Carla A. Sousa

An estimation of the entomological inoculation rate for Ifakara: a semi-urban area in a region of intense malaria transmission in Tanzania.

Summary An entomological study on vectors of malaria and their relative contribution to Plasmodium falciparum transmission in the semi‐urban area of Ifakara, south‐eastern Tanzania, was conducted. A total of 32 houses were randomly sampled from the area and light trap catches (LTC) performed in one room in each house every 2 weeks for 1 year. A total of 147 448 mosquitoes were caught from 789 LTC; 26 134 Anopheles gambiae s.l., 615 A. funestus, 718 other anophelines and 119 981 culicines. More than 60% of the total A. gambiae s.l. were found in five (0.6%) LTCs, with a maximum of 5889 caught in a single trap. Of 505 A. gambiae s.l. speciated by polymerase chain reaction, 91.5% were found to be A. arabiensis. Plasmodium falciparum sporozoite enzyme‐linked immunosorbent assay tests were performed on 10 108 anopheles mosquitoes and 39 (0.38%) were positive. Entomological inoculation rate (EIR) estimates were generated using a standard method and an alternative method that allows the calculation of confidence intervals based on a negative binomial distribution of sporozoite positive mosquitoes. Overall EIR estimates were similar; 31 vs. 29 [95% confidence interval (CI): 19, 44] infectious bites per annum, respectively. The EIR ranged from 4 (95% CI: 1, 17) in the cool season to 108 (95% CI: 69, 170) in the wet season and from 54 (95% CI: 30, 97) in the east of the town to 15 (95% CI: 8, 30) in the town centre. These estimates show large variations over short distances in time and space. They are all markedly lower than those reported from nearby rural areas and for other parts of Tanzania.

Asymmetric introgression between sympatric molestus and pipiens forms of Culex pipiens (Diptera: Culicidae) in the Comporta region, Portugal

BackgroundCulex pipiens L. is the most widespread mosquito vector in temperate regions. This species consists of two forms, denoted molestus and pipiens, that exhibit important behavioural and physiological differences. The evolutionary relationships and taxonomic status of these forms remain unclear. In northern European latitudes molestus and pipiens populations occupy different habitats (underground vs. aboveground), a separation that most likely promotes genetic isolation between forms. However, the same does not hold in southern Europe where both forms occur aboveground in sympatry. In these southern habitats, the extent of hybridisation and its impact on the extent of genetic divergence between forms under sympatric conditions has not been clarified. For this purpose, we have used phenotypic and genetic data to characterise Cx. pipiens collected aboveground in Portugal. Our aims were to determine levels of genetic differentiation and the degree of hybridisation between forms occurring in sympatry, and to relate these with both evolutionary and epidemiological tenets of this biological group.ResultsAutogeny and stenogamy was evaluated in the F1 progeny of 145 individual Cx. pipiens females. Bayesian clustering analysis based on the genotypes of 13 microsatellites revealed two distinct genetic clusters that were highly correlated with the alternative traits that define pipiens and molestus. Admixture analysis yielded hybrid rate estimates of 8-10%. Higher proportions of admixture were observed in pipiens individuals suggesting that more molestus genes are being introgressed into the pipiens form than the opposite.ConclusionBoth physiological/behavioural and genetic data provide evidence for the sympatric occurrence of molestus and pipiens forms of Cx. pipiens in the study area. In spite of the significant genetic differentiation between forms, hybridisation occurs at considerable levels. The observed pattern of asymmetric introgression probably relates to the different mating strategies adopted by each form. Furthermore, the differential introgression of molestus genes into the pipiens form may induce a more opportunistic biting behaviour in the latter thus potentiating its capacity to act as a bridge-vector for the transmission of arboviral infections.

Bioecology and Vectorial Capacity of Aedes albopictus (Diptera: Culicidae) in Macao, China, in Relation to Dengue Virus Transmission

Abstract Until 2001, the Chinese Territory of Macao had not registered any autochthonous dengue cases, despite the abundance of Aedes albopictus (Skuse), a known vector. This work describes a bioecological characterization of the local Ae. albopictus adult population, with the purpose of estimating the receptivity of Macao to dengue introduction. In the wet seasons of 1997 and 1998 and the dry season of 1998, Ae. albopictus was the most abundant human-biting mosquito. Daily biting rates of 314 mosquitoes per person were recorded in the wet season with a reduction to 94 in the dry season. Ae. albopictus was mainly exophagic and exophilic and had a human blood index of 44%. The parity rate of field-collected mosquitoes was 57%. Daily survival rate ranged from 91 to 97%. Estimates of vectorial capacity ranged from 144 to 880, depending on what parameter values were used. These estimates indicated a great receptivity for the introduction of dengue viruses, as the 2001 outbreak came to prove.

