Malaria vectors in the municipality of Serra do Navio, State of Amapá, Amazon Region, Brazil

We conducted a survey to determine the vectors of malaria in six localities of Serra do Navio municipality, State of Amapá, from 1990 to 1991. Malaria infection rates of 29.3%, 6.2% and 20.4% were detected by human blood smears in Colônia Água Branca, Porto Terezinha and Arrependido, respectively. There was no malaria infection detected in Serra do Navio. Fifteen species were identified among 3,053 anopheline mosquitoes collected by human bait and 64.4% were identified as Anopheles albitarsis s.l., 16.7% An. braziliensis, 9.5% An. nuneztovari and 5.8% An. triannulatus. An. darlingi, the main vector of malaria in the Amazon region of Brazil, was scare. Using enzyme-linked immunosorbent assay (ELISA), a total positive rate of 0.8% (23/2876) was found for six species: fifteen An. albitarsis s.l., four An. nuneztovari, and one of each: An. braziliensis, An. triannulatus, An. oswaldoi and An. rangeli. Nine of 23 positive mosquitoes were infected with Plasmodium malariae, eight with P. vivax VK210, three with P. vivax VK247 and three with P. falciparum. Since An. albitarsis s.l. was collected feeding on humans, was present in the highest density and was positive by ELISA for malaria sporozoites, it probably plays an important role in malaria transmission in this area.

Biting Indices, Host-seeking Activity and Natural Infection Rates of Anopheline Species in Boa Vista, Roraima, Brazil from 1996 to 1998

The epidemiology of the transmission of malaria parasites varies ecologically. To observe some entomological aspects of the malaria transmission in an urban environment, a longitudinal survey of anopheline fauna was performed in Boa Vista, Roraima, Brazil. A total of 7,263 anophelines was collected in human bait at 13 de Setembro and Caranã districts: Anopheles albitarsis sensu lato (82.8%), An. darlingi (10.3%), An. braziliensis (5.5%), An. peryassui (0.9%) and An. nuneztovari (0.5%). Nightly 12 h collections showed that An. albitarsis was actively biting throughout the night with peak activities at sunset and at midnight. An. darlingi bit during all night and did not demonstrate a defined biting peak. Highest biting indices, entomological inoculation rates and malaria cases were observed seasonally during the rainy season (April-November). Hourly collections showed host seek activity for all mosquitoes peaked during the first hour after sunset. An. darlingi showed the highest plasmodial malaria infection rate followed by An. albitarsis, An. braziliensis and An. nuneztovari (8.5%, 4.6%, 3% and 2.6%, respectively). An. albitarsis was the most frequently collected anopheline, presented the highest biting index and it was the second most frequently collected infected species infected with malaria parasites. An. albitarsis and An. darlingi respectively, are the primary vectors of malaria throughout Boa Vista.

Morphometric discrimination of females of five species of Anopheles of the subgenus Nyssorhynchus from Southern and Northwest Colombia

The most important vectors of human Plasmodium in the neotropics belong to the subgenus Nyssorhynchus. These species are generally sympatric in terms of their geographical distributions. Some are difficult to identify based solely on examination of adult females using the available morphological keys, in these cases examination of immature stages and male genitalia is required to make correct determinations. However, in epidemiological studies it is necessary to identify the species of adult females which are found near humans, i.e. in studies of malaria transmission or evaluation of control measures. The purpose of the present study was to evaluate the discrimination of adult females of different species of Nyssorhynchus isolated mainly from Southern Colombia (department of Putumayo), using morphometric analysis. Adult females were obtained after rearing larvae collected in natural breeding places and from the progeny of females collected on humans. The morphological characteristics of the immature stages allowed the identification of four species of the subgroup Oswaldoi from Southern Colombia: Anopheles rangeli Gabaldon, Cova Garcia & Lopez, An. oswaldoi (Peryassu), An. benarrochi Gabaldon, Cova Garcia & Lopez and An. triannulatus (Neiva & Pinto). The species An. nuneztovari (Gabaldon) from the Northwest of Colombia was included for comparison. Morphometric analysis allowed differentiation of the females of all species to a confidence level approaching 90% using principal components analysis of 10 wing and leg variables, followed by canonical variate analysis of the first four principal components. We conclude that morphometrics may represent a useful taxonomic tool for this group and that its use should be further studied.

