Comparison of sampling anopheline mosquitoes by light-trap and human-bait collections indoors at Bagamoyo, Tanzania

Abstract. The mosquito sampling efficiency of CDC miniature light-traps, relative to night-biting collections, was evaluated indoors at two sites in coastal Tanzania. We found that the total number of anophelines captured overnight by light-traps (hung beside a bednet in use) was 1.23 times the number of anophelines captured by human-bait collections. This relationship was not affected significantly by changes in the mosquito density, order of trapping method, date of sampling, or number of household occupants. Malaria sporozoite rates were twice as high among mosquitoes captured by light-trap as compared to those captured by night-biting collection. This was attributed to the tendency of light-traps to capture a larger proportion of gravid mosquitoes, which also had high sporozoite rates. The differences in sporozoites rates according to abdominal stage indicates that unfed mosquitoes captured by light-traps may define more precisely the human-biting activity and sporozoite rates as seen by night-biting collections. Our study shows that light-traps, when used in combination with night-biting collections, can be an effective and sensitive means for measuring human-biting activity and the sporozoite rate.     

Anopheles arabiensis and An. funestus are equally important vectors of malaria in Matola coastal suburb of Maputo, southern Mozambique

Transmission characteristics of malaria were studied in Matola, a coastal suburb of Maputo, the capital City, in southern Mozambique, from November 1994 to April 1996. The local climate alternates between cool dry season (May-October) and hot rainy season (November-April) with mean annual rainfall 650-850 mm. Saltmarsh and freshwater pools provide mosquito breeding sites in Matola. Malaria prevalence reached approximately 60% among people living nearest to the main breeding sites of the vectors. Plasmodium falciparum caused 97% of malaria cases, others being P. malariae and P. ovale. Potential malaria vector mosquitoes (Diptera: Culicidae) collected at Matola during daytime indoor-resting (n = 1021) and on human bait at night (n = 5893) comprised 12% Anopheles coustani Laveran (93% biting outdoors), 46% An. funestus Giles (68% biting indoors) and 42% An. gambiae Giles sensu lato (60% biting outdoors). All 215 specimens of An. gambiae s.l. identified genetically were An. arabiensis Patton. Anopheles funestus populations remained stable throughout the year, whereas densities of the An. gambiae complex fluctuated considerably, with An. arabiensis peaking during the rainy season. No concomitant rise in malaria incidence was observed. Human landing indices of An. funestus and An. arabiensis averaged 1.8 and 3.8 per man-night, respectively. Overall Plasmodium sporozoite rates were 2.42+/-1.24% in 2181 An. funestus and 1.11+/-1.25% in 1689 An. arabiensis dissected and examined microscopically. Mean daily survival rates were 0.79 for both vector species. Estimated infective bites/person/year were 15 An. funestus and 12 An. arabiensis. Biting rates were greatest at 2100-24.00 hours for An. funestus (68% endophagic) and 21.00-03.00 hours for An. arabiensis (40% endophagic). The entomological inoculation rate (EIR) declined sharply over very short distances (50% per 90m) away from breeding-sites of the vectors. Consequently, P. falciparum prevalence among Matola residents was halved 350 m within the town. Implications for the protective effectiveness of a 'cordon sanitaire' by residual house-spraying and/or the use of insecticide-treated bednets are discussed.     

Malaria transmission potential of Anopheles mosquitoes in the Mwea-Tebere irrigation scheme, Kenya

1. Anopheles arabiensis Patton and An. funestus Giles were identified as vectors of Plasmodium falciparum malaria in the Mwea-Tebere irrigation scheme, Kenya. An. arabiensis was the only member of the An. gambiae complex identified from chromosome characteristics. Other Anopheles species found included An. pharoensis Theobald, An. rufipes Gough and An. coustani Laveran. Survival rates per gonotrophic cycle for An. arabiensis averaged 0.37 during the short rains (October-November), 0.49 during the dry season (February) and 0.78 during the long rains (May-June). Vectorial capacities were correspondingly low due to low survival rates and a high degree of zoophily. The average duration of infective life for P. falciparum was 0.2 days for both An. arabiensis and An. funestus. In contrast, entomological inoculation rates were comparatively high: 6-8 infective bites/man/month. An. pharoensis averaged 110 bites/man/night during the short rains; 1/999 (0.1%) was positive by ELISA for P. falciparum circumsporozoite antigen, but the ELISA evidence is not conclusive for vector incrimination. In correspondence with clinical observations, the transmission of P. malariae and P. ovale is unlikely due to the low vector survival rates. The observed anomaly between low vectorial capacities and high entomological inoculation rates demonstrates the importance of accurately estimating vector sporozoite rates to monitor unstable malaria transmission in irrigated areas.     

