Low coverage of species constrains the use of DNA barcoding to assess mosquito biodiversity.

Mosquitoes (Culicidae) represent the main vector insects globally, and they also inhabit many of the terrestrial and aquatic habitats of the world. DNA barcoding and metabarcoding are now widely used in both research and routine practices involving mosquitoes. However, these methodologies rely on information available in databases consisting of barcode sequences representing taxonomically identified voucher specimens. In this study, we assess the availability of public data for mosquitoes in the main online databases, focusing specifically on the two most widely used DNA barcoding markers in Culicidae: COI and ITS2. In addition, we test hypotheses on possible factors affecting species coverage (i.e., the percentage of species covered in the online databases) for COI in different countries and the occurrence of the DNA barcode gap for COI. Our findings showed differences in the data publicly available in the repositories, with a taxonomic or species coverage of 28.4-30.11% for COI in BOLD + GenBank, and 12.32% for ITS2 in GenBank. Afrotropical, Australian and Oriental biogeographic regions had the lowest coverages, while Nearctic, Palearctic and Oceanian had the highest. The Neotropical region had an intermediate coverage. In general, countries with a higher diversity of mosquitoes and higher numbers of medically important species had lower coverage. Moreover, countries with a higher number of endemic species tended to have a higher coverage. Although our DNA barcode gap analyses suggested that the species boundaries need to be revised in half of the mosquito species available in the databases, additional data must be gathered to confirm these results and to allow explaining the occurrence of the DNA barcode gap. We hope this study can help guide regional species inventories of mosquitoes and the completion of a publicly available reference library of DNA barcodes for all mosquito species.

Inaccurate recording of Mansonella perstans in free-ranging primates outside its endemic area in Brazil?

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.

Change in the faunal composition of mosquitoes (Diptera: Culicidae) along a heterogeneous landscape gradient in the Brazilian Amazon.

This study aimed to evaluate the influence of different anthropic landscape profiles on the diversity and distribution of mosquito species in a rural settlement of the Brazilian Amazon. Eight field collections were conducted at 18 sampling points interspersed throughout 2020-2021. Plastic containers, bamboo internodes, and tires were used as traps to capture immature mosquitoes in three distinct habitats: forest, forest edge, and peridomicile. A total of 15,547 individuals, distributed in 26 species of culicids, were collected. The most abundant species were Culex urichii (8,376 specimens), Culex (Melanoconion) (2,473 specimens), and Aedes albopictus (1,252 specimens). Forest habitat showed the highest abundance, and forest edge showed the highest species richness. Different types of environments influenced both the abundance and richness of mosquitoes. The species composition was also significantly different between the analyzed sites, mainly between forest and peridomicile environments. The change in species dominance could largely explain this change in mosquito community composition. Haemagogus janthinomys, an important sylvatic arbovirus vector, was found in peridomicile habitats and Ae. albopictus, a vector associated with human environments, was found in forest habitats, thus providing evidence of species spillover. Our results indicated that landscape changes affect mosquito communities, influencing their richness and abundance. These changes may have implications for future arboviral outbreaks in this rural settlement due to the possible establishment of sylvatic vector species in anthropic environments.

Dirofilaria immitis is endemic in rural areas of the Brazilian Amazonas state capital, Manaus.

The canine filarial parasite Dirofilaria immitis has not been reported in Brazil´s Amazonas state capital, Manaus, for over a century. Here, we report one imported and 27 autochthonous D. immitis infections from a microfilarial survey of 766 domestic dog blood samples collected between 2017 and 2021 in Manaus. An Overall prevalence estimate of 15.44% (23/149) was calculated from our two rural collection sites; a prevalence of 1.22% (4/328) was estimated at our periurban collection site, and an overall prevalence of 0.35% (1/289) was calculated from our two urban clinic collections. Our data suggest that in the urban areas of Manaus, where the parasites are very likely vectored by the same species of mosquito that historically vectored Wuchereria bancrofti (Culex quinquefasciatus), prevalence levels are very low and possibly maintained by an influx from rural areas where sylvatic reservoirs and/or more favorable vector transmission dynamics maintain high prevalences.

Influence of meteorological and seasonal parameters on the activity of Culicoides paraensis (Diptera: Ceratopogonidae), an annoying anthropophilic biting midge and putative vector of Oropouche Virus in Rondônia, Brazilian Amazon.

