Identification of Anopheles (Nyssorhynchus) albitarsis complex species (Diptera: Culicidae) using rDNA internal transcribed spacer 2-based polymerase chain reaction primes

Anopheles (Nyssorhynchus) marajoara is a proven primary vector of malaria parasites in Northeast Brazil, and An. deaneorum is a suspected vector in Western Brazil. Both are members of the morphologically similar Albitarsis Complex, which also includes An. albitarsis and an undescribed species, An. albitarsis "B". These four species were recognized and can be identified using random amplified polymorphic DNA (RAPD) markers, but various other methodologies also point to multiple species under the name An. albitarsis. We describe here a technique for identification of these species employing polymerase chain reaction (PCR) primers based on ribosomal DNA internal transcribed spacer 2 (rDNA ITS2) sequence. Since this method is based on known sequence it is simpler than the sometimes problematical RAPD-PCR. Primers were tested on samples previously identified using RAPD markers with complete correlation.

Comportamento exófilo de Anopheles darlingi Root, em região meridional do Brasil

Observou-se o comportamento da população "Araraquara" de Anopheles darlingi, em seu ambiente original e em relação à sua atividade exófila com isca humana. As coletas foram realizadas às margens do rio Jacaré-Pepira, no Município de Dourado, Estado de São Paulo, Brasil. O ciclo nictemeral caracteriza-se pelo aspecto bimodal, com os dois picos correspondentes a cada crepúsculo, ou seja, vespertino e matutino. O seu detalhamento permitiu detectar bimodalidade secundária, subdividindo cada pico em um eocrepuscular precedendo ao intracrepuscular propriamente dito. A variação sazonal revelou aumento do número de mosquitos na estação chuvosa e quente e nítido declínio na seca e fria. Embora com dados insuficientes, houve indícios de que o An. albitarsis local, apresente também ritmo bimodal em seu ciclo diário de atividade.

Espécies de Anopheles (Culicidae, Anophelinae) em área endêmica de malária, Maranhão, Brasil

OBJETIVO: Estudar a flutuação sazonal, freqüência horária, abundância relativa e riqueza de espécies de Anopheles em ambiente antrópico, para entender a bioecologia do grupo e para a monitorização do programa de controle da malária. MÉTODOS: Os anofelinos foram estudados durante um ano, de outubro de 1996 a setembro de 1997, das 18 às 6 horas, a cada 30 dias, no Município da Raposa, Ilha de São Luís, Maranhão. Utilizou-se o método de captura de fêmeas em iscas humanas no intra e peridomicílio, com tubo de sucção e foco luminoso orientado. RESULTADOS: Foram coletados 1.407 espécimes assim distribuídos: Anopheles aquasalis (82%), Anopheles galvaoi (10,2%) e Anopheles albitarsis (6,4%). As demais espécies, Anopheles evansae, Anopheles nuneztovari e Anopheles triannulatus davisi representaram juntas 1,4%. Os anofelinos ocorreram o ano inteiro, principalmente no período chuvoso, sendo mais freqüentes no intra (75,3%) do que no peridomicílio (24,7%), preferindo sugar sangue no crepúsculo vespertino e nas primeiras horas da noite. CONCLUSÃO: As variações observadas no comportamento do anofelino mostram que as diferentes espécies vêm adaptando-se, em maior ou menor grau, ao convívio com o homem nas suas habitações.

A survey of the Anopheles Mosquitoes (Diptera: Culicidae) along the Acre And Purus Rivers, Amazon Basin, Brazil

From January 19 to February 25, 1997 an entomological survey of the seringais and larger towns along the Acre and Purus rivers was made as part of the project “Revisitando a Amazônia de Carlos Chagas; da Borracha a Biodiversidad”. Eleven anopheline species and 1285 specimens were collected landing on human baits. The four most abundant species were Anopheles albitarsis s.l. (n=778), A. darlingi (n=359), A. rangeli (n=69) and A. oswaldoi (n=60). A total of 252 larvae were collected of which 10 anopheline species were identified. The most abundant species collected were A. albitarsis s.l. (n=88), A. deaneorum (n=45) and A. triannulatus (n=40). The low numbers of Anopheles collected and the absence of the principal malaria vector A. darlingi at the seringais sites suggests that they arc not high risk malarious areas. Other Diptera collected were Culex sp., Mansonia titillans, Mansonia pseudotitillans, Psorophora ciliata, Psorophora sp., Coquillettidia (Rhynchotaenia) sp., Simulium amazonicum and S. sanguineum.

