ABSTRACT The present study aimed identifying the behavioral patterns of Anopheles species as well as to estimate the parity rate and natural infection analysis for Plasmodium species in the district of Ilha de Santana, state of Amapá, Brazil. The samples were obtained in four and 12-hours collections. In the intradomiciliary and peridomiciliary conditions and also in environments with the presence of animals from January/2017 to December/2018. The entomological parameters evaluated were human biting rate (HBR); Indexes of Anthropophily (IA) and Zoophily (IZ); Parity Rate (PR); Natural Infection Rate (NIR); Monthly and annual entomological inoculation rate (EIR). A total of 1,330 Anopheles specimens were collected, distributed in nine species. All captured species showed preference biting in outdoor enviroment. Anopheles darlingi was the most frequent species collected in indoor environment and the most anthropophilic (IA = 0.39) compared with the remaining species captured. It was also the unique species positive for Plasmodium vivax, had the highest anthropophily degree, highest biting activity and HBR in the first hours with a hight rate of parous females. Anopheles nuneztovari s.l. was the most zoophilic species (IZ = 0.65). These findings suggest that A. darlingi is the main malaria vector in the studied area. Anopheles albitarsis s.l. was the second species more anthropophilic (IA = 0.31) and revealed a stable pattern with a biting activity peak after sunset, consequently this species may contribute with malaria transmission in area.

ABSTRACT The objective of this study was to evaluate the effect of the construction of the Jirau hydroelectric plant on the anopheline species, in Porto Velho, Rondônia, Brazil. For this, sampling was carried out in 23 locations in the pre (2004) and 23 after construction (2018) phases in the hydroelectric dam's coverage area, performed by human landing catch between 6:00 pm and 10:00 pm. We found 14 anopheline species. In the pre-construction phase, An. darlingi (73%) was the most abundant and after construction An. braziliensis (31.5%) was the most abundant. The T test indicated that there were no differences in the indexes – diversity (H '), richness (Sjack 1), equitability (J’) and dominance (d) of anophelines species - evaluated between the phases (p > 0.05). The spatio-temporal distribution of An. darlingi proved to be wide, with the highest density values observed in March ( x ¯w = 27.1) in the pre-construction phase and in the months of May ( x ¯w = 4) and July ( x ¯w = 3.1), post-construction. Differences in hematophagic activity peaks were observed between species and between phases. According to the data obtained in this study, the construction of the hydroelectric plant had little influence on the composition of anopheline species, however changes were observed in the spatio-temporal distribution of the An. darlingi and in the pattern of hematophagic activity of the species, which directly influenced the dynamics of malaria in the region.

RESUMO Relatamos o primeiro registro de Aedes albopictus (Diptera: Culicidae) no Estado do Amapá, no norte do Brasil. Dois espécimes foram coletados em uma armadilha de Shannon na localidade de “Ilha de Santana”, município de Santana, no Amapá. O registro foi confirmado com a instalação de dez ovitrampas. Aedes albopictus é uma espécie com importância epidemiológica na transmissão de arboviroses. O Amapá era um de dois estados brasileiros sem registro dessa espécie.

ABSTRACT We report the first record of Aedes albopictus (Diptera: Culicidae) in the state of Amapá, northern Brazil. Two female specimens were collected from a Shannon trap in the locality of Ilha de Santana, municipality of Santana, Amapá. The occurrence of the species was confirmed with the installation of ten ovitraps. Aedes albopictus is a species of epidemiological importance for the transmission of arboviruses. Amapá was one of two Brazilian states without any record of this species.

Abstract INTRODUCTION: The muscoid fly Dermatobia hominis causes cutaneous myiases in mammals. Females of this species use a vector to carry their eggs to the host. This note describes Anopheles konderi acting as phoretic vector for D. hominis. METHODS: A female A. konderi carrying D. hominis was collected using light traps in Oriximiná, Pará, Brazil. The A. konderi specimen was identified at morphological and molecular levels. RESULTS: Eight eggs of D. hominis were observed on the Anopheles konderi female. CONCLUSIONS: Anopheles konderi, only the third Anopheles species recorded as a phoretic vector, may be a potential vector of D. hominis.

