Forest cover percentage drives the peak biting time of Nyssorhynchus darlingi (Diptera: Culicidae) in the Brazilian Amazon.

BACKGROUND: Deforestation is an important driver of malaria dynamics, with a relevant impact on mosquito ecology, including larval habitat availability, blood-feeding behaviour, and peak biting time. The latter is one of several entomological metrics to evaluate vectorial capacity and effectiveness of disease control. This study aimed to test the effect of forest cover percentage on the peak biting time of Plasmodium-uninfected and infected Nyssorhynchus darlingi females. METHODS: Mosquitoes were captured utilizing human landing catch (HLC) in the peridomestic habitat in field collections carried out in the wet, wet-dry transition, and dry seasons from 2014 to 2017 in areas with active malaria transmission in Amazonian Brazil. The study locations were in rural settlements in areas with the mean annual malaria parasite incidence (Annual Parasite Incidence, API ≥ 30). All Ny. darlingi females were tested for Plasmodium spp. infection using real time PCR technique. Forest cover percentage was calculated for each collection site using QGIS v. 2.8 and was categorized in three distinct deforestation scenarios: (1) degraded, < 30% forest cover, (2) intermediate, 30-70% forest cover, and (3) preserved, > 70% forest cover. RESULTS: The highest number of uninfected female Ny. darlingi was found in degraded landscape-sites with forest cover < 30% in any peak biting time between 18:00 and 0:00. Partially degraded landscape-sites, with (30-70%) forest cover, showed the highest number of vivax-infected females, with a peak biting time of 21:00-23:00. The number of P. falciparum-infected mosquitoes was highest in preserved sites with > 70% forest cover, a peak biting at 19:00-20:00, and in sites with 30-70% forest cover at 22:00-23:00. CONCLUSIONS: Results of this study show empirically that degraded landscapes favour uninfected Ny. darlingi with a peak biting time at dusk (18:00-19:00), whereas partially degraded landscapes affect the behaviour of Plasmodium-infected Ny. darlingi by shifting its peak biting time towards hours after dark (21:00-23:00). In preserved sites, Plasmodium-infected Ny. darlingi bite around dusk (18:00-19:00) and shortly after (19:00-20:00).

The COVID-19 crisis and Amazonia's indigenous people: Implications for conservation and global health.

Ecosystem health and zoonotic diseases are closely interwoven. Even as we grapple with the SARS-Coronavirus-2 pandemic, which may have its origins in wildlife, weakening environmental policies in the Brazilian Amazon are elevating the risk of additional zoonotic spillover events. We examine the links between deforestation and disease emergence in the Amazon, as illustrated by outbreaks of yellow fever virus, Venezuelan equine encephalitis virus, and Oropouche virus. It has been well established that in Brazil, indigenous territories exhibit lower rates of forest conversion and degradation than in areas designated for sustainable use. In this way, Amazonia's indigenous tribes promote public health while sustaining ecosystem services. However, indigenous land rights are under attack due to current policies enabling illegal land grabbing, mining and logging. Further adding to the existential struggle of indigenous tribes, malaria and SARS-Coronavirus-2 are wreaking havoc on these vulnerable populations. There is a critical need for protection of indigenous people's rights and health, as well as a sustained effort to support the study of mechanisms underlying anthropogenic land use change and zoonotic disease risk.

Bacterial diversity associated with the abdomens of naturally Plasmodium-infected and non-infected Nyssorhynchus darlingi.