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.

Notes on the holotype of Anopheles marajoara Galvão & Damasceno (Diptera, Culicidae)

During studies on the dynamics of malaria transmission in Marajó Island, State of Pará, Brazil, Galvão & Damasceno (1942) collected a single specimen of a new species that they named Anopheles (Nyssorhynchus) marajoara Galvão & Damasceno, 1942. Now, examining genitalia slide associated to the holotype, we observed that the ventral claspette of the male genitalia is distinct from those of all other species of the Argyritarsis Section and consequently from members of the complex Anopheles albitarsis Lynch Arribalzaga, 1878. The male genitalia of the slide belong to a specimen of Anopheles aquasalis Curry, 1932, nevertheless, it was originally labeled as Anopheles marajoara. To solve this problem, we are setting aside the male genitalia slide associated with the holotype of Anopheles marajoara and excluding it from the type material. Illustrations of the male genitalia and adult male are included.

Studies on mosquitoes (Diptera: Culicidae) and anthropic environment: 2 - Immature stages research at a rice irrigation system location in South-Eastern Brazil

A relation between a rice irrigation system and mosquito breeding was established in a study undertaken at the Ribeira Valley Experimental Station, from January through December 1992. Flooding favoured Anopheles (Nyssorhynchus) and Culex (Melanoconion) species, while empty paddies condition were propitious to Aedes scapularis and Culex (Culex) species. Compared with a more primitive area of the same region, several species showed high a degree of adaptation to the anthropic environment. Among them, Anopheles albitarsis, a potential malaria vector that breeds in the irrigation system, has shown immature stage production thirteen times higher than at the natural breeding sites. In addition, Ae. scapularis, An. oswaldoi, Cx. bastagarius, and Cx. chidesteri presented high levels of synanthropy.

Mosquitos de ambientes peri e extradomiciliares na região sul do Brasil

Durante um ano de coletas de culicídeos no Município de Querência do Norte, no Estado do Paraná, Brasil, utilizando-se isca humana e armadilhas de Falcão, investigou-se a composição faunística, a sazonalidade, o horário de maior densidade, a afinidade ao hospedeiro humano c a presença desses dípteros em abrigos de animais domésticos. De junho de 1989 a maio de 1990 foram coletados 5.923 mosquitos dos gêneros Aedes, Aedomyia, Coquillettidia, Culex, Mansonia, Psorophora, Sabethes e Uranotaenia. Identificaram-se 32 espécies de culícideos, dentre as quais Aedes scapularis, Anopheles albitarsis, Aedomyia squamipennis, Coquillettidia lynchi, Mansonia titillans e Coquillettidia venezuelensis tiveram maior prevalência, tendo sido capturados em grande número em isca humana, exceto Aedomyia squamipennis que compareceu sobretudo em abrigos de animais domésticos. O horário de maior atividade foi entre 18 e 19 horas e o mês de maior densidade foi o de abril, considerando-se o conjunto dos insetos capturados.

