Moderate transmission but high prevalence of malaria in Madagascar

Malaria transmission remains poorly documented in areas of low transmission. A study has been carried out over two consecutive years in Analamiranga, a village located at an altitude of 885m on the western edge of the Malagasy highlands, with the aim of generating and updating malariometric indexes for both mosquitoes and schoolchildren. In this village, no vector control measures were performed during the study period nor during previous decades. Mosquitoes were collected monthly when landing on human volunteers and in various resting-places. Blood samples were taken every 3 months from schoolchildren aged 6-12 years and microscopically examined. Of 7,480 mosquitoes collected on human subjects, 5,790 were anophelines. Ten anopheline species were represented and three of these, Anopheles funestus, Anopheles arabiensis and Anopheles mascarensis, accounted for 59.2% of the collection. Of these three species 4,640 were also collected in resting places. The proportion of mosquitoes fed on bovids was high; conversely, the anthropophilic rate (mosquitoes fed on human beings) was especially low: 31%, 7% and 1%, respectively, for A. funestus, A. arabiensis and A. mascarensis. The only confirmed malaria vector was A. funestus with a low sporozoite index (of 6,830 A. funestus, five were positive for Plasmodium falciparum and four for Plasmodium vivax). The annual entomological inoculation rate (number of bites of infected anophelines per adult person) was estimated at 2.49 with low variation over the 2 years. Overall, 909 thick blood smears were tested from blood samples taken from schoolchildren with 30.3% being malaria-positive. The four Plasmodium species infecting human subjects were detected in the following proportions: P. falciparum 78.9%, P. vivax 19.4%, Plasmodium malariae 1.0% and Plasmodium ovale 0.7%. The proportions of children who were infected with any Plasmodium ranged from 10.7% in February to 51.0% in September. Parasitemic children with fever (axillary temperature >37.5 degrees C) accounted for 16.4% of the children sampled. This study demonstrates that there are substantial parasitological consequences of even a relatively low entomological transmission and also recommends including exterior resting-places of mosquitoes in future spraying campaigns in the highlands of Madagascar.     

Year to year and seasonal variations in vector bionomics and malaria transmission in a humid savannah village in west Burkina Faso

A longitudinal entomological study was carried out from 1999 to 2001 in Lena, a humid savannah village in the western region of Burkina Faso in order to establish malaria vector bionomics and the dynamics of malaria transmission. In the first year, malaria transmission was mainly due to An. gambiae s.s., but during the two later years was due to An. funestus, which were observed in high frequency towards the end of the rainy season. PCR identification of samples of An. gambiae s.1. showed 93% to be An. gambiae s.s. and 7% An. arabiensis. An. funestus constituting more than 60% of the vectors were identified in PCR as An. funestus s.s. The persistence of intense vectorial activity in this village was probably due to the road building in a swampy area creating a semi-permanent swamp that provided large sites for larval mosquitoes. These swampy sites seemed to be more favorable for An. funestus than for An. gambiae s.s. Thus, land development must be monitored and subjected to planning to minimize vector proliferation. Such a system of planning could lead to the restriction or even elimination of the swamp that is the source of larvae developing in the heart of the village.     

Transmission of malaria in the Tesseney area of Eritrea: parasite prevalence in children, and vector density, host preferences, and sporozoite rate.

Malaria transmission was studied from July to September, 2002 in three villages of the Tesseney sub-zone, in the western lowlands of Eritrea. The three methods used for mosquito collection were light traps, pyrethrum spray catches, and pit shelter collections. All anopheline mosquitoes that were collected belonged to the Anopheles gambiae complex and they were identified by PCR as the sibling species Anopheles arabiensis (Patton). Apart from An. arabiensis, the only other mosquitoes caught were culicines. The vector population increased greatly for about a month after the start of the rains. The anthropophilic indices obtained from the blood-fed An. arabiensis resting indoors and outdoors were only 20% and 25%, respectively, with most of the other meals on goats. ELISA for P. falciparum circumsporozoite protein revealed only one positive out of 1,026 tested. The malaria prevalence among children <10 years was only 3.3% (all P.falciparum) from 300 slides examined. These low rates seem to reflect recent success in malaria control in Eritrea.     