BACKGROUND: The bacterial community present in the abdomen in Anophelinae mosquitoes can influence mosquito susceptibility to Plasmodium infection. Little is known about the bacteria associated with Nyssorhynchus darlingi, a primary malaria vector in the Amazon basin. We investigated the abdominal bacterial community compositions of naturally Plasmodium-infected (P-positive, n = 9) and non-infected (P-negative, n = 7) Ny. darlingi from the Brazilian Amazon region through massive parallel sequencing of the bacterial V4 variable region of the 16S rRNA gene. RESULTS: Bacterial richness of Ny. darlingi encompassed 379 operational taxonomic units (OTUs), the majority of them belonging to the Proteobacteria, Firmicutes and Bacteroides phyla. Escherichia/Shigella and Pseudomonas were more abundant in the P-positive and P-negative groups, respectively, than in the opposite groups. Enterobacter was found only in the P-negative group. The results of statistical analyses conducted to compare bacterial abundance and diversity between Plasmodium-infected and Plasmodium-non-infected mosquitoes were not significant. CONCLUSIONS: This study increased knowledge about bacterial composition in Ny. darlingi and revealed that Plasmodium-positive and Plasmodium-negative groups share a common core of bacteria. The genera Prevotella 9, Sphingomonas, Bacteroides, and Bacillus were reported for the first time in Ny. darlingi.

First record of translocation in Culicidae (Diptera) mitogenomes: evidence from the tribe Sabethini.

BACKGROUND: The tribe Sabethini (Diptera: Culicidae) contains important vectors of the yellow fever virus and presents remarkable morphological and ecological diversity unequalled in other mosquito groups. However, there is limited information about mitochondrial genomes (mitogenomes) from these species. As mitochondrial genetics has been fundamental for posing evolutionary hypotheses and identifying taxonomical markers, in this study we sequenced the first sabethine mitogenomes: Sabethes undosus, Trichoprosopon pallidiventer, Runchomyia reversa, Limatus flavisetosus, and Wyeomyia confusa. In addition, we performed phylogenetic analyses of Sabethini within Culicidae and compared its mitogenomic architecture to that of other insects. RESULTS: Similar to other insects, the Sabethini mitogenome contains 13 protein-coding genes, 22 transfer RNA genes, two ribosomal RNA genes, and a control region. However, the gene order is not the same as that in other mosquitoes; the tyrosine (Y) and cysteine (C) tRNA genes have translocated. In general, mitogenome rearrangements within insects are uncommon events; the translocation reported here is unparalleled among Culicidae and can be considered an autapomorphy for the Neotropical sabethines. CONCLUSIONS: Our study provides clear evidence of gene rearrangements in the mitogenomes of these Neotropical genera in the tribe Sabethini. Gene order can be informative at the taxonomic level of tribe. The translocations found, along with the mitogenomic sequence data and other recently published findings, reinforce the status of Sabethini as a well-supported monophyletic taxon. Furthermore, T. pallidiventer was recovered as sister to R. reversa, and both were placed as sisters of other Sabethini genera (Sabethes, Wyeomyia, and Limatus).

Comparison of malaria incidence rates and socioeconomic-environmental factors between the states of Acre and Rondônia: a spatio-temporal modelling study.

BACKGROUND: Plasmodium falciparum malaria is a threat to public health, but Plasmodium vivax malaria is most prevalent in Latin America, where the incidence rate has been increasing since 2016, particularly in Venezuela and Brazil. The Brazilian Amazon reported 193,000 cases in 2017, which were mostly confirmed as P. vivax (~ 90%). Herein, the relationships among malaria incidence rates and the proportion of accumulated deforestation were contrasted using data from the states of Acre and Rondônia in the south-western Brazilian Amazon. The main purpose is to test the hypothesis that the observed difference in incidence rates is associated with the proportion of accumulated deforestation. METHODS: An ecological study using spatial and temporal models for mapping and modelling malaria risk was performed. The municipalities of Acre and Rondônia were the spatial units of analysis, whereas month and year were the temporal units. The number of reported malaria cases from 2009 until 2015 were used to calculate the incidence rate per 1000 people at risk. Accumulated deforestation was calculated using publicly available satellite images. Geographically weighted regression was applied to provide a local model of the spatial heterogeneity of incidence rates. Time-series dynamic regression was applied to test the correlation of incidence rates and accumulated deforestation, adjusted by climate and socioeconomic factors. RESULTS: The malaria incidence rate declined in Rondônia but remained stable in Acre. There was a high and positive correlation between the decline in malaria and higher proportions of accumulated deforestation in Rondônia. Geographically weighted regression showed a complex relationship. As deforestation increased, malaria incidence also increased in Acre, while as deforestation increased, malaria incidence decreased in Rondônia. Time-series dynamic regression showed a positive association between malaria incidence and precipitation and accumulated deforestation, whereas the association was negative with the human development index in the westernmost areas of Acre. CONCLUSION: Landscape modification caused by accumulated deforestation is an important driver of malaria incidence in the Brazilian Amazon. However, this relationship is not linearly correlated because it depends on the overall proportion of the land covered by forest. For regions that are partially degraded, forest cover becomes a less representative component in the landscape, causing the abovementioned non-linear relationship. In such a scenario, accumulated deforestation can lead to a decline in malaria incidence.