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.

Tabanidae (Diptera) captured on horses in Amazon Forest fragments of the state of Rondônia, Brazil.

Tabanidae is one of the most diverse families of hematophagous dipterans. Tabanids, in general, are mechanical vectors of some pathogens. Given the vector importance and the lack of knowledge of the tabanid fauna in horses in Amazon Forest fragments of the state of Rondônia, this work aimed to determine the season that the different species of horse flies prefer to carryout hematophagy on horses and verify whether the horse fly community remains the same throughout the year. The sampling areas for tabanid captures were in the municipality of Monte Negro, Western Amazon, Brazil. Four new occurrences were recorded for the state of Rondônia: Stenotabanus albilinearis, Tabanus fuscofasciatus, T. macquarti and T. restrepoensis, which increases the number of species for the state to 109. The horse flies were most frequently collected on the hind leg (43.15%) and front leg (31.11%), followed by the belly (7.41%) and the ear (5.18%). In the other anatomical regions, the collection frequency was 13.15% of the remaining individuals.

Blood-stage antiplasmodial activity and oocyst formation-blockage of metallo copper-cinchonine complex.

In the fight against malaria, the key is early treatment with antimalarial chemotherapy, such as artemisinin-based combination treatments (ACTs). However, Plasmodium has acquired multidrug resistance, including the emergence of P. falciparum strains with resistance to ACT. The development of novel antimalarial molecules, that are capable of interfering in the asexual and sexual blood stages, is important to slow down the transmission in endemic areas. In this work, we studied the ability of the mettalo copper-cinchonine complex to interfere in the sexual and asexual stages of Plasmodium. The tested compound in the in vitro assay was a cinchonine derivative, named CinCu (Bis[Cinchoninium Tetrachlorocuprate(II)]trihydrate). Its biological functions were assessed by antiplasmodial activity in vitro against chloroquine-resistant P. falciparum W2 strain. The mice model of P. berghei ANKA infection was used to analyze the antimalarial activity of CinCu and chloroquine and their acute toxicity. The oocyst formation-blocking assay was performed by experimental infection of Anopheles aquasalis with P. vivax infected blood, which was treated with different concentrations of CinCu, cinchonine, and primaquine. We found that CinCu was able to suppress as high as 81.58% of parasitemia in vitro, being considered a molecule with high antiplasmodial activity and low toxicity. The in vivo analysis showed that CinCu suppressed parasitemia at 34% up to 87.19%, being a partially active molecule against the blood-stage forms of P. berghei ANKA, without inducing severe clinical signs in the treated groups. The transmission-blocking assay revealed that both cinchonine and primaquine were able to reduce the infection intensity of P. vivax in A. aquasalis, leading to a decrease in the number of oocysts recovered from the mosquitoes' midgut. Regarding the effect of CinCu, the copper-complex was not able to induce inhibition of P. vivax infection; however, it was able to induce an important reduction in the intensity of oocyst formation by about 2.4 times. It is plausible that the metallo-compound also be able to interfere with the differentiation of parasite stages and/or ookinete-secreted chitinase into the peritrophic matrix of mosquitoes, promoting a reduction in the number of oocysts formed. Taken together, the results suggest that this compound is promising as a prototype for the development of new antimalarial drugs. Furthermore, our study can draw a new pathway for repositioning already-known antimalarial drugs by editing their chemical structure to improve the antimalarial activity against the asexual and sexual stages of the parasite.

Amazonian Anopheles with low numbers of oocysts transmit Plasmodium vivax sporozoites during a blood meal.

Anopheles darlingi is the main malarial vector in the Brazilian Amazon region. An. nuneztovari s.l., An. triannulatus s.l., An. evansae, and An. benarrochi s.l. do not have a defined role as malarial vectors, although they have been found to be naturally infected with Plasmodium vivax, and some develop oocysts. In this study, we evaluated the importance of low numbers of oocysts in sporozoite salivary gland invasion and transmission. Field-collected mosquitoes were experimentally infected with P. vivax. The infection rates and oocyst and sporozoite infection intensities were evaluated and compared with those of An. aquasalis. We found the highest number of oocysts in An. darlingi (mean = 39.47) and the lowest in An. nuneztovari s.l. (mean = 2). The highest number of sporozoites was observed in An. darlingi (mean = 610) and lowest in An. benarrochi s.l. (mean = 30). Plasmodium vivax DNA was detected in the saliva of all mosquito species after a blood meal. Regardless of the number of oocysts, all species transmitted sporozoites during blood meals. Considering the abundance of these mosquitoes and transmission of sporozoites, it is logical to assume that An. nuneztovari s.l. and An. triannulatus s.l. are involved in the transmission of P. vivax.

