Bacterial diversity on larval and female Mansonia spp. from different localities of Porto Velho, Rondonia, Brazil.

Studies based on the bacterial diversity present in Mansonia spp. are limited; therefore, the aim of this study was to investigate the bacterial diversity in females and larvae of Mansonia spp., describe the differences between the groups identified, and compare the microbiota of larvae from different collection sites. Sequences of the 16S rRNA region from the larvae and females of Mansonia spp. were analyzed. Diversity analyzes were performed to verify the possible bacterial differences between the groups and the collection sites. The results showed Pseudomonas was the most abundant genus in both females and larvae, followed by Wolbachia in females and Rikenellaceae and Desulfovibrio in larvae. Desulfovibrio and Sulfurospirillum, sulfate- and sulfur-reducing bacteria, respectively, were abundant on the larvae. Aminomonas, an amino acid-degrading bacterium, was found only in larvae, whereas Rickettsia was identified in females. Bacterial diversity was observed between females and larvae of Mansonia and between larvae from different collection sites. In addition, the results suggest that the environment influenced bacterial diversity.

Evaluation of Long-lasting insecticidal nets (LLINs) for malaria control in an endemic area in Brazil.

BACKGROUND: Most cases of malaria in Brazil are concentrated in the Amazon region. One of the vector control alternatives recommended by the WHO is the long-lasting insecticidal net (LLIN). This tool is used in the nine federal states of the Brazilian Legal Amazon, where LLINs are essential for reducing vector density and disease transmission as they prevent contact between the mosquito and the individual. The objective of this study was to evaluate the residuality and use of LLIN insecticides in different health regions in a city located in the Brazilian Amazon. METHODS: A total of 17,027 LLINs were installed in the third, fifth and ninth health regions of the municipality of Porto Velho, Rondonia State, Brazil. The LLINs were of two types: Olyset (permethrin), for around the bed, and Interceptor (alphacypermethrin), for around hammocks. The residuality of 172 LLINs was evaluated using cone bioassays to verify the mortality rate of the mosquito Nyssorhynchus darlingi, over a period of 2 years. Structured questionnaires on the acceptance and use of LLINs were distributed to the participating population (n = 391), covering a total sample of 1147 mosquito nets. The mortality rate was evaluated both in terms of days after LLIN installation and the type of insecticide used. Statistical analyses were based on analysis of variance (ANOVA) and Chi-square and were performed using the SPSS statistical program. RESULTS: For the Ny. darlingi mosquito, Interceptor-type LLINs showed residual efficacy, with mortality rates ≥ 80% during the 2-year study period, as determined by the WHO. In contrast, Olyset-type LLINs were associated with a reduction in mortality rates, with 76% and 45% mortality rates in the last two assessments, which occurred during the last 6 months of the study period. Based on the structured questionnaires, the acceptance rate, i.e. percentage of individuals accepting the permanence of the 1147 LLINs sampled, in the three health regions of Porto Velho was 93.8% (of 1076 LLINs). CONCLUSION: The alphacypermethrin-impregnated LLIN was more effective than the LLIN impregnated with permethrin. The results indicate that the correct use of mosquito nets-and consequently the protection of the population-needs to be supported by health promotion actions. These initiatives are considered to be essential for the success of this vector control strategy. New studies that consider the monitoring of the placement of mosquito nets are necessary to provide effective support in the correct use of this methodology.

Oviposition Activity of Mansonia Species in Areas Along the Madeira River, Brazilian Amazon Rainforest.

Mansonia are aggressive mosquito species that are abundant in aquatic ecosystems where the macrophyte plants occur. These mosquitoes are commonly found across the Amazon/Solimões River basin. However, little is known about the oviposition behavior of these species. In the present study, we registered observations on the oviposition activity of 3 species: Mansonia amazonensis, Ma. humeralis, and Ma. cf. titillans, in 5 species of macrophytes in the vicinities of the Madeira River, Porto Velho, State of Rondônia, Brazil. Overall, 197 egg batches were collected. A greater amount of egg batches was found in Salvinia molesta as compared with other macrophytes sampled. In addition, 2 new oviposition habitats were noted in Ludwigia helmintorrhiza and Limnobium spongia. These findings will be important to understand the reproductive dynamics of these mosquitoes in the Brazilian Amazon basin.