Nyssorhynchus dunhami: bionomics and natural infection by Plasmodium falciparum and P. vivax in the Peruvian Amazon.

BACKGROUND Nyssorhynchus dunhami, a member of the Nuneztovari Complex, has been collected in Brazil, Colombia, and Peru and described as zoophilic. Although to date Ny. dunhami has not been documented to be naturally infected by Plasmodium, it is frequently misidentified as other Oswaldoi subgroup species that are local or regional malaria vectors. OBJECTIVES The current study seeks to verify the morphological identification of Nuneztovari Complex species collected in the peri-Iquitos region of Amazonian Peru, to determine their Plasmodium infection status, and to describe ecological characteristics of their larval habitats. METHODS We collected Ny. nuneztovari s.l. adults in 2011-2012, and Ny. nuneztovari s.l. larvae and adults in 2016-2017. When possible, samples were identified molecularly using cytochrome c oxidase subunit I (COI) barcode sequencing. Adult Ny. nuneztovari s.l. from 2011-2012 were tested for Plasmodium using real-time PCR. Environmental characteristics associated with Ny. nuneztovari s.l. larvae-positive water bodies were evaluated. FINDINGS We collected 590 Ny. nuneztovari s.l. adults and 116 larvae from eight villages in peri-Iquitos. Of these, 191 adults and 111 larvae were identified by COI sequencing; all were Ny. dunhami. Three Ny. dunhami were infected with P. falciparum, and one with P. vivax, all collected from one village on one night. Ny. dunhami larvae were collected from natural and artificial water bodies, and their presence was positively associated with other Anophelinae larvae and amphibians, and negatively associated with people living within 250m. MAIN CONCLUSIONS Of Nuneztovari Complex species, we identified only Ny. dunhami across multiple years in eight peri-Iquitos localities. This study is, to our knowledge, the first report of natural infection of molecularly identified Ny. dunhami with Plasmodium. We advocate the use of molecular identification methods in this region to monitor Ny. dunhami and other putative secondary malaria vectors to more precisely evaluate their importance in malaria transmission.

Anophelines species and the receptivity and vulnerability to malaria transmission in the Pantanal wetlands, Central Brazil.

BACKGROUND Studies on malaria vectors in the Pantanal biome, Central Brazil, were conducted more than half a century ago. OBJECTIVES To update anopheline records and assess receptivity and vulnerability to malaria transmission. METHODS Five-day anopheline collections were conducted bimonthly in Salobra, Mato Grosso do Sul state, for one year. Indoors, mosquitoes were collected from their resting places, while in open fields, they were captured using protected human-baited and horse-baited traps near the house and at the Miranda River margin, respectively. Hourly biting activity outdoors was also assessed. Secondary data were collected on the arrival of tourists, economic projects, and malaria cases. FINDINGS A total of 24,894 anophelines belonging to 13 species were caught. The main Brazilian malaria vector Anopheles darlingi was the predominant species, followed by An. triannulatus s.l. Hourly variation in anopheline biting showed three main peaks occurring at sunset, around midnight, and at sunrise, the first and last being the most prominent. The highest density of all species was recorded near the river margin and during the transition period between the rainy and early dry seasons. This coincides with the time of main influx of outsider workers and tourists, whose activities mostly occur in the open fields and frequently start before sunrise and last until sunset. Some of these individuals originate from neighbouring malaria-endemic countries and states, and are likely responsible for the recorded imported and introduced malaria cases. MAIN CONCLUSION Pantanal is a malaria-prone area in Brazil. Surveillance and anopheline control measures must be applied to avoid malaria re-emergence in the region.