The impact of permethrin-impregnated bednets on malaria vectors of the Kenyan coast

Abstract. The effects of introducing permethrin-impregnated bednets on local populations of the malaria vector mosquitoes Anopheles funestus and the An.gambiae complex was monitored during a randomized controlled trial at Kilifi on the Kenyan coast. Pyrethrum spray collections inside 762 households were conducted between May 1994 and April 1995 after the introduction of bednets in half of the study area. All-night human bait collections were performed in two zones (one control and one intervention) for two nights each month during the same period. PCR identifications of An.gambiae sensu lato showed that proportions of sibling species were An.gambiae sensu stricto > An.merus > An.arabiensis.
Indoor-resting densities of An.gambiae s.l. and the proportion of engorged females decreased significantly in intervention zones as compared to control zones. However, the human blood index and Plasmodium falciparum sporozoite rate remained unaffected. Also vector parous rates were unaltered by the intervention, implying that survival rates of malaria vectors were not affected. The human-biting density of An.gambiae s.l. , the predominant vector, was consistently higher in the intervention zone compared to the control zone, but showed 8% reduction compared to pre-intervention biting rates - versus 94% increase in the control zone.
Bioassay, susceptibility and high-performance liquid chromatography results all indicated that the permethrin content applied to the nets was sufficient to maintain high mortality of susceptible vectors throughout the trial. Increased rates of early outdoor-biting, as opposed to indoor-biting later during the night, were behavioural or vector composition changes associated with this intervention, which would require further monitoring during control programmes employing insecticide-treated bednets.     

Malaria in urban and rural Kinshasa: the entomological input

Abstract. Mosquitoes were collected on human bait over a 16-month period (September 1988 to December 1989) in an urban and a rural area of Kinshasa, Zaïre. P.falciparum malaria sporozoite rates were determined by ELISA. In the urban area Culex quinquefasciatus accounts for 96% of the 121 bites/ person/night (b/p/n). The only anopheline is Anopheles gambiae, sensu stricto, with an average of 5.1 b/p/n and a sporozoite rate of 1.86%. The entomological inoculation rate (EIR) averages 0.08 infective b/p/n. Malaria transmission is almost interrupted at the end of the dry season. In the rural area mosquito nuisance is small (20b/p/n), almost entirely due to six species of Anopheles including four vectors of malaria: An.gambiae (13.3 b/p/n), An.funestus (2.4b/p/n), An.nili (0.4b/p/n) and An.brunnipes (0.7b/p/n) with mean sporozoite rates of 7.85%, 6.60%, 6.63% and 0.53% respectively. An.paludis (0.4b/p/n) and An.hancocki (0.2b/p/n) were not found infective. Malaria transmission is intense and perennial: the overall EIR varies monthly between 0.60 and 3.29 infective b/p/n. The specific contributions of An.gambiae, An.funestus and An.nili average 1.07, 0.14 and 0.03 infective b/p/n respectively. Malaria transmission peaks during the rainy season in both study areas. The daily mean survival rates for An.gambiae were 0.91 and 0.78 in the rural and urban area, respectively. All An.gambiae examined belonged to the forest cytotype (Coluzzi et al., 1979). Through its effect on the sporozoite rate, the higher vector survival rate in the rural environment appears to be the major determinant of the greater malaria transmission rate in the rural area as compared to urban Kinshasa.     