Host Feeding Patterns of Mansonia (Diptera, Culicidae) in Rural Settlements near Porto Velho, State of Rondonia, Brazil.

Mosquito females of the genus Mansonia (Blanchard) can be a nuisance to humans and animals since they are voraciously hematophagous and feed on the blood of a variety of vertebrates. Despite their relevance, there is a lack of investigation into the blood-feeding patterns of the Mansonia species. Knowledge of the host preference is crucial in establishing the public health importance of a mosquito species and its potential to be involved in the transmission dynamics of pathogens. Species that are primarily anthropophilic can be more effective in spreading vector-borne pathogens to humans. In this study, we used an Illumina Nextera sequencing protocol and the QIIME2 workflow to assess the diversity of DNA sequences extracted in the ingested blood of mosquito species to evaluate the overall and local host choices for three species: Ma. titillans, Ma. Amazonensis, and Ma. humeralis, in rural areas alongside the Madeira River in the vicinities of the Santo Antonio Energia (SAE) reservoir in the municipality of Porto Velho, Rondônia, Western Brazil. By performing our analysis pipeline, we have found that host diversity per collection site showed a significant heterogeneity across the sample sites. In addition, in rural areas, Ma. amazonensis present a high affinity for B. taurus, Ma. humeralis shows an overall preference for C. familiaris and B. taurus, but also H. sapiens and E. caballus in urban areas, and Ma. titillans showed more opportunistic behavior in rural areas, feeding on wild animals and G. gallus, though with an overall preference for H. sapiens.

Next-Generation High-Throughput Sequencing to Evaluate Bacterial Communities in Freshwater Ecosystem in Hydroelectric Reservoirs.

The quality of aquatic ecosystems is a major public health concern. The assessment and management of a freshwater system and the ecological monitoring of microorganisms that are present in it can provide indicators of the environment and water quality to protect human and animal health. with bacteria is. It is a major challenge to monitor the microbiological bacterial contamination status of surface water associated with anthropogenic activities within rivers and freshwater reservoirs. Understanding the composition of aquatic microbial communities can be beneficial for the early detection of pathogens, improving our knowledge of their ecological niches, and characterizing the assemblages of microbiota responsible for the degradation of contaminants and microbial substrates. The present study aimed to characterize the bacterial microbiota of water samples collected alongside the Madeira River and its small tributaries in rural areas near the Santo Antonio Energia hydroelectric power plant (SAE) reservoir in the municipality of Porto Velho, Rondonia state, Western Brazil. An Illumina 16s rRNA metagenomic approach was employed and the physicochemical characteristics of the water sample were assessed. We hypothesized that both water metagenomics and physicochemical parameters would vary across sampling sites. The most abundant genera found in the study were Acinetobacter, Deinococcus, and Pseudomonas. PERMANOVA and ANCOM analysis revealed that collection points sampled at the G4 location presented a significantly different microbiome compared to any other group, with the Chlamidomonadaceae family and Enhydrobacter genus being significantly more abundant. Our findings support the use of metagenomics to assess water quality standards for the protection of human and animal health in this microgeographic region.

Mansonia spp. population genetics based on mitochondrion whole-genome sequencing alongside the Madeira River near Porto Velho, Rondonia, Brazil.

