysis showed a significant effect only for tDCS over LTA for loudness (SMD=-0.46, 95%CI=-0.80 to -0.12, p = 0.009), and no other area resulted in significant change. There was no significant effect of treatment on psychiatric symptoms., Conclusion: tDCS may improve tinnitus loudness and distress with a small to moderate effect size. Despite the overall positive effect, only LTA tDCS yielded a significant effect. Further well-controlled studies with larger sample sizes and broader exploration of tDCS montages and doses are warranted., Competing Interests: Declaration of Competing Interest None., (Copyright © 2021 Elsevier Masson SAS. All rights reserved.)

There is a lack of studies regarding tick-associated Rickettsia in the Amazon biome. Aiming to contribute to this knowledge, our research group collected ticks in the Western Amazon to better understand the tick fauna and their associated Rickettsia. In this study, we detected Rickettsia parkeri strain Atlantic rainforest DNA in the tick Amblyomma ovale Koch, 1844 in Rio Branco municipality, Acre state, northern Brazil. This is the first time that the R. parkeri strain Atlantic rainforest has been reported in the Amazon biome and is the first evidence of the circulation of a pathogenic spotted fever group (SFG) Rickettsia in this biome. This finding provides substantial information to help public health authorities understand which species of Rickettsia may be related to Amazon spotted fever cases., (Copyright © 2021. Published by Elsevier GmbH.)

Cutaneous Leishmaniasis (CL) is a disease caused by Leishmania species and transmitted by the bites of infected female sand fly species. The diversity of these insects in Rondônia State (where CL is the predominant form) is large but unexplored, and consequently the vector species are unknown. The purpose of this study was to evaluate the sand fly fauna in two environments (forest fragment and peridomicile) in rural areas of four municipalities of the state, DNA amplification of Leishmania species and the presence of blood meal sources for these insects. After identifying the species, sample coverage was applied to estimate the fauna coverage in each environment. Females were used to amplify and detect Leishmania DNA and blood meal sources, then posteriorly identified by sequencing. A total of 1706 individuals were included in 61 species, which was a sample coverage of 97% for the forest fragments (56 species), whereas 98% was observed (32 species) in peridomiciles. Next, 41 pools were prepared from 1227 females, and none were positive for Leishmania DNA. We observed 160 engorged females (forest fragments: 21, peridomiciles: 139) belonging to females of the Antunesi complex and another eight species. Two of these females were positive for Leishmania braziliensis-DNA: one in the Antunesi complex and one in Psychodopygus hirsutus. Seven blood meal sources were identified by sequencing analysis: Bos taurus and Sus scrofa from the peridomiciles, and Dasypus novemcinctus, Pecari tajacu, Philander canus, Plecturocebus bernhardi, and Tamandua tetradactyla from the forest fragments. Our data confirmed the feeding behavior of field-caught sand flies and could contribute to our understanding about local vectors and possible reservoirs in the transmission of Leishmania spp., (Copyright © 2021. Published by Elsevier B.V.)

Human malaria continues to be a public health problem and an important cause of morbidity and mortality in the world. Malaria control is achieved through both individual protection against mosquito bites and drug treatment, which is hampered by the spread of Plasmodium falciparum resistance to most antimalarials, including artemisinin derivatives. One of the key pharmacological strategies for controlling malaria is to block transmission of the parasites to their mosquito vectors. Following this rational, MEFAS, a synthetic hybrid salt derived from artesunate (AS) and mefloquine has been previously reported for its activity against asexual P. falciparum parasites in vitro, in addition to a pronounced reduction in the viability of mature gametocytes. Herein, MEFAS was tested against asexual forms of Plasmodium vivax and for its ability to block malaria transmission in Anopheles darlingi mosquitoes in a membrane feeding assay using P. vivax field isolates. MEFAS demonstrated high potency, with a IC50 of 6.5 nM against asexual forms of P. vivax. At 50 μM, MEFAS completely blocked oocyst formation in mosquitoes, regardless of the oocyst number in the control group. At lower doses, MEFAS reduced oocyst prevalence by greater than 20%. At equivalent doses, AS irregularly reduced oocyst formation and caused only slight inhibition of mosquito infections. These results highlight the potential of MEFAS as a novel transmission-blocking molecule, as well as its high blood schizonticidal activity against P. vivax and P. falciparum field isolates, representing a starting point for further development of a new drug with dual antimalarial activity., (Copyright © 2021 The Authors. Published by Elsevier Ltd.. All rights reserved.)