DNA Barcoding of genus Culicoides biting midges (Diptera: Ceratopogonidae) in the Brazilian Amazon.

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.

Definition of the main vector of cutaneous leishmaniasis: Ecology and mapping in endemic area of Northeast Brazil.

Cutaneous leishmaniasis is endemic in Pernambuco. Aiming to determine the vector species of cutaneous leishmaniasis in an endemic area of the Northeast region of Brazil, this study aimed to use the spatial mapping of human cases of CL and correlate with ecological studies of the vectors in the municipality of Timbaúba, Pernambuco, Brazil. Individuals infected with CL were recruited through active search in their homes and clinically and serologically diagnosed during the period from 2018 to 2019. Sandflies were captured with CDC-type light traps in peridomiciliary environments and these were identified at the species level. Females were separated for DNA extraction and subsequent analysis by PCR. The points of collection of phlebotomes and the residences of individuals with lesions were marked with GPS. During the study period, 60 cases of CL were diagnosed. A higher concentration of CL cases was observed in proximity to Atlantic forest remnants confirmed by heat map. A total of 3744 sandflies was captured and five distinct species were identified, with the predominance of Nyssomyia whitmani. From the females separated for the identification of Leishmania braziliensis DNA, a rate of 0.68% of infected sandflies was obtained. It was concluded that cutaneous leishmaniasis continues to be a rural feature of the area. And from this study, it is concluded that Ny. whitmani is the carrier species of CL in the municipality of Timbaúba, Pernambuco. This is due to abundance in catching, specialization of species and PCR positivity for Leishmania braziliensis.

First record of antibodies to the bluetongue virus in ewe (Ovis aries) in the state of Amazonas, Brazil / Primeiro registro de anticorpos para o vírus da língua azul em ovino (Ovis aries) no estado do Amazonas, Brasil

The objective of this work was to describe the first record of antibodies to the Bluetongue Virus (BTV) in ewe, in the state of Amazonas. The ewe, which was in twin pregnancy, gave birth on May 9, 2015, but a lamb died hours after delivery. Veterinary service was then requested by the owner, where emaciation, loss of wool, pyrexia, apathy, dyspnea, mucoid nasal secretion, facial, lingual and submandibular edema were observed. There was a visit by the Agricultural Defense Agency of the State of Amazonas to the property and blood samples were collected from the animal. The whole blood and serum were sent to the National Agricultural Laboratory, where it was possible to detect the presence of specific antibodies to BTV, through the Agar Gel Double Immunodiffusion. The ewe was submitted to a new blood collection, following the same protocols and the samples were sent to the Biological Institute of São Paulo, confirmed diagnosis. The animal in a serious clinical condition, could not resist and died in July 2015. The occurrence of an allochthonous case, in an area where vector insects occur, can trigger an endemic process in the Amazon region. With this, the epidemiological control of these occurrences is necessary, in order to avoid the spread of the disease in the country.

O objetivo do trabalho foi descrever o primeiro registro de anticorpos para o Vírus da Língua Azul (VLA) em ovino, no estado do Amazonas. A ovelha, que se encontrava em gestação gemelar, pariu no dia 9 de maio de 2015, porém um cordeiro faleceu horas após o parto. Foi então solicitado serviço veterinário por parte do proprietário, onde foi observado emaciação, perda de lã, pirexia, apatia, dispneia, secreção nasal mucoide, edema facial, lingual e submandibular. Houve visita da Agência de Defesa Agropecuária do Estado do Amazonas na propriedade e coletadas amostras de sangue do animal. O sangue total e soro foram enviados ao Laboratório Nacional Agropecuário, no qual foi possível detectar a presença de anticorpos específicos para VLA, através do teste de Imunodifusão Dupla em Gel de Ágar. A ovelha foi submetida a uma nova coleta de sangue, seguindo os mesmos protocolos e as amostras foram enviadas ao Instituto Biológico de São Paulo, confirmando diagnóstico. O animal em estado clínico grave, não resistiu e veio a óbito em julho de 2015. A ocorrência de um caso alóctone, em uma área de ocorrência de insetos vetores, pode desencadear um processo de endemia na região amazônica. Com isso, o controle epidemiológico destas ocorrências, se fazem necessários, afim de se evitar a disseminação da doença no país.

