Intra-population plasticity of Anopheles darlingi's (Diptera, Culicidae) biting activity patterns in the state of Amapá, Brazil

OBJECTIVE: To assess the variation in Anopheles darlingi's biting activity compared to An. marajoara in the same locality and to biting activity data from other regions. METHODS: Using human bait, eight observations of the biting activity of An. darlingi and An. marajoara were carried out during 1999 and 2000 in the municipality of São Raimundo do Pirativa, state of Amapá, Brazil. Each observation consisted of three consecutive 13-hour collections, close to full moon. There were shifts of collectors in the observation points and nocturnal periods. RESULTS: An. darlingi revealed considerable plasticity of biting activity in contrast to An. marajoara, which showed well-defined crepuscular biting peaks. No significant correlation between density and biting activity was found, but a significant correlation existed between time and proportional crepuscular activity, indicating underlying ecological processes not yet understood. Two of the four available data sets having multiple observations at one locality showed considerable plasticity of this species' biting patterns as well. CONCLUSION: Intra-population variation of biting activity can be as significant as inter-population variation. Some implications in malaria vector control and specific studies are also discussed.

Infection of Anopheles darlingi fed on patients infected with Plasmodium vivax before and during treatment with chloroquine plus primaquine in Costa Marques, Rondônia, Brazil

Five patients with asexual and sexual parasites of Plasmodium vivax were treated orally with 600 mg chloroquine diphosphate (hour 0) followed with 300 mg at 8, 24 and 48 h later. Primaquine phospate, 15 mg, was administered concurrently at h 0 and 24 h intervals for 14 days. Anopheles darlingi were fed before the first dose (h-0.5) and 0.5, 1, 2, 4, 6, 8, 10, 12, 16, 20, 24, 36, 48, 60 and 72 h later. Mosquitoes were examined for oocysts on day 8 and for sporozoites on day 15 after infection. Four of the five patients studied were still infective to mosquitoes from 1-5 h after the first dose of chloroquine plus primaquine. One of these and one other patient, who vomited 15 min after the first dose, became inffective again at hours 10 and 12, respectively. Once produced, oocysts in mosquitoes fed on patients before, during and after chloroquine plus primaquine treatment appeared normal and produced sporozoite infected salivary glands. In view of these data , it is concluded that primaquine demonstrated rapid gametocytocidal activity and should be administred concurrently with chloroquine to reduce vivax malaria transmission.

Is there an efficient trap or collection method for sampling Anopheles darlingi and other malaria vectors that can describe the essential parameters affecting transmission dynamics as effectively as human landing catches? - A Review

Distribution, abundance, feeding behaviour, host preference, parity status and human-biting and infection rates are among the medical entomological parameters evaluated when determining the vector capacity of mosquito species. To evaluate these parameters, mosquitoes must be collected using an appropriate method. Malaria is primarily transmitted by anthropophilic and synanthropic anophelines. Thus, collection methods must result in the identification of the anthropophilic species and efficiently evaluate the parameters involved in malaria transmission dynamics. Consequently, human landing catches would be the most appropriate method if not for their inherent risk. The choice of alternative anopheline collection methods, such as traps, must consider their effectiveness in reproducing the efficiency of human attraction. Collection methods lure mosquitoes by using a mixture of olfactory, visual and thermal cues. Here, we reviewed, classified and compared the efficiency of anopheline collection methods, with an emphasis on Neotropical anthropophilic species, especially Anopheles darlingi, in distinct malaria epidemiological conditions in Brazil.

A importância do Anopheles darlingi Root, 1926 e Anopheles marajoara Galvão e Damasceno, 1942 na transmissão de malária no município de Macapá/AP - Brasil