Leishmaniases are zoonotic diseases caused by protozoa of the genus Leishmania. The disease has two clinical manifestations described in humans: visceral (VL) and cutaneous (CL) leishmaniasis. In Brazil, there has been an expansion of human VL. The participation of the dog as a reservoir of Leishmania infantum, the agent of VL, is important for the epidemiology of the disease since canine cases generally precede human cases. The present study aimed to evaluate the occurrence of Leishmania spp. infection in dogs in the municipality of Ji-Paraná by PCR assays using blood samples. Leishmania DNA was detected in two of the 105 studied dogs. The PCR products were sequenced and confirmed that the two samples (1.90%) correspond to L. infantum. The dogs had allochthonous history. Therefore, the positive results found here should serve as a warning to public health agencies. This is because Ji-Paraná is the third municipality to register cases of canine leishmaniasis (CanL) in Rondônia state. Thus, reinforcing the importance of expanding studies on the epidemiology and surveillance of VL in the region.

Individuals with asymptomatic infection due to Plasmodium vivax are posited to be important reservoirs of malaria transmission in endemic regions. Here we studied a cohort of P. vivax malaria patients in a suburban area in the Brazilian Amazon. Overall 1,120 individuals were screened for P. vivax infection and 108 (9.6%) had parasitemia detected by qPCR but not by microscopy. Asymptomatic individuals had higher levels of antibodies against P. vivax and similar hematological and biochemical parameters compared to uninfected controls. Blood from asymptomatic individuals with very low parasitemia transmitted P. vivax to the main local vector, Nyssorhynchus darlingi. Lower mosquito infectivity rates were observed when blood from asymptomatic individuals was used in the membrane feeding assay. While blood from symptomatic patients infected 43.4% (199/458) of the mosquitoes, blood from asymptomatic infected 2.5% (43/1,719). However, several asymptomatic individuals maintained parasitemia for several weeks indicating their potential role as an infectious reservoir. These results suggest that asymptomatic individuals are an important source of malaria parasites and Science and Technology for Vaccines granted by Conselho Nacional de may contribute to the transmission of P. vivax in low-endemicity areas of malaria., Competing Interests: The authors have declared that no competing interests exist.

Culicoides biting midges are insects involved in the transmission of filarial nematodes, protozoans, and viruses. Greater knowledge of Culicoides blood meal sources could improve our understanding of parasite transmission cycles. Our study used molecular tools to evaluate the blood meal sources of Culicoides biting midges from sylvatic environments. This study was conducted in Jamari National Forest, Rondônia, Brazil. Culicoides were captured using HP light traps positioned at ground level (1.5 m above ground) and in canopy (15 m above ground). To identify blood meal sources, females were subjected to DNA extraction and PCR targeting the cytb gene fragment, and the obtained sequences were analyzed and compared with sequences from GenBank. DNA extraction and PCR were performed on 455 Culicoides females, and blood meal sources were identified in 186 females. Thirty Culicoides specimens were collected from the Potosi trail and 156 were collected from the Santa Maria trail. A total of 22 species were captured; all 22 species were collected in canopy (100%) but only three species were collected at ground level (13.6%). The cytb fragment was amplified in 162 of 186 samples. Sample sequencing identified cytb DNA from nine blood-meal sources: Pauxi sp., Psophia viridis, Ramphastos tucanus tucanus, Choloepus didactylus, Choloepus hoffmanni, Tamandua tetradactyla, Ateles chamek, Homo sapiens and Pithecia irrorata. We observed that several different blood meal sources were utilized by a high diversity of Culicoides species. The abundance of Culicoides in the canopy may be related to the fact that the majority of blood meal hosts feed in treetops. We observed that C. (Hoffmania) sp. and C. coutinhoi tend to be more generalist, feeding on a range of mammals and piciform, gruiform and galliform birds. This data improves our knowledge of the feeding profile of biting midges from forest environments and should serve as a future basis for defining zoonotic transmission cycles., (Copyright © 2021. Published by Elsevier B.V.)

Introduction: This study evaluated the epidemiology of American cutaneous leishmaniasis in the immediate region of Ji-Paraná, Rondônia State., Methods: Samples and epidemiological data were collected from 105 patients., Results: Leishmania infection was observed in 58 (55.2%) patients, and Leishmania braziliensis was present in 82.9% of the 41 sequenced samples. Infected patients were predominantly male (93.1%). Leishmania infection was twice as prevalent among rural inhabitants versus urban inhabitants. Lesions were more frequent in the upper limbs (arms/hands, 41.82%)., Conclusions: The present data corroborate the zoonotic profile of cutaneous leishmaniasis; this information could help to improve surveillance and control strategies.