Introduction The diverse and complex environmental conditions of the Amazon Basin favor the breeding and development of Anopheles species. This study aimed to describe the composition, abundance and temporal frequency of Anopheles species and to correlate these factors with precipitation, temperature and relative humidity. Methods The study was conducted in the District of Coração, State of Amapá, Brazil. Samples were collected monthly during three consecutive nights, from 6:00 PM to 10:00 PM, from December 2010 to November 2011. In addition, four 12-hour collections (i.e., 6:00 PM to 6:00 AM) were performed during this period. Results A total of 1,230 Anopheles specimens were collected. In the monthly collections, Anopheles darlingi was the predominant species, followed by An. braziliensis and An. albitarsis s.l., whereas An. darlingi, An. peryassui and An. braziliensis were the most frequent species collected in the 12-hour collections. The greatest number of anophelines was collected in September (the dry season). The highest frequency of anophelines was observed for An. darlingi during September, when there were the least rainfalls of the year, along with lower relative humidity and higher temperatures. There was little variation in the abundance of this species in other months, with the exception of slight increases in February, July and August. Conclusions The major malaria vectors, An. darlingi and An. albitarsis s.l. (likely An. marajoara), were the most abundant species collected in the study area. Consequently, prevention and control measures should be taken to prevent malaria outbreaks in the District of Coração.

Genetic variation was estimated in ten samples populations of Aedes aegypti from the Brazilian Amazon, by using a 380 bp fragment of the mitochocondrial NADH dehydrogenase subunit 4 (ND4) gene. A total of 123 individuals were analyzed, whereby 13 haplotypes were found. Mean genetic diversity was slightly high (h = 0.666 ± 0.029; π = 0.0115 ± 0.0010). Two AMOVA analyses indicated that most of the variation (~70%-72%) occurred within populations. The variation found among and between populations within the groups disclosed lower, but even so, highly significant values. F ST values were not significant in most of the comparisons, except for the samples from Pacaraima and Rio Branco. The isolation by distance (IBD) model was not significant (r = 0.2880; p = 0.097) when the samples from Pacaraima and Rio Branco were excluded from the analyses, this indicating that genetic distance is not related to geographic distance. This result may be explained either by passive dispersal patterns (via human migrations and commercial exchange) or be due to the recent expansion of this mosquito in the Brazilian Amazon. Phylogenetic relationship analysis showed two genetically distinct groups (lineages) within the Brazilian Amazon, each sharing haplotypes with populations from West Africa and Asia.

The population genetic structure of Anopheles darlingi, the major human malaria vector in the Neotropics, was examined using seven microsatellite loci from nine localities in central and western Amazonian Brazil. High levels of genetic variability were detected (5-25 alleles per locus; H E = 0.519-0.949). There was deviation from Hardy-Weinberg Equilibrium for 59.79% of the tests due to heterozygote deficits, while the analysis of linkage disequilibrium was significant for only two of 189 (1.05%) tests, most likely caused by null alleles. Genetic differentiation (F ST = 0.001-0.095; Nm = 4.7-363.8) indicates that gene flow is extensive among locations < 152 km apart (with two exceptions) and reduced, but not absent, at a larger geographic scale. Genetic and geographic distances were significantly correlated (R² = 0.893, P < 0.0002), supporting the isolation by distance (IBD) model. The overall estimate of Ne was 202.4 individuals under the linkage disequilibrium model, and 8 under the heterozygote excess model. Analysis of molecular variance showed that nearly all variation (~ 94%) was within sample locations. The UPGMA phenogram clustered the samples geographically, with one branch including 5/6 of the state of Amazonas localities and the other branch the Acre, Rondônia, and remaining Amazonas localities. Taken together, these data suggest little genetic structure for An. darlingi from central and western Amazonian Brazil. These findings also imply that the IBD model explains nearly all of the differentiation detected. In practical terms, populations of An. darlingi at distances < 152 km should respond similarly to vector control measures, because of high gene flow.

Electrophoretic studies of isozymes were done with four closely related species of the Simulium perflavum species group (Diptera: Simuliidade) in the Brazilian Amazon, using last-instar larvae collected in the field. Ten enzymes were studied, which yielded 11 loci. Diagnostic loci were not found between Simulium maroniense cytotype D and Simulium rorotaense. Simulium maroniense and S. rorotaense differed from Simulium trombetense by two diagnostic loci (Me and Xdh), and Simulium perflavum differed from the other three species by four diagnostic loci (Me, Xdh, Mdh, and Got). The mean number of alleles per locus ranged from 1.30 to 2.30, the percentage of polymorphic loci ranged from 18.2 to 63.6% and the mean heterozygosity values observed ranged from 0.062 to 0.108. Genetic distances among the species ranged from 0.010 to 0.581. The lowest value was obtained between S. maroniense and S. rorotaense, and the highest between S. perflavum and S. trombetense. The genetic relationships among the four S. perflavum group species indicate that they are closely related. The high similarity at the isozyme level, allied to previous studies of morphology and polytene chromosomes, may suggest that the divergence time since the separation of S. maroniense and S. rorotaense is still too recent for diagnostic loci to have evolved.