In high abundance, females of the genus Mansonia (Blanchard) can be a nuisance to humans and animals because they are voraciously hematophagous and feed on the blood of a myriad of vertebrates. The spatial-temporal distribution pattern of Mansonia species is associated with the presence of their host plants, usually Eichhornia crassipes, E. azurea, Ceratopteris pteridoides, Limnobium laevigatum, Pistia stratiotes, and Salvinia sp. Despite their importance, there is a lack of investigation on the dispersion and population genetics of Mansonia species. Such studies are pivotal to evaluating the genetic structuring, which ultimately reflects populational expansion-retraction patterns and dispersal dynamics of the mosquito, particularly in areas with a history of recent introduction and establishment. The knowledge obtained could lead to better understanding of how anthropogenic changes to the environment can modulate the population structure of Mansonia species, which in turn impacts mosquito population density, disturbance to humans and domestic animals, and putative vector-borne disease transmission patterns. In this study, we present an Illumina NGS sequencing protocol to obtain whole-mitogenome sequences of Mansonia spp. to assess the microgeographic genetic diversity and dispersion of field-collected adults. The specimens were collected in rural environments in the vicinities of the Santo Antônio Energia (SAE) hydroelectric reservoir on the Madeira River.

Seasonality and Biting Behavior of Mansonia (Diptera, Culicidae) in Rural Settlements Near Porto Velho, State of Rondônia, Brazil.

Mansonia (Diptera: Culicidae) are known to cause discomfort to the local populations of Amazon. Considering the fact that the effective control of these mosquitoes can only be obtained by understanding their ecology and behavior, entomological monitoring becomes essential. In view of this, mosquitoes of the genus Mansonia were collected by human landing catches (HLC) from 2015 to 2019, in four locations of Porto Velho, Rondônia, Brazil. The collections were performed inside and outside the homes, once in every four months, uninterrupted for 24 hr. Human bite indices/hour was used to analyze the hourly activity of the species for different seasons and environment (indoor and outdoor). Moreover, nonparametric Mann-Whitney tests were conducted to indicate if there were differences between exophagic and endophagic behavior. The seasonality of Mansonia species was also analyzed. Overall, 96,766 specimens were collected over five years of sampling. Mansonia titillans (Walker) was found to be the most abundant species (76.9%). The highest percentage of mosquitoes was collected in February (48.4%), followed by October (39.6%) and June (12.0%). The biting activity of the two most abundant species showed peak host seeking activity/behavior during twilight and night, more perceptible in the outdoor environment (peridomiciliary). In general, seasonality showed a tendency towards a reduction in the abundance of Mansonia in the years after 2015. Our results will be essential in the formulation of effective control methodology for Mansonia in the studied area.

Efficient Monitoring of Adult and Immature Mosquitoes Through Metabarcoding of Bulk Samples: A Case Study for Non-Model Culicids With Unique Ecologies.

The rapid and economical monitoring of mosquitos is imperative to understanding the dynamics of both disease vectors and nuisance species. In light of technological advances in mosquito sampling and DNA sequencing, health agencies can now utilize the full potential of metabarcoding pipelines for rapid and standardizable surveillance. Here, we describe mosquito spatial and temporal variation, with particular focus on Mansonia Blanchard species, in the Madeira (Rondônia State) and the Ribeira (São Paulo) watersheds, Brazil using metabarcoding of the D2 rDNA marker. Sampling and molecular pipelines were used to evaluate the taxonomic contribution of mosquitos in pools of culicids collected en masse from macrophyte-roots (immatures) and from Mosquito Magnet traps and protected human landings (adults). Results for adult captures are comparable to morphological diagnoses and clarify previously unknown temporal and spatial species turnover. Metabarcoding of immature stages also confirmed the extent of the geographical distribution of some species and each taxon's association with macrophyte species. Given the benefits of metabarcoding, such as taxonomic acuity, high throughput processing, and objectivity, we suggest such techniques should be more fully incorporated into culicid monitoring schemes. The metabarcoding protocol described herein paired with standardized field sampling schemes, when used by mosquito monitoring professionals, offers substantial improvements in terms of practicality, speed and cost.

Dynamics of Hg and MeHg in the Madeira River basin (Western Amazon) before and after impoundment of a run-of-river hydroelectric dam.