Molecular detection of Mansonella mariae incriminates Simulium oyapockense as a potentially important bridge vector for Amazon-region zoonoses.

OBJECTIVE: To assess the emergent zoonotic disease risk posed by the voracious human-biting blackfly species Simulium oyapockense in the peripheral regions of an expanding urban centre situated deep in the Brazilian Amazon rainforest. METHODS: We performed nine human landing catches at three periurban sites surrounding the Brazilian Amazon town of São Gabriel da Cachoeira. Using the detection of non-human primate filarial parasites as an indicator of the zoonotic disease threat posed by a biting insect, we screened 3328 S. oyapockense blackflies for the presence of zoonotic filarial DNA with an ITS-1 PCR assay and Sanger sequencing. RESULTS: Between 98 and 100% of the biting insects captured during our nine collections were identified as S. oyapockense; at our three collection sites and during our three seasonally-distinct collections this species was captured at rates between 28 and 294 blackflies per hour. PCR screening of the march-collected S. oyapockense detected infectious-stage (L3) Mansonella mariae parasites (which are only known to infect non-human primates) in >0.15% of the tested head samples. CONCLUSIONS: Our results show that residents of the periurban regions of São Gabriel da Cachoeira are routinely exposed to the bites of S. oyapockense blackflies which have previously fed on non-human primates.

Ticks on reptiles and amphibians in Central Amazonia, with notes on rickettsial infections.

Reptiles and amphibians are exceptional hosts for different ectoparasites, including mites and ticks. In this study, we investigated tick infestations on reptiles and amphibians trapped in Central Amazonia, and also assessed the presence of rickettsial infections in the collected ticks. From September 2016 to September 2019, 385 reptiles (350 lizards, 20 snakes, 12 tortoises, and three caimans) and 120 amphibians (119 anurans and one caecilian) were captured and examined for ectoparasites. Overall, 35 (10%) lizards, three (25%) tortoises and one (0.8%) toad were parasitized by ticks (124 larvae, 32 nymphs, and 22 adults). In lizards, tick infestation varied significantly according to landscape category and age group. Based on combined morphological and molecular analyses, these ticks were identified as Amblyomma humerale (14 larvae, 12 nymphs, 19 males, and one female), Amblyomma nodosum (three larvae, one nymph, and one female), and Amblyomma rotundatum (four larvae, three nymphs, and one female), and Amblyomma spp. (103 larvae and 16 nymphs). Our study presents the first records of A. nodosum in the Amazonas state and suggests that teiid lizards are important hosts for larvae and nymphs of A. humerale in Central Amazonia. Moreover, a nymph of A. humerale collected from a common tegu (Tupinambis teguixin) was found positive for Rickettsia amblyommatis, which agrees with previous reports, suggesting that the A. humerale-R. amblyommatis relationship may be more common than currently recognized.

Genetic structure of allopatric populations of Lutzomyia longipalpis sensu lato in Brazil.