Plasmodium falciparum in the southeastern Atlantic forest: a challenge to the bromeliad-malaria paradigm?

BACKGROUND: Recently an unexpectedly high prevalence of Plasmodium falciparum was found in asymptomatic blood donors living in the southeastern Brazilian Atlantic forest. The bromeliad-malaria paradigm assumes that transmission of Plasmodium vivax and Plasmodium malariae involves species of the subgenus Kerteszia of Anopheles and only a few cases of P. vivax malaria are reported annually in this region. The expectations of this paradigm are a low prevalence of P. vivax and a null prevalence of P. falciparum. Therefore, the aim of this study was to verify if P. falciparum is actively circulating in the southeastern Brazilian Atlantic forest remains. METHODS: In this study, anophelines were collected with Shannon and CDC-light traps in seven distinct Atlantic forest landscapes over a 4-month period. Field-collected Anopheles mosquitoes were tested by real-time PCR assay in pools of ten, and then each mosquito from every positive pool, separately for P. falciparum and P. vivax. Genomic DNA of P. falciparum or P. vivax from positive anophelines was then amplified by traditional PCR for sequencing of the 18S ribosomal DNA to confirm Plasmodium species. Binomial probabilities were calculated to identify non-random results of the P. falciparum-infected anopheline findings. RESULTS: The overall proportion of anophelines naturally infected with P. falciparum was 4.4% (21/480) and only 0.8% (4/480) with P. vivax. All of the infected mosquitoes were found in intermixed natural and human-modified environments and most were Anopheles cruzii (22/25 = 88%, 18 P. falciparum plus 4 P. vivax). Plasmodium falciparum was confirmed by sequencing in 76% (16/21) of positive mosquitoes, whereas P. vivax was confirmed in only 25% (1/4). Binomial probabilities suggest that P. falciparum actively circulates throughout the region and that there may be a threshold of the forested over human-modified environment ratio upon which the proportion of P. falciparum-infected anophelines increases significantly. CONCLUSIONS: These results show that P. falciparum actively circulates, in higher proportion than P. vivax, among Anopheles mosquitoes of fragments of the southeastern Brazilian Atlantic forest. This finding challenges the classical bromeliad-malaria paradigm, which considers P. vivax circulation as the driver for the dynamics of residual malaria transmission in this region.

Finding connections in the unexpected detection of Plasmodium vivax and Plasmodium falciparum DNA in asymptomatic blood donors: a fact in the Atlantic Forest.

A recent paper in Malaria Journal reported the observation of unexpected prevalence rates of healthy individuals carrying Plasmodium falciparum (5.14%) or Plasmodium vivax (2.26%) DNA among blood donors from the main transfusion centre in the metropolitan São Paulo, a non-endemic area for malaria. The article has been challenged by a group of authors who argued that the percentages reported were higher than those found in blood banks of the endemic Amazon Region and also that that paper had not considered the literature on the classical dynamics of malaria transmission in the Atlantic Forest, which involves Anopheles (Kerteszia) cruzii and bromeliad malaria, due to P. vivax and Plasmodium malariae parasites, but not P. falciparum. The present commentary paper responds to this challenge and brings evidence and literature data supporting that the observed prevalence ratios may indicate a proportion of individuals that are exposed to Plasmodium transmission in permissive environments; that blood carrying parasite DNA may not be necessarily infective if used in transfusion; and that in the literature, there are examples supporting the circulation of P. falciparum in the area.