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 infection potential of anopheline mosquitoes sampled by light trapping indoors in coastal Tanzanian villages

Abstract. Anopheline mosquito populations were studied during 1992 in seven villages south of Bagamoyo, coastal Tanzania, prior to malaria control intervention using insecticide treated bednets. To collect mosquitoes, CDC light traps were used in ten houses per village fortnightly for 12 months. Anopheles females were identified and checked by ELISA for the presence of malaria sporozoite antigen and source of bloodmeal. An. funestus peaked in June-July after the long rains. Three members of the An. gambiae complex had different seasonality: An. arabiensis, An. gambiae and small numbers of An. merus were collected.
In most villages transmission was extremely high and perennial with the entomological inoculation rate reaching three to eleven infective bites per person per night in July and persisting at around 0.1 and 1 for most of the remainder of the year. Sporozoite infection rates within the An. gambiae complex ranged from 2% to 25%, with the peaks in January and July following the two rainy periods. An. funestus showed a similar pattern. The light traps were reliable, simple to operate, and proved to be satisfactory to study the mosquito vector population.     

Use of an electrostatic sprayer for control of anopheline mosquitoes

The Electrodyn sprayer was compared with a compression sprayer (Hudson X-pert) for residual application of cypermethrin, a pyrethroid insecticide, to control the malaria vectors Anopheles arabiensis Patton and An. funestus Giles in experimental huts at Magugu in Tanzania. The time taken for hut spraying, 2-2.5 min per hut, was similar for both types of sprayer. Two or three huts were treated internally with cypermethrin at 40 or 80 mg a.i./m2 using Electrodyn formulation for comparison with 80 mg a.i./m2 using wettable powder formulation. Each of the twelve huts (including five untreated controls) was fitted with window exit traps and either louvre or verandah traps for mosquito sampling. The Electrodyn sprayer was fitted with a pair of elbowed deflectrodes to direct the positively charged spray droplets onto walls and ceiling. All treatments gave 94-100% mortality-rates of indoor-resting anophelines throughout the evaluation period of 11 weeks post-spray. Reductions of An.arabiensis and An.funestus females by 10-42% and 62-91%, respectively, in rooms and by 72% and 51% in exit traps indicated that cypermethrin deterred mosquitoes from entering the huts. Overall mortality-rates of mosquitoes were 66% of both species in huts treated with 40 mg/m2 Electrodyn, 43% An.funestus and 71% An.arabiensis due to 80 mg/m2 Electrodyn formulation and 49% An.funestus and 64% An.arabiensis due to 80 mg/m2 WP formulation (no significant differences). It is concluded that the Electrodyn sprayer with deflectrodes is a convenient and effective means of residual house-spraying with pyrethroid insecticide for malaria vector control.     

Is the Mbita trap a reliable tool for evaluating the density of anopheline vectors in the highlands of Madagascar?

BACKGROUND: One method of collecting mosquitoes is to use human beings as bait. This is called human landing collection and is a reference method for evaluating mosquito density per person. The Mbita trap, described by Mathenge et al in the literature, consists of an entry-no return device whereby humans are used as bait but cannot be bitten. We compared the Mbita trap and human landing collection in field conditions to estimate mosquito density and malaria transmission. METHODS: Our study was carried out in the highlands of Madagascar in three traditional villages, for 28 nights distributed over six months, with a final comparison between 448 men-nights for human landing and 84 men-nights for Mbita trap, resulting in 6,881 and 85 collected mosquitoes, respectively. RESULTS: The number of mosquitoes collected was 15.4 per human-night and 1.0 per trap-night, i.e. an efficiency of 0.066 for Mbita trap vs. human landing. The number of anophelines was 10.30 per human-night and 0.55 per trap-night, i.e. an efficiency of 0.053. This efficiency was 0.10 for indoor Anopheles funestus, 0.24 for outdoor An. funestus, and 0.03 for Anopheles arabiensis. Large and unexplained variations in efficiency were observed between villages and months. CONCLUSION: In the highlands of Madagascar with its unique, highly zoophilic malaria vectors, Mbita trap collection was poor and unreliable compared to human landing collections, which remains the reference method for evaluating mosquito density and malaria transmission. This conclusion, however, should not be extrapolated directly to other areas such as tropical Africa, where malaria vectors are consistently endophilic.     