Malaria vector composition and insecticide susceptibility status in Guinea Conakry,West Africa

This study provides data on malaria vector species composition and insecticide susceptibility status from three localities in Guinea Conakry. A total of 497 mosquitoes were collected resting indoors and morphologically identified as belonging to the Anopheles gambiae complex. The majority of these were An. gambiae s.s. (99.6%), but a small percentage (0.4%) were identified as Anopheles arabiensis. Thirty‐four Anopheles funestus s.s. were also collected. The molecular S form of An. gambiae s.s. was predominant over the M form in Siguiri (95%) and Boffa (97.4%), whereas at Mt Nimba the M form was more abundant (61.4%) than the S form (38.1%). One hybrid M/S specimen was recorded from Mt Nimba. Siguiri populations showed high levels of resistance to DDT, dieldrin and bendiocarb. Anopheles gambiae from Boffa were largely susceptible to the insecticides tested. At Mt Nimba, resistance to DDT and bendicocarb was detected. Biochemical enzyme analysis showed that an altered acetylcholinesterase is operating in the field at low levels. The frequency of the 1014F kdr allele in the An. gambiae S form was 0.24 at Siguiri and 0.14 at Mt Nimba. A single RR specimen was found in the M form. The heterogeneity in species composition and resistance profiles between sites requires vector control interventions to be tailored to each site based on the data collected from ongoing monitoring and surveillance.     

DNA probes for species identification of mosquitoes in the Anopheles gambiae complex

Abstract. Identification of species within the Anopheles gambiae Giles species complex is essential for the correct evaluation of malaria vector ecology studies and control programmes. The development of DNA probes to distinguish species of the An.gambiae complex is described. Genomic libraries were prepared for four members of the An.gambiae complex. These were screened using radiolabeled DNA from different species of An. gambiae sensu lato and a number of clones selected on the basis of their species specificity. These clones could be divided into two groups, each containing homologous sequences. Sequences homologous to group 1 inserts are highly reiterated in the genomes of Anopheles arabiensis Patton and Anopheles merus Dönitz, present in low copy number in Anopheles melas Theobald, but were not detected in Anopheles gambiae sensu stricto. Studies on the organization of this sequence in the genome of An.arabiensis show that homologous sequences are male specific and interspersed within the chromatin. Sequences homologous to group 2 inserts are highly repeated in the genomes of An.merus and An.melas, but present in low copy number in An.gambiae s.s. and An.arabiensis. Group 2 homologous sequences are not sex-specific in the species tested and appear to be tandemly repeated. When used as hybridization probes, these sequences provide a sensitive means for the identification of species within the Anopheles gambiae complex.     

Malaria transmission risk variations derived from different agricultural practices in an irrigated area of northern Tanzania

Malaria vector Anopheles and other mosquitoes (Diptera: Culicidae) were monitored for 12 months during 1994-95 in villages of Lower Moshi irrigation area (37 degrees 20' E, 3 degrees 21' S; approximately 700 m a.s.l.) south of Mount Kilimanjaro in northern Tanzania. Adult mosquito populations were sampled fortnightly by five methods: human bait collection indoors (18.00-06.00 hours) and outdoors (18.00-24.00 hours); from daytime resting-sites indoors and outdoors; by CDC light-traps over sleepers. Anopheles densities and rates of survival, anthropophily and malaria infection were compared between three villages representing different agro-ecosystems: irrigated sugarcane plantation; smallholder rice irrigation scheme, and savannah with subsistence crops. Respective study villages were Mvuleni (population 2200), Chekereni (population 3200) and Kisangasangeni (population approximately/= 1000), at least 7 km apart. Anopheles arabiensis Patton was found to be the principal malaria vector throughout the study area, with An. funestus Giles sensu lato of secondary importance in the sugarcane and savannah villages. Irrigated sugarcane cultivation resulted in water pooling, but this did not produce more vectors. Anopheles arabiensis densities averaged four-fold higher in the ricefield village, although their human blood-index was significantly less (48%) than in the sugarcane (68%) or savannah (66%) villages, despite similar proportions of humans and cows (ratio 1:1.1-1.4) as the main hosts at all sites. Parous rates, duration of the gonotrophic cycle and survival rates of An. arabiensis were similar in villages of all three agro-ecosystems. The potential risk of malaria, based on measurements of vectorial capacity of An. arabiensis and An.funestus combined, was four-fold higher in the ricefield village than in the sugarcane or savannah villages nearby. However, the more realistic estimate of malaria risk, based on entomological inoculation rates, indicated that exposure to infective vectors was 61-68% less for people in the ricefield village, due to the much lower sporozoite rate in An. arabiensis (ricefield 0.01%, sugarcane 0.1%, savannah 0.12%). This contrast was attributed to better socio-economic conditions of rice farmers, facilitating relatively more use of antimalarials and bednets for their families. Our findings show that, for a combination of reasons, the malaria challenge is lower for villagers associated with an irrigated rice-growing scheme (despite greater malaria vector potential), than for adjacent communities with other agro-ecosystems bringing less socio-economic benefits to health. This encourages the development of agro-irrigation schemes in African savannahs, provided that residents have ready access to antimalaria materials (i.e. effective antimalaria drugs and insecticidal bednets) that they may better afford for protection against the greater vectorial capacity of An. arabiensis from the ricefield agro-ecosystem.     

