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.

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.

BACKGROUND The longstanding application of insecticides for vector control without periodic monitoring of the population response to these chemicals can directly drive the selection of resistant populations of vector mosquitoes. Tocantins is the newest state of the Brazilian Federation. Despite a historically high number of dengue cases, studies and monitoring data concerning insecticide resistance in the state are lacking.OBJECTIVES To verify the resistance status of Aedes aegypti from 10 populations distributed throughout the state connected by rivers and roads.METHODS Between 50 and 150 ovitraps were installed in house gardens within each municipality. Collection points were established based on the importance of the towns and on geographic aspects. Dose response bioassays were performed in accordance with World Health Organization guidelines. Molecular assays were conducted to detect kdr mutations, which are related to pyrethroid resistance.FINDINGS Of the 3,200 ovitrap paddles analysed, 25.8% contained eggs, with a total of 55,687 eggs collected. With the exception of Caseara, all evaluated populations were considered to be resistant to temephos. The data showed different levels of resistance to deltamethrin among the samples. Caseara and Guaraí showed the lowest RR95 values. On average, the NaVR1 kdr allele was most frequent (40.3%), followed by NaVS (38.1%), and NaVR2 (21.6%). Palmas, the capital of the state, had the highest frequency of kdr alleles (87.5%).MAIN CONCLUSIONS With the exception of Palmas, the towns with the highest indexes (ovitrap positivity, number and density of eggs), as well with high levels of resistance and kdr alleles were located along the BR-153 road, indicating that the flow of people and cargo can contribute to the dispersion of the vector and potentially resistance. This study contributes substantially to knowledge regarding the insecticide resistance profile of Tocantins mosquito populations; the data generated via the study could facilitate the judicious use of insecticides by vector control programs.

Background Despite the importance of some species of Culex (Melanoconion) (Diptera: Culicidae) as vectors of several arboviruses that cause diseases in humans and other animals, there are few taxonomic studies focusing on species of the subgenus, especially providing morphological keys for species identification. Results Thirteen species of the Atratus Group of Culex (Melanoconion) were reviewed, five new species are described, and two taxonomic changes are proposed: Cx. (Mel.) exedrus Root, 1927 and Cx. (Mel.) loturus Dyar, 1925 are resurrected from synonymy with Cx. (Mel.) dunni Dyar, 1918 and Cx. (Mel.) zeteki Dyar, 1918, respectively. The Atratus Group now includes fourteen species: Cx. (Mel.) atratus Theobald, 1901; Cx. (Mel.) caribeanus Galindo & Blanton, 1954; Cx. (Mel.) columnaris Sá & Hutchings n. sp.; Cx. (Mel.) commevynensis Bonne-Wepster & Bonne, 1919; Cx. (Mel.) comptus Sá & Sallum n. sp.; Cx. (Mel.) dunni; Cx. (Mel.) ensiformis Bonne-Wepster & Bonne, 1919; Cx. (Mel.) exedrus; Cx. (Mel.) longisetosus Sá & Sallum n. sp.; Cx. (Mel.) longistylus Sá & Sallum n. sp.; Cx. (Mel.) loturus; Cx. (Mel.) spinifer Sá & Sallum n. sp.; Cx. (Mel.) trigeminatus Clastrier, 1970; and Cx. (Mel.) zeteki. Keys, descriptions and illustrations for the identification of the male, female, pupal and fourth-instar larval stages of each species are provided. The treatment of each species includes a complete synonymy, descriptions of available life stages, a taxonomic discussion, updated bionomics and geographical distribution, and a list of material examined. Conclusions The taxonomy of the Atratus Group of Culex (Melanoconion) is updated, including descriptions of five new species. The number of valid species is greater than the number recognized in the previous taxonomic study of the group, increasing from seven to 14 species. Distributional and bionomical data are updated. Morphology-based identification keys for females, males, fourth-instar larvae and pupae provided in this study will facilitate species identification.

On the ecological scale of an organism, a homogeneous geographical landscape can represent a mosaic of heterogeneous landscapes. The bionomy of Kerteszia mosquitoes can contribute to foundation landscape ecology by virtue of in the role of the configuration and composition of the habitat played in the distribution of mosquito species. Thus, this study aimed: to compare the abundance of Kerteszia in dense tropical rainforest, restinga and rural area, to assess the bioecological characteristics of the main bromeliads hosting Kerteszia, and to associate the bioecological arrangement of the bromeliads with Kerteszia distribution. Field collections were conducted in a monthly schedule from December of 2010 to November 2011. The vegetation of landscapes was characterized on the basis of a digital cartographic database, the manual of the Brazilian vegetation, environmental atlas information, satellite images and visits to the sites. Multivariate generalized linear models were employed using the R-project statistical program. The results were: Anopheles cruzii was the most frequent species in dense tropical rainforest (67.42%), with a positive association (deviance=25.8; P=0.002). Anopheles bellator was more abundant in the Restinga area (78.97%), with a positive association (deviance=10.4, P=0.018). There was a positive aggregation of Restinga with An. bellator (RR=2.42) but a lower level with An. cruzii (RR=0.31). Thus we can conclude that landscape characteristics influence the distribution of Kerteszia mosquitoes. An. bellator has a higher prevalence in Restinga areas, whereas An. cruzii was the most prevalent in the dense tropical rainforest.