Feeding and indoor resting behaviour of the mosquito Anopheles longipalpis in an area of hyperendemic malaria transmission in southern Zambia

Anopheles longipalpis (Theobald) (Diptera: Culicidae) is a predominantly zoophilic mosquito that has not been implicated in malaria transmission. However, this species was collected indoors with An. funestus s.l. in southern Zambia, where transmission of Plasmodium falciparum is hyperendemic, and we initially misidentified it morphologically and molecularly as An. funestus s.l. The indoor resting density and blood-feeding behaviour of An. longipalpis were investigated during the 2004-05 and 2005-06 transmission seasons in Mufwafwi village in southern Zambia. Numbers of endophilic An. longipalpis increased towards the end of the rainy season. Although specimens were collected during human landing catches, the feeding behaviour of An. longipalpis was significantly biased towards cattle (88.7%), with other bloodmeals originating from dogs, goats and chickens. None of the 177 specimens of An. longipalpis were infected with P. falciparum. These data are consistent with existing reports that An. longipalpis is not involved in malaria transmission. However, more extensive sampling is necessary. Importantly, the correct identification of An. longipalpis is crucial for malaria control programmes in areas where An. funestus s.l and An. longipalpis exist sympatrically so that scarce resources are not wasted on the control of a non-vector.     

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 Caran? 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.     

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.     

Anopheles funestus resistant to pyrethroid insecticides in South Africa

Northern Kwazulu/Natal (KZN) Province of South Africa borders on southern Mozambique, between Swaziland and the Indian Ocean. To control malaria vectors in KZN, houses were sprayed annually with residual DDT 2 g/ m2 until 1996 when the treatment changed to deltamethrin 20-25 mg/m2. At Ndumu (27 degrees 02'S, 32 degrees 19'E) the recorded malaria incidence increased more than six-fold between 1995 and 1999. Entomological surveys during late 1999 found mosquitoes of the Anopheles funestus group (Diptera: Culicidae) resting in sprayed houses in some sectors of Ndumu area. This very endophilic-vector of malaria had been eliminated from South Africa by DDT spraying in the 1950s, leaving the less endophilic An. arabiensis Patton as the only vector of known importance in KZN. Deltamethrin-sprayed houses at Ndumu were checked for insecticide efficacy by bioassay using susceptible An. arabiensis (laboratory-reared) that demonstrated 100% mortality. Members of the An. funestus group from Ndumu houses (29 males, 116 females) were identified by the rDNA PCR method and four species were found: 74 An. funestus Giles sensu stricto, 34 An. parensis Gillies, seven An. rivulorum Leeson and one An. leesoni Evans. Among An. funestus s.s. females, 5.4% (4/74) were positive for Plasmodium falciparum by ELISA and PCR tests. To test for pyrethroid resistance, mosquito adults were exposed to permethrin discriminating dosage and mortality scored 24h post-exposure: survival rates of wild-caught healthy males were 5/10 An. funestus, 1/9 An. rivulorum and 0/2 An. parensis; survival rates of laboratory-reared adult progeny from 19 An. funestus females averaged 14% (after 1h exposure to 1% permethrin 25:75cis:trans on papers in WHO test kits) and 27% (after 30 min in a bottle with 25 microg permethrin 40:60cis:trans). Anopheles funestus families showing >20% survival in these two resistance test procedures numbered 5/19 and 12/19, respectively. Progeny from 15 of the families were tested on 4% DDT impregnated papers and gave 100% mortality. Finding these proportions of pyrethroid-resistant An. funestus, associated with a malaria upsurge at Ndumu, has serious implications for malaria vector control operations in southern Africa.     

Blood-feeding behaviour of the malarial mosquito Anopheles arabiensis: implications for vector control