Male size does not affect mating success (of Anopheles gambiae in São Tomé)

Abstract For malaria control, the utility of transgenic vector Anopheles mosquitoes (Diptera: Culicidae) refractory to Plasmodium transmission, will depend on their interbreeding with the wild vector population. In many species, larger males are more successful in obtaining mates. In São Tomé island, we determined that size did not affect mating success of male Anopheles gambiae Giles sensu stricto, the main malaria vector in tropical Africa. Also we showed that larval intraspecific competition is probably insignificant in this population of An. gambiae. Thus, the potential success of transgenic An. gambiae is unlikely to be affected by size selection under field conditions.

Potential mosquito vectors of arboviruses in Portugal: species, distribution, abundance and West Nile infection.

Circulation of West Nile virus in Portugal was demonstrated by serological surveys, and the virus was isolated in 1969 from Anopheles maculipennis s.l. A survey of the whole country was carried out (2001-2004) to assess the abundance of mosquito species and to screen them for arbovirus infection. A total of 770 collections yielded 32460 mosquitoes of 15 species. The regions with the highest abundance of mosquitoes were the coastal and estuarine districts of Santarém, Setúbal and Faro. Culex pipiens s.l., An. maculipennis s.l., Cx. theileri and Ochlerotatus caspius were the most abundant and widespread, accounting for 92% of mosquitoes caught. Anopheles maculipennis s.l. and Cx. pipiens s.l. were present all over the country. Culex theileri and. Oc. caspius were more abundant in the southern and coastal areas, respectively. A total of 2355 mosquito pools were screened by RT-PCR for flaviviruses, of which 987 pools were also screened for bunyaviruses. Culex pipiens s.l. and Cx. univittatus collected in 2004 in the southern district of Faro were found to be infected with West Nile virus. The density and proximity of these mosquitoes to the human populations may constitute a public health threat in the case of involvement in arbovirus transmission cycles.

Malaria in São Tomé and Príncipe: parasite prevalences and vector densities.

A cross-sectional survey was carried out in 16 localities on the island of São Tomé and three on the island of Príncipe, at the end of the rainy season of 1997, to determine malaria prevalence and vector densities. Blood samples from 664 inhabitants of all ages were examined by optical microscopy (OM) and PCR. Mosquito collections were made by outdoor landing captures from 21:00-23:00 h. Great differences were found between OM and PCR readings. OM had a sensitivity of 66%, a specificity of 79% and failed to reveal any mixed-infections. Overall prevalence, determined by PCR, was higher in São Tomé (53%) than in Príncipe (35%). It was highest in children below 16 years-old. All four human Plasmodium species occurred in São Tomé but P. ovale was not detected in Príncipe. The human population was largely asymptomatic. Bednet users had lower prevalence than did non-users. The FOREST form of Anopheles gambiae s.s., identified by PCR and cytogenetics, was the only vector on the islands. The sporozoite rate in São Tomé, assessed by ELISA, was 0.5%. Parasite prevalence and vector densities were positively correlated in São Tomé, where malaria transmission must occur predominantly in the more populated coastal areas.

A clonal Plasmodium falciparum population in an isolated outbreak of malaria in the Republic of Cabo Verde

We present the first parasitological, molecular and longitudinal analysis of an isolated outbreak of malaria. This outbreak occurred on Santiago Island (Republic of Cabo Verde), a region where malaria is hypoendemic and controlled, and thus the population is considered non-immune. Blood samples were collected from the inhabitants over 1 month and during cross-sectional surveys in the following year. The presence and nature of the parasites was determined by PCR. Plasmodium falciparum was the only species detected. Genetic analysis revealed that the circulating parasites were genetically homogeneous, and probably clonal. Gametocytes were found throughout this period. Our data suggest that this represented a focal outbreak, resulting in the infection of at least 40% of the villagers with a clonal parasite line. Thus, P. falciparum infections can persist for at least 1 year in a substantial proportion (10%) of the hosts. Implications for malaria control and the interpretation of epidemiological data are discussed.