Coexistence mechanisms at multiple scales in mosquito assemblages.

BACKGROUND: Species coexistence in mosquito assemblages may depend on mechanisms related to interspecific resource partitioning occurring at multiple scales. In the present work we investigated co-occurrence or spatial segregation in mosquito assemblages sharing resources at micro-habitat, habitat and landscape scales. Environmental characteristics, mosquito fauna as adults and larvae were assessed along vegetation gradient in a natural landscape of tropical rainforest. Huisman-Olff-Fresco (HOF) and Generalized Additive (GAM) models were employed to explore relationships between abundances of potential competitors in mosquito assemblages and vegetation gradient (e.g., scrublands, mixed arboreal vegetation and dense ombrophilous forest). We tested hypotheses concerning mosquito species co-occurrence or spatial segregation employing binomial logistic regression models. RESULTS: Co-occurrences and spatial segregation of mosquito species showed evidences of three scales of coexistence mechanisms: 1) micro-habitat--scale 1: different behaviors in response to food availability in specific vertical strata within larval container; 2) habitat--scale 2: specialized strategies related to heterogeneity of resource availability among larval containers and 3) landscape--scale 3: asymmetrical competition dependent upon the context of abiotic and biotic variables. CONCLUSION: Results of the present work suggest that coexistence mechanisms can concomitantly work at multiple scales.

Effectiveness of Mosquito Magnet® trap in rural areas in the southeastern tropical Atlantic Forest.

Traps are widely employed for sampling and monitoring mosquito populations for surveillance, ecological and fauna studies. Considering the importance of assessing other technologies for sampling mosquitoes, we addressed the effectiveness of Mosquito Magnet® Independence (MMI) in comparison with those of the CDC trap with CO2 and Lurex3® (CDC-A) and the CDC light trap (CDC-LT). Field collections were performed in a rural area within the Atlantic Forest biome, southeastern state of São Paulo, Brazil. The MMI sampled 53.84% of the total number of mosquitoes, the CDC-A (26.43%) and CDC-LT (19.73%). Results of the Pearson chi-squared test (χ2) showed a positive association between CDC-LT and species of Culicini and Uranotaeniini tribes. Additionally, our results suggested a positive association between CDC-A and representatives of the Culicini and Aedini tribes, whereas the MMI was positively associated with the Mansoniini and Sabethini as well as with Anophelinae species. The MMI sampled a greater proportion (78.27%) of individuals of Anopheles than either the CDC-LT (0.82%) or the CDC-A traps (20.91%). Results of the present study showed that MMI performed better than CDC-LT or CDC-A in sampling mosquitoes in large numbers, medically important species and assessing diversity parameters in rural southeastern Atlantic Forest.

Revision of the Strodei Subgroup of Nyssorhynchus (Diptera: Culicidae), with descriptions of 2 new species.

Nyssorhynchus (Nyssorhynchus) ibiapabaensis (Sant'Ana & Sallum n. sp.) and Ny. (Nys.) untii (Sant'Ana & Sallum n. sp.) are new species of the Arthuri Complex of the Strodei Subgroup. The new species are described and validated using morphological characters of the male, female, and immature stages. The description of the male, female, fourth-instar larva and pupa of Ny. arthuri (Unti, 1941) and Ny. albertoi (Unti, 1941) are provided for the first time. To avoid nomenclature instability, neotypes are designated for both species. All life stages of Ny. strodei (Root, 1926) employing specimens collected in the Agua Limpa District, Juiz de Fora, Minas Gerais, Brazil are redescribed.

Comparison of automatic traps to capture mosquitoes (Diptera: Culicidae) in rural areas in the tropical Atlantic rainforest.