Neste estudo, apresentamos a variabilidade no DNA mitocondrial de três populações da Amazônia brasileira e uma do oeste da Colômbia (Sitronela) de A. nuneztovari, utilizando-se quatro endonucleases de restrição (BclI, ClaI, HindIII, SstI). A diversidade haplotípica (h) foi ligeiramente elevada em todas as populações (0.5000 a 0.6765), enquanto que a diversidade de nucleotídeo (pi) foi menor na população de Sitronela (0.0029) e maior nas populações da Amazônia brasileira (0.0056 a 0.0098). O grau de divergência de seqüência (delta) estimado entre as populações da Amazônia brasileira e Sitronela (0.0329 a 0.0371) sugere que estas populações geográficas de A. nuneztovari podem eventualmente representar espécies distintas. Os valores baixos de divergência de seqüência entre as três populações da Amazônia brasileira (0.0012 a 0.0031) indicam que estas populações são geneticamente semelhantes. Estes resultados são consistentes com os reportados recentemente para aloenzimas nestas mesmas populações.

In the present study, we have examined the variability in Anopheles nuneztovari mitochondrial DNA of three populations from the Brazilian Amazon and one from western Colombia (Sitronela), using four restriction endonucleases (BclI, ClaI, HindIII, SstI). The haplotype diversity (h) was slightly elevated in all populations (0.5000 to 0.6765), whereas the nucleotide diversity (pi) was lower in the Sitronela population (0.0029) and higher in populations from the Brazilian Amazon (0.0056 to 0.0098). The degree of sequence divergence (delta) estimated within the Brazilian Amazon and that in Sitronela (0.0329 to 0.0371) suggests that these geographic populations of A. nuneztovari may eventually constitute separate species. The low sequence divergence values among the three Brazilian Amazon populations (0.0012 to 0.0031) indicate that these populations are genetically similar. These results are consistent with those recently reported for allozymes of these same populations.

Dados sobre a leucina aminopeptidase durante o desenvolvimento ontogenético em Anopheles nuñez-tovarìevidenciaram seis zonae de atividade, eendo a LAP1, LAP2, LAP4 e LAP5 presentes nos estádios larvais, pupae e adultos e a LAP3 e LAP6 características desses dois últimos estágios. Diferenças na intensidade de coloração foram detectadas conforme o estágio considerado. A LAP1 e LAP5 apresentam intensidade fraca em todos os estágios e a LAP2 e LAP4 mostram atividade intensa nos estádios larvais, diminuindo nos dois estágios subseqüentes. Considerando-se as funções das enzimas, foi admitido que a LAP3, no estágio de pupa, possa estar relacionada com a histólise dos tecidos larvais. Dos seis locidetectados, variação alélica foi constatada apenas para o locus LAP5,sendo detectados dois alelos de ação codominante. As freqüências genotípicas de progênies de fêmeas inseminadas naturalmente desviam-se significativamente do equilíbrio de Hardy-Weinberg.

Data on the leucine aminopeptidase system during ontogenetic growth in Anopheles nuñez-tovarirevealed six areas of activity, of which LAP1, LAP2, LAP4 and LAP5 were present in the larval, pupal and adult stages and LAP3 and LAP6 were characteristics of the last two stages. Pifferences in intensity were detected according to the stage considered. LAW and LAP5 show a weak intensity in all the stages and LAP2 and LAP4 show intense activity in the larval stages, decreasing in the pupa and adult. Considering the functions of the enzymes, admitted that LAP3 in the pupae stage may be related to hystolysis of the larval tissues. Of the six loci detected, allelic variation was observed only for the LAP5 locus, with of codomi-nant action. The frequencies of genotypes among the progeny of females inseminated naturally deviate significantly from the Hardy-Weinberg equilibrium.

No estudo do ciclo biológico de Anopheles nuñez-tovari foram analisadas 108 posturas, totalizando 13.999 ovos. A média de ovos postos por postura foi de 128,83 + 4,85 e uma taxa de eclosão elevada de 84,06%. Foi constatado qe 89% dos ovos eclodem com 48 horas após a oviposição, mostrando, no total, um período médio de eclosão de 1,86 + 0,004 dias. A média de ovos eclodidos por postura foi de 109,39 + 5,26. Os dados de correlação entre ovos postos e ovos eclodidos evidenciaram que à medida que aumenta o número de ovos por postura há também um aumento correspondente na taxa de eclosão . Contudo, quando as posturas são reunidas em intervalos de classe de 40 ovos, constatou-se que não há correlação para algumas delas. Para as fases imaturas, verificou-se que existem diferenças no tempo de desenvolvimento, sendo o estádio 4o - pupa - o mais longo, apresentando uma média de 3,15 + 0,03 dias. A menor média foi observada para os estádios 2o - 3o com 1,46 + 0,02 dias. Valores intermediários foram constatados para os estádios 1o - 2o, 3o - 4oe estágio de pupa - adulto (2,34 + 0,02; 1,67 + 0,02 e 1,54 + 0,01 dias, respetivamente). Para o ciclo completo ovo-adulto observou-se um tempo médio de desenvolvimento de 12,02 + 0,04 dias. As maiores taxas de mortalidade foram verificadas para o 4o. estádio (19,6%0) e para o estádio até a emergência do imago, foi de 71,40%, sendo um valor elevado segundo os dados da literatura. A comparação dos períodos do desenvolvimento entre A. nuñez-tovari com A. darlingi e A. trannulatuspossibilitou considerar A. nuñez-tovari como mais próxima da primeira espécie.