A malária no município de Macapá é principalmente peri urbana, áreas estas caracterizadas por ressaca, pela presença de fragmentos de floresta e assentamentos desordenados (invasões). O objetivo deste estudo foi verificar a importância dos Anopheles darlingi e Anopheles marajoara na transmissão de malária em Macapá. O estudo foi realizado de outubro de 2007 a setembro de 2008, na comunidade de Lagoa dos Índios, Macapá. Foram coletados 4.601 mosquitos, dos quais 3.029 foram Anopheles marajoara (65,8%), 917 Anopheles darlingi (19,9%), 429 Anopheles braziliensis (9,3%), 208 Anopheles triannulatus (4,5%), 18 Anopheles peryassui (0,4%) e cinco Anopheles nuneztovari (0,1%). Apenas 32,8% dos espécimes foram coletadas no intradomicilio (1.511) e 67,2% no peridomicilio (3.090). O Indice de Picada Homem Hora do An. darlingi no intradomicilio variou entre 0 a 6,5 e no peridomicilio de 0 a 22 picadas homem hora. Já para o An. marajoara a variaçao foi de 0 a 22 no intradomicilio e de 0 a 175,5 no peridomicilio. A analise das exúvias e da dissecção de genitália resultaram na confirmaçao das duas espécies estudadas, An. darlingi e An. marajoara, e que o An. marajoara é a única espécie do complexo albitarsis circulante na área. A abundância dos vetores flutuou associada com o padrão sazonal das chuvas. An. darlingi é mais abundante no final e inicio das chuvas, enquanto o An. marajoara esteve presente em alta densidade durante todo o período de chuvas. Dos 4.601 mosquitos testados, 100 foram positivos para plasmódios humanos pelo método de ELISA, resultando em uma taxa de infecção de 2,17%. Dos 3.029 An. marajoara testados 71 (2,34%) foram positivos e dos 917 An. darlingi, 28 (3,05%). Este estudo demonstrou que as duas espécies estudadas mantêm a transmissão de malária durante todo o ano, ratificando assim a importância das mesmas.

Development of the BG-Malaria trap as an alternative to human-landing catches for the capture of Anopheles darlingi

Although the human-landing catch (HLC) method is the most effective for collecting anthropophilic anophelines, it has been increasingly abandoned, primarily for ethical considerations. The objective of the present study was to develop a new trap for the collection of Anopheles darlingi . The initial trials were conducted using the BG-Sentinel trap as a standard for further trap development based on colour, airflow direction and illumination. The performance of the trap was then compared with those of the CDC, Fay-Prince, counterflow geometry trap (CFG) and HLC. All trials were conducted outdoors between 06:00 pm-08:00 pm. Female specimens of An. darlingi were dissected to determine their parity. A total of 8,334 anophelines were captured, of which 4,945 were identified as An. darlingi . The best trap configuration was an all-white version, with an upward airflow and no required light source. This configuration was subsequently named BG-Malaria (BGM). The BGM captured significantly more anophelines than any of the other traps tested and was similar to HLC with respect to the number and parity of anophelines. The BGM trap can be used as an alternative to HLC for collecting anophelines.

Susceptibility of Anopheles aquasalis and An. darlingi to Plasmodium vivax VK210 and VK247

The susceptibility of Anopheles aquasalis (F₃ generation) and An. darlingi (F₁ generation) to Plasmodium vivax circumsporozoite protein phenotypes from a limited number of blood samples of malaria patients in Belém, state of Pará, Brazil, was examined. A polymerase chain reaction was used to determine the P. vivax phenotypes in blood samples and the blood-fed infected mosquitoes were dissected and tested by ELISA. In all patient infections, more infected An. aquasalis and An. darlingi were positive for VK210 compared with VK247.

Ecology of malaria vectors in the Americas and future direction

The resurgence of malaria in the Americas has renewed interest in Anopheles biology. Anopheles darlingi, An. albimanus, An. nuneztovai and An aquasalis are reconfirmed as major malaria vectors and other species are playing important roles in regional malaria transmission. Adultbiting activity and larval ecology are discussed in detail. Seasonal abundance and daily biting activity of Anopheles vary considerably among species and geographically for the same species. Anopheles albimanus has the least amount of variation in biting activity over its range and An. darlingi has the greatest. All species studied are more exophilic and exophagic than endophilic and endophagic. Anopheles darling is more antropophilic, endophilic and endophagic than other Anophelines. Larval studies remain more descriptive than comprehensive. Research on Anophelines is becoming more integrated and biologists are using new biochemical techniques and ecological principles to answer critical questions. This "pluralization" will help us understand species complexes, population dynamics and malaria transmission. integrated control programs will require more regional, in-depth ecological studies.

Primeiro registro de Anopheles (Kerteszia) homunculus Komp (Diptera, Culicidae) no Estado do Espírito Santo, Brasil

Em pesquisa de campo realizada no interior do Estado do Espírito Santo, Brasil, em janeiro de 2006, como parte de projeto sobre a transmissão de Plasmodium, foram coletadas larvas de anofelinos em bromélias. Os imaturos foram mantidos no laboratório até a obtenção dos adultos machos e fêmeas associados com as exúvias das larvas e das pupas, para serem identificados. Conseqüentemente, verificou-se que dois espécimes pertenciam a Anopheles (Kerteszia) homunculus Komp, 1937. Este é o primeiro registro dessa espécie de Kerteszia no Espírito Santo. O encontro evidencia a importância de estudos adicionais de modo a estabelecer a distribuição geográfica do An. homunculus, bem como o status taxonômico e a importância epidemiológica da espécie na dinâmica da transmissão da malária em áreas de Mata Atlântica.