Lutzomyia longipalpis sensu lato is a complex of phlebotomine sand fly species, which are widespread in the Neotropics. They have a great medico-veterinary importance due their role as vectors of Leishmania infantum, the causative agent of visceral leishmaniasis. Morphological variations of Lu. longipalpis s.l. males were reported in the late 1960s in Brazil. Male populations can present either one pair of spots on third abdominal tergites or two pairs on third and fourth ones, namely 1S and 2S phenotypes, respectively. Since then, there has been much interest on the taxonomic status of Lu. longipalpis s.l. Thereafter, several lines of evidence have been congruent in suggesting the existence of an uncertain number of cryptic species within Lu. longipalpis s.l. in Brazil. Herein, a 525 bp-fragment of the period gene was used for assessing the genetic structure and phylogenetic relationship of Lu. longipalpis s.l. populations in Brazil. We performed two set of analyses, first we originally sequenced three populations (Passira, Santarém and Teresina) of Lu. longipalpis s.l. and compared them. Thereafter, we performed a global analysis including in our dataset other three pairs of sympatric populations of Lu. longipalpis s.l. from three Brazilian localities available in GenBank. Fixed single nucleotide polymorphisms (SNPs) sharing, maximum likelihood inference, genetic structure and haplotype analyses revealed the presence of two genetic groups, one composed of Teresina population, and the other encompassing Passira and Santarém populations. The global analysis reflected the first of its kind, and two prominent groups were observed: the clade I comprising Teresina 1S, Bodocó 1S, Caririaçu 1S and Sobral 1S; and the clade II encompassing Passira 2S, Santarém 1S, Bodocó 2S, Caririaçu 2S and Sobral 2S. Genetic differentiation data suggested a limited gene flow between populations of the clade I versus clade II. Our results disclosed the presence of two prominent genetic groups, which could reasonably represent populations of Lu. longipalpis s.l. whose males produce the same courtship song.

Presence of trypanosomatids, with emphasis on Leishmania, in Rodentia and Didelphimorphia mammals of a rural settlement in the central Amazon region.

BACKGROUND: Trypanosomatids are widespread and cause diseases - such as trypanosomiasis, sleeping sickness, Chagas disease, and cutaneous and visceral leishmaniasis - in animals and humans. These diseases occur in both rural and urban regions due to unplanned growth and deforestation. Thus, wild and synanthropic reservoir hosts living in residential areas are risk factors. OBJECTIVE: We aimed to evaluate the diversity of small mammals (rodents and marsupials), and the occurrence of trypanosomatids, especially Leishmania, in the rural settlement of Presidente Figueiredo, Amazonas. METHODS: Animals were collected using Sherman, Tomahawk, and Pitfall traps along 16 trails in four landscapes: continuous forest, forest with planting, planting, and peridomiciliar. Leishmania sp. was detected in liver samples by polymerase chain reaction targeting kDNA. FINDINGS: Diversity was higher in forests with planting and lower around residences. In total, 135 mammals (81 rodents and 54 marsupials covering 14 genera) were captured. Rodents presented infection rates (IR) of 74% and marsupials of 48%. Rodents in domicile landscapes presented a higher IR (92.9%), while marsupials showed a higher IR in forests (53.3%). MAIN CONCLUSIONS: The results suggest high prevalence of trypanosomatids across 12 mammalian genera possibly involved as reservoir hosts in the enzootic transmission of leishmaniasis in the Amazon's rural, peridomiciliar landscape.

DNA-based blood meal analysis of Culicoides (Diptera: Ceratopogonidae) species from Jamari National Forest, Southwestern Amazon, Brazil.

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.

Description of three new species of biting midge of the genus emAtrichopogon/em Kieffer (Diptera: Ceratopogonidae) from Brazilian Amazon.

Three new species of Atrichopogon with spotted wings of the family Ceratopogonidae (Diptera: Culicomorpha) from the Amazonas State of Brazil are described and illustrated. Male and female specimens of Atrichopogon janseni sp. nov., A. riopardensis sp. nov., and A. sergioluzi sp. nov. were associated by wing pigmentation patterns: with two darker spots, one over r-m and the other one in cell r3, posterior to the apex of R3. Male A. janseni sp. nov. have tergite 9 that is 2.5 × as broad as long, not extending to the apex of gonocoxite, and sternite 9 stout, sub-trapezoidal, with distal margin with a row of stout spines; gonostylus of A. janseni sp. nov. is spatulate, with distinct middle notch. Atrichopogon riopardensis sp. nov. is separated from other species with similar wing patterns and forked gonostylus, with gonostylus forked near midlength, with longitudinal furrow, inner portion short and fingernail-like, outer portion elongate and apically curved directed mesally; outer portion is 4 × longer than inner, with one long seta in basal 1/3. Contrastingly, A. sergioluzi sp. nov. has a small fork at the apex of gonostylus, and cercus broad basally with rounded tip. Females of the three new species are very similar (as are other females in this group), and it is probably not possible to distinguish them using the morphological features described. With the addition of the three new species described here, there are now known 15 Neotropical species of Atrichopogon with pigmented wings.