In several countries, surveillance of insect vectors is accomplished with automatic traps. This study addressed the performance of Mosquito Magnet® Independence (MMI) in comparison with those of CDC with CO2 and lactic acid (CDC-A) and CDC light trap (CDC-LT). The collection sites were in a rural region located in a fragment of secondary tropical Atlantic rainforest, southeastern Brazil. Limatus durhami and Limatus flavisetosus were the dominant species in the MMI, whereas Ochlerotatus scapularis was most abundant in CDC-A. Culex ribeirensis and Culex sacchettae were dominant species in the CDC-LT. Comparisons among traps were based on diversity indices. Results from the diversity analyses showed that the MMI captured a higher abundance of mosquitoes and that the species richness estimated with it was higher than with CDC-LT. Contrasting, difference between MMI and CDC-A was not statistically significant. Consequently, the latter trap seems to be both an alternative for the MMI and complementary to it for ecological studies and entomological surveillance.

COI barcode versus morphological identification of Culex (Culex) (Diptera: Culicidae) species: a case study using samples from Argentina and Brazil.

Sequences of the cytochrome c oxidase subunit I (COI) mitochondrial gene from adults of 22 Culex (Culex) species from Argentina and Brazil were employed to assess species identification and to test the usefulness of COI for barcoding using the best close match (BCM) algorithm. A pairwise Kimura two-parameter distance matrix including the mean intra and interspecific distances for 71 COI barcode sequences was constructed. Of the 12 COI lineages recovered in the Neighbour-joining topology, five confirmed recognised morphological species (Cx. acharistus, Cx. chidesteri, Cx. dolosus, Cx. lygrus and Cx. saltanensis) with intraspecific divergences lower than 1.75%. Cx. bilineatus is formally resurrected from the synonymy of Cx. dolosus. Cx. maxi , Cx. surinamensis and the Coronator group species included were clustered into an unresolved lineage. The intraspecific distance of Cx. pipiens (3%) was almost twice the interspecific between it and Cx. quinquefasciatus (1.6%). Regarding the BCM criteria, the COI barcode successfully identified 69% of all species. The rest of the sequences, approximately 10%, 18% and 3%, remained as ambiguously, mis and unidentified, respectively. The COI barcode does not contain enough information to distinguish Culex (Cux.) species.

Entomological Reference Collection: 85 years of contributions to public health.

The Department of Hygiene of the Faculty of Medicine of São Paulo (FMUSP), organized with the support of the Rockefeller Foundation, became the Institute of Hygiene, with the inaugural class taught by Samuel Darling in 1918. The history of Public Health Entomology is mixed with that of the Institute itself, which became the Faculty of Hygiene and Public Health in 1945. Still in the 1930s, Paulo César de Azevedo Antunes and John Lane organized Public Health Entomology within the Medical Parasitology area of the then Institute of Hygiene. During this period, the entomology laboratory came to be recognized for its research in the systematics of hematophagous insects, as well as in the ecology, biology and behavior of vectors. The Entomological Reference Collection (CER) originated naturally from the research of Paulo César Antunes and John Lane and is a national and international heritage covering primary and secondary types of insect species that are of interest to public health. Over the years, it has been consolidated with the efforts of Augusto Ayroza Galvão, Renato Corrêa, José Coutinho, Nelson Cerqueira, Ernesto Rabello, Oswaldo Forattini and others. In its over eighty years of activities, CER has enabled the training of several scientists able to act in programs of surveillance and control of endemic diseases associated with insect vectors throughout Latin America, in addition to training taxonomists focused on insects of interest in Public Health. Researchers from other Brazilian institutes and abroad joined the entomology laboratory because of its importance and the research developed in it. The growing scientific production made it possible for entomological studies developed at the Faculty of Public Health (FSP) to gain international visibility, contributing to the development of disease prevention and epidemic control actions in the country.

DNA Barcodes of Mansonia (Mansonia) Blanchard, 1901 (Diptera, Culicidae).