The impact of a hydroelectric run-of-river (RoR) dam construction on the dynamics of total mercury (THg) and methylmercury (MeHg) is of interest to the environment and health of human and wild life. We monitored (May 2010 to October 2018) THg and MeHg in the waters and in the suspended particulate matter (SPM) of the Madeira River and its tributaries (before and after dam construction) to evaluate changes upstream and downstream from the Santo Antonio Hydroelectric Dam (SAHD). We collected 2826 samples of water and SPM at sampling stations upstream (UPMD-01, UPMD-02 and UPMD-03) and downstream the Madeira River (DWMD-04, DWMD-05 and DWMD-06), and Tributaries upstream (Branco River, Jaci-Parana River, Jatuarana-I Igarapé, Contra River, Caripunas Igarapé, Ceara Igarapé, and Teotonio Igarapé) and downstream (Jatuarana-II Igarapé and Belmont Igarapé) from the SAHD and monitored water and the total load of SPM. SPM was significantly higher in the Madeira River (median: 178.63 mg.L-1) than in upstream and downstream tributaries (median: 15.30 mg.L-1); however, the THg and MeHg concentrations were significantly higher in the tributaries (median: 161.14 ng g-1 and 9.03 ng g-1, respectively) than in the mainstream Madeira River (median = 57.06 ng g-1 and 1.63 ng g-1, respectively). THg concentration in the water was significantly higher in the mainstream Madeira River (median = 6.51 ng.L-1) than in the tributaries (median = 2.57 ng.L-1). However, the percentage of methylation in the tributaries (median = 4.9%) was 4-times higher than in the Madeira River (median: 1.3%). The significantly higher MeHg percentages in the tributaries may indicate natural (hydro-bio-geochemical factors) still predominant in this changing landscape of the Western Amazon. So far, the data suggest that this RoR has not yet impacted the THg and MeHg concentrations.

Culicidae-centric metabarcoding through targeted use of D2 ribosomal DNA primers.

A practical limitation to many metabarcoding initiatives is that sampling methods tend to collect many non-target taxa, which become "amplicon noise" that can saturate Next Generation Sequencing results and lead to both financial and resource inefficiencies. An available molecular tool that can significantly decrease these non-target amplicons and decrease the need for pre-DNA-extraction sorting of bycatch is the design of PCR primers tailored to the taxa under investigation. We assessed whether the D2 extension segment of the 28S ribosomal operon can limit this shortcoming within the context of mosquito (Culicidae) monitoring. We designed PCR primers that are fully conserved across mosquitos and exclude from amplification most other taxa likely to be collected with current sampling apparatuses. We show that, given enough sequencing depth, D2 is an effective marker for the detection of mosquito sequences within mock genomic DNA pools. As few as 3,050 quality-filtered Illumina reads were able to recover all 17 species in a bulk pool containing as little as 0.2% of constituent DNA from single taxa. We also mixed these mosquito DNA pools with high concentrations of non-Culicidae bycatch DNA and show that the component mosquito species are generally still recoverable and faithful to their original relative frequencies. Finally, we show that there is little loss of fidelity in abundance parameters when pools from degraded DNA samples were sequenced using the D2 primers.

Efficacy of insecticides used in indoor residual spraying for malaria control: an experimental trial on various surfaces in a "test house".