Feeding behaviour of the malaria vector Anopheles arabiensis Patton (Diptera: Culicidae) was monitored for 12 months (March 2003-February 2004) in the Konso District of southern Ethiopia (5 degrees 15'N, 37 degrees 28'E). More than 45 000 An. arabiensis females were collected by host-baited sampling methods (light-traps, human landing catches, cattle-baited traps) and from resting sites (huts and pit shelters). In the village of Fuchucha, where the ratio of cattle : humans was 0.6 : 1, 51% of outdoor-resting mosquitoes and 66% of those collected indoors had fed on humans, human baits outdoors caught > 2.5 times more mosquitoes than those indoors and the mean catch of mosquitoes from pit shelters was about five times that from huts. Overall, the vast majority of feeding and resting occurred outdoors. In the cattle camps of Konso, where humans slept outdoors close to their cattle, approximately 46% of resting mosquitoes collected outdoors had fed on humans despite the high cattle : human ratio (17 : 1). In both places, relatively high proportions of bloodmeals were mixed cow + human: 22-25% at Fuchucha and 37% in the cattle camps. Anthropophily was also gauged experimentally by comparing the numbers of mosquitoes caught in odour-baited entry traps baited with either human or cattle odour. The human-baited trap caught about five times as many mosquitoes as the cattle-baited one. Notwithstanding the potential pitfalls of using standard sampling devices to analyse mosquito behaviour, the results suggest that the An. arabiensis population is inherently anthropophagic, but this is counterbalanced by exophagic and postprandial exophilic tendencies. Consequently, the population feeds sufficiently on humans to transmit malaria (sporozoite rates: 0.3% for Plasmodium falciparum and 0.5% for P. vivax, by detection of circumsporozoite antigen) but also takes a high proportion of meals from non-human hosts, with 59-91% of resting mosquitoes containing blood from cattle. Hence, classical zooprophylaxis is unlikely to have a significant impact on the malaria vectorial capacity of An. arabiensis in Konso, whereas treating cattle with insecticide might do.     

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.     

Village-scale evaluation of mosquito nets treated with a tablet formulation of deltamethrin against malaria vectors

A field trial was carried out in the Sundargarh district of Orissa, India on the efficacy of mosquito nets treated with a tablet formulation of deltamethrin (K-O TAB) against malaria vectors. Treated nets were used in one village, and in the two control villages, one used untreated nets and the other used indoor spraying with DDT, without nets. In this area the primary malaria vectors are Anopheles culicifacies Giles sensu lato (Diptera: Culicidae) and An. fluviatilis James s.l., which are both endophagic and endophilic, and fully susceptible to deltamethrin. Treatment of a 10-m(2) mosquito net with one of the tablets gave a deltamethrin deposit of 25 mg/m(2). Bioassays repeated on domestically used nets over 7 months showed persistence of almost 100% mortality of An. fluviatilis, whereas An. culicifacies showed a decline from 100% to 71% mortality over this period, after which the nets were re-treated and bioassays were not continued. The sum of collections of mosquitoes resting in village houses and those in exit traps and dead on floor sheets showed a reduction in the numbers of the two vector species due to the treated nets, compared with untreated or no nets, but no reduction in other anophelines or Culex species. Large proportions of the collections of the vector and non-vector anophelines were dead on the floor sheets, but among Culex, mortality was delayed. Treated and untreated nets reduced the proportion of anophelines that had blood-fed; the treated nets did so more effectively than the untreated in the case of An. culicifacies and of Culex mosquitoes. In rooms with treated nets a larger proportion of the total collections [dead + live] were in the exit traps, which can be attributed to the excito-repellent effect of deltamethrin. It is easier to pack and handle tablets of insecticide than liquid concentrate and the use of one tablet per net may be preferable to making up a large volume of diluted insecticide and dipping many nets at a time.     

Repellent effects on Anopheles arabiensis biting humans in Kruger Park, South Africa

Abstract. Distribution of biting sites on the human body by the malaria vector Anopheles arabiensis Patton (Diptera: Culicidae) was investigated near a source of mosquitoes in the Kruger National Park, South Africa. Eight adult male volunteers (2 teams × 2 pairs of subjects) conducted human bait collections while seated on camp chairs in the open-air, wearing only short trousers (no shirt, socks or shoes). Mosquito collections during 18.30–22.30 hours on five consecutive nights in April 1998 yielded a total of 679 An. arabiensis females biting subjects with or without their ankles and feet treated with deet insect repellent (15% diethyl-3-methylbenzamide, Tabard™ lotion). On subjects whose feet and ankles were smeared with repellent, 160 An. arabiensis females were captured biting in 60 man-hours: 88.1% on the legs, 1.4% on the arms and 1.2% on other parts of the body, but none on the repellent-treated feet or ankles. On subjects without repellent treatment, 519 An. arabiensis were caught biting in 60 man-hours: 81.1% on feet and ankles, 16.4% on legs, 1.4% on arms and 1.2% on the rest of the body. For individual subjects, the reduction of An. arabiensis bites ranged from 36.4 to 78.2% (mean protection 69.2%). Results of this study confirm previous findings that, in this part of South Africa − inhabited only by wildlife − when people sit outside during the evening An. arabiensis prefers to bite their lower limbs: 97.5% below the knees. Overall, the number of bites by the malaria vector An. arabiensis was reduced more than three-fold (from 26 to 8/person/evening), simply by treating ankles and feet with a consumer brand of deet repellent. Whether or not this provides a satisfactory degree of protection against malaria risk would depend on the malaria sporozoite rate in the malaria vector population.     