Culicídeos do lago de Itaipu, no rio Paraná, Sul do Brasil

De janeiro a dezembro de 1991 foram realizadas capturas mensais de culicídeos, no Município de Guaíra, Estado do Paraná (Brasil), na margem esquerda do lago de Itaipu, ao lado da via de acesso que conduzia aos Saltos das Sete Quedas, usando-se armadilha luminosa de Shannon, e isca humana em área urbanizada. Obtiveram-se informações sobre a fauna culicidiana, as espécies prevalentes, a variação sazonal, o horário de maior densidade e a afinidade dos mosquitos em relação ao hospedeiro humano. Foram identificadas 41 espécies de culicídeos dos gêneros Anopheles, Aedes, Aedomyia, Coquillettidia, Culex, Mansonia, Psorophora e Uranotaenia. Capturaram-se 21.280 mosquitos em armadilha de Shannon e 1.010 em isca humana. As espécies mais freqüentes em armadilha de Shannon - Coquillettidia shannoni, Mansonia humeralis, Anopheles triannulatus, Aedes scapularis e Anopheles albitarsis - perfizeram 82,78% dos mosquitos capturados. Em isca humana Aedes scapularís, Mansonia humeralis e Anopheles albitarsis, somaram 91,21% dos insetos capturados. Em armadilha de Shannon o horário de maior atividade das espécies prevalentes foi entre 19 e 22 h. Em isca humana Aedes scapularis teve maior densidade das 20 às 21 h, enquanto Mansonia humeralis e Anopheles albitarsis prevaleceram das 18 às 19 h. Dentre as espécies prevalentes em armadilha de Shannon, Coquillettidia shannoni, Mansonia humeralis e Anopheles albitarsis tiveram pico de atividade no mês de abril, Anopheles triannulatus em janeiro e Aedes scapularis em fevereiro.

Studies on entomological monitoring: mosquito species frequency in riverine habitats of the Igarapava Dam, Southern Region, Brazil

Diversity of mosquito species was evaluated in different habitats before and after the Igarapava reservoir flooding in the Grande River, Southern Cerrado of Brazil. We aimed at verifying changes in these mosquito populations in consequence of the lake formation. Four habitats were selected as sampling stations: peridomiciliary habitat, pasture, "veredas" and gallery forest patch. Bimonthly collections were made with the Shannon trap and human bait, including diurnal, crepuscular and nocturnal period of mosquito activity. The Shannon Index results from the potential vectors were compared using Student t-test. Aedes scapularis, Anopheles darlingi and An. albitarsis senso latu seasonal abundance were described with moving average and compared using chi2 test. There were changes in the mosquito frequency in the habitats, except for the "veredas" that was 13 km away from the catchment area. The altering in mosquito species seasonal abundance suggests breeding places expansion. Diversity indexes can be used to monitor changes in mosquito vector population in environments where abrupt disturbance can alter disease transmission cycles.

Affinity and diversity indices for anopheline immature forms

As for the entire Amazon Region, malaria continues to be a major health public problem in Roraima that presented an Annual Parasitic Index of 85.4 in 2005, the highest in Brazil. Information on anopheline breeding sites is an essential component in malaria control strategies. Aiming to contribute to the limited knowledge on anopheline immature forms in Roraima, collections and breeding site observations were performed in 10 breeding sites around the capital city Boa Vista. Collections were carried out in the rainy and dry season periods between April 2004 and January 2005. Breeding sites comprised natural and artificial water reservoirs. A total of 623 immature forms were collected belonging to Anopheles albitarsis s.l., An.triannulatus s.l., An. nuneztovari/dunhami, An. braziliensis, An. evansae, An. oswaldoi s.l., An. strodei and An. darlingi. An. albitarsis and An. braziliensis were the most frequently found species. Eight larvae of An. darlingi were found in only one breeding site located in the forest. An. triannulatus/An. nuneztovari and An. albitarsis/An. braziliensis were the pairs of species that mostly occurred together. Both pair of species displayed the highest affinity index what might indicate a high compatibility for the same breeding conditions and/or a synergistic co-occurrence. Species diversity index was higher for the dry season.

The influence of the area of the Serra da Mesa Hydroelectric Plant, State of Goiás, on the frequency and diversity of anophelines (Diptera: Culicidae): a study on the effect of a reservoir

INTRODUCTION: Bioecological aspects of anophelines (Diptera: Culicidae) near areas under the direct influence of the hydroelectric plant reservoir of Serra da Mesa in Goiás, Brazil, were analyzed. METHODS: Samples were collected at the surrounding dam area during the phases before and after reservoir impoundment. The influence of climatic and environmental factors on the occurrence of Anopheles darlingi, Anopheles albitarsis, Anopheles triannulatus, Anopheles oswaldoi and Anopheles evansae was assessed using Pearson's correlations with indicators for richness and diversity as well as the index of species abundance (ISA) and the standardized index of species abundance (SISA). RESULTS: The highest anopheline density occurred during the phase after filling the tank; however, no direct correlation with the climatic factors was observed during this stage. The reservoir formation determined the incidence of the anopheline species. An. darlingi was the predominant species (SISA = 1.00). CONCLUSIONS: The significant difference (p < 0.05) observed between the species incidence during the different reservoir phases demonstrates the environmental effect of the reservoir on anophelines.