Multi-country field trials comparing wash-resistance of PermaNet and conventional insecticide-treated nets against anopheline and culicine mosquitoes

Insecticide-treated bednets (ITNs) are commonly used as a means of personal protection from malaria transmission by anopheline mosquitoes (Diptera: Culicidae). Long-lasting insecticidal nets (LLINs) have special treatments intended to remain effective after many washes. The present trials assessed the efficacy and wash-resistance of several production batches of PermaNet (polyester net coated with polymer resin containing pyrethroid insecticide deltamethrin 55 mg ai/m2) against malaria vectors in Pakistan, Iran and Tanzania compared to ITNs conventionally treated with alphacypermethrin 15 or 20 mg ai/m2, or deltamethrin 25 or 50 mg ai/m2. Insecticidal efficacy of the nets before and after repeated washing (using W.H.O. recommended and traditional local washing procedures) was monitored through contact bioassays with Anopheles and by experimental hut and outdoor platform tests. Local washing regimes gradually reduced the insecticidal efficacy of conventionally treated nets, but they were not exhausted, even after 21 washes. Using a more rigorous laboratory washing method, insecticide was more readily stripped from conventionally treated nets. PermaNet retained high efficacy after 21 washes, giving more than 97% mortality of Anopheles in contact bioassays with 3-min exposure. Using the more sensitive bioassay criterion of 'median time to knockdown', PermaNet showed no loss of insecticidal activity against Anopheles after washing repeatedly in 2 out of 6 trials; whereas in a further three trials knockdown activity of PermaNet and conventional ITNs declined at comparable rates. Higher mortality levels of Anopheles in contact bioassays did not always translate to superiority in experimental hut or enclosed platform trials. In only one of four comparative field trials did PermaNet out-perform conventional ITNs after washing: this was in the trial of PermaNet 2.0--the product with improved quality assurance. Because PermaNet and conventionally treated nets were both quite tolerant of local washing procedures, it is important in field trials to compare LLINs with conventional ITNs washed an equivalent number of times. Our comparison of PermaNet 2.0 against conventionally treated deltamethrin nets (CTDN) in Pakistan demonstrated superior performance of the LLIN after 20 washes in phase I and phase II bioassays, and this was corroborated by chemical assays of residual deltamethrin. Although PermaNet 2.0 has received WHOPES interim recommendation for malaria control purposes, its performance should be monitored in everyday use throughout its lifespan in various cultural settings to assess its durability and long-term effectiveness for malaria prevention and control. As many millions of conventionally treated nets are already in routine use, and these will require regular re-treatment, programme strategies should be careful to preserve the effectiveness of ITNS before and after establishing the reliability of LLINs in long-term use.     

Longitudinal follow‐up of malaria transmission dynamics in two villages in a Sahelian area of Niger during a nationwide insecticide‐treated bednet distribution programme

Malaria transmission was monitored in two villages in the Sahel zone of Niger over 4 years. During this period, a nationwide vector control programme was carried out in which insecticide‐treated bednets were distributed free to mothers of children aged <5 years. Anopheles gambiae and Anopheles arabiensis (Diptera: Culicidae) were found to be the major malaria vectors. The dynamics of An. gambiae s.l. did not vary dramatically over the study period although the proportion of female mosquitoes found resting indoors decreased in both villages and, in one village, the parity rate and sporozoite index were significantly reduced after bednet distribution. By contrast with An. gambiae, the dynamics of Anopheles funestus altered greatly after the bednet distribution period, when adult density, endophagous rate and sporozoite rates decreased dramatically. Our observations highlight the importance of quantifying and monitoring the dynamics and infections of malaria vectors during large‐scale vector control interventions.     