The importance of Anopheles albitarsis E and An. darlingi in human malaria transmission in Boa Vista, state of Roraima, Brazil

In several districts of Boa Vista, state of Roraima, Brazil we found Anopheles (Nyssorhynchus) albitarsis E to be the primary vector of human malaria parasites, and during 2001-2002 it was significantly more abundant than An. darlingi (p < 0.001). Other species sampled were An. (Nys.) braziliensis, An. (Ano.) peryassui, An. (Nys.) nuneztovari, An. (Nys.) oswaldoi s.l., and An. (Nys.) triannulatus. As determined by the ELISA technique An. darlingi had a higher overall infection rate (2.1%) compared with An. albitarsis E (1.2%). However, a marginally higher proportion of An. albitarsis E was infected with Plasmodium vivax compared with An. darlingi, and the An. albitarsis E biting index was also much higher. These results suggest the importance of An. albitarsis E in malaria transmission in a savannah ecoregion of northern Amazonian Brazil, and reconfirm the importance of An. darlingi even if at lower abundance.

Systematic notes on Anopheles Meigen (Diptera: Culicidae) species in the state of Amapá, Brazil

Identification of Anopheles nuneztovari Gabaldón and An. goeldii Rozeboom and Gabaldón based on the male genitalia traits is discussed. An. goeldii is in the synonymy of An. nuneztovari, however, characters of the aedeagus of male genitalia distinguish both species. We hypothesize that An. goeldii may be a valid species, however, further studies using molecular characters, especially ITS2 rDNA sequences will be necessary to elucidate the taxonomic status of the species. An. konderi Galvão and Damasceno and An. forattinii Wilkerson and Sallum are registered for the first time in the state of Amapá.

Species composition and natural infectivity of anthropophilic Anopheles (Diptera: Culicidae) in the states of Córdoba and Antioquia, Northwestern Colombia

Malaria is a serious health problem in the states of Córdoba and Antioquia, Northwestern Colombia, where 64.4% of total Colombian cases were reported in 2007. Because little entomological information is available in this region, the aim of this work was to identify the Anopheles species composition and natural infectivity of mosquitoes distributed in seven localities with highest malaria transmission. A total of 1,768 Anopheles mosquitoes were collected using human landing catches from March 2007-July 2008. Ten species were identified; overall, Anopheles nuneztovari s.l. was the most widespread (62%) and showed the highest average human biting rates. There were six other species of the Nyssorhynchus subgenus: Anopheles albimanus (11.6%), Anopheles darlingi (9.8%), Anopheles braziliensis (6.6%), Anopheles triannulatus s.l. (3.5%), Anopheles albitarsis s.l. and Anopheles oswaldoi s.l. at < 1%; and three of the Anopheles subgenus: Anopheles punctimacula, Anopheles pseudopunctipennis s.l. and Anopheles neomaculipalpusat < 1% each. Two species from Córdoba, An. nuneztovari and An. darlingi, were found to be naturally infected by Plasmodium vivax VK247, as determined by ELISA and confirmed by nested PCR. All species were active indoors and outdoors. These results provide basic information for targeted vector control strategies in these localities.