Introdução: As diversas armadilhas apresentam rendimento diferenciado na captura, havendo a necessidade de buscar instrumentos que amostrem porções representativas das populações de mosquitos. Levanta-se a hipótese de que as armadilhas Mosquito Magnet ® Independence e CDC com CO2+Lurex3® sejam mais eficazes na captura de culicídeos do que a armadilha CDC luminosa. Objetivos: Avaliar a eficácia das armadilhas Mosquito Magnet ® Independence e CDC+CO2+Lurex3 ® , em relação à CDC-luminosa, para a captura de culicídeos adultos em área agrícola na Fazenda Experimental em Pariquera-Açu, Vale do Ribeira, São Paulo, Brasil. Material e Métodos: Capturas mensais, das 15h00 às 21h00, por meio do delineamento em quadrado latino 3X3, no período de dezembro/2010 a novembro/2011. Perfis de diversidade de Rényi e índices de riqueza, dominância, diversidade, equabilidade e similaridade foram estimados para cada armadilha para compará-las. Resultados: Capturou-se 6.055 de 70 espécies e 12 gêneros. A CDC-luminosa amostrou 990 indivíduos de 42 espécies e 10 gêneros, a CDC+CO2+lurex3 ® 1.419 indivíduos em 41 espécies e 10 gêneros e a Mosquito Magnet ® 3151 indivíduos de 46 espécies e 11 gêneros. A riqueza, pelo índice de Margalef e Perfil de Rényi, não apresentou diferença estatística para nenhuma armadilha no período dez/mai. A Mosquito Magnet ® obteve maior rendimento na mensuração da diversidade, segundos os índices de Shannon e Simpson, observados no perfil de Rényi. A dominância, pelo índice de Berger-Parker isolado, foi maior na Mosquito Magnet ® do que nas demais, nos dois períodos de capturas. O perfil de Rényi não apontou diferença significante para dominância. Índice de Pielou não mostrou significância em relação à CDC-luminosa para nenhuma das armadilhas nos dois períodos avaliados. Índices de Jaccard e Sorensen apontaram maior similaridade na composição de espécies entre a Mosquito Magnet ® e CDC+CO2+lurex3 ®. Conclusões: A Mosquito Magnet ® Independence e CDC+CO2+Lurex3 ® apresentaram rendimento semelhante, nos meses mais frios e secos do ano. A utilização dessas armadilhas para a captura de determinados táxons pode ser importante ponto na estratégia de vigilância de espécies vetoras. Estudos com maior esforço amostral devem ser realizados para avaliar o rendimento das armadilhas Mosquito Magnet ® Independence e CDC+CO 2 +lurex3 ® em áreas de ambiente alterado de Floresta Atlântica.

High abundance of hematophagous mosquitoes of the genus Mansonia Blanchard, 1901 (Diptera: Culicidae) threatens human and domestic animal health and well-being. Knowledge of the biology of nuisance mosquito species is necessary to understand specific ecological and biological factors to enable rapid and effective monitoring measures for sustainable control programs. The establishment and dispersion of Mansonia species are associated with the occurrence of aquatic macrophytes species, which are indispensable for the development of larvae and pupae. To increase knowledge of the host plants for Mansonia immature stages in Porto Velho, Rondonia State, Brazil, specimens of four plant species, which occur across the tributaries of the Madeira River were sampled and inspected for the presence of egg batches, larvae, and pupae. A total of 1,386 larvae and pupae of Mansonia spp. were collected attached to the roots of Eichhornia crassipes (Mart.) Solms (Commelinales: Pontederiaceae), Pistia stratiotes L. (Alismatales: Araceae), and Limnobium laevigatum (Humb. and Bonpl. Ex Willd.) Heine (Alismatales: Hydrocharitaceae). The novel association of Mansonia species with L. laevigatum is presented. Egg batches of Mansonia spp. were found only on Salvinia molesta D.S. Mitch. (Salviniales: Salviniaceae). Possible differences in the roles played by E. crassipes and S. molesta in the reproductive cycle of Mansonia spp. in the surveyed area are discussed. All species of host plants including E. crassipes, P. stratiotes, S. molesta, and L. laevigatum should be considered when planning macrophyte management for the control of Mansonia species.

The Educator Group of the Melanoconion Section of Culex (Melanoconion) (Diptera, Culicidae) is reviewed. Currently, the group includes the following valid species: Culex (Melanoconion) aphyllus  Talaga, 2020, Culex (Melanoconion) cristovaoi Duret, 1968, Culex (Melanoconion) educator  Dyar & Knab, 1906, Culex (Melanoconion) eknomios  Forattini & Sallum, 1992, Culex (Melanoconion) inadmirabilis  Dyar, 1928, Culex (Melanoconion) rachoui Duret, 1968, Culex (Melanoconion) theobaldi (Lutz), 1904, Culex (Melanoconion) vaxus  Dyar, 1920, Culex (Melanoconion) angularis Sá & Sallum n. sp., Culex (Melanoconion) spiniformis Sá & Hutchings n. sp., Culex (Melanoconion) longistriatus Sá & Hutchings n. sp., Culex (Melanoconion) aneles  Dyar & Ludlow, 1922, Culex (Melanoconion) apeteticus Howard, Dyar & Ludlow, 1913, and Culex (Melanoconion) bibulus  Dyar, 1920. The last three species were resurrected from synonymy of either Cx. educator or Cx. vaxus. Descriptions, differential diagnoses, bionomics, and geographical data are provided for each species when available.