Malaria transmission dynamics in central Côte d'Ivoire: the influence of changing patterns of irrigated rice agriculture

The dynamics of malaria transmission was studied comparatively in the villages of Zatta and Tiemelekro, central Cote d'Ivoire, from February 2002 to August 2003. Prominent agroecosystems in these villages are irrigated rice growing and vegetable farming, respectively. Mosquitoes (Diptera: Culicidae) were collected on human bait at night and by pyrethrum knock-down spray sheet collections at four randomly selected sentinel sites in each village. In 2002, for a total of 96 man-nights per village, 7716 mosquitoes were collected in Zatta and 3308 in Tiemelekro. In 2003, with half the sampling effort, 859 and 2056 mosquitoes were collected in Zatta and Tiemelekro, respectively. Anopheles gambiae Giles s.l. was the predominant mosquito and the key malaria vector throughout, followed by An. funestus Giles. Anthropophily among adult female Anopheles exceeded 95% in both villages. Comparison between years revealed that the biting rate of An. gambiae s.l. in Zatta decreased several-fold from 49.3 bites per person per night (b/p/n) in 2002 to 7.9 b/p/n in 2003 (likelihood ratio test (LRT) = 1072.66; P < 0.001). Although the biting rate remained fairly constant in Tiemelekro, the difference between years was significant (16.1 vs. 18.2 b/p/n; LRT = 148.06; P < 0.001). These observations were paralleled by a marked decrease in the infective rate of An. gambiae s.l. in Zatta (4.6-1.2%), and an increase in Tiemelekro (3.1-7.6%). Meanwhile, the entomological inoculation rate of An. gambiae s.l. decreased 21-fold in Zatta, from 789 to 38 infective bites per person per year (ib/p/y), whereas it remained high in Tiemelekro (233 vs. 342 ib/p/y). The interruption of irrigated rice growing in Zatta in 2003, consequential to a farmers' conflict over land, might be the underlying cause for the significant reduction in malaria transmission, whereas more stable conditions occurred in Tiemelekro.     

Malaria transmission risk by the mosquito Anopheles baimaii (formerly known as An. dirus species D) at different hours of the night in North-east India

The risk of acquiring malaria transmitted by Anopheles baimaii Sallum & Peyton, 2005, formerly known as An. dirus species D (Sallum et al., 2005) (Diptera: Culicidae), at different hours of the night in a forest-fringed village of Assam, North-east India was assessed through all-night mosquito landing catches during 1995-2000. An estimated overall mean biting rate of 36.1 bites/person/night (95% CI = 26.2-45.8), a sporozoite rate of 1.9% (95% CI = 1.1-2.9%) and a parous rate of 58.7% (95% CI = 55.3-62.0%) were recorded. Parous and sporozoite-positive females tended to be caught mainly before midnight. The effective entomological inoculation rate was the highest (0.249 positive bites/person/night) from 21.00 to 24.00 hours, suggesting that the second quartile of the night is the most risky period for malaria transmission by An. baimaii. Considering that approximately 21% of mean inoculations take place before 21.00 hours, it appears that there is a need for appropriate protective measures during the pre-bed time period to supplement the impact of insecticide-treated nets against An. baimaii in north-east India.     

Host choice of anopheline mosquitoes in a malaria endemic area of western Venezuela

Abstract. Bloodmeals of exophilic anopheline mosquitoes collected resting on vegetation in a malaria endemic area in western Venezuela were identified by ELISA. Using a TMB peroxidase substrate in the ELISA, human bloodmeals were readily identified up to 40 h after ingestion in all laboratory-fed mosquitoes tested. Assay sensitivity declined to 75% identifiable 44 h post-feeding.
The Human Blood Index and the Feeding Index of each species differed between the three villages studied. An.triannulatus was generally more anthropophilic than An.nuneztovari and An.oswaldoi. These contrasting results emphasize the difficulties of interpreting host choice data.     