Molecular comparison of topotypic specimens confirms Anopheles (Nyssorhynchus) dunhami Causey (Diptera: Culicidae) in the Colombian Amazon

The presence of Anopheles (Nyssorhynchus) dunhami Causey in Colombia (Department of Amazonas) is confirmed for the first time through direct comparison of mtDNA cytochrome c oxidase I (COI) barcodes and nuclear rDNA second internal transcribed spacer (ITS2) sequences with topotypic specimens of An. dunhami from Tefé, Brazil. An. dunhami was identified through retrospective correlation of DNA sequences following misidentification as Anopheles nuneztovari s.l. using available morphological keys for Colombian mosquitoes. That An. dunhami occurs in Colombia and also possibly throughout the Amazon Basin, is of importance to vector control programs, as this non-vector species is morphologically similar to known malaria vectors including An. nuneztovari, Anopheles oswaldoi and Anopheles trinkae. Species identification of An. dunhami and differentiation from these closely related species are highly robust using either DNA ITS2 sequences or COI DNA barcode. DNA methods are advocated for future differentiation of these often sympatric taxa in South America.

Malaria vector species in Colombia: a review

Here we present a comprehensive review of the literature on the vectorial importance of the major Anopheles malaria vectors in Colombia. We provide basic information on the geographical distribution, altitudinal range, immature habitats, adult behaviour, feeding preferences and anthropophily, endophily and infectivity rates. We additionally review information on the life cycle, longevity and population fluctuation of Colombian Anopheles species. Emphasis was placed on the primary vectors that have been epidemiologically incriminated in malaria transmission: Anopheles darlingi, Anopheles albimanus and Anopheles nuneztovari. The role of a selection of local, regional or secondary vectors (e.g., Anopheles pseudopunctipennis and Anopheles neivai) is also discussed. We highlight the importance of combining biological, morphological and molecular data for the correct taxonomical determination of a given species, particularly for members of the species complexes. We likewise emphasise the importance of studying the bionomics of primary and secondary vectors along with an examination of the local conditions affecting the transmission of malaria. The presence and spread of the major vectors and the emergence of secondary species capable of transmitting human Plasmodia are of great interest. When selecting control measures, the anopheline diversity in the region must be considered. Variation in macroclimate conditions over a species' geographical range must be well understood and targeted to plan effective control measures based on the population dynamics of the local Anopheles species.

Can Mosquito Magnet® substitute for human-landing catches to sample anopheline populations?

The efficiency of the Mosquito Magnet Liberty PlusTM (MMLP) trap was evaluated in comparison to human-landing catches (HLCs) to sample anopheline populations in Jabillal, state of Bolivar, southern Venezuela. The village comprised 37 houses and a population of 101; malaria in this village is primarily due to Plasmodium vivax and the Annual Parasite Index is 316.8 per 1,000 population. A longitudinal study was conducted between June 2008-January 2009 for three nights per month every two months between 17:30 pm-21:30 pm, a time when biting mosquitoes are most active. Anopheles darlingi and Anopheles nuneztovari were the most common species collected by both methods, whereas Anopheles marajoara was more abundant according to the HLC method. The MMLP trap was more efficient for collecting An. nuneztovari [63%, confidence interval (CI): 2.53] than for collecting An. darlingi (31%, CI: 1.57). There were significant correlations (p < 0.01) between the two methods for An. darlingi [Pearson correlation (R²) = 0.65] and An. nuneztovari (R² = 0.48). These preliminary results are encouraging for further investigations of the use of the MMLP trap for monitoring anopheline populations in remote malaria-endemic areas in the Amazon Basin.

Malaria entomological risk factors in relation to land cover in the Lower Caura River Basin, Venezuela

To explore the effects of deforestation and resulting differences in vegetation and land cover on entomological parameters, such as anopheline species composition, abundance, biting rate, parity and entomological inoculation rate (EIR), three villages were selected in the Lower Caura River Basin, state of Bolívar, Venezuela. All-night mosquito collections were conducted between March 2008-January 2009 using CDC light traps and Mosquito Magnet(r) Liberty Plus. Human landing catches were performed between 06:00 pm-10:00 pm, when anophelines were most active. Four types of vegetation were identified. The Annual Parasite Index was not correlated with the type of vegetation. The least abundantly forested village had the highest anopheline abundance, biting rate and species diversity. Anopheles darlingi and Anopheles nuneztovari were the most abundant species and were collected in all three villages. Both species showed unique biting cycles. The more abundantly forested village of El Palmar reported the highest EIR. The results confirmed previous observations that the impacts of deforestation and resulting changes in vegetation cover on malaria transmission are complex and vary locally.