Molecular phylogeny of the Myzorhynchella Section of Anopheles (Nyssorhynchus) (Diptera: Culicidae): genetic support for recently described and resurrected species

Phylogenetic relationships among species of the Myzorhynchella Section of Anopheles (Nyssorhynchus) were investigated using the nuclear ribosomal DNA second internal transcribed spacer (ITS2), the nuclear whitegene and mitochondrial cytochrome oxidase subunit I (COI) regions. The recently described Anopheles pristinus and resurrected Anopheles guarani were also included in the study. Bayesian phylogenetic analyses found Anopheles parvus to be the most distantly related species within the Section, a finding that is consistent with morphology. An. pristinus and An. guarani were clearly resolved from Anopheles antunesi and Anopheles lutzii, respectively. An. lutzii collected in the same mountain range as the type locality were found within a strongly supported clade, whereas individuals from the southern state of Rio Grande do Sul, tentatively identified as An. lutzii based on adult female external morphology, were distinct from An. lutzii, An. antunesi and from each other, and may therefore represent two new sympatric species. A more detailed examination of An. lutzii sensu latoalong its known geographic range is recommended to resolve these anomalous relationships.

Habitat suitability of Anopheles vector species and association with human malaria in the Atlantic Forest in south-eastern Brazil

Every year, autochthonous cases of Plasmodium vivax malaria occur in low-endemicity areas of Vale do Ribeira in the south-eastern part of the Atlantic Forest, state of São Paulo, where Anopheles cruzii and Anopheles bellator are considered the primary vectors. However, other species in the subgenus Nyssorhynchus of Anopheles (e.g., Anopheles marajoara) are abundant and may participate in the dynamics of malarial transmission in that region. The objectives of the present study were to assess the spatial distribution of An. cruzii, An. bellator and An. marajoara and to associate the presence of these species with malaria cases in the municipalities of the Vale do Ribeira. Potential habitat suitability modelling was applied to determine both the spatial distribution of An. cruzii, An. bellator and An. marajoara and to establish the density of each species. Poisson regression was utilized to associate malaria cases with estimated vector densities. As a result, An. cruzii was correlated with the forested slopes of the Serra do Mar, An. bellator with the coastal plain and An. marajoara with the deforested areas. Moreover, both An. marajoara and An. cruzii were positively associated with malaria cases. Considering that An. marajoara was demonstrated to be a primary vector of human Plasmodium in the rural areas of the state of Amapá, more attention should be given to the species in the deforested areas of the Atlantic Forest, where it might be a secondary vector.

Anopheles (Nyssorhynchus) atacamensis (Diptera: Culicidae), a new species from Northern Chile

Anopheles (Nyssorhynchus) atacamensis, a new species in the subgenus Nyssorhynchus, is described and validated using morphological characters of the male and female adult, male genitalia and immature stages. Molecular characterization employing sequences of the ITS2 rDNA and COI mtDNA are provided. The new taxon is compared with Anopheles (Nyssorhynchus) pictipennis (Philippi) from central Chile based on morphological features of the adults, male genitalia and larva. Illustrations of the diagnostic characteristics of the male genitalia, fourth-instar larva and pupa are provided.

Morphological analysis of three populations of Anopheles (Nyssorhynchus) nuneztovari Gabaldón (Diptera: Culicidae) from Colombia

Based on the results of comparative analyses of 1,039 specimens of several progenies of Anopheles nuneztovarifrom three localities in Colombia, eight costal wing spot patterns were observed. Patterns I and III were the most frequent: 77.96% and 11.36%, respectively. Using the diagnostic characters ratio of the length of the basal dark area of hind tarsomere II/length of hind tarsomere II, ratio of the length of the humeral pale spot/length of the pre-humeral dark spot, and the ratio of the length of the subcostal pale spot/length of the distal sector dark spot (DS-III2/Ta-III2, HP/PHD, SCP/DSD) approximately 5% of the adult females were misidentified as a species of Nyssorhynchus, different from An. nuneztovari. Approximately 5% of the specimens showed DS-III2/Ta-III2 ratio less than 0.25 (range 0.21 - 0.24), and among them 3.34% shared a HP/PHD ratio less than 1.50. Consequently, 1.52% of An. nuneztovari individuals can be misidentified as Anopheles oswaldoi. In those specimens with the DS-III2/Ta-III2 ratios higher than 0.25, 34.45% displayed SCP/DSD values greater than 0.50 and of these, 3.65% displayed HP/PHD values greater than 1.8. This combination of characters could lead one to misidentify samples of An. nuneztovari as Anopheles rangeli. Similarly, 2.43% of the females could be identified erroneously as either Anopheles aquasalis or Anopheles benarrochi. Individuals with a HP/PHD ratio greater than 2.0, could be misidentified as Anopheles trinkae, Anopheles strodei or Anopheles evansae. A distinct combination of diagnostic characters for An. nuneztovari from Colombia is proposed.