Alguns aspectos da ecologia dos mosquitos (Diptera: Culicidae) de uma área de planície (Granjas Calábria), em Jacarepaguá, Rio de Janeiro: III. Preferência horária das fêmeas para o hematofagismo

Apresentamos os resultados de observação sobre o ciclo circadiano de atividade hematofágica dos mosquitos, em Granja Calábria, Jacarepaguá, na planície litorânea do Rio de Janeiro, onde realizamos, em isca humana, ao ar livre, capturas semanais, de 8 às 10, de 13 às 15 e 18 às 20 horas, de agosto de 1981 a julho de 1982, além de três capturas horárias de 24 horas seguidas. A maioria das espécies locais revelou caráter crepuscular vespertino e noturno. Contudo Limatus durhami, Phoniomyia davisi, Wyeomyia leucostigma e Wyeomyia (Dendromyia) sp. foram essencialmente diurnas, enquanto Anopheles albitarsis, Culex chidesteri e Culex quinquefasciatus foram obtidas somente no crepúsculo vespertino e à noite. Embora Anopheles aquasalis, Culex coronator, Culex saltanensis, Culex crybda e Coquillettidia venezuelensis fossem preponderantemente noturnas e Phoniomyia deanei e Phoniomyia theobaldi principalmente diurnas, obtivemô-las algumas vezes, fora do horário preferencial, sendo que Phoniomyia deanei teve nítido incremento pré-crepuscular vespertino. Aedes scapularis, Aedes taeniorhynchus e Mansonia titillans, espécies mais ecléticas, picaram durante todo o nictêmero, mas com flagrante acentuação crepuscular vespertina.

Immature stages of mosquitoes (Diptera: Culicidae) Collected during the autumn-winter period in Cordoba Province, Argentina

Eggs, immature mosquito collections were made at Cosquin, La Calera ( Chaco phytogeographic region), Villa Allende and Villa del Rosario (Espinal phytogeographic region) during April/September of two consecutive years 1989, 1990. Specific immature habitats in each locality were identified and sampled monthly. Eggs and/or immatures of Aedes albifasciatus, Ae. fluviatilis, Anopheles albitarsis, culex acharistus, Cx. apicinus, Cx. bidens, Cx. coronator, Cx. chidesteri, Cx. dolosus, Cx. maxi, Cx. quinquefasciatus, Cx. saltanensis, Psorophora ciliata and Uranotaenia lowii were collected. Three species (Cx. acharistus, Cx. dolosus and Cx. quinquefasciatus) were collected during the sampling period for all developmental stages. This suggests that immature of these species do not overwinter but continue to develop throughout the cold autumn and winter seasons.

Classification of immature stage habitats of Culicidae (Diptera) collected in Córdoba, Argentina

In order to classify mosquito immature stage habitats, samples were taken in 42 localities of Córdoba Province, Argentina, representing the phytogeographic regions of Chaco, Espinal and Pampa. Immature stage habitats were described and classified according to the following criteria: natural or artificial; size; location related to light and neighboring houses; vegetation; water: permanence, movement, turbidity and pH. Four groups of species were associated based on the habitat similarity by means of cluster analysis: Aedes albifasciatus, Culex saltanensis, Cx. mollis, Cx. brethesi, Psorophora ciliata, Anopheles albitarsis, and Uranotaenia lowii (Group A); Cx. acharistus, Cx. quinquefasciatus, Cx. bidens, Cx. dolosus, Cx. maxi and Cx. apicinus (Group B); Cx. coronator, Cx. chidesteri, Mansonia titillans and Ps. ferox (Group C); Ae. fluviatilis and Ae. milleri (Group D). The principal component analysis (ordination method) pointed out that the different types of habitats, their nature (natural or artificial), plant species, water movement and depth are the main characters explaining the observed variation among the mosquito species. The distribution of mosquito species by phytogeographic region did not affect the species groups, since species belonging to different groups were collected in the same region.