In a study of the developmental cycle of Anopheles nuñez-tovari, 108 egg hatches with a total of 13.999 eggs were analyzed. The mean number of eggs per hatch was 128, 83 4,85 and there was a high eclosing rate of 84,06%. 89% out of the total number of eggs that ecloded within 240 hours, did so within the first 48 hours with a mean time of 1,86 + 0,004 days. The mean number of eclosions per hatch was 109,39 + 5,26. The proportion of eggs ecloding is positively correlated with the number of eggs per hatch. However, when eggs hatches are grouped in class intervals of 40 eggs there is no correlation for some of them. The developmental time of larval stages varied, the longest being the fourth (pre-pupal) stage with a mean of 3,15 + 0,03 days. The shortest mean interval was from the second to third instar (mean 1,46 + 0,02 days). Itermediate values were observed for the stages 1st + 2nd instar, 3rd - 4th instar and pupal-adult instar (2,34 + 0,02; 1,67 + 0,02 and 1,54 + 0,01 days, respectively). The mean developmental time from oviposition to adult emergencewas 12,02 + 0,004 days. The highest mortality was observed in the fourth larval (19,6%) and pupal (6,0%) stages. 71,4% of first instar larvae reached the adult stage, which is a high survival compared to previoous reports. The developmental time of A. nuñez-tovari appears to be closer to that of A. darlingi than A. triannulatus.

Dados sobre o grau de incidência e distribuição de espécies Anopheles, em Ariquemes (RO), evidenciaram que a diversidade é maior na periferia da cidade e que Anopheles darlingi é registrada em praticamente todas as localidades de coleta. O inquérito entomológico revelou níveis diferentes de penetração da espécie na área urbana, podendo-se constatar que os Setores 1 e 3 são áreas livres de malária; Setores 2 e 4 mostram riscos na periferia; e a Área Industrial e Setor de Áreas Especiais, Conjunto BNH, Setor 5 e Vila Velha constituem áreas de alto risco da malária. Nestes últimos, os índices de mosquitos por homem/hora foram os mais elevados, observando-se variações no decorrer das amostragens e conforme a localização da área urbana. Medidas de densidade populacional revelaram mudanças estacionais, sendo os menores valores registrados no período de inverno. A transmissão da malária é discutida, considerando-se: 1) o papel da estrutura física da cidade, na época da fundação, 2) os igarapés que margeam a área urbana e suas relações com o ciclo de desenvolvimento dos anofelinos, 3) os padrões comportamentais da atividade de picar das espécies correlacionados a ambientes naturais e às áreas ecologicamente alteradas, e 4) a importância do manuseio ambiental no controle da malária, para redução da densidade populacional. Para conter o processo migratório do vetor é proposto um cinturão de proteção à cidade, constituído de mata não densa, incluindo também proteção biológica para incentivar a zoofilia dos anofelinos. Os resultados de infecção natural, obtidos em áreas de autoctonia da malária, permitem citar A. darlingi como vetor, sendo discutida a possibilidade de que outras espécies estejam envolvidas na transmissão.

Data on the prevalence and distribution of species of Anopheles in Ariquemes (State of Rondonia) show, that the diversity is greater at the periphery of the town and that Anopheles darlingi is recorded at nearly all of the sampling points. The entomological survey revealed different levels of penetration by A. darlingi in to the urban area: Sectors 1 and 3 are free of malaria transmission, Sectors 2 and 4 show risks at the periphery, and the Industrial Sector, Sector of Special Areas, BNH housing estate, Sector 5 and the Old Town are areas of high risk for malaria. These areas had the highest indices of mosquitoes per man-hour, with variations of this index during the sampling period and depending on locality. Measures of population density showed seasonal variation, with the lowest indices during the rainy season. Malaria transmission is discussed in relation to 1) the physical structure of the town at the time it was jounded ; 2) the streams that border the urban area and their relation to the life cycle of the anophelines; 3) biting activity of the species colected in natural environments and areas altered by man; and 4) the importance of environmental management for malaria control by reducing the vector population density. To reduce the penetration of urban areas by mosquitoes from the surrounding countryside, a protective belt around the town is proposed. This belt should be of sparse forest and include alternative hosts to encourage the zoophilic tendencies of the anophelines. Infection rates among anophelines in areas authocthonous for malaria incriminate A. darlingi as a vector. The possibility of transmission by other species is discussed.