Approaches to passive mosquito surveillance in the EU

The recent emergence in Europe of invasive mosquitoes and mosquito-borne disease associated with both invasive and native mosquito species has prompted intensified mosquito vector research in most European countries. Central to the efforts are mosquito monitoring and surveillance activities in order to assess the current species occurrence, distribution and, when possible, abundance, in order to permit the early detection of invasive species and the spread of competent vectors. As active mosquito collection, e.g. by trapping adults, dipping preimaginal developmental stages or ovitrapping, is usually cost-, time- and labour-intensive and can cover only small parts of a country, passive data collection approaches are gradually being integrated into monitoring programmes. Thus, scientists in several EU member states have recently initiated programmes for mosquito data collection and analysis that make use of sources other than targeted mosquito collection. While some of them extract mosquito distribution data from zoological databases established in other contexts, community-based approaches built upon the recognition, reporting, collection and submission of mosquito specimens by citizens are becoming more and more popular and increasingly support scientific research. Based on such reports and submissions, new populations, extended or new distribution areas and temporal activity patterns of invasive and native mosquito species were found. In all cases, extensive media work and communication with the participating individuals or groups was fundamental for success. The presented projects demonstrate that passive approaches are powerful tools to survey the mosquito fauna in order to supplement active mosquito surveillance strategies and render them more focused. Their ability to continuously produce biological data permits the early recognition of changes in the mosquito fauna that may have an impact on biting nuisance and the risk of pathogen transmission associated with mosquitoes. International coordination to explore synergies and increase efficiency of passive surveillance programmes across borders needs to be established.

Raised houses reduce mosquito bites

BackgroundIn many parts of continental Africa house construction does not appear to impede entry of malaria vectors and, given their generally late biting cycle, the great majority of transmission takes place indoors. In contrast, many houses in São Tomé, 140 km off the coast of Gabon, are raised on stilts and built of wooden planks. Building on stilts is a time-honoured, but largely untested, way of avoiding mosquito bites. Exposure may also be affected by mosquito activity times and age composition of host-seeking females. A study was therefore undertaken on the island of São Tomé to determine if exposure to Anopheles gambiae, the only vector on the island, varied with house construction or time of the night.MethodsA series of all-night landing collections were undertaken out of doors at ground level, inside houses at ground level, on the verandas of, and inside houses built on stilts. The gonotrophic age of an unselected sample of insects from the first three hours of landing collection (18:00–21:00) was determined by dissection. In addition, 1,149 miniature light-trap collections were obtained from 125 houses in the study area. Numbers collected were related to house construction.ResultsBiting of An. gambiae took place primarily outside at ground level. Less than one third of biting occurred inside houses. Houses built on stilts had half the number of An. gambiae in them compared to those built at ground level. Conversely houses with an eaves gap had more An. gambiae in them than houses without such a gap. Gonotrophic age did not affect house entry rates in An. gambiae. House construction affected Culex quinquefasciatus less than An. gambiae. Mean density per house, derived from a series of 1,490 randomly assigned light-trap collections, was over-dispersed with 18% of houses having 70% of the vectors.ConclusionHouse construction plays an important role in determining exposure to malaria vectors in São Tomé. Neighbours can have very different exposure levels. Recommendations for improvement in control are given.

Feeding patterns of molestus and pipiens forms of Culex pipiens (Diptera: Culicidae) in a region of high hybridization

BackgroundTwo biological forms of the mosquito Culex pipiens s.s., denoted pipiens and molestus, display behavioural differences that may affect their role as vectors of arboviruses. In this study, the feeding patterns of molestus and pipiens forms were investigated in Comporta (Portugal), where high levels of inter-form admixture have been recorded.MethodsIndoor and outdoor mosquito collections were performed in the summer of 2010. Collected Cx. pipiens s.l. females were molecularly identified to species and form by PCR and genotyped for six microsatellites. The source of the blood meal in post-fed females was determined by ELISA and mitochondrial DNA sequencing.ResultsThe distribution of the forms differed according to the collection method. The molestus form was present only in indoor collections, whereas pipiens and admixed individuals were sampled both indoors and outdoors. In both forms, over 90% of blood meals were made on avian hosts. These included blood meals taken from Passeriformes (Passer domesticus and Turdus merula) by females caught resting inside domestic shelters.ConclusionGenetic structure and blood meal analyses suggest the presence of a bird biting molestus population in the study area. Both forms were found to rest indoors, mainly in avian shelters, but at least a proportion of females of the pipiens form may bite outdoors in sylvan habitats and then search for anthropogenic resting sites to complete their gonotrophic cycle. This behaviour may potentiate the accidental transmission of arboviruses to humans in the region.