The sandfly fauna, anthropophily and the seasonal activities of Pintomyia spinicrassa (Diptera: Psychodidae: Phlebotominae) in a focus of cutaneous leishmaniasis in northeastern Colombia

This study was conducted to identify the sandfly fauna and the anthropophilic species in a coffee-growing area of Villanueva, Norte de Santander, Colombia, a focus of American cutaneous leishmaniasis, and to analyse the relationship between the most frequent species and rainfall, relative humidity and temperature, with the aim of contributing to epidemiological surveillance in the area. Sandfly collections were performed fortnightly between February 2006-September 2007 using automatic light traps, Shannon traps, protected human bait and aspiration in resting places. A total of 7,051 sandflies belonging to 12 species were captured. Pintomyia spinicrassa (95.7%) predominated. Pintomyia oresbia and Lutzomyia sp. of Pichinde were found in the state of Norte de Santander for the first time. Pi. spinicrassa, Pintomyia nuneztovari, Micropygomyia venezuelensis, Lutzomyia (Helcocyrtomyia) scorzai and Lu. (Helcocyrtomyia) sp. were captured on the protected human bait. A significant association between Pi. spinicrassa abundance and the total rainfall and the average temperature and humidity 10 days before the collection was observed. The dominance of Pi. spinicrassa, a recognised vector of Leishmania braziliensis, especially during the dry periods, indicates that the risk of parasite transmission may increase.

Anthropophilic Anopheles species composition and malaria in Tierradentro, Córdoba, Colombia

Malaria is still a primary health problem in Colombia. The locality of Tierradentro is situated in the municipality of Montelíbano, Córdoba, in the northwest of Colombia, and has one of the highest annual parasite index of malaria nationwide. However, the vectors involved in malaria transmission in this locality have not yet been identified. In this study, the local anthropophilic Anopheles composition and natural infectivity with Plasmodium were investigated. In August 2009, 927 female Anopheles mosquitoes were collected in eight localities using the human landing catch method and identified based on their morphology. Cryptic species were determined by restriction fragment length polymorphism-internal transcribed spacer (ITS)2 molecular analysis. Eight species [Anopheles nuneztovari s.l. (92.8%), Anopheles darlingi (5.1%), Anopheles triannulatus s.l. (1.8%), Anopheles pseudopunctipennis s.l. (0.2%), Anopheles punctimacula s.l. (0.2%), Anopheles apicimacula (0.1%), Anopheles albimanus (0.1%) and Anopheles rangeli (0.1%)] were identified and species identity was confirmed by ITS2 sequencing. This is the first report of An. albimanus, An. rangeli and An. apicimacula in Tierradentro. Natural infectivity with Plasmodium was determined by ELISA. None of the mosquitoes was infectious for Plasmodium. An. nuneztovari s.l. was the predominant species and is considered the primary malaria vector; An. darlingi and An. triannulatus s.l. could serve as secondary vectors.

Phylogenetic relationships among species of Anopheles (Nyssorhynchus) (Diptera, Culicidae) based on nuclear and mitochondrial gene sequences

Phylogenetic relationships among 21 species of mosquitoes in subgenus Nyssorhynchus were inferred from the nuclear white and mitochondrial NADH dehydrogenase subunit 6 (ND6) genes. Bayestan phylogenetic methods found that none of the three Sections within Nyssorhynchus (Albimanus, Argyritarsis, Myzorhynchella) were supported in all analyses, although Myzorhynchella was found to be monophyletic at the combined genes Within the Albimanus Section the monophyly of the Stroder Subgroup was strongly supported and within the Myzorhynchella Section Anopheles anrunesi and An lutzu formed a strongly supported monophyletic group The epidemiologically significant Albitarsis Complex showed evidence of paraphyly (relative to An lanet-Myzorhynchella) and discordance across gene trees, and the previously synonomized species of An. dunhami and An goeldii were recovered as sister species Finally, there was evidence of complexes in several species, including An antunesi, An deaneorum, and An. strodei (c) 2010 Elsevier B.V. All rights reserved