The majority of malaria cases in South America occur in rural areas of the Amazon region. Although these areas have a significant impact on malaria cases, few entomological studies have been carried out there. This study aimed to describe entomological parameters in settlements in Rondonia State, Brazil. Collections of anopheles were carried out using the Protected Human Attraction Technique (PHAT). The risk and the potential for malaria transmission were assessed using the human biting rate (HBR), the sporozoite rate (SR) and the entomological inoculation rate (EIR). The results confirmed that Nyssorhynchus darlingi is the predominant species in the two studied locations. Although settlement in the two study sites has occurred at different times, the species richness found was low, showing that environmental changes caused by anthropological actions have probably favor the adaptation of Ny. darlingi species. From the total of 615 anopheline mosquitoes assessed, seven (1.1%) were positive for Plasmodium sp. infections. The EIR revealed that Ny. darlingi contributes to malaria transmission in both locations, as it was responsible for 0.05 infectious bites in humans at night in the old settlement and 0.02 in the recent occupation. In the two study sites, the biting occurred more frequently at dusk. Nyssorhynchus darlingi was prevalent in areas of recent colonization but, even when present in a low density, this species could maintain the transmission of malaria in the older settlement. The entomological information obtained in this study is important and may aid the selection of vector control actions in these locations.

The taxonomic identity of two species of sand flies, Psathyromyia pradobarrientosi (Le Pont, Matias, Martinez & Dujardin, 2004) and Psathyromyia runoides (Fairchild & Hertig, 1953) (Diptera, Psychodidae), was evaluated morphologically and molecularly based upon specimens collected in Brazilian states. The morphological component compared collected specimens with paratypes of Pa. runoides and Pa. pradobarrientosi and their descriptions. Phylogenetic analysis of coI sequences of Pa. pradobarrientosi showed a well-supported group distinct from Pa. runoides. Morphologically, Psathyromyia runoides and Pa. pradobarrientosi males are distinguished by characteristics of the aedeagal ducts and parameral sheath in the genitalia; females are distinguished by the number and shape of the teeth in the cibarium and by the shape of the spermathecae. Given the morphological similarity between the males and the absence of the description of the female of Pa. pradobarrientosi, it is possible that specimens previously identified as Pa. runoides in Brazil are in fact Pa. pradobarrientosi., (© The Author(s) 2020. Published by Oxford University Press on behalf of Entomological Society of America.All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

Rondônia is the only state in the North Region of Brazil to have registered confirmed cases of Brazilian Spotted Fever (BSF). The present study investigated the epidemiological cycle of Rickettsia spp. by surveying free-living ixodofauna and tick parasitism of dogs in the municipality of Porto Velho, Rondônia State. Ticks and dogs were tested for the presence of Rickettsia spp. DNA and dog serum was tested for reactivity to anti-Rickettsia spp. antibodies. Tick collection and dog blood sampling were performed in peri-urban and rural environments at 11 locations. Eight free-living Amblyomma species and one Haemaphysalis species were collected: A. scalpturatum, A. naponense, A. oblongoguttatum, A. coelebs, A. latepunctatum, A. pacae, A. ovale, Amblyomma sp., and H. juxtakochi. Three tick species were found parasitizing dogs: Rhipicephalus sanguineus sensu lato, A. oblongoguttatum and A. ovale. Molecular analysis did not identify the presence of the gltA gene fragment in any tick specimen. Results from an indirect immunofluorescent assay (IFA) showed that 20.8% of peri-urban and 15.4% of rural dog sera exhibited reactivity to Rickettsia rhipicephali, Rickettsia amblyommatis, Rickettsia bellii and Rickettsia parkeri antigens. Antibody prevalence in dogs was 16.4%. This study is the first to describe the prevalence of Rickettsia spp. infection in dogs from Porto Velho municipality. Our findings enhance current knowledge of Rickettsia spp. circulation in the Western Amazon.

Culicoides biting midges are small hematophagous flies, some species of which transmit pathogens to livestock and humans. In the Brazilian Amazon Basin, Culicoides diversity has remained largely unexamined. Jamari National Forest (JNF) is a conservation area in Rondônia State where research and environmental education are conducted in conjunction with the protection of natural resources, and the recovery of degraded areas. The present study augments our knowledge of Culicoides diversity in JNF. The survey collected 640 individuals from 41 species; 632 individuals were collected by HP light trap and 8 were collected by the BG Sentinel trap. Twelve species are first-time records for Rondônia: Culicoides acotylus Lutz, Culicoides albuquerquei Wirth & Blanton, Culicoides baniwa Felippe-Bauer, Culicoides bricenoi Ortiz, Culicoides brownei Spinelli, Culicoides carsiomelas Wirth & Blanton, Culicoides debilipalpis Lutz, Culicoides forattinii Ortiz, Culicoides ginesi Ortiz, Culicoides profundus Santarém, Felippe-Bauer & Trindade, Culicoides pseudoreticulatus Santarém, Felippe-Bauer & Castellón, and Culicoides spurius Santarém, Felippe-Bauer & Trindade. The most abundant species were Culicoides glabrior Macfie, Culicoides pseudodiabolicus Fox, Culicoides debilipalpis, Culicoides quasiparaensis Clastrier, and Culicoides brownei Spinelli. The present study updates the Culicoides species count in Rondônia to 55, and highlights the importance of JNF as a hotbed for biting midge diversity., (© The Author(s) 2020. Published by Oxford University Press on behalf of Entomological Society of America. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)