BACKGROUND: Malaria is a public health problem in the Brazilian Amazon region. In integrated vector management for malaria (anopheline) control, indoor residual spraying (IRS) represents one of the main tools in the basic strategy applied in the Amazonian states. It is essential to understand the residual efficacy of insecticides on different surfaces to determine spray cycles, ensure their rational use, and prevent wastage. This study aimed to evaluate the residual efficacy of six insecticide formulations used in the National Malaria Control Programme on four different types of walls in a field simulation at a "test house". METHODS: The tests were performed as a field-simulating evaluation at a "test house" built in the municipality of Macapá. Six insecticide formulations comprising four pyrethroids, a carbamate, and an organophosphate were used, and evaluated when applied on different wall surfaces: painted wood, unpainted wood, plastered cement, and unplastered cement. The insecticides were applied to the interior walls of the "test house" by a trained technician. RESULTS: In the bioassays performed with pyrethroids, deltamethrin water-dispersible granules (WG) performed particularly well, presenting residual bioefficacy of 8 months on both wood surfaces after the IRS, whereas alpha-cypermethrin suspension concentrate (SC) and etofenprox wettable powder (WP) demonstrated residual bioefficacy of 4 months on at least one of the wood surfaces; however, the pyrethroid lambda-cyhalothrin WP showed a low residual bioefficacy (< 3 months) on all tested surfaces, demonstrating its inefficiency for areas with a long transmission cycle of malaria. For the carbamate-bendiocarb WP, residual bioefficacy for 3 months was achieved only on wood surfaces. In general, the organophosphate pirimifos-methyl capsule suspension (CS) demonstrated the best result, with a mortality rate < 80% over a period of 6 months on all surfaces tested. CONCLUSION: Insecticide efficiency varies among different types of surface; therefore, a "test house" is a valuable evaluation tool. This work highlights the usefulness of associating the residual efficacy of insecticides on the surfaces commonly found in houses in endemic areas, together with knowledge about the transmission cycle duration of the transmission cycle and the insecticide susceptibility of the vector. This association helps in the decision-making for the malaria control intervention regarding.

Phlebotominae sand flies (Diptera: Psychodidae): potential vectors of American cutaneous leishmaniasis agents in the area associated with the Santo Antônio Hydroelectric System in Western Amazonian Brazil.

INTRODUCTION: An entomological study was conducted as part of a vector-monitoring program in the area associated with the Santo Antônio hydroelectric system in State of Rondônia, Western Amazonian Brazil. METHODS: Fourteen sampling sites were surveyed to obtain data on the potential vectors of Leishmania spp. in the area. Sand flies were collected from 2011 to 2014 during the months of January/February (rainy season), May/June (dry season), and September/October (intermediary season) using light traps arranged in three vertical strata (0.5, 1, and 20m). RESULTS: A total of 7,575 individuals belonging to 62 species/subspecies were collected. The five most frequently collected sand flies were Psychodopygus davisi (Root) (36.67%), Trichophoromyia ubiquitalis (Mangabeira) (8.51%), Nyssomyia umbratilis (Ward & Fraiha) (6.14%), Bichromomyia flaviscutellata (Mangabeira) (5.74%), and Psychodopygus complexus (Mangabeira) (5.25%). These species have been implicated in the transmission of American cutaneous leishmaniasis agents in the Brazilian Amazon region and described as potential vectors of this disease in the study area. CONCLUSIONS: Additional surveillance is needed, especially in areas where these five species of sand fly are found.

Phlebotominae sand flies (Diptera: Psychodidae): potential vectors of American cutaneous leishmaniasis agents in the area associated with the Santo Antônio Hydroelectric System in Western Amazonian Brazil

INTRODUCTION: An entomological study was conducted as part of a vector-monitoring program in the area associated with the Santo Antônio hydroelectric system in State of Rondônia, Western Amazonian Brazil. METHODS: Fourteen sampling sites were surveyed to obtain data on the potential vectors of Leishmania spp. in the area. Sand flies were collected from 2011 to 2014 during the months of January/February (rainy season), May/June (dry season), and September/October (intermediary season) using light traps arranged in three vertical strata (0.5, 1, and 20m). RESULTS : A total of 7,575 individuals belonging to 62 species/subspecies were collected. The five most frequently collected sand flies were Psychodopygus davisi (Root) (36.67%), Trichophoromyia ubiquitalis (Mangabeira) (8.51%), Nyssomyia umbratilis (Ward & Fraiha) (6.14%), Bichromomyia flaviscutellata (Mangabeira) (5.74%), and Psychodopygus complexus (Mangabeira) (5.25%). These species have been implicated in the transmission of American cutaneous leishmaniasis agents in the Brazilian Amazon region and described as potential vectors of this disease in the study area. CONCLUSIONS: Additional surveillance is needed, especially in areas where these five species of sand fly are found. .