Dynamics of immature stages of Anopheles arabiensis and other mosquito species (Diptera: Culicidae) in relation to rice cropping in a rice agro-ecosystem in Kenya.

We determined changes in species composition and densities of immature stages of Anopheles arabiensis mosquitoes in relation to rice growth cycle in order to generate data for developing larval control strategies in rice ecosystems. Experimental rice paddies (6.3m x 3.15m) exposed to natural colonization of mosquitoes were sampled weekly for two rice growing cycles between February 2004 and March 2005. Overall, 21,325 Anopheles larvae were collected, of which 91.9% were 1st and 2nd instars and 8.1% were 3rd and 4th instars. An. arabiensis was the predominant species (84.1%) with other species, An. pharoensis (13.5%), An. funestus (2.1%), An. coustani (0.3%), and An. maculipalpis (0.1%) accounting for only a small proportion of the anophelines collected. Culex quinquefasciatus (65.7%) was the predominant species among the non-anopheline species. Others species collected included: C. annulioris (9.9%), C. poicilipes (7.3%), C. tigripes (7.2%), C. duttoni (0.6%), Aedes aegypti (5.3%), Ae. cumminsii (3.5%), and Ae. vittatus (0.7%). The densities of the major anopheline species were closely related to rice stage and condition of the rice field. An. arabiensis, the predominant species, was most abundant over a three-week period after transplanting. Low densities of larvae were collected during the late vegetative, reproductive, and ripening phases of rice. An increase in larval density ten days post-transplanting was found to correlate with the application of fertilizer (sulphate of ammonia). Culicine and aedine species densities were significantly higher during the post-harvesting period. Our results suggest that the transplanting stage is favorable for the growth of immature stages of An. arabiensis and provides a narrow window for targeted larval intervention in rice.     

Multimodal pyrethroid resistance in malaria vectors, Anopheles gambiae s.s., Anopheles arabiensis, and Anopheles funestus s.s. in western Kenya

Anopheles gambiae s.s., Anopheles arabiensis, and Anopheles funestus s.s. are the most important species for malaria transmission. Pyrethroid resistance of these vector mosquitoes is one of the main obstacles against effective vector control. The objective of the present study was to monitor the pyrethroid susceptibility in the 3 major malaria vectors in a highly malaria endemic area in western Kenya and to elucidate the mechanisms of pyrethroid resistance in these species. Gembe East and West, Mbita Division, and 4 main western islands in the Suba district of the Nyanza province in western Kenya were used as the study area. Larval and adult collection and bioassay were conducted, as well as the detection of point mutation in the voltage-gated sodium channel (1014L) by using direct DNA sequencing. A high level of pyrethroid resistance caused by the high frequency of point mutations (L1014S) was detected in An. gambiae s.s. In contrast, P450-related pyrethroid resistance seemed to be widespread in both An. arabiensis and An. funestus s.s. Not a single L1014S mutation was detected in these 2 species. A lack of cross-resistance between DDT and permethrin was also found in An. arabiensis and An. funestus s.s., while An. gambiae s.s. was resistant to both insecticides. It is noteworthy that the above species in the same area are found to be resistant to pyrethroids by their unique resistance mechanisms. Furthermore, it is interesting that 2 different resistance mechanisms have developed in the 2 sibling species in the same area individually. The cross resistance between permethrin and DDT in An. gambiae s.s. may be attributed to the high frequency of kdr mutation, which might be selected by the frequent exposure to ITNs. Similarly, the metabolic pyrethroid resistance in An. arabiensis and An. funestus s.s. is thought to develop without strong selection by DDT.     