Prevalence of obesity, high blood pressure, dyslipidemia and their associated factors in children and adolescents in a municipality in the Brazilian Amazon region / Prevalência de obesidade, pressão arterial elevada e dislipidemia e seus fatores associados em crianças e adolescentes de um município amazônico, Brasil

INTRODUCTION: The incidence of chronic noncommunicable disease (CND) are rocketting over the world, including in young adults. The WHO estimates that more than half of the deaths in the world, even in underdeveloped countries, are caused by CND. OBJECTIVES: The study aimed to estimate the prevalence of obesity, high blood pressure (HBP) and dyslipidemia and its associated factors. METHODS: The authors carried out a cross-sectional study of 1,431 schools in the public-school system of Monte, Brazilian Western Amazon, with children and adolescents aged 6 to 15 years. A random sampling of 496 individuals was carried out. The OpenEpi platform was used to calculate the sample size, considering p<0.05 and a presumed prevalence of CND of 50%. The authors applied a clinical-epidemiological questionnaire, made anthropometric measurements and laboratory tests. Diagnostic parameters recommended by the recent guidelines of the Ministry of Health in Brazil were used. RESULTS: Prevalence of CND was: Obesity 11.8%, HBP of 6.7% and dyslipidemia of 25.4%. After multivariate log-binomial analysis of the dependent variables, the statistically significant risk factors were overweight 18.4%, sedentary lifestyle 32.2%, family history of cardiovascular disease 23.4%, family history of HBP 84.2%, family dyslipidemia 55.8%, family obesity 38.7% and family chronic renal disease 40.6%. CONCLUSION: The findings pointed out to a context with a relatively high prevalence of CND, as well as their associated factors. Intervention measures such as health education, food education, stimulation of physical exercise, better school feeding and an improvement of the public health system are needed to mitigate the occurrence of CND.

INTRODUÇÃO: A incidência de Doenças Crônicas Não Transmissíveis (DCNT) está aumentando em todo o mundo, inclusive em adultos jovens. A OMS estima que mais da metade das mortes no mundo, mesmo em países subdesenvolvidos, são causadas por DCNT. OBJETIVO: O estudo teve como objetivo estimar a prevalência de obesidade, pressão arterial elevada (PAE) e dislipidemia e seus fatores associados. MÉTODO: Os autores realizaram um estudo transversal com uma amostra randomizada de 496 de 1.431 alunos das escolas da rede pública de ensino de Monte Negro, Amazônia Ocidental, com crianças e adolescentes de 6 a 15 anos. Foi realizada uma amostragem aleatória de 496 indivíduos. Para o cálculo do tamanho da amostra foi utilizada a plataforma OpenEpi, considerando p <0,05 e prevalência presumida de DCNT de 50%. Os autores aplicaram um questionário clínico-epidemiológico, realizaram medidas antropométricas e exames laboratoriais. Foram utilizados parâmetros diagnósticos recomendados pelas diretrizes recentes do Ministério da Saúde do Brasil. Os dados foram analisados por por tetes estatísticos univariados e depois, multivariados, para se detectar associação entre causas e desfechos. RESULTADOS: A prevalência de DCNT foi: Obesidade 11,8%, Pressão Arterial Elevada de 6,7% e dislipidemia de 25,4%. Após análise log-binomial multivariada das variáveis dependentes, os fatores associados estatisticamente significativos foram sobrepeso 18,4%, sedentarismo 32,2%, história familiar de doença cardiovascular 23,4%, história familiar de hipertensão arterial sistêmica 84,2%, dislipidemia familiar 55,8%, obesidade familiar 38,7% e doença renal crônica familiar 40,6%. CONCLUSÃO: Os achados apontam para um contexto com prevalência relativamente elevada de DCNT, bem como seus fatores associados em crianças/adolescentes. Medidas de intervenção como educação em saúde, educação alimentar, estímulo à prática de exercícios físicos, melhor alimentação escolar e melhoria do sistema público de saúde são necessárias para mitigar a ocorrência de DCNT.

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

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

Biting Midges in Jamari National Forest, in the Brazilian Amazon, With 12 New Records of Culicoides Species (Diptera: Ceratopogonidae) for the State of Rondônia.

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.