Malaria outbreak control in an African village by community application of 'deet' mosquito repellent to ankles and feet

The malaria vector Anopheles arabiensis Patton (Diptera: Culicidae) shows a marked predilection (> 80%) for biting the ankles and feet of human subjects, as revealed by our previous observations at Malahlapanga in the Kruger National Park, South Africa. Topical application of insect repellent, 15% deet (N,N-diethyl-3-methylbenzamide), to feet and ankles reduced the overall biting rate of An. arabiensis by 69%. A focal malaria epidemic in Albertsnek village (25 degrees 33'S, 31 degrees 59' E) near the Mozambique border, following flooding during February 2000, provided an opportunity to apply these findings of operational research for outbreak containment. Twice-nightly topical application of deet to ankles and feet of Albertsnek inhabitants was followed by rapid restoration of preepidemic malaria incidence levels after one incubation period. This encouraging outcome should be attempted in other outbreak-prone settings where infective mosquito bites are sporadic and malaria has unstable endemicity.     

Location of genes on chromosome arms in the Anopheles gambiae group of species and their correlation to linkage data for other anopheline mosquitoes

The use of paracentric inversions as genetic markers in the Anopheles gambiae group of mosquitoes is described. The gene for dieldrin resistance is assigned to chromosome 2 which in turn is correlated to the previous assignment of the gene to linkage group II. The locus of the enzyme phosphoglucomutase 2 (Pgm 2) is similarly assigned to chromosome 2 and evidence is presented for possible linkage between Pgm 2 and dieldrin resistance. There was no linkage or correlation of chromosome 2 and loci of the enzymes superoxide dismutase (Sod) and octanol dehydrogenase (Odh). These genes are therefore assumed to be on chromosome 3 (linkage group III). Evidence that such gene linkage group/chromosome correlations may extend to other species for which chromosome maps and homologies have been worked out is discussed.     

Anatomical dissemination of circumsporozoite protein in wild Afrotropical Anopheles affects malaria sporozoite rate determination by ELISA

The head, thorax, wings, legs and abdomen of 320 wild-caught Anopheles gambiae Giles sensu lato and 115 An.funestus Giles were tested by an enzyme-linked immunosorbent assay (ELISA) for Plasmodium falciparum Welch to determine how anatomical dissemination of circumsporozoite (CS) protein could affect the estimation of malaria sporozoite rates by ELISA. Of fifty-three Anopheles with CS protein detected in any body part, positive reactions were observed for 58.5% of heads, 67.0% of thoraces, 39.6% of wings, 52.8% of legs and 60.4% of abdomens. Mean absorbance values (range 0-2.00) were highest in thorax samples (1.17), followed by heads (0.80), abdomens (0.67), wings (0.48) and legs (0.46). Circumsporozoite protein was present in the wings or legs, but not in the head or thorax, in 11.3% (6/53) of the infected Anopheles. The ELISA infection rate of 12.8% (41/320) for An.gambiae would have increased to 14.7% (47/320) by inclusion of six mosquitoes with CS protein in wings or legs alone. The slight overestimation of the proportion of infective mosquitoes due to disseminated CS protein would have little effect on estimates of relative infection rates by ELISA for field-collected Anopheles, with abdomens removed prior to testing. However, the widespread dissemination of CS protein indicates that sporozoite load estimates by ELISA, for mosquitoes without abdomens, may not provide adequate measurements of the numbers of sporozoites in the salivary glands. Operationally, careful processing of mosquito samples for the determination of infectivity rates by ELISA is necessary to prevent the mixing of wings or legs among samples representing individual mosquitoes.     

Use of rDNA-PCR to investigate the ecological distribution of Anopheles bwambae in relation to other members of the An.gambiae complex of mosquitoes in Bwamba County, Uganda

Abstract Environmental relationships were investigated among three species of the Anopheles gambiae complex of mosquitoes associated with the geothermal springs located in Bwamba County, Uganda. The degree of ecological isolation between An.gambiae and An. bwambae, a sibling species known only from the geothermal springs environment, was assessed on the basis of adult distribution and abundance as well as differences in larval habitats. Field data were gathered during June 1995 without knowing which of the species were being collected. Specimens identified subsequently by rDNA-PCR were used to interpret the ecological data. Ten of twenty aquatic sites sampled were found positive for immature stages of the An.gambiae complex. Larvae of An.bwambae were associated with ‘springwater’ habitats having much higher conductivity, much greater concentrations of dissolved solids and slightly higher temperature and pH than ‘normal’ fresh water sites inhabited by larvae of An.gambiae. Larval habitats of both species were unshaded: An.bwambae occurred among dense sedge (Cyperus laevigatus) whereas those of An.gambiae were almost devoid of vegetation. One mixed sample showed that larvae of both species occur together in peripheral aquatic sites with intermediate physical and ecological characteristics. In water preference tests, free-flying females were reluctant to lay eggs on bowls of water in cages; gravid females (with one wing amputated) placed on the surface of water in a cup laid eggs on seasoned rainwater (12/51 An.bwambae; 2/3 An.gambiae) as well as spring-water (39/51 An.bwambae; 1/3 An.gambiae). All three An.gambiae oviposited on the first water option, whereas 86% of An. bwambae witheld oviposition until being moved to the other type of water after 5–6 h, and 82% (36/44) of these laid eggs on geothermal water in preference to rainwater. Larval and adult collections showed that An.gambiae occurs sympatrically with An.bwambae throughout its range in the humid foothill environment of the geothermal springs, whereas the distribution of An.arabiensis overlaps only slightly with An.bwambae towards the savanna environment north of the springs.     