Identificação molecular de espécies do complexo Anopheles albitarsis (Diptera, Culicidae, Anophelinae), coletadas em dois municípios da Amazônia brasileira, com análise de suscetibilidade natural a plasmódios humanos

Anofelinos membros de complexos de espécies crípticas podem exibir diferenças comportamentais, de susceptibilidade a infecção malárica, e resistência a inseticidas. Assim, a identificação de espécies vetoras tem relevância epidemiológica, o que nem sempre é possível por critérios morfológicos. Métodos alternativos têm sido empregados para tal, como os que analisam regiões altamente conservadas do DNA ribossômico, variável entre as espécies, conhecidas como espaçadoras internas transcritas (ITS). Considera-se atualmente que o complexo Anopheles c seja composto por seis espécies: An. albitarsis s.s., An. oryzalimnetes, An. albitarsis F, An. marajoara, An. deaneorum, e An. janconnae. Destas, pelo menos as três últimas são incriminadas como vetores de malária na Amazônia brasileira. O objetivo deste estudo foi realizar identificação molecular de espécies do complexo An. albitarsis, por análise da seqüências do ITS2 do rDNA, com vistas a analisar sua importância na transmissão de malária nos municípios de Macapá, Amapá e Peixe-Boi, Pará, inclusive investigando pela primeira vez a ocorrência do An. albitarsis F nestas duas áreas epidemiologicamente distintas: a primeira com histórico de alto risco de transmissão de malária e a segunda não. O estudo foi realizado entre janeiro de 2009 e abril de 2010, e consistiu de capturas de anofelinos de 12 horas de duração (ecostofase) no peridomicílio. Todas as fêmeas coletadas foram morfologicamente identificadas e apenas os An. albitarsis s.l. tiveram cabeça e tórax separadas para análise da infecção natural por ELISA; ovários para análise de paridade e patas, asas e carcaça para identificação molecular. Em Macapá foram realizadas seis coletas, obtendo-se um total de 584 anofelinos, sendo 366 An. albitarsis s.l. (62,7%), 167 An. darlingi (28,6%), 33 An. triannulatus s.l (5,6%), 15 An. braziliensis (2,6%) e 3 An. nuneztovari (0,5%). Pela PCR foi possível visualizar a banda específica de An. marajoara em 320 espécimes dos An. albitarsis s.l testados. Do restante, 33 foram negativos e 13 amplificaram um fragmento de ~490 pb nos iniciadores empregados, não permitindo chegar ao diagnóstico específico. O An. marajoara apresentou características biológicas e comportamentais que ratificam sua importância epidemiológica na transmissão de malária em Macapá, tais como: ser a espécie mais prevalente, com maior proporção de fêmeas paridas (73,0%), e portanto com maiores chances de se infectarem com o plasmódio, ocorrer tanto na estação menos quanto na mais chuvosa, e apresentar atividade hematofágica durante toda a ecostofase, alem disso, foi encontrado naturalmente infectado por P. vivax e P. falciparum (taxa de infecção natural de 3,1%). Em Peixe-Boi, foram capturados 43 anofelinos: An. triannulatus s.l (20 espécimes, 46,5 %), An. albitarsis s.l. (13: 30,2 %), An. darlingi (8: 18,6%), e An. nuneztovari (2: 4,7%). Todos os An. albitarsis s.l. coletados foram identificados pela ITS2 como An. oryzalimnetes. Nenhum deles foi encontrado infectado pelos plasmódios testados, e a maioria das fêmeas era parida (84,6%). São necessários levantamentos entomológicos sistemáticos que analisem a importância deste anofelino na transmissão de malária na cidade. O An. albitarsis F não foi encontrado nas duas áreas estudadas. Nossos resultados contribuem para o entendimento da epidemiologia da malária na região Amazônica brasileira.

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.