Effects of flooding of the River Paraná on the temporal activity of Anopheles (Nyssorhynchus) darlingi Root (Diptera: Culicidae), at the border state of Mato Grosso do Sul and São Paulo, Brazil

INTRODUCTION: Study of the temporal activity of malaria vectors during the implantation of a hydroelectric power station on the River Paraná, intended to generate electrical energy. The river separates the States of São Paulo and Mato Grosso do Sul, in Brazil. The objective was to verify whether alterations occurred in the wealth and diversity indices of Anopheles, following two successive floods, extended to the temporal activity and nycthemeral rhythm followed over a five year period. METHODS: Mosquito capture was performed monthly using the Human Attraction Technique and Shannon Traps. The first, executed for 24h, provided the nycthemeral rhythm and the second, lasting 15h, permitted the tracking of Anopheles during the two floods. RESULTS: The bimodal pattern of Anopheles darlingi defined before these floods was modified throughout the environment interventions. The same effect had repercussions on the populations of An albitarsis s.l., An triannulatus and An galvaoi. Activity prior to twilight was less affected by the environment alterations. CONCLUSIONS: The dam construction provoked changes in Anopheles temporal activity patterns, permitting classification of the area as an ecologically steady and unstable situation. Differences observed in Anopheles behavior due to the capture methods revealed the influence of solo and multiple attractiveness inside the populations studied.

Ecologia de Anopheles (Nyssorhynchus) darlingi Root em área de implantação de empreendimento hidrelétrico, na divisa dos Estados do Mato Grosso do Sul e São Paulo

INTRODUÇÃO: Hidrelétricas alteram o fluxo das águas e provocam impactos sobre a composição de mosquitos, justificando-se essa pesquisa. O objetivo da pesquisa foi estudar anofelinos de área sob a influência de um novo lago e avaliar a vulnerabilidade relativa à malária. MÉTODOS: Foram feitas coletas de Anopheles nas margens da Represa Porto Primavera, durante as fases do alagamento até sua cota máxima. Utilizaram-se as técnicas: atrativa humana, de armadilha de Shannon e concha entomológica. Os indicadores Riqueza e Diversidade foram utilizados para medir o impacto. A análise das distribuições temporais foi realizada pelo teste Mann-Whitney, considerando localidade, cota e método de captura como variáveis independentes (α=0,05). RESULTADOS: A densidade de Anopheles darlingi oscilou entre as localidades A, B e C, sendo que os maiores picos foram para B e C. Com a estabilidade do lago, no último nível, evidenciou-se a tendência de redução da densidade de Anopheles darlingi. CONCLUSÕES: Sugere-se que o risco de autoctonia de malária nas proximidades do lago permanece inalterado, ficando o alerta para esporádicas infecções humanas.

Encontro de imaturos de Anopheles cruzii em bromélias de área urbana, litoral de São Paulo

Primeiro relato da ocorrência de larvas de Anopheles (Kerteszia) cruzii, mosquito essencialmente silvestre, em bromélias de solo em área urbana do município de Ilhabela, litoral norte do estado de São Paulo. De março de 1998 a julho de 1999 foram capturadas 312 formas imaturas de An. cruzii, sendo 8,6% em bromélias do ambiente urbano, 40,1% em bromélias do periurbano e 51,3% na mata. O número médio de bromélias com An. cruzii foi de 4,0% dentre o total de pesquisadas, com valores próximos de positividade para ambiente periurbano e mata. A presença de An. cruzii no ambiente urbano provavelmente é resultante da sua ocorrência prévia na mata, aliada à elevada presença desse criadouro na área urbana, de fonte alimentar e abrigos disponíveis. Alerta-se para a possibilidade de transferência de infecções entre esses ambientes