Zoophily of Anopheles arabiensis and An. gambiae in Madagascar demonstrated by odour-baited entry traps

In Madagascar we used odour-baited entry traps (OBETs) for host choice tests of wild female anopheline mosquitoes (Diptera: Culicidae) at representative localities on the East and West sides of the island (villages Fenoarivo and Tsararano, respectively) and at the southern margin of the central plateau (Zazafotsy village, 800 m altitude). No insecticide house-spraying operations have been undertaken at these villages. Odours from a man and a calf of similar mass, concealed in different tents, were drawn by fans into separate OBETs set side by side. Traps were alternated to compensate for position effects, and different pairs of individual baits were employed for successive replicates. Totals of 266 An. funestus Giles sensu stricto and 362 An. gambiae Giles sensu lato were collected in 48 trap nights during March-June 1999. For each mosquito species the 'index of anthropophily' was defined as the proportion of females caught in the human-baited trap. For An. funestus this index was found to be consistently greater than 0.5 (value for random choice between traps/hosts), indicating that this species 'preferred' human to calf odour (index=0.83). Conversely, the index of anthropophily for An. gambiae s.l. indicated they 'chose' calf in preference to human odour (index=0.26). No significant differences of relative preference for calf or man were detected between villages; geographical variance accounted for <8% of the total experimental variance. Molecular identifications of 181 specimens of the An. gambiae complex (approximately 50% of the samples) revealed only An. arabiensis Patton at Tsararano and Zazafotsy, but >97% An. gambiae Giles sensu stricto at Fenoarivo, in accordance with prior knowledge of the differential distributions of these sibling species on the island. Predominant zoophily (i.e. intrinsic 'preference' for cattle odours) by both An. arabiensis and An. gambiae s.s. in Madagascar contrasts with their greater anthropophily in continental Africa.     

Development of an exposure-free bednet trap for sampling Afrotropical malaria vectors

An exposure-free bednet trap (the 'Mbita trap') for sampling of Afrotropical malaria vectors was developed during preliminary studies of mosquito behaviour around human-occupied bednets. Its mosquito sampling efficiency was compared to the CDC miniature light-trap and human landing catches under semi-field conditions in a screen-walled greenhouse using laboratory-reared Anopheles gambiae Giles sensu stricto (Diptera: Culicidae). When compared in a competitive manner (side by side), the Mbita trap caught 4.1+/-0.5 times as many mosquitoes as the CDC light-trap, hung beside an occupied bednet (P < 0.000 1) and 43.2+/-10% the number caught by human landing catches (P < 0.0001). The ratio of Mbita trap catches to those of the CDC light trap increased with decreasing mosquito density. Mosquito density did not affect the ratio of Mbita trap to human-landing catches. In a non-competitive comparison (each method independent of the other), the Mbita trap caught 89.7+/-10% the number of mosquitoes caught by human landing catches (P < 0.0001) and 1.2+/-0.1 times more mosquitoes than the CDC light trap (P = 0.0008). Differences in Mbita trap performance relative to the human landing catch under noncompetitive vs. competitive conditions were explained by the rate at which each method captured mosquitoes. Such bednet traps do not expose people to potentially infectious mosquito bites and operate passively all night without the need for skilled personnel. This trap is specifically designed to catch host-seeking mosquitoes only and may be an effective, sensitive, user-friendly and economic alternative to existing methods for mosquito surveillance in Africa.