Adult anopheline ecology and malaria transmission in irrigated areas of South Punjab, Pakistan

Surface irrigation in the Punjab province of Pakistan has been carried out on a large scale since the development of the Indus Basin Irrigation System in the late 19th century. The objective of our study was to understand how the population dynamics of adult anopheline mosquitoes (Diptera: Culicidae) could be related to malaria transmission in rural areas with intensive irrigation and a history of malaria epidemics. In this paper we present our observations from three villages located along an irrigation canal in South Punjab. The study was carried out from 1 April 1999 to 31 March 2000. Mosquitoes were collected from bedrooms using the pyrethroid spraycatch method and from vegetation and animal sheds using backpack aspirators. Overall, Anopheles subpictus Grassi sensu lato predominated (55.6%), followed by An. stephensi Liston s.l. (41.4%), An. culicifacies Giles s.l. (2.0%), An. pulcherrimus Theobald (1.0%) and An. peditaeniatus Leicester (0.1%). Most mosquitoes (98.8%) were collected from indoor resting-sites whereas collections from potential resting-sites outdoors accounted for only 1.2% of total anopheline densities, confirming the endophilic behaviour of anophelines in Pakistan. Anopheles stephensi, An. culicifacies and An. subpictus populations peaked in August, September and October, respectively. High temperatures and low rainfall negatively affected seasonal abundance in our area. There were interesting differences in anopheline fauna between villages, with An. culicifacies occurring almost exclusively in the village at the head of the irrigation canal, where waterlogged and irrigated fields prevailed. Monthly house-to-house fever surveys showed that malaria transmission remained low with an overall slide positivity rate of 2.4% and all cases were due to infection with Plasmodium vivax. The most plausible explanation for low transmission in our study area seems to be the low density of Pakistan's primary malaria vector, An. culicifacies. The role of other species such as An. stephensi is not clear. Our observations indicate that, in South Punjab, irrigation-related sites support the breeding of anopheline mosquitoes, including the vectors of malaria. As our study was carried out during a year with exceptionally hot and dry climatic conditions, densities and longevity of mosquitoes would probably be higher in other years and could result in more significant malaria transmission than we observed. To assess the overall importance of irrigation-related sites in the epidemiology of malaria in the Punjab, more studies are needed to compare irrigated and non-irrigated areas.     

Mosquito responses to carbon dioxide in B West African Sudan savanna village

Abstract. Mosquito responses to carbon dioxide were investigated in Noungou village, 30 km northeast of Ouagadougou in the Sudan savanna belt of Burkina Faso, West Africa. Species of primary interest were the main malaria vectors Anopheles gambiae S.S. and An.arabiensis, sibling species belonging to the An.gambiae complex. Data forAn.finestus, An.pharoensis, Culex quinquefasciatus and Mansonia uniformis were also analysed. Carbon dioxide was used at concentrations of 0.04-0.6% (cf. 0.03% ambient concentration) for attracting mosquitoes to odour-baited entry traps (OBETs). The ‘attractiveness’ of whole human odour was also compared with CO₂, emitted at a rate equivalent to that released by the human bait. In a direct choice test with two OBETs placed side-by-side, the number of An.gambiae s. I. entering the trap with human odour was double the number trapped with CO₂, alone (at the human equivalent rate), but there was no significant difference between OBETs for the other species of mosquitoes. When OBETs were positioned 20 m apart, again CO₂, alone attracted half as many An.gambiae s.l. and only 40% Anlfunestus, 65% Ma.uniformis but twice as many An.pharoensis compared to the number trapped with human odour. The dose-response for all mosquito species was essentially similar: a linear increase in catch with increasing dose on a log-log scale. The slopes of the dose-response curves were not significantly different between species, although there were significant differences in the relative numbers caught. If the dose-response data are considered in relation to a standard human bait collection (HBC), however, the behaviour of each species was quite different. At one extreme, even the highest dose of CO₂, did not catch more An.gambiae s.1. than one HBC. At the other extreme, the three highest doses of CO₂, caught significantly more Ma.unifonnis than did one HBC. An.pharoensis and Cx quinquefasciatus showed a threshold response to CO₂, responding only at doses above that normally released by one man. An.funestus did not respond to CO₂, alone at any dose in sufficient numbers to assess the dose response. Within the An.gambiae complex, An.arabiensis 'chose' the CO₂,-baited trap with a higher probability than An.gambiae S.S. Also An.arabiensis, the less anthropophilic of the two species, was more abundant in CO₂,-baited OBETs than in human bait collections.