Females of the genus Mansonia feed on the blood of humans, livestock, and other vertebrates to develop their eggs. The females' biting behavior may cause severe disturbance to blood hosts, with a negative impact on public health and economics. Certain species have been identified as potential or effective disease vectors. The accurate species identification of field-collected specimens is of paramount importance for the success of monitoring and control strategies. Mansonia (Mansonia) morphological species boundaries are blurred by patterns of intraspecific heteromorphism and interspecific isomorphism. DNA barcodes can help to solve taxonomic controversies, especially if combined with other molecular tools. We used cytochrome c oxidase subunit I (COI) gene 5' end (DNA barcode) sequences to identify 327 field-collected specimens of Mansonia (Mansonia) spp. The sampling encompassed males and females collected from three Brazilian regions and previously assigned to species based on their morphological characteristics. Eleven GenBank and BOLD sequences were added to the DNA barcode analyses. Initial morphospecies assignments were mostly corroborated by the results of five clustering methods based on Kimura two-parameter distance and maximum likelihood phylogeny. Five to eight molecular operational taxonomic units may represent taxonomically unknown species. The first DNA barcode records for Mansonia fonsecai, Mansonia iguassuensis, and Mansonia pseudotitillans are presented.

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.

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.

Mutations Linked to Insecticide Resistance Not Detected in the Ace-1 or VGSC Genes in Nyssorhynchus darlingi from Multiple Localities in Amazonian Brazil and Peru.

Indoor residual spray (IRS), mainly employing pyrethroid insecticides, is the most common intervention for preventing malaria transmission in many regions of Latin America; the use of long-lasting insecticidal nets (LLINs) has been more limited. Knockdown resistance (kdr) is a well-characterized target-site resistance mechanism associated with pyrethroid and DDT resistance. Most mutations detected in acetylcholinesterase-1 (Ace-1) and voltage-gated sodium channel (VGSC) genes are non-synonymous, resulting in a change in amino acid, leading to the non-binding of the insecticide. In the present study, we analyzed target-site resistance in Nyssorhynchus darlingi, the primary malaria vector in the Amazon, in multiple malaria endemic localities. We screened 988 wild-caught specimens of Ny. darlingi from three localities in Amazonian Peru and four in Amazonian Brazil. Collections were conducted between 2014 and 2021. The criteria were Amazonian localities with a recent history as malaria hotspots, primary transmission by Ny. darlingi, and the use of both IRS and LLINs as interventions. Fragments of Ace-1 (456 bp) and VGSC (228 bp) were amplified, sequenced, and aligned with Ny. darlingi sequences available in GenBank. We detected only synonymous mutations in the frequently reported Ace-1 codon 280 known to confer resistance to organophosphates and carbamates, but detected three non-synonymous mutations in other regions of the gene. Similarly, no mutations linked to insecticide resistance were detected in the frequently reported codon (995) at the S6 segment of domain II of VGSC. The lack of genotypic detection of insecticide resistance mutations by sequencing the Ace-1 and VGSC genes from multiple Ny. darlingi populations in Brazil and Peru could be associated with low-intensity resistance, or possibly the main resistance mechanism is metabolic.

A new species of the Arthuri Complex of the Strodei Subgroup of Nyssorhynchus (Diptera: Culicidae).

Nyssorhynchus (Nyssorhynchus) rondoniensis, a new species of the Arthuri Complex of the Strodei Subgroup, is described and validated using morphological characters of the adult male and female, male genitalia, fourth-instar larva and pupa. The new species is recorded in the municipalities of Campo Novo de Rondnia and Monte Negro, Rondnia State, Brazil. Based on DNA sequence data, the new species (as Ny. arthuri species C) was found to belong to a separate lineage within the Arthuri Complex. Morphological characteristics of the male genitalia and fourth-instar larva confirmed that the new species shared morphological similarities with other species of the Arthuri Complex, but it can be distinguished by characteristics of the male genitalia, adult female and larva. Nyssorhynchus rondoniensis may be involved in malaria transmission because females can be easily misidentified as Ny. oswaldoi (Peryass, 1922) s.l. or Ny. konderi (Galvo Damasceno, 1942) s.l. Both species were previously hypothesized to be local vectors in